PMID- 18674468 TI - Fractured internal carotid artery stent. AB - Carotid artery stenting is gaining wider acceptance as an alternative option in the treatment of carotid artery stenosis. Conventional complications such as stroke and recurrent in-stent restenosis are well documented in the literature. However, we believe that carotid stent complications are underreported. The carotid artery segment has features that are relatively smilar to the femoropopliteal segment with respect to forces affecting the stented segment, which can contribute to stent complications such as fracture. In this article, we present a case of internal carotid artery stent fracture and thrombus formation after the patient was exposed to direct trauma to the chest and neck. PMID- 18674469 TI - Case of ventricular noncompaction: the crypts and the blood. PMID- 18674470 TI - Intensive lipid intervention in the post-ENHANCE era. PMID- 18674471 TI - Intensive lipid lowering with atorvastatin in patients with coronary artery disease, diabetes, and chronic kidney disease. AB - OBJECTIVE: To investigate the effect of intensive lipid lowering with high-dose atorvastatin on the incidence of major cardiovascular events compared with low dose atorvastatin in patients with coronary artery disease and type 2 diabetes, with and without chronic kidney disease (CKD). PATIENTS AND METHODS: Following 8 weeks' open-label therapy with atorvastatin (10 mg/d), 10,001 patients with coronary artery disease were randomized to receive double-blind therapy with either 80 mg/d or 10 mg/d of atorvastatin between July 1, 1998, and December 31, 1999. Of 1501 patients with diabetes, renal data were available for 1431. Patients with CKD were defined as having a baseline estimated glomerular filtration rate (eGFR) below 60 mL/min per 1.73 m2, using the Modification of Diet in Renal Disease equation. RESULTS: After a median follow-up of 4.8 years, 95 (17.4%) of 546 patients with diabetes and CKD experienced a major cardiovascular event vs 119 (13.4%) of 885 patients with diabetes and normal eGFRs (hazard ratio [HR], 1.32; 95% confidence interval [CI], 1.00-1.72; P<.05). Compared with 10 mg of atorvastatin, 80 mg of atorvastatin reduced the relative risk of major cardiovascular events by 35% in patients with diabetes and CKD (20.9% [57/273] vs 13.9% [38/273]; HR, 0.65; 95% CI, 0.43-0.98; P=.04) and by 10% in patients with diabetes and normal eGFR (14.1% [62/441] vs 12.8% [57/444]; HR, 0.90; 95% CI, 0.63-1.29; P=.56). The absolute risk reduction in patients with diabetes and CKD was substantial, yielding a number needed to treat of 14 to prevent 1 major cardiovascular event over 4.8 years. Both treatments were well tolerated. CONCLUSION: Patients with diabetes, stable coronary artery disease, and mild to moderate CKD experience marked reduction in cardiovascular events with intensive lipid lowering, in contrast to previous observations in patients with diabetes and end-stage renal disease. TRIAL REGISTRATION: clinicaltrials.gov identifier: NCT00327691. PMID- 18674472 TI - Effectiveness and safety of short-course vs long-course antibiotic therapy for group a beta hemolytic streptococcal tonsillopharyngitis: a meta-analysis of randomized trials. AB - OBJECTIVE: To evaluate the effectiveness and safety of short-course antibiotic treatment of group A beta-hemolytic streptococcal (GAS) tonsillopharyngitis. METHODS: We performed a meta-analysis of randomized controlled trials (RCTs) retrieved from PubMed and the Cochrane Central Register of Controlled Trials using a structured search strategy. The last date either database was accessed was November 14, 2007. We included RCTs that involved patients of any age with GAS tonsillopharyngitis, comparing short-course (< or =7 days) vs long-course (at least 2 days longer than short-course) treatment with the same antibiotics. The primary analysis compared 5 to 7 days with 10 days of treatment, using a random effects model. RESULTS: Eleven RCTs comparing short-course vs long-course treatment (5 with penicillin V, 4 with oral cephalosporins, 1 with intramuscular ceftriaxone, and 1 with clindamycin; 6 of the 11 were open label) were included. In the primary analysis, microbiological eradication rates of GAS were inferior for short-course vs long-course treatment (8 RCTs, 1607 patients; odds ratio [OR], 0.49; 95% confidence interval [CI], 0.32-0.74). This association was noted with penicillin V treatment (3 RCTs, 500 patients; OR, 0.36; 95% CI, 0.13-0.99) but was nonsignificant with cephalosporin treatment (4 RCTs, 1018 patients; OR, 0.62; 95% CI, 0.38-1.03). Microbiological eradication was less likely with short course treatment in trials involving primarily children and adolescents (aged <18 years) (6 RCTs, 1258 patients; OR, 0.63; 95% CI, 0.40-0.98). Clinical success was inferior in patients who received short-course treatment (5 RCTs, 1217 patients; OR, 0.49; 95% CI, 0.25-0.96). Adverse events did not differ between compared groups. The above associations were consistent in the analyses involving all included RCTs. CONCLUSION: Short-course treatment for GAS tonsillopharyngitis, particularly with penicillin V, is associated with inferior bacteriological eradication rates. PMID- 18674473 TI - Safety of prescribing PDE-5 inhibitors via e-medicine vs traditional medicine. AB - OBJECTIVE: To determine the safety of a US-based, state-regulated Internet system vs a multispecialty primary care system for prescribing phosphodiesterase type 5 (PDE-5) inhibitors for erectile dysfunction. PATIENTS AND METHODS: From January 1, 2001, through December 31, 2005, 500 e-medicine clients (mean+/-SD age, 47+/ 11 years; hypertension, 60%; type 2 diabetes mellitus, 2%; mean+/-SD number of medications, 0.4+/-0.8) vs 500 traditional medicine patients (mean+/-SD age, 57+/ 12 years; hypertension, 50%; type 2 diabetes mellitus, 23%; mean+/-SD number of medications, 5.1+/-3.1) with erectile dysfunction symptoms were assessed. Noninferiority safety was assessed in this retrospective, cross-sectional study with stratified random sampling by identification of prescribing in the presence of clinically important PDE-5 inhibitor drug interactions with or without high risk cardiovascular disease, by asking about diagnostic symptoms specific to erectile dysfunction, and by determining frequency of patient counseling. RESULTS: Noninferiority of the e-medicine system was shown for the 6 safety end points, relative to a traditional medicine system. Numbers of inappropriate prescriptions, after correction for disease and medication covariates, did not differ between systems. Medication counseling showed superiority of the e medicine system. Standard diagnostic questions were required for e-medicine prescribing but were infrequently asked in traditional medicine. CONCLUSION: Safety in prescribing PDE-5 inhibitors for erectile dysfunction was similar between a US-based, state-regulated Internet prescribing system and a multispecialty primary care system. PMID- 18674475 TI - Obesity as a risk factor for unanticipated admissions after ambulatory surgery. AB - OBJECTIVE: To test the hypothesis that obesity is an independent risk factor for unplanned hospital admission or readmission among patients scheduled for ambulatory surgery in a tertiary medical center. PATIENTS AND METHODS: Existing databases were used to identify 235 obese patients (body mass index [BMI] >40) scheduled for ambulatory surgery from January 2, 2002, through December 31, 2003, at Mayo Clinic's site in Rochester, MN. Each patient was matched to a normal weight control (BMI <25) by age, sex, surgical procedure, type of anesthesia, and date of surgery, and the medical records of all patients were reviewed. Conditional logistic regression analysis was performed to assess whether obesity is an independent risk factor for unplanned postoperative hospital admission. In all cases, 2-sided tests were performed. P<.05 was considered statistically significant. RESULTS: Obese patients (mean +/- SD BMI, 44+/-4) were matched with control patients (mean +/- SD BMI, 23+/-2). Comorbidity was more frequent in the obese cohort. The frequency of unplanned hospital admission did not differ between groups: 61 obese patients (26.0%) and 52 control patients (22.1%) were admitted (odds ratio, 1.3; 95% confidence interval, 0.8-2.0; P=.30). CONCLUSION: Obesity is not a significant independent risk factor for unplanned admission after ambulatory surgery, suggesting that obesity per se should not prevent ambulatory surgery from being scheduled. PMID- 18674474 TI - Effect of bariatric surgery on the metabolic syndrome: a population-based, long term controlled study. AB - OBJECTIVE: To assess the effect of weight loss by bariatric surgery on the prevalence of the metabolic syndrome (MetS) and to examine predictors of MetS resolution. PATIENTS AND METHODS: We performed a population-based, retrospective study of patients evaluated for bariatric surgery between January 1, 1990, and December 31, 2003, who had MetS as defined by the American Heart Association/National Heart, Lung, and Blood Institute (increased triglycerides, low high-density lipoprotein, increased blood pressure, increased fasting glucose, and a measure of obesity). Of these patients, 180 underwent Roux-en-Y gastric bypass, and 157 were assessed in a weight-reduction program but did not undergo surgery. We determined the change in MetS prevalence and used logistic regression models to determine predictors of MetS resolution. Mean follow-up was 3.4 years. RESULTS: In the surgical group, all MetS components improved, and medication use decreased. Nonsurgical patients showed improvements in high density lipoprotein cholesterol levels. After bariatric surgery, the number of patients with MetS decreased from 156 (87%) of 180 patients to 53 (29%); of the 157 nonsurgical patients, MetS prevalence decreased from 133 patients (85%) to 117 (75%). A relative risk reduction of 0.59 (95% confidence interval [CI], 0.48 0.67; P<.001) was observed in patients who underwent bariatric surgery and had MetS at follow-up. The number needed to treat with surgery to resolve 1 case of MetS was 2.1. Results were similar after excluding patients with diabetes or cardiovascular disease or after using diagnostic criteria other than body mass index for MetS. Significant predictors of MetS resolution included a 5% loss in excess weight (odds ratio, 1.26; 95% CI, 1.19-1.34; P<.001) and diabetes mellitus (odds ratio, 0.32; 95% CI, 0.15-0.68; P=.003). CONCLUSION: Roux-en-Y gastric bypass induces considerable and persistent improvement in MetS prevalence. Our results suggest that reversibility of MetS depends more on the amount of excess weight lost than on other parameters. PMID- 18674476 TI - Occult malignancy in patients with suspected paraneoplastic neurologic syndromes: value of positron emission tomography in diagnosis. AB - OBJECTIVE: To determine the value of positron emission tomography (PET) in diagnosing occult malignancies in patients with paraneoplastic neurologic syndromes (PNSs) at Mayo Clinic's site in Rochester, MN. PATIENTS AND METHODS: We retrospectively reviewed the medical charts of all 107 patients who underwent PET from January 1, 2000, to July 31, 2006, for the indication of suspected PNS. Three patients did not meet inclusion criteria. PET results were considered positive if increased fludeoxyglucose F 18 uptake indicated malignancy (24 patients). Results from computed tomography were interpreted as positive if any suspect lesion was consistent with malignancy (26 patients). RESULTS: One hundred four patients with PNS were identified from the PET central database; 73 patients had at least 1 positive result for paraneoplastic antibody, and 31 had antibody negative PNS. Malignancy was confirmed pathologically in 10 patients, of whom 8 had positive PET results. There were 2 cases of confirmed malignancy (fallopian tube adenocarcinoma and spindle cell uterine carcinoma) for which PET results were negative. Two patients with positive PET results declined biopsy. Computed tomography was able to identify 3 of the 10 malignancies detected. Five cases of malignancy were detected only by PET. All patients with confirmed malignancy had positive results for at least 1 paraneoplastic antibody. One patient with positive results for PNS antibody and negative PET results was diagnosed as having small cell carcinoma on a follow-up PET scan after 27 months. PET had sensitivity, specificity, positive predictive value, and negative predictive value of 80%, 67%, 53%, and 88%, respectively. CONCLUSION: PET scan was shown to be more sensitive than computed tomography for detecting occult malignancy (confirmed by positive test results for autoantibody) among patients with suspected PNS. The greatest clinical utility of PET could be in its high negative predictive value. PMID- 18674477 TI - Design of the REVEAL registry for US patients with pulmonary arterial hypertension. AB - The Registry to Evaluate Early and Long-term PAH Disease Management (REVEAL) has been designed to meet the need for current information about patients with pulmonary arterial hypertension (PAH). The main objectives of REVEAL are to better define and understand PAH and to assess the consequences of treatment strategies. REVEAL is collecting clinically relevant data from 3500 consecutively enrolled patients with confirmed PAH diagnoses. Outcomes will be evaluated longitudinally and compared according to the baseline classification of PAH. The primary outcome for group comparisons will be survival. Collected data include World Health Organization functional class, 6-minute walk distance, cardiopulmonary exercise testing, pulmonary function test results, hemodynamic measurements, functional status, hospitalizations, and death. REVEAL will be the richest source of data on patients with World Health Organization group I PAH. PMID- 18674478 TI - A practical "ABCDE" approach to the metabolic syndrome. AB - The metabolic syndrome comprises a cluster of risk factors for cardiovascular disease and type 2 diabetes mellitus that are due to abdominal obesity and insulin resistance. This increasingly important proinflammatory condition remains both underrecognized and undertreated. To aid physicians in their approach to the metabolic syndrome, we assessed and synthesized the literature on cardiovascular risk assessment and early intervention for risk reduction. We performed a comprehensive search of MEDLINE and the Cochrane database for peer-reviewed clinical studies published from January 1, 1988, to December 31, 2007, augmented by consultation with content experts. We used the search terms metabolic syndrome, abdominal obesity, waist circumference, insulin resistance, cardiovascular disease, prediabetes, diabetes, treatment, prevention, aspirin, hypertension, cholesterol, atherogenic dyslipidemia, lifestyle therapy, diet, and exercise. Criteria used for study review were controlled study design, English language, relevance to clinicians, and validity based on experimental design and appropriateness of conclusions. Although growing evidence supports early intervention in patients with the metabolic syndrome, many physicians do not recognize the risk associated with this condition and fail to initiate early treatment. A comprehensive management plan can be assembled through an "ABCDE" approach: "A" for assessment of cardiovascular risk and aspirin therapy, "B" for blood pressure control, "C" for cholesterol management, "D" for diabetes prevention and diet therapy, and "E" for exercise therapy. This ABCDE approach provides a practical and systematic framework for encouraging metabolic syndrome recognition and for implementing a comprehensive, evidence-based management plan for the reduction of cardiovascular risk. PMID- 18674479 TI - Peripheral arterial disease: diagnosis and management. AB - Peripheral arterial disease is a common but underdiagnosed and undertreated disorder with substantial morbidity and mortality. The pathophysiology of peripheral arterial disease and the risk factors for developing it are similar to those for atherosclerotic disease occurring at other sites. Peripheral arterial disease can be diagnosed accurately with simple, noninvasive, office-based tests that measure the severity of the disease and provide valuable prognostic information. Optimal medical therapy includes a supervised exercise program, tobacco cessation, and modification of treatable risk factors. Cilostazol can improve pain-free and peak walking distances in patients with intermittent claudication. As a general rule, patients with lifestyle-limiting claudication who do not respond to medical management or those with critical limb ischemia should be referred to a vascular specialist for consideration of revascularization. PMID- 18674480 TI - 45-year-old woman with recurrent headache and photophobia. PMID- 18674481 TI - Increased use of beta-agonists leading to non-ST-segment elevation myocardial infarction in a patient with hemodynamically significant myocardial bridging. AB - Myocardial bridging is not uncommon. However, hemodynamic instability in the setting of myocardial bridging is rare. We describe an unusual case of a 47-year old woman whose increased use of inhaled albuterol led to a non-ST-segment elevation myocardial infarction and unmasking of hemodynamically significant myocardial bridging. We detail its diagnosis through myocardial perfusion imaging and coronary angiography. To our knowledge, this is the first time previously undiagnosed myocardial bridging has led to a myocardial infarction in the setting of inhaled albuterol use. PMID- 18674482 TI - Delayed clinician responses to elevated prostate-specific antigen values. PMID- 18674483 TI - Nocturnal pagophagia complicating gastric bypass. PMID- 18674484 TI - [Prognostic value of the left ventricular ejection fraction in patients with acute heart failure]. AB - BACKGROUND AND OBJECTIVE: The relation between left ventricular ejection fraction (LVEF) and prognosis in patients with heart failure is controversial. The aim of this study was to determine the relation of LVEF in long-term mortality and readmissions for acute heart failure in a non-selected population of patients admitted with acute heart failure (AHF). PATIENTS AND METHOD: We included 507 patients admitted consecutively for AHF in a cardiology department of a single centre. LVEF was assessed with transthoracic echocardiography during hospitalization. All-cause mortality and readmission for AHF were selected as primary and secondary endpoints, respectively. The independent association between LVEF and endpoints was assessed with traditional Cox regression analysis for all-cause mortality and Cox regression for competing risks for readmission for AHF. RESULTS: 47% of patients exhibited LVEF > or = 50%. During a median follow-up of one year, 151 (30%) deaths and 139 (27%) readmissions for AHF were observed. Mortality rates were higher in patients with LVEF < 50% (34 vs 25%; p = 0.028) and no differences were observed for readmissions for AHF (26 vs 29%, p = 0.510). In multivariate analysis, after adjustment for traditional risk factors, patients with LVEF < 50% did not show higher risk of mortality (hazard ratio [HR] = 1.08; 95% confidence interval [CI], 0.76-1.57; p = 0.645) or readmissions for AHF (HR = 1.00; 95% CI, 0.68-1.47; p = 1). CONCLUSIONS: Patients with preserved LVEF constitute a substantial proportion of patients with AHF, exhibiting similar mortality and readmissions risks compared with patients with depressed LVEF. PMID- 18674485 TI - Functional status as a risk factor for mortality in very elderly patients with pneumonia. AB - BACKGROUND AND OBJECTIVE: The most commonly used prognostic mortality indexes for pneumonia take into account several variables including comorbidities, physical examination results, and laboratory test results, as well as age. Other factors such as functional status are not included. The objective of this study was to know whether the preadmission functional status was related to 30-day mortality in old or very old patients who were hospitalized for pneumonia. PATIENTS AND METHOD: This was a prospective study including all patients who were hospitalized for pneumonia in the Acute Geriatric Unit of Hospital de Mataro, Barcelona. We calculated the Pneumonia Severity Index (PSI), preadmission and admission Barthel Index (BI), Charlson Comorbidity Index and Mini Nutritional Assessment (MNA). Patients were assessed during hospitalisation and until death or 30 days after admission. RESULTS: We studied 117 patients, 69 (59%) were men. The mean age (standard deviation) was 84.7 (6.5) years. The 30-day mortality was 16.2%. The PSI score was 134.2 (31.8) on admission, and the BI scores on preadmission and admission were 60.3 (35.8) and 37.1 (33.5), respectively. In a multiple logistic regression model, using all variables statistically significant in the univariate analysis, those independently associated with 30-day mortality were: preadmission BI lower than 60 points (odds ratio = 4.89; 95% confidence interval, 1.27-18.9) and lymphopenia (odds ratio = 7.11; 95% confidence interval, 1.7-30.2). CONCLUSIONS: In very old patients who were hospitalized for pneumonia, preadmission functional status was an independent predictor of mortality. Functional status should be included in the severity indices of pneumonia in this population. PMID- 18674486 TI - [Heart failure with normal systolic function: relevance of a clinical problem]. PMID- 18674487 TI - [Functional status of hospitalized old patients: an important prognostic factor]. PMID- 18674488 TI - [Neurological manifestations in patients with Gaucher disease and in their relatives]. AB - BACKGROUND AND OBJECTIVE: Gaucher disease (GD) is characterized by a wide spectrum of manifestations. Previous reports indicate that GD relatives could develop neurological abnormalities more frequently than the general population. We aimed to know the presence of neurological symptoms (NS) in GD patients and their relatives. PATIENTS AND METHOD: From January to December 2006 we performed a postal survey contacting 42 physicians and 92 families to evaluate NS and correlate them with genetic characteristics. Statistical analysis using descriptive parameters, ANOVA, t-test and a correlation study including Pearson coefficient were performed. RESULTS: Information from 72 families (78.3% responses) including 99 patients and 266 relatives was obtained. Thirty type 1 GD (32.6%) reported NS: tremor 8 (8.7%), uncoordinated movements 9 (9.8%), concentration defects 11 (11.9%), strabism 7 (7.6%), deafness 8 (8.7%), Parkinson disease (PD) 7 (7.6%) and peripheral neuropathy 10 (10.9%). Thirty-six (13.5%) first or second degrees relatives presented the following NS: PD 14 (4.9%), epilepsy 8 (3.0%), tremor 7 (2.6%), deafness 2 (0.7%) and others 5 (1.9%). 17.3% of carriers had NS versus 5.7% in non-carriers (p = 0.0096). Patients with PD had mutations in S364R, D409H, L444P, [IVS4-2a ==> g; c.(-203)A ==> G], c.500insT and L336P. In relatives with PD a wide spectrum of mutations was observed: L444P, N370S, V398I, G202R, c.1439-1445del7, [E326K; N188S] and c.953delT. In other NS, predominant mutations were D409H, G195W, R120W, R147X, L336P and G377S. CONCLUSIONS: A higher incidence than expected of PD and other NS in GD type 1 patients and relatives was observed. These manifestations appear frequently in L444P or rare mutations carriers. It is important to perform a systematic neurological exam in type 1 GD patients and carriers with risk mutations. PMID- 18674489 TI - [Determinants of physician compensation]. PMID- 18674490 TI - [Pharmacogenetics in oncology]. AB - Pharmacogenetics studies the relationship between genetic polymorphisms and individual responses to drugs. In recent years, there has been a great progress in our knowledge of the effects of drug-metabolizing enzymes and molecular target genetic polymorfisms on cancer chemotherapy. Pharmacogenetics focuses on the prediction of drug efficacy and toxicity based on a patient's genetic profile with routinely applicable genetic tests to select the most appropriate medication at optimal doses for each individual patient. Two years ago the FDA approved one genetic test to detect patients with increased risk of severe toxicity associated with irinotecan therapy. There have also been commercialized genetic chips to genotyping two cytochrome P450 enzymes at the same time. Prospectively, stratifying patients based on genotype may identify subpopulations likely to experience severe toxicity or to derive benefit from a particular treatment strategy, helping us move toward the ultimate goal of individualized therapy. In this review, we describe the clinical effects of polymorphisms that may influence cancer chemotherapy. PMID- 18674492 TI - [Therapeutic inertia: importance of the perception in the blood pressure control]. PMID- 18674491 TI - [Prognostic marker of serum alpha-fetoprotein levels in hepatocellular carcinoma: retrospective analysis of 230 patients]. PMID- 18674493 TI - [Methemoglobinemia induced by local anesthetic administration previous to laser depilation]. PMID- 18674494 TI - [Rare diseases: inadequeate data for a serious problem]. PMID- 18674495 TI - [Mycotic aneurysms in lower limbs due to infectious endocarditis]. PMID- 18674496 TI - [Reflex sympathetic dystrophy]. PMID- 18674497 TI - Simple criteria to assess mortality in patients with community-acquired pneumonia. AB - BACKGROUND AND OBJECTIVES: The Pneumonia Patient Outcomes Research Team (PORT) developed a prediction rule to identify patients with community-acquired pneumonia (CAP) who are at risk for death and other adverse outcomes. Simpler criteria are needed to evaluate risk of mortality in CAP. PATIENTS AND METHODS: Observational study of patients with CAP admitted to a tertiary care university hospital. Epidemiological, clinical, radiological and laboratory data associated with mortality were analysed. RESULTS: A cohort of 211 patients with CAP was studied. Severity distribution according to PORT score was 12.3%, 15.6%, 19%, 35.5% and 17.5% in groups I, II, III, IV and V, respectively; mean age was 63 years (range, 13 to 100 years); in 43.6% age was < 65 years; 61.5% patients had > or = 1 underlying disease (congestive heart failure in 33.6% and chronic lung disease in 29.9%). Mortality rate was 0% in groups I-II, 2.5% in group III, 5.3% in group IV and 27% in group V. All variables considered in PORT-score were included in a mortality predicting model; factors significantly associated with death were: altered mental status, respiratory rate > or = 30/min, pH < 7.35, glucose > or = 250 mg/dl and age > 65 years; 99% of patients who had none of these abnormalities survived. CONCLUSIONS: Simpler criteria to assess mortality in CAP were identified. Non-existence of altered mental status, respiratory rate > or = 30/min, pH < 7.35, glucose > or = 250 mg/dl and age > 65 years predicted a non-fatal outcome in 99% of patients. These clinical or laboratory findings should be considered as mortality predictors, can be used as severity adjustment measure and may help physicians make more rational decisions about hospitalization in CAP. PMID- 18674498 TI - [Prevalence and related factors of overweight and obesity in Spanish population aged 60 years-old or older. The PREV-ICTUS study]. AB - BACKGROUND AND OBJECTIVE: Data on the prevalence of obesity in elderly population in Spain are scarce. The objective of this work was to describe the prevalence of obesity and the related factors in a random sample of Spanish population aged 60 years-old or more, stratified by autonomous communities. SUBJECTS AND METHOD: We analyzed the PREV-ICTUS study, a population-based study carried out between September and December 2005 in a random sample stratified by autonomous communities according to the census of inhabitants and the habitat in each community. Subjects were classified as with normal weight (body mass index [BMI] < 25 kg), overweight (BMI 25-29.9) and obesity (BMI > or = 30 kg). RESULTS: In 6,843 subjects -mean age (standard deviation): 71.9 (7.1) years-old; 53.3% women , prevalence of obesity was 34.5% (95% confidence interval [CI], 33.3-35.5%), higher in women (38.4%; 95% CI, 36.8-39.9%) than in men (30.0%; 95% CI, 28.4 31.6%; p < 0.001), with a tendency to decrease in the older population. In total, 81.7% of the population showed overweight (BMI > or = 25). Prevalence differed among communities from 46.4% to 20.7% (p < 0.001). In the multivariate analysis, obesity was inversely related to age (odds ratio [OR] = 0.98 per each year increment), and directly to female gender (OR = 1.25), rural (OR = 1.50) or semi urban habitat (OR = 1.38), sedentary lifestyle (OR = 1.86) and non-smoking history (OR = 1.19). CONCLUSIONS: Obesity was present in one out of 3 individuals of this reasonably representative sample of the Spanish population in an age range, 60 years-old or more, which concentrates a high cardiovascular risk. Differences in the prevalence among autonomus communities were detected. PMID- 18674499 TI - [Factors related with clinical evolution in whiplash]. AB - BACKGROUND AND OBJECTIVE: Factors of poor clinical recovery in acute whiplash are not conclusive. The goal of this prospective longitudinal study was to identify factors with influence in clinical evolution allowing us to identify patients with risk for developing chronic symptoms and disabilities after an acute whiplash. PATIENTS AND METHOD: We included 226 patients who suffered acute whiplash after road traffic accident and met the Department of Physical Medicine and Rehabilitation for medical evaluation and physiotherapy treatment. We collected variables following a protocol designed for the study. All patients were assessed through the visual analogue scale (VAS) for the intensity of neck pain, the Goldberg Depression and Anxiety Scale and the Northwick Park Neck Pain Questionnaire (NPH) for cervical column functionality, at initial evaluation and at discharge of treatment. RESULTS: Factors related with poor recovery of NPH at discharge were: number of days of cervical column immobilization with collar, presence of headache, dizziness, and dorsal pain at initial evaluation and initial evaluation of VAS score and Goldberg Depression and Anxiety Scale. In the multivaried analysis we found that variables with influence on NPH at discharge were statistically significant for VAS, Goldberg depression subscale and NPH scores at initial evaluation. CONCLUSIONS: The most important factors that determine the evolution of patients with acute whiplash are the initial evaluation of the neck pain with the Visual Analogue Scale, the cervical column functionality with NPH and Goldberg Depression subscale. PMID- 18674500 TI - [Hospital admission in community-acquired pneumonia]. PMID- 18674501 TI - [Cathepsin K: biological aspects and therapeutic possibilities]. PMID- 18674502 TI - [Genetic abnormalities of digestive tract adenocarcinomas and correlation with the histologic sequence of their development]. AB - Over 90% of digestive tract malignancies are adenocarcinomas (ADC) and almost 95% of them have gastric (G), colorectal (CR) or pancreatic (P) localizations. The objectives of this work are to review the genetic abnormalities of ADC in these locations and their potential coincidences, along with the histogenetic correlation of their emergence. Genetic abnormalities affecting over 50% of cases include: in G-ADC, inactivation of suppressor genes of p53, APC and DCC tumor in its intestinal variant, hypoexpression of of caderine E in the diffuse variant and hyperexpression of cyclooxygenase-2 and cyclyn D in the intestinal form; in in CR-ADC, inactivation of of genes p53, APC and DCC together with mutational activation of k-ras oncogen, and in P-ADC, the inactivation of suppressor genes p53, p16 and DPC4 along with mutational activation of k-ras oncogen. P-ADC is the one showing a more characteristic and exclusive genetic mark, followed by CR-ADC. Finally, the histogenetic correlation in the tumorigenic sequence is more evident in CR-ADC, followed by P-ADC. The complex biologic reality of G-ADC makes it more difficult to draw its genetic profile and its histogenetic correlation. In order to understand better the arguments of this work, the authors comment on the genetic-molecular basis governing the life and death of normal somatic cells and the biologic profile of the groups of genes mainly involved in tumorigenesis. PMID- 18674503 TI - [Angiotensin converting enzyme 2 and its emerging role in the regulation of the renin angiotensin system]. AB - The renin-angiotensin system (RAS) plays a key role in the regulation of cardiovascular and renal function. Thus, RAS blockade with an angiotensin converting enzyme (ACE) and/or angiotensin receptor blocker decreases blood pressure, cardiovascular events, and delays the progression of kidney disease. The discovery of ACE2, a homologue of ACE, capable of degrading angiotensin II to angiotensin 1-7, may offer new insights into the RAS. In this review we discuss the possible protective role of ACE2 in different organs, namely heart, lungs and kidneys. The role of this enzyme is inferred from recent studies performed using genetically manipulated mice that lack the ACE2 gene and also mice treated with pharmacological ACE2 inhibitors. These results suggest that ACE2 might be a new therapeutic target within the RAS. PMID- 18674504 TI - [Characteristics of patients with tuberculosis admitted from emergency department in a University Hospital: comparative study 1996-2006]. PMID- 18674505 TI - [Isolated abdominal cystic angiomatosis without soft tissue or visceral involvement]. PMID- 18674506 TI - [Bilateral adrenal hemorrhage and chronic myeloproliferative syndrome]. PMID- 18674507 TI - [Evaluation of research activity by means of the Hirsch h index]. PMID- 18674508 TI - [Diseminated histoplasmosis in a HIV-infected patient]. PMID- 18674509 TI - An enzyme-linked immunosorbent poly(ADP-ribose) polymerase biomarker assay for clinical trials of PARP inhibitors. AB - Many established cancer therapies involve DNA-damaging chemotherapy or radiotherapy. The DNA repair capacity of the tumor represents a common mechanism used by cancer cells to survive DNA-damaging therapy. Poly(ADP-ribose) polymerase (PARP) is a nuclear enzyme that is activated by DNA damage and has critical roles in DNA repair. Inhibition of PARP potentiates the activity of DNA-damaging agents such as temozolomide, topoisomerase inhibitors and radiation in both in vitro and in vivo preclinical models. Recently, several PARP inhibitors have entered clinical trials either as single agents or in combination with DNA-damaging chemotherapy. Because PARP inhibitors are not cytotoxic, a biomarker assay is useful to guide the selection of an optimal biological dose. We set out to develop an assay that enables us to detect 50% PAR reduction in human tumors with 80% power in a single-plate assay while assuring no more than a 10% false positive rate. We have developed and optimized an enzyme-linked immunosorbent assay (ELISA) to measure PARP activity that meets the above-mentioned criterion. This robust assay is able to detect PAR levels of 30-2000 pg/ml in both tumor and peripheral blood monocyte samples. In a B16F10 mouse syngeneic tumor model, PARP inhibitor ABT-888 potentiates the effect of temozolomide in suppressing tumor growth, and PARP activity is greatly reduced by ABT-888 at efficacious doses. In summary, the ELISA assay described here is suitable for biomarker studies in clinical trials of PARP inhibitors. PMID- 18674510 TI - A noncommercial polymerase chain reaction-based method to approach one hundred percent recombinant clone selection efficiency. AB - Molecular cloning is an important procedure in molecular biology, but this is often a rate-limiting step and can be very time-consuming, possibly due to low ligation efficiency. Here, we describe a simple polymerase chain reaction (PCR) based strategy to approach 100% selection efficiency. The post-ligation mixture containing the recombinant was subjected to insert-specific primer-mediated PCR amplification using a high-fidelity DNA Pfu polymerase generating a plasmid containing staggered nicks. The PCR mixture was then digested with endonuclease DpnI, which digests the methylated and hemimethylated parental DNA template. The nicked vector was transformed into XL1 blue supercompetent cells where the nicks were repaired, thus amplifying and selecting only the newly amplified recombinant clones. PMID- 18674511 TI - Electrochemical assay for deoxyribonuclease I activity. AB - A thiolated oligonucleotide having three ferrocenes was immobilized on a gold electrode through the sulfur-gold linkage. This electrode showed a current response based on the redox reaction of the ferrocene moieties and this response was decreased after treatment with deoxyribonuclease I (DNase I), suggesting the disappearance of the ferrocene moieties on the electrode by the DNase I digestion. A linear correlation between i(0) and i, which are current peaks before and after DNase I treatment, respectively, was observed and this slope was decreased with increase in the amount of DNase I. No current decrease was observed in the presence of EDTA or RNase A instead of DNase I. These results suggested that the current decrease responded specifically to the amount of DNase I and this electrode could be used for an electrochemical DNase I assay. Under the optimum conditions of DNase I digestion at 37 degrees C for 30 min, a quantitative analysis could be achieved in the range of 10(-4)-10(-2)units/microl of DNase I. PMID- 18674512 TI - A high-throughput genome-walking method and its use for cloning unknown flanking sequences. AB - We developed a PCR-based high-throughput genome-walking protocol. The novelty of this protocol is in the random introduction of unique walker primer binding sites into different regions of the genome efficiently by taking advantage of the rolling circle mode of DNA synthesis by Phi29 DNA polymerase after annealing the partially degenerate primers to the denatured genomic DNA. The inherent strand displacement activity of the Phi29 DNA polymerase displaces the 5' ends of downstream strands and DNA synthesis continues, resulting in a large number of overlapping fragments that cover the whole genome with the unique walker adapter attached to the 5' end of all the genomic DNA fragments. The directional genome walking can be performed using a locus-specific primer and the walker primer and Phi29 DNA polymerase-amplified genomic DNA fragments as template. The locus specific primer will determine the position and direction of the genome walk. Two rounds of successive PCR amplifications by locus-specific and walker primers and their corresponding nested primers effectively amplify the flanking DNA fragments. The desired PCR fragment can be either cloned or sequenced directly using another nested, locus-specific primer. We successfully used this protocol to isolate and sequence 5' flanking regions/promoters of selected plant genes. PMID- 18674513 TI - Neurite responses to ephrin-A5 modulated by BDNF: evidence for TrkB-EphA interactions. AB - In the developing visual system, growing retinal ganglion cell (RGC) axons are exposed to multiple guidance and growth factors. Furthermore, the relative levels of these factors are differentially regulated as topography is roughly established and then refined. We have shown that during the establishment of rough topography (P3), growth cones of pure and explanted RGCs treated with combinations of BDNF and ephrin-A5-Fc responded differently than RGCs treated with BDNF or ephrin-A5-Fc alone (p=0.0083). The response to the combined treatment mimicked that of RGCs cultured with ephrin-A5-Fc alone once topography refines. The guidance cue receptors EphA and TrkB were shown to co-localise in RGCs in vitro. Furthermore, EphA and TrkB receptors interacted directly in in vitro binding assays. Our results suggest that the conversion of growth cone responses from collapse to stabilisation as topography refines, occurs as a result of interactions between EphA and TrkB receptors. PMID- 18674514 TI - Identification and characterization of a new Leishmania major specific 3'nucleotidase/nuclease protein. AB - We report the characterization of a new Leishmania major gene, lmaj3'nt/nu, encoding a 382 amino acids protein, Lmaj3'NT/NU, that belongs to the 3'nucleotidase/nuclease family. Interestingly, sequence and phylogenetic analysis show that this protein is Leishmania major specific and thus constitutes a new 3'nucleotidase/nuclease subgroup. Lmaj3'NT/NU displays nuclease enzymatic activity and Western blot analysis shows that it is exclusively expressed in promastigotes. Immunofluorescence microscopy using a specific anti-Lmaj3'NT/NU shows that the protein has a plasma membrane localization. Surprisingly, contrary to the previously described Leishmania mexicana 3'NT/NU, lmaj3'nt/nu is not up regulated when parasites are cultured under purine starvation conditions. Together, these findings suggest Lmaj3'NT/NU may constitute a new important compound of the L. major purine scavenging pathway and could be involved in sandfly parasite survival and colonization. PMID- 18674515 TI - The human UDP-glucuronosyltransferase UGT1A3 is highly selective towards N2 in the tetrazole ring of losartan, candesartan, and zolarsartan. AB - Losartan, candesartan, and zolarsartan are AT(1) receptor antagonists that inhibit the effect of angiotensin II. We have examined their glucuronidation by liver microsomes from several animals and by recombinant human UDP glucuronosyltransferases (UGTs). Large differences in the production of different glucuronide regioisomers of the three sartans were observed among liver microsomes from human (HLM), rabbit, rat, pig, moose, and bovine. However, all the liver microsomes produced one or two N-glucuronides in which either N1 or N2 of the tetrazole ring were conjugated. O-Glucuronides were also detected, including acyl glucuronides of zolarsartan and candesartan. Examination of individual human UGTs of subfamilies 1A and 2B revealed that N-glucuronidation activity is widespread, along with variable regioselectivity with respect to the tetrazole nitrogens of these sartans. Interestingly, UGT1A3 exhibited a strong regioselectivity towards the N2 position of the tetrazole ring in all three sartans. Moreover, the tetrazole-N2 of zolarsartan was only conjugated by UGT1A3, whereas the tetrazole-N1 of this aglycone was accessible to other enzymes, including UGT1A5. Zolarsartan O-glucuronide was mainly produced by UGTs 1A10 and 2B7. UGT2B7, alongside UGT1A3, glucuronidated candesartan at the tetrazole-N2 position, whereas UGTs 1A7-1A10 mainly yielded candesartan O-glucuronide. In the case of losartan, no O-glucuronide was generated by any tested human enzyme. Nevertheless, UGTs 1A1, 1A3, 1A10, 2B7, and 2B17 glucuronidated losartan at the tetrazole-N2, while UGT1A10 also yielded the respective N1-glucuronide. Kinetic analyses revealed that the main contributors to losartan glucuronidation in HLM are UGT1A1 and UGT2B7. The results provide ample new data on substrate specificity in drug glucuronidation. PMID- 18674516 TI - Transcellular transport of organic cations in double-transfected MDCK cells expressing human organic cation transporters hOCT1/hMATE1 and hOCT2/hMATE1. AB - To clarify the transcellular transport of organic cations via basolateral and apical transporters, we established double-transfected Madin-Darby canine kidney (MDCK) cells expressing both human organic cation transporter hOCT1 and hMATE1 (MDCK-hOCT1/hMATE1), and hOCT2 and hMATE1 (MDCK-hOCT2/hMATE1) as models of human hepatocytes and renal epithelial cells, respectively. Using the specific antibodies, hOCT1 and hMATE1 or hOCT2 and hMATE1 were found to be localized in the basolateral and apical membranes of MDCK-hOCT1/hMATE1 or MDCK-hOCT2/hMATE1 cells, respectively. A representative substrate, [14C]tetraethylammonium, was transported unidirectionally from the basolateral to apical side in these double transfectants. The optimal pH was showed to be 6.5 for the transcellular transport of [14C]tetraethylammonium, when the pH of the incubation medium on the apical side was varied from 5.5 to 8.5. The basolateral-to-apical transport also decreased in the presence of 10 mM 1-methyl-4-phenylpyridinium or 1 mM levofloxacin on the basolateral side of both double transfectants. In MDCK hOCT2/hMATE1 cell monolayers, but not in MDCK-hOCT1/hMATE1 cell monolayers, the accumulation of [14C]tetraethylammonium was decreased in the presence of 10 mM 1 methyl-4-phenylpyridinium, but significantly increased in the presence of 1 mM levofloxacin. The uptake of [14C]tetraethylammonium, [3H]1-methyl-4 phenylpyridinium, [14C]metformin and [3H]cimetidine, but not of [14C]procainamide and [3H]quinidine, by HEK293 cells was stimulated by expression of the hOCT1, hOCT2 or hMATE1 compared to control cells. However, transcellular transport of [14C]procainamide and [3H]quinidine was clearly observed in both double transfectants. These cells could be useful for examining the routes by which compounds are eliminated, or predicting transporter-mediated drug interaction. PMID- 18674517 TI - Characterization of eltenac and novel COX-2 selective thiopheneacetic acid analogues in vitro and in vivo. AB - We assessed the effect of novel selective thiopheneacetic acids on cyclooxygenase isoenzymes in vitro and in vivo. Thiopheneacetic acid Eltenac and derivatives were investigated in this study. In human whole blood experiments these derivatives were potent inhibitors of COX-2 (IC(50)=0.02-0.4 microM) with less pronounced effect on COX-1 (IC(50)=0.15-5.6 microM). With COX-1/COX-2 ratios between 7.5- and 16-fold they are in the range of Celecoxib (13-fold). The parent drug Eltenac demonstrated no selectivity for COX-2. In a rat paw edema model, these compounds showed reduction of edema volume in the range of 36-45% at 10 mg/kg (Eltenac 52%, Diclofenac 51%). However, the compounds were superior to Diclofenac and Eltenac with respect to their ulcerogenic and gastrointestinal properties. Introduction of a nitrate-ester moiety to either Eltenac or a derivative did neither improve selectivity or potency in vitro, nor ulcerogenicity in vivo. Molecular modeling of selective thiopheneacetic acid derivatives to the active site of human COX-2 suggested similar binding properties as Lumiracoxib and Diclofenac. In summary, modification of Eltenac generates moderately selective COX-2 drugs in the range of Celecoxib with respect to potency and selectivity. The drugs showed potent anti-inflammatory properties and significant improvement of animal survival in a sub-chronical experimental set up. Thiopheneacetic derivatives are characterized by low pK(a) values, short microsomal half-lives and binding mode to COX-2 similar to Diclofenac and Lumiracoxib. These properties may also have an impact on the transient inhibition of COX-2-dependent prostacyclin, thereby being less associated with vascular complications. PMID- 18674518 TI - Dopaminergic and non-dopaminergic value systems in conditioning and outcome specific revaluation. AB - Animals are motivated to choose environmental options that can best satisfy current needs. To explain such choices, this paper introduces the MOTIVATOR (Matching Objects To Internal VAlues Triggers Option Revaluations) neural model. MOTIVATOR describes cognitive-emotional interactions between higher-order sensory cortices and an evaluative neuraxis composed of the hypothalamus, amygdala, and orbitofrontal cortex. Given a conditioned stimulus (CS), the model amygdala and lateral hypothalamus interact to calculate the expected current value of the subjective outcome that the CS predicts, constrained by the current state of deprivation or satiation. The amygdala relays the expected value information to orbitofrontal cells that receive inputs from anterior inferotemporal cells, and medial orbitofrontal cells that receive inputs from rhinal cortex. The activations of these orbitofrontal cells code the subjective values of objects. These values guide behavioral choices. The model basal ganglia detect errors in CS-specific predictions of the value and timing of rewards. Excitatory inputs from the pedunculopontine nucleus interact with timed inhibitory inputs from model striosomes in the ventral striatum to regulate dopamine burst and dip responses from cells in the substantia nigra pars compacta and ventral tegmental area. Learning in cortical and striatal regions is strongly modulated by dopamine. The model is used to address tasks that examine food-specific satiety, Pavlovian conditioning, reinforcer devaluation, and simultaneous visual discrimination. Model simulations successfully reproduce discharge dynamics of known cell types, including signals that predict saccadic reaction times and CS dependent changes in systolic blood pressure. PMID- 18674519 TI - Paradoxical sleep deprivation impairs spatial learning and affects membrane excitability and mitochondrial protein in the hippocampus. AB - Previous research has demonstrated that paradoxical sleep has a key role in learning and memory, and sleep deprivation interferes with learning and memory. However, the mechanism of memory impairment induced by sleep deprivation is poorly understood. The present study investigated the effect of paradoxical sleep deprivation (PSD) on spatial learning and memory using the Morris Water Maze. Effects of PSD on CA1 pyramidal neurons in hippocampus were also examined. PSD impaired spatial learning of rats. PSD induced translocation of Bax to mitochondria and cytochrome c release into the cytoplasm, and decreased the membrane excitability of CA1 pyramidal neurons, effects which may contribute to the deficits in learning behavior. These results may partially explain the mechanism of the effect of PSD on learning. Modulating the excitability of hippocampal neurons and protecting mitochondrial function are possible targets for preventing the effects of paradoxical sleep deprivation. PMID- 18674520 TI - The influence of strongly focused visual attention on the detection of change in an auditory pattern. AB - The mismatch negativity, an ERP that reflects the detection of change in the auditory environment, is considered to be a relatively automatic process. Its automaticity has by in large been studied using the oddball paradigm, in which a physical feature of a frequently presented standard stimulus is changed. In the present study, the automaticity of the MMN is tested using a MMN elicited by a violation of a more abstract auditory pattern. Fourteen subjects were presented with an alternating pattern of two tones (ABABAB) that was occasionally broken by deviant repetitions (e.g., ABABABBBAB). The alternating tones were separated by 1 or 6 semitones in different conditions. The subjects were engaged in a continuous multiple object tracking (MOT) task and thus ignored the auditory stimuli. Difficulty of the MOT task was manipulated by increasing the number of objects to be tracked. Subjects were also asked to read a text and ignore the auditory stimuli in another condition. A much larger MMN was elicited by pattern violations in the 6 than in the 1 semitone condition. The difficult visual task should have presumably required greater attentional focus than the easy task, and performance did deteriorate during the difficult MOT. The MMN, however, was not affected by the demands of the MOT task. This finding suggests that the MMN elicited by the violation of a pattern is not affected by the presumed attentional demands of a difficult continuous task such as multiple object tracking. PMID- 18674523 TI - Brain-derived neurotrophic factor (BDNF) and TrkB in the piglet brainstem after post-natal nicotine and intermittent hypercapnic hypoxia. AB - Brain-derived neurotrophic factor (BDNF) and its receptor TrkB play a significant role in the regulation of cell growth, survival and death during central nervous system development. The expression of BDNF and TrkB is affected by noxious insults. Two insults during the early post-natal period that are of interest to our laboratory are exposure to nicotine and to intermittent hypercapnic hypoxia (IHH). Piglet models were used to mimic the conditions associated with the risk factors for the sudden infant death syndrome (SIDS) including post-natal cigarette smoke exposure (nicotine model) and prone sleeping where the infant is subjected to re-breathing of expired gases (IHH model). We aimed to determine the effects of nicotine and IHH, alone or in combination, on pro- and rhBDNF and TrkB expression in the developing piglet brainstem. Four piglet groups were studied, with equal gender ratios in each: control (n=14), nicotine (n=14), IHH (n=10) and nic+IHH (n=14). Applying immunohistochemistry, and studying six nuclei of the caudal medulla, we found that compared to controls, TrkB was the only protein significantly decreased after nicotine and nic+IHH exposure regardless of gender. For pro-BDNF and rhBDNF however, observed changes were more evident in males than females exposed to nicotine and nic+IHH. The implications of these findings are that a prior nicotine exposure makes the developing brainstem susceptible to greater changes in the neurotrophic effects of BDNF and its receptor TrkB in the face of a hypoxic insult, and that the effects are greater in males than females. PMID- 18674521 TI - PDE5 inhibitors enhance tumor permeability and efficacy of chemotherapy in a rat brain tumor model. AB - The blood-brain tumor barrier (BTB) significantly limits delivery of therapeutic concentrations of chemotherapy to brain tumors. A novel approach to selectively increase drug delivery is pharmacologic modulation of signaling molecules that regulate BTB permeability, such as those in cGMP signaling. Here we show that oral administration of sildenafil (Viagra) and vardenafil (Levitra), inhibitors of cGMP-specific PDE5, selectively increased tumor capillary permeability in 9L gliosarcoma-bearing rats with no significant increase in normal brain capillaries. Tumor-bearing rats treated with the chemotherapy agent, adriamycin, in combination with vardenafil survived significantly longer than rats treated with adriamycin alone. The selective increase in tumor capillary permeability appears to be mediated by a selective increase in tumor cGMP levels and increased vesicular transport through tumor capillaries, and could be attenuated by iberiotoxin, a selective inhibitor for calcium-dependent potassium (K(Ca)) channels, that are effectors in cGMP signaling. The effect by sildenafil could be further increased by simultaneously using another BTB "opener", bradykinin. Collectively, this data demonstrates that oral administration of PDE5 inhibitors selectively increases BTB permeability and enhances anti-tumor efficacy for a chemotherapeutic agent. These findings have significant implications for improving delivery of anti-tumor agents to brain tumors. PMID- 18674522 TI - Neonatal intrahippocampal injection of the HIV-1 proteins gp120 and Tat: differential effects on behavior and the relationship to stereological hippocampal measures. AB - HIV-1 proteins, such as Tat and gp120, are believed to play a crucial role in the central nervous system (CNS) pathology of acquired immune deficiency syndrome (AIDS). The present study sought to determine the potential role of Tat and/or gp120 on behavioral development and the relationship to the long-term effects of the HIV-1 proteins on the rat hippocampus. Male pups of 13 Sprague-Dawley litters were bilaterally injected on postnatal day (P)1. Every litter contributed an animal to each of four treatment condition: VEH (0.5 microl sterile buffer), gp120 (100 ng), Tat (25 microg) or combined gp120+Tat (100 ng+25 microg). Body weight was not affected by either protein treatment. Tat revealed a transient effect on many of the behavioral assessments early in development as well as on preattentive processes and spatial memory in adulthood. Gp120 had more selective effects on negative geotaxis (P8-P10) and on locomotor activity (P94-P96). Combined gp120+Tat effects were noted for eye opening with potential interactive effects of gp120 and Tat on negative geotaxis. Anatomical assessment at approximately 7 1/2 months of age was conducted by using design-based stereology to quantify the total cell number in five hippocampal subregions [granule layer (GL), hilus of the dentate gyrus (DGH), cornu ammonis fields (CA)2/3, CA1, and subiculum (SUB)] [Fitting, S., Booze, R.M., Hasselrot, U., Mactutus, C.F., 2007a. Differential long-term neurotoxicity of HIV-1 proteins in the rat hippocampal formation: a design-based stereological study. Hippocampus 18(2), 135-147]. A relationship between early reflex development and estimated cell number in the adult hippocampus was indicated by simple regression analyses. In addition, estimated number of neurons and astrocytes in the DGH explained 81% of the variance of the distribution of searching behavior in the probe test. Collectively, these data indicate that the DGH may participate in the spatial memory alterations observed in adulthood consequent to neonatal exposure to HIV-1 proteins. PMID- 18674524 TI - Arsenic-induced neurotoxicity in relation to toxicokinetics: effects on sciatic nerve proteins. AB - In our previous study in rats acutely exposed to As, we observed an effect of As on neurofilaments in the sciatic nerve. This study deals with the effects of inorganic As in Wistar rats on the cytoskeletal protein composition of the sciatic nerve after subchronic intoxication. Sodium meta-arsenite (NaAsO2) dissolved in phosphate-buffered saline (PBS) was administered daily in doses of 0, 3 and 10 mg/kg body weight/day (n=9 rats/group) by intragastric route for 4, 8 and 12 week periods. Toxicokinetic measurements revealed a saturation of blood As in the 3- and 10-mg/kg dose groups at approximately 14 microg/ml, with an increase in renal clearance of As at increasing doses. After exsanguination, sciatic nerves were excised and the protein composition was analyzed. Analysis of the sciatic nerves showed compositional changes in their proteins. Protein expression of neurofilament Medium (NF-M) and High (NF-H) was unchanged. Neurofilament protein Low (NF-L) expression was reduced, while mu- and m-calpain protein expression was increased, both in a dose/time pattern. Furthermore, NF-H protein was hypophosphorylated, while NF-L and microtubule-associated protein tau (MAP-tau) proteins were (hyper)-phosphorylated. In conclusion, we show that expression of mu- and m-calpain protein is increased by exposure to As, possibly leading to increased NF-L degradation. In addition, hyperphosphorylation of NF-L and MAP-tau by As also contribute to destabilization and disruption of the cytoskeletal framework, which eventually may lead to axonal degeneration. PMID- 18674525 TI - The clinical implications of increased OX40 ligand expression in patients with acute coronary syndrome. AB - BACKGROUND: Increasing evidence show that OX40 ligand (OX40L), also known as tumor necrosis factor superfamily member 4 (TNFSF4), plays an important role in the pathogenesis of atherosclerosis. We investigated whether expression levels of soluble OX40L in serum and of membrane OX40L on platelets were related to serum concentrations of matrix metalloproteinases (MMPs) and stability of coronary atherosclerotic plaque in patients with acute coronary syndrome (ACS). METHODS: We included healthy controls (n=30), patients with stable angina (SA) (n=40) and patients with ACS, including unstable angina (UA) (n=70) and acute myocardial infarction (AMI) (n=40). The expression of OX40L on platelets (pOX40L) was analyzed with flow cytometry whereas serum concentrations of soluble OX40L (sOX40L), MMP-9 and MMP-3 were determined with ELISA. All coronary stenoses with >or=30% diameter reduction were assessed by angiographic coronary stenosis morphology. RESULTS: The expression of OX40L on platelets were significantly higher in patients with ACS (61.5+/-11.5) compared with healthy controls (28.9+/ 7.4) or with the group of patients with SA (31.2+/-8.1) (mean fluorescence intensity+/-SD) (p<0.001). Similarly, we observed higher sOX40L concentrations in patients with ACS (34.6+/-9.3) compared with controls (10.2+/-4.7) or patients with SA (11.4+/-5.8) (ng/ml+/-SD) (p<0.001). Serum MMP-3 and MMP-9 levels in patients were two times greater than those in the control group. A positive correlation was observed between OX40L expression on platelets and MMP-9 and MMP 3 serum concentrations. OX40L expression on platelets were furthermore correlated with soluble OX40L in serum and with complex coronary stenoses (r1=0.61, r2=0.57, p<0.001). CONCLUSION: Patients with ACS show increased OX40L system (pOX40L and sOX40L) expression which may create a proinflammatory milieu for aggravating the development of atherosclerosis, and may be a valuable marker for predicting the severity of ACS. PMID- 18674526 TI - Evaluation of adsorption of urine cystatin C to the polymer materials on the microplate by an antigen capture enzyme-linked immunosorbent assay. AB - BACKGROUND: Cystatin C is a low molecular weight protein of 13 kDa with an isoelectric point of 9.3. Its adsorption on the urine sampling containers may cause the underestimation of cystatin C levels. We newly developed an antigen capture enzyme-linked immunosorbent assay (ELISA) of sandwich method for measurement of adsorbed level. METHODS: We used a polystyrene microplates with 3 different polymers. These include high hydrophobic, low hydrophobic, and hydrophilic materials. Using the same microplate, the absorbed protein was measured by an antigen Capture ELISA, and calibration was conducted by an ordinary ELISA. RESULTS: In normal urine the concentrations of absorbed cystatin C levels to the 3 materials at day 1 were 0.50, 0.32-0.84 microg/l (median, interquartile range), 0.28, 0.21-0.37 microg/l, and <0.08, <0.08-0.09 microg/l in high hydrophobic, low hydrophobic, and high hydrophilic material, respectively. The absorption rate was 6%, 3%, and 1%, respectively. The adsorption is dependent on urine pH. It changes reciprocally with urine protein concentration. In pathologic urine, the absolute absorption level was <0.08 microg/l on the median, and the adsorption ratio (absorption level/urine level) was much less than 0.5% of that in normal urine. CONCLUSION: In the clinical setting, the absorption of cystatin C to sample containers is negligible since the rate of adsorption is low both in normal and pathologic urine. The material with high hydrophilic surface processing may be used for other proteins when interaction of the proteins with surface material affects the value to clinical decision. PMID- 18674527 TI - Patient selection has a strong impact on cystatin C and Modification of Diet in Renal Disease (MDRD) estimated glomerular filtration rate. AB - OBJECTIVE: Estimation of the glomerular filtration rate (GFR) is essential for the evaluation of patients with kidney disease, and for correct dosage of drugs that are eliminated from the circulation by the kidneys. In most cases GFR is estimated based on serum creatinine and the Modification of Diet in Renal Disease (MDRD) formula. As both cystatin C and creatinine are used for the determination of GFR it is important to investigate if estimated GFR by the two methods differ in various patient groups. DESIGN AND METHODS: We have compared cystatin C and MDRD estimated GFR calculated from the same request from primary care units (n=488), a cardiology ward (n=826), the cardiointensive care unit (n=1026), two oncology wards (n=919 and 1021), and the neurosurgical intensive care unit (n=1515) in an observational cross-sectional study. RESULTS: We found better agreement between the two GFR estimates in samples from primary care patients and patients in the cardiology wards, than in samples from oncology wards or the neurosurgical intensive care unit. In the latter settings there was a pronounced difference between the two GFR estimates. CONCLUSION: The comparisons show that differences in patient selections have a strong impact on the agreement between cystatin C and MDRD estimated glomerular filtration rate. PMID- 18674528 TI - Sensory mechanisms controlling the timing of larval developmental and behavioral transitions require the Drosophila DEG/ENaC subunit, Pickpocket1. AB - Growth of multicellular organisms proceeds through a series of precisely timed developmental events requiring coordination between gene expression, behavioral changes, and environmental conditions. In Drosophila melanogaster larvae, the essential midthird instar transition from foraging (feeding) to wandering (non feeding) behavior occurs prior to pupariation and metamorphosis. The timing of this key transition is coordinated with larval growth and size, but physiological mechanisms regulating this process are poorly understood. Results presented here show that Drosophila larvae associate specific environmental conditions, such as temperature, with food in order to enact appropriate foraging strategies. The transition from foraging to wandering behavior is associated with a striking reversal in the behavioral responses to food-associated stimuli that begins early in the third instar, well before food exit. Genetic manipulations disrupting expression of the Degenerin/Epithelial Sodium Channel subunit, Pickpocket1(PPK1) or function of PPK1 peripheral sensory neurons caused defects in the timing of these behavioral transitions. Transient inactivation experiments demonstrated that sensory input from PPK1 neurons is required during a critical period early in the third instar to influence this developmental transition. Results demonstrate a key role for the PPK1 sensory neurons in regulation of important behavioral transitions associated with developmental progression of larvae from foraging to wandering stage. PMID- 18674529 TI - Spatial and temporal segregation of auditory and vestibular neurons in the otic placode. AB - The otic placode generates the auditory and vestibular sense organs and their afferent neurons; however, how auditory and vestibular fates are specified is unknown. We have generated a fate map of the otic placode and show that precursors for vestibular and auditory cells are regionally segregated in the otic epithelium. The anterior-lateral portion of the otic placode generates vestibular neurons, whereas the posterior-medial region gives rise to auditory neurons. Precursors for vestibular and auditory sense organs show the same distribution. Thus, different regions of the otic placode correspond to particular sense organs and their innervating neurons. Neurons from contiguous domains rarely intermingle suggesting that the regional organisation of the otic placode dictates positional cues to otic neurons. But, in addition, vestibular and cochlear neurogenesis also follows a stereotyped temporal pattern. Precursors from the anterior-lateral otic placode delaminate earlier than those from its medial-posterior portion. The expression of the proneural genes NeuroM and NeuroD reflects the sequence of neuroblast formation and differentiation. Both genes are transiently expressed in vestibular and then in cochlear neuroblasts, while differentiated neurons express Islet1, Tuj1 and TrkC, but not NeuroM or NeuroD. Together, our results indicate that the position of precursors within the otic placode confers identity to sensory organs and to the corresponding otic neurons. In addition, positional information is integrated with temporal cues that coordinate neurogenesis and sensory differentiation. PMID- 18674530 TI - Effect of propofol on calcium homeostasis in hypoxia-reoxygenated neonatal rat cardiomyocytes. AB - Intracellular Ca2+ overload induced by hypoxia-reoxygenation alters Ca2+ homeostasis, which plays an important role in myocardial cell injury. Even though propofol is known as a radical scavenger with Ca2+ channel blocking properties, little is known about cardioprotective effect associated with Ca2+ homeostasis in cardiomyocytes. In the present study, we showed that propofol protects cardiomyocytes against hypoxia-reoxygenation injury. In propofol-treated cardiomyocytes, we observed a decrease in the expression of pro-apoptotic protein Bax, cytochrome c, caspase-3 activation and intracellular Ca2+ content. We also found that propofol treatment enhanced expression of anti-apoptotic protein Bcl-2 and activation of ERK concerned with survival. Propofol attenuated alterations of genes involving Ca2+-regulatory mechanism and significantly modulated abnormal changes of SERCA2a genes in hypoxia-reoxygenated neonatal cardiomyocytes. These results suggest that propofol modulates the expression of genes involved in Ca2+ homeostasis, thereby producing cardioprotective effects through a reduction in apoptotic cell death. PMID- 18674531 TI - Melatonin attenuates tyrosine hydroxylase loss and hypolocomotion in MPTP lesioned rats. AB - Parkinson's disease is a chronic neurological disease characterized by dopaminergic neuron degeneration in the substantia nigra pars compacta. Melatonin is a powerful antioxidant agent secreted by the pineal gland which has numerous physiological functions and seems to exert an important neuroprotective effect. The 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model has been used to understand the pathophysiology of the disease because of its capacity to mimic biochemical and histological features observed in Parkinson's disease. This study investigated the effect of pretreatment with melatonin (50 mg/kg) on MPTP lesioned animals 24 h and 7 days after neurotoxin infusion using the open-field test, two-way avoidance task and immunohistochemistry. Twenty-four hours after lesioning, the MPTP+vehicle group exhibited hypolocomotion and significant loss of tyrosine hydroxylase-immunoreactive cells, whereas no differences in these parameters were observed in lesioned animals receiving melatonin. Seven days after surgery, the MPTP-lesioned rats did not show hypolocomotion compared to control animals, while there was a significant dopaminergic neuronal loss. In the two-way avoidance task, MPTP-treated animals presented a cognitive deficit compared to the control groups and melatonin administration did not repair this defect. The present results suggest that melatonin reduces neuronal loss in the MPTP animal model of Parkinson's disease. PMID- 18674532 TI - Nitric oxide and glutamate in the dorsal facial area regulate common carotid blood flow in the cat. AB - Nitric oxide (NO) or glutamate stimulation of dorsal facial area (DFA) increases blood flow in the common carotid artery (CCA), which supplies intra-and extra cranial tissues. Nitrergic fibers and neurons as well as preganglionic cholinergic neurons are present in the DFA. We hypothesized the presence of nitrergic-glutamatergic fibers and preganglionic nitrergic-cholinergic neurons in the DFA that are involved in the regulation of CCA blood flow. In microdialysis studies, perfusion of the DFA with S-nitroso-N-acetylpenicillamine (SNAP, an NO donor) increased the glutamate concentration in the dialysate. This effect was abolished by co-perfusion of methylene blue (a guanylyl cyclase inhibitor). Intra DFA injection of l-arginine (an NO precursor) or glutamate increased CCA blood flow. The l-arginine-induced flow increase was reduced by prior administration of NG-nitro-arginine methyl ester (l-NAME, a non-specific NO synthase inhibitor), 7 nitroindazole (7-NI, a relatively selective neuronal NO synthase inhibitor), d-2 amino-5-phosphonopentanoate (d-AP5, a competitive NMDA receptor antagonist), or glutamate diethylester (GDEE, a competitive AMPA receptor antagonist). The glutamate-induced blood flow increase was reduced by prior administration of l NAME, 7-NI, or methylene blue. The induced increase in CCA blood flow, however, was not affected by endothelial NO synthase inhibitor. The findings indicate that NO-signal transduction within the DFA might cause glutamate release from presynaptic nitrergic-glutamatergic fibres and that the released glutamate activates NMDA/AMPA receptors on preganglionic nitrergic-cholinergic neurons in the nucleus to activate neuronal NO synthase and guanylyl cyclase in the neurons, leading to an increase in CCA blood flow. These findings may be important for developing therapeutic strategies for the diseases associated with CCA blood flow. PMID- 18674533 TI - Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei. AB - SR proteins are multidomain splicing factors which are important for spliceosome assembly and for regulation of alternative splicing. In mammalian nuclei these proteins localise to speckles from where they are recruited to transcription sites. By using fluorescent protein fusion technology and different experimental approaches it has been shown that Arabidopsis SR proteins, in addition to diffuse nucleoplasmic staining, localise into an irregular nucleoplasmic network resembling speckles in mammalian cells. As Arabidopsis SR proteins fall into seven conserved sub-families we investigated co-localisation of members of the different sub-families in transiently transformed tobacco protoplast. Here we demonstrate the new finding that members of different SR protein sub-families localise into distinct populations of nuclear speckles with no, partial or complete co-localisation. This is particularly interesting as we also show that these proteins do interact in a yeast two-hybrid assay as well as in pull-down and in co-immunopreciptiation assays. Our data raise the interesting possibility that SR proteins are partitioned into distinct populations of nuclear speckles to allow a more specific recruitment to the transcription/pre-mRNA processing sites of particular genes depending on cell type and developmental stage. PMID- 18674534 TI - Disruption of cell-matrix interactions by heparin enhances mesenchymal progenitor adipocyte differentiation. AB - Differentiation of marrow-derived mesenchymal progenitors to either the osteoblast or adipocyte lineage is reciprocally regulated. Factors that promote osteoblastogenesis inhibit adipogenesis, while adipogenic factors are inhibitory to osteoblast differentiation. Heparin, a soluble glycosaminoglycan, inhibits bone formation in vivo and osteoblast cell differentiation and function in vitro, and has been shown to promote adipocyte differentiation. To elucidate the role that heparin plays in the adipogenic induction of murine mesenchymal progenitors, we studied immortalized marrow stromal cells (IM-MSC), the MSC cell line, ST2, and 3T3L1 pre-adipocytes. Heparin alone was not sufficient to induce adipogenesis, but enhanced the induction under a variety of adipogenic cocktails. This effect was both dose- and time-dependent. Heparin showed a positive effect at concentrations > 0.1 microg/ml when applied before day 3 during the induction course. Heparin's effect on adipogenesis was independent of cell proliferation, cell density, and extracellular lipid. This effect is likely related to the unique structure of heparin because another polyanionic glycosaminoglycan, dextran sulfate, did not promote adipogenic differentiation. Heparin treatment altered morphology and adhesion characteristics of progenitor cells, resulting in cell rounding and aggregation. As well, heparin counteracted the known inhibitory effect of fibronectin on adipogenesis and decreased basal focal adhesion kinase and paxillin phosphorylation. We conclude that heparin-mediated disruption of cell-matrix adhesion enhances adipogenic potential. PMID- 18674535 TI - Characterization of monoclonal antibodies against the glaucoma-associated protein myocilin. AB - Although the glaucoma-associated protein myocilin has been the focus of intensive research, its biological function is still unknown. One of the limiting factors has been the lack of well-characterized antibodies, particularly monoclonal antibodies. We describe the development of six monoclonal antibodies specific to myocilin and characterize their suitability in Western blot and immunohistochemical applications. Three of the six monoclonal antibodies recognize the N-terminus of myocilin (amino acids 33-214), two antibodies recognize the middle third of the protein (amino acids 215-368), and one antibody recognizes the C-terminus (amino acids 369-504). Isotyping revealed that all antibodies are of the IgG1 kappa class except one, which is IgG2b kappa. Purified myocilin monoclonal antibodies were able to recognize myocilin in human aqueous humor separated on denatured/reduced and native gels, and human trabecular meshwork lysate by Western blot. Myocilin was also detected by immunohistochemistry in trabecular meshwork, ciliary body, iris, cornea, sclera, choroid, and retinal pigment epithelial cells. PMID- 18674536 TI - Haplorchis taichui: worm recovery rate and immune responses in infected rats (Rattus norvegicus). AB - Worm recovery rate, mucosal mast cells (MMCs), eosinophils and serum IgE concentration in rats were investigated after orally feeding 300 Haplorchis taichui metacercariae to male rats. The duodenal, jejunal and ileal tissue sections were stained with 1% alcian blue and 0.5% safranin-O for MMC count. Eosinophil count and the serum IgE concentration assay were measured from cardiac puncture blood. The average worm recovery rates were 20.00%, 13.00%, 0.67%, 1.67% and 0.00% on day 3, 7, 14, 21 and 28 post-infection (PI), respectively. The number of MMCs in the infected rats were significantly higher than in the controls (P<0.01), reaching a peak on day 21 PI. They decreased thereafter, with the decline in worm recovery. Eosinophil count and Serum IgE concentration were also increased but not significantly higher than the controls. However, they showed a positive relationship to worm recovery. It could be concluded from the results that MMCs, eosinophils and IgE may play an important role in the expulsion of H. taichui from rat intestine. However, the mechanism by which the MMC result in the helminth expulsion still need to be understood, and it is recommended that other cells such as goblet cells be studied further. PMID- 18674537 TI - The [4Fe-4S] cluster of quinolinate synthase from Escherichia coli: investigation of cluster ligands. AB - Nicotinamide adenine dinucleotide (NAD) derives from quinolinic acid which is synthesized in Escherichia coli from l-aspartate and dihydroxyacetone phosphate through the concerted action of l-aspartate oxidase and the [4Fe-4S] quinolinate synthase (NadA). Here, we addressed the question of the identity of the cluster ligands. We performed in vivo complementation experiments as well as enzymatic, spectroscopic and structural in vitro studies using wild-type vs. Cys-to-Ala mutated NadA proteins. These studies reveal that only three cysteine residues, the conserved Cys113, Cys200 and Cys297, are ligands of the cluster. This result is in contrast to the previous proposal that pointed the three cysteines of the C(291)XXC(294)XXC(297) motif. Interestingly, we demonstrated that Cys291 and Cys294 form a disulfide bridge and are important for activity. PMID- 18674538 TI - Myeloid-derived suppressor cells in inflammatory bowel disease: a new immunoregulatory pathway. AB - BACKGROUND & AIMS: CD11b(+)Gr-1(+) myeloid-derived suppressor cells (MDSCs) have been shown to cause T-cell tolerance in tumor-bearing mice; however, little is known about the role of MDSCs in chronic inflammation. Here, for the first time, we have identified and analyzed their role in inflammatory bowel disease (IBD). METHODS: Repetitive adoptive transfer of clone 4/T-cell receptor (CL4-TCR) transgenic CD8(+) T cells into VILLIN-hemagglutinin (HA) transgenic mice was performed on days 1, 12, and 27. Recipient mice were analyzed for immunopathology, HA-specific CD8(+) T-cell responses, and CD11b(+)Gr-1(+) MDSCs (frequency, phenotype, expression analysis, and in vitro as well as in vivo function). In addition, peripheral blood from patients with active Crohn's disease and ulcerative colitis was examined for the presence and function of human MDSCs denoted as CD14(+)HLA-DR(-/low) cells. RESULTS: Repetitive transfer of HA-specific CD8(+) T cells prevented VILLIN-HA recipient mice from development of severe enterocolitis, which is seen after a single transfer of T cells. Repeated transfer of antigen-specific T cells led to an increase in the frequency of nitric oxide synthase 2 and arginase-expressing CD11b(+)Gr-1(+) MDSCs in spleen and intestine of VILLIN-HA mice with immunosuppressive function. Cotransfer of MDSCs with HA-specific CD8(+) T cells into naive VILLIN-HA mice ameliorated enterocolitis, indicating a direct immune regulatory effect of MDSCs on induction of IBD by antigen-specific T cells. Finally, an increase in the frequency of human MDSCs with suppressor function was observed in peripheral blood from patients with IBD. CONCLUSIONS: These results identify MDSCs as a new immune regulatory pathway in IBD. PMID- 18674539 TI - Late-stage alterations in myofibrillar contractile function in a transgenic mouse model of dilated cardiomyopathy (Tgalphaq*44). AB - Mechanical and biochemical alterations were investigated in permeabilized cardiomyocytes along with the progression of dilated cardiomyopathy (DCM) in a transgenic mouse line overexpressing the activated Galphaq protein (Tgalphaq*44). The isometric force, its Ca(2+) sensitivity (pCa(50)) and the turnover rate of the actin-myosin cycle (k(tr)) were determined at sarcomere lengths (SLs) of 1.9 mum and 2.3 mum before (at 4 and 10 months of age) and after hemodynamic decompensation (at 14 and 18 months of age) in Tgalphaq*44 cardiomyocytes and in age-matched control cardiomyocytes. The SL-dependence of pCa(50) was not different in Tgalphaq*44 and control hearts. In contrast, a significant increase in pCa(50) was observed in the Tgalphaq*44 cardiomyocytes (DeltapCa(50): 0.10 0.15 vs. the controls) after 10 months of age that could be diminished by exposures to the catalytic subunit of protein kinase A (PKA). Accordingly, a decline in endogenous PKA activity and decreased troponin I phosphorylation were detected after 10 months in the Tgalphaq*44 hearts. Finally, the maximal Ca(2+) activated force (F(o)) and k(tr) were lower and the passive force (F(passive)) was higher at 18 months in the Tgalphaq*44 cardiomyocytes compared to the control. These mechanical alterations were paralleled by a robust increase in beta-myosin heavy chain expression in the Tgalphaq*44 hearts. In conclusion, our data suggested that an initial decrease of PKA signaling and subsequent changes in myofilament protein expression may contribute to the development of dilated cardiomyopathy in Tgalphaq*44 hearts. PMID- 18674540 TI - Specific heteromeric association of four transmembrane peptides derived from platelet glycoprotein Ib-IX complex. AB - As the receptor on the platelet surface for von Willebrand factor, glycoprotein (GP) Ib-IX complex is critically involved in hemostasis and thrombosis. How the complex is assembled from GP Ibalpha, GP Ib beta and GP IX subunits, all of which are type I transmembrane proteins, is not entirely clear. Genetic and mutational analyses have identified the transmembrane (TM) domains of these subunits as active participants in assembly of the complex. In this study, peptides containing the transmembrane domain of each subunit have been produced and their interaction with one another characterized. Only the Ib beta TM sequence, but not the Ibalpha and IX counterparts, can form homo-oligomers in SDS-PAGE and TOXCAT assays. Following up on our earlier observation that a Ib beta-Ibalpha-Ib beta peptide complex (alphabeta(2)) linked through native juxtamembrane disulfide bonds could be produced from isolated Ibalpha and Ib beta TM peptides in detergent micelles, we show here that addition of the IX TM peptide facilitates formation of the native alphabeta(2) complex, reproducing the same effect by the IX subunit in cells expressing the GP Ib-IX complex. Specific fluorescence resonance energy transfer was observed between donor-labeled alphabeta(2) peptide complex and acceptor-conjugated IX TM peptide in micelles. Finally, the mutation D135K in the IX TM peptide could hamper both the formation of the alphabeta(2) complex and the energy transfer, consistent with its reported effect in the full length complex. Overall, our results have demonstrated directly the native-like heteromeric interaction among the isolated Ibalpha, Ib beta and IX TM peptides, which provides support for the four-helix bundle model of the TM domains in the GP Ib-IX complex and paves the way for further structural analysis. The methods developed in this study may be applicable to other studies of heteromeric interaction among multiple TM helices. PMID- 18674541 TI - Experimental evidence for the existence of a stable half-barrel subdomain in the (beta/alpha)8-barrel fold. AB - The (beta/alpha)(8)-barrel is one of the most common folds functioning as enzymes. The emergence of two (beta/alpha)(8)-barrel enzymes involved in histidine biosynthesis, each of which has a twofold symmetric structure, has been proposed to be a consequence of tandem duplication and fusion of a (beta/alpha)(4)-half-barrel. However, little evidence has been found for the existence of an ancestral half-barrel in the evolution of other (beta/alpha)(8) barrel proteins. In order to detect remnants of an ancestral half-barrel in the (beta/alpha)(8)-barrel structure of Escherichia coli N-(5' phosphoribosyl)anthranilate isomerase, we engineered three potential half-barrel units, (beta/alpha)(1-4), (beta/alpha)(3-6), and (beta/alpha)(5-8). Among these three arrangements, only (beta/alpha)(3-6) is stable; it exists in equilibrium between monomeric and dimeric forms. Thus, the central segment of N-(5' phosphoribosyl)anthranilate isomerase from E. coli can serve as a half-barrel precursor. A tandem duplication of (beta/alpha)(3-6) yielded predominantly monomeric structures that were quite stable. This result exemplified that the structural characteristics of noncovalently assembled half-barrels could be improved by duplication and fusion. Moreover, our results may provide information regarding the local structural units that encompass interactions important for the early folding events of this ubiquitous protein conformation. PMID- 18674542 TI - Crystal structure and mutational analysis of Ca2+-independent type II antifreeze protein from longsnout poacher, Brachyopsis rostratus. AB - We recently found that longsnout poacher (Brachyosis rostratus) produces a Ca(2+) independent type II antifreeze protein (lpAFP) and succeeded in expressing recombinant lpAFP using Phichia pastoris. Here, we report, for the first time, the X-ray crystal structure of lpAFP at 1.34 A resolution. The lpAFP structure displayed a relatively planar surface, which encompasses two loop regions (Cys86 Lys89 and Asn91-Cys97) and a short beta-strand (Trp109-Leu112) with three unstructured segments (Gly57-Ile58, Ala103-Ala104, and Pro113-His118). Electrostatic calculation of the protein surface showed that the relatively planar surface was divided roughly into a hydrophobic area (composed of the three unstructured segments lacking secondary structure) and a hydrophilic area (composed of the loops and beta-strand). Site-directed mutation of Ile58 with Phe at the center of the hydrophobic area decreased activity significantly, whereas mutation of Leu112 with Phe at an intermediate area between the hydrophobic and hydrophilic areas retained complete activity. In the hydrophilic area, a peptide swap mutant in the loops retained 60% activity despite simultaneous mutations of eight residues. We conclude that the epicenter of the ice-binding site of lpAFP is the hydrophobic region, which is centered by Ile58, in the relatively planar surface. We built an ice-binding model for lpAFP on the basis of a lattice match of ice and constrained water oxygen atoms surrounding the hydrophobic area in the lpAFP structure. The model in which lpAFP has been docked to a secondary prism (2 1-10) plane, which is different from the one determined for Ca(2+)-independent type II AFP from sea raven (11-21), appears to explain the results of the mutagenesis analysis. PMID- 18674543 TI - Cotranslational folding promotes beta-helix formation and avoids aggregation in vivo. AB - Newly synthesized proteins must form their native structures in the crowded environment of the cell, while avoiding non-native conformations that can lead to aggregation. Yet, remarkably little is known about the progressive folding of polypeptide chains during chain synthesis by the ribosome or of the influence of this folding environment on productive folding in vivo. P22 tailspike is a homotrimeric protein that is prone to aggregation via misfolding of its central beta-helix domain in vitro. We have produced stalled ribosome:tailspike nascent chain complexes of four fixed lengths in vivo, in order to assess cotranslational folding of newly synthesized tailspike chains as a function of chain length. Partially synthesized, ribosome-bound nascent tailspike chains populate stable conformations with some native-state structural features even prior to the appearance of the entire beta-helix domain, regardless of the presence of the chaperone trigger factor, yet these conformations are distinct from the conformations of released, refolded tailspike truncations. These results suggest that organization of the aggregation-prone beta-helix domain occurs cotranslationally, prior to chain release, to a conformation that is distinct from the accessible energy minimum conformation for the truncated free chain in solution. PMID- 18674544 TI - The Cys3-Cys4 loop of the hydrophobin EAS is not required for rodlet formation and surface activity. AB - Class I hydrophobins are fungal proteins that self-assemble into robust amphipathic rodlet monolayers on the surface of aerial structures such as spores and fruiting bodies. These layers share many structural characteristics with amyloid fibrils and belong to the growing family of functional amyloid-like materials produced by microorganisms. Although the three-dimensional structure of the soluble monomeric form of a class I hydrophobin has been determined, little is known about the molecular structure of the rodlets or their assembly mechanism. Several models have been proposed, some of which suggest that the Cys3 Cys4 loop has a critical role in the initiation of assembly or in the polymeric structure. In order to provide insight into the relationship between hydrophobin sequence and rodlet assembly, we investigated the role of the Cys3-Cys4 loop in EAS, a class I hydrophobin from Neurospora crassa. Remarkably, deletion of up to 15 residues from this 25-residue loop does not impair rodlet formation or reduce the surface activity of the protein, and the physicochemical properties of rodlets formed by this mutant are indistinguishable from those of its full-length counterpart. In addition, the core structure of the truncation mutant is essentially unchanged. Molecular dynamics simulations carried out on the full length protein and this truncation mutant binding to an air-water interface show that, although it is hydrophobic, the loop does not play a role in positioning the protein at the surface. These results demonstrate that the Cys3-Cys4 loop does not have an integral role in the formation or structure of the rodlets and that the major determinant of the unique properties of these proteins is the amphipathic core structure, which is likely to be preserved in all hydrophobins despite the high degree of sequence variation across the family. PMID- 18674545 TI - High-throughput analysis of the protein sequence-stability landscape using a quantitative yeast surface two-hybrid system and fragment reconstitution. AB - Stability evaluation of many mutants can lead to a better understanding of the sequence determinants of a structural motif and of factors governing protein stability and protein evolution. The traditional biophysical analysis of protein stability is low throughput, limiting our ability to widely explore sequence space in a quantitative manner. In this study, we have developed a high throughput library screening method for quantifying stability changes, which is based on protein fragment reconstitution and yeast surface display. Our method exploits the thermodynamic linkage between protein stability and fragment reconstitution and the ability of the yeast surface display technique to quantitatively evaluate protein-protein interactions. The method was applied to a fibronectin type III (FN3) domain. Characterization of fragment reconstitution was facilitated by the co-expression of two FN3 fragments, thus establishing a yeast surface two-hybrid method. Importantly, our method does not rely on competition between clones and thus eliminates a common limitation of high throughput selection methods in which the most stable variants are recovered predominantly. Thus, it allows for the isolation of sequences that exhibit a desired level of stability. We identified more than 100 unique sequences for a beta-bulge motif, which was significantly more informative than natural sequences of the FN3 family in revealing the sequence determinants for the beta-bulge. Our method provides a powerful means for the rapid assessment of the stability of many variants, for the systematic assessment of the contribution of different factors to protein stability, and for enhancement of the protein stability. PMID- 18674546 TI - A stochastic model of evolutionary dynamics with deterministic large-population asymptotics. AB - An evolutionary birth-death process is proposed as a model of evolutionary dynamics. Agents residing in a continuous spatial environment X, play a game G, with a continuous strategy set S, against other agents in the environment. The agents' positions and strategies continuously change in response to other agents and to random effects. Agents spawn asexually at rates that depend on their current fitness, and agents die at rates that depend on their local population density. Agents' individual evolutionary trajectories in X and S are governed by a system of stochastic ODEs. When the number of agents is large and distributed in a smooth density on (X,S), the collective dynamics of the entire population is governed by a certain (deterministic) PDE, which we call a fitness-diffusion equation. PMID- 18674548 TI - Liver Kupffer cells control the magnitude of the inflammatory response in the injured brain and spinal cord. AB - The CNS inflammatory response is regulated by hepatic chemokine synthesis, which promotes leukocytosis and facilitates leukocyte recruitment to the site of injury. To understand the role of the individual cell populations in the liver during the hepatic response to acute brain injury, we selectively depleted Kupffer cells (KC), using clodronate-filled liposomes, and assessed the inflammatory response following a microinjection of IL-1beta into the rat brain or after a compression injury in the spinal cord. We show by immunohistochemistry that KC depletion reduces neutrophil infiltration into the IL-1beta-injected brain by 70% and by 50% into the contusion-injured spinal cord. qRT-PCR analysis of hepatic chemokine mRNA expression showed that chemokine expression in the liver after brain injury is not restricted to a single cell population. In non depleted rats, CXCL-10, IL-1beta, CCL-2, and MIP-1alpha mRNAs were increased up to sixfold more than in KC depleted rats. However, CXCL-1 and MIP-1beta were not significantly affected by KC depletion. The reduction in chemokine mRNA expression by the liver was not associated with decreased neutrophil mobilisation as might have been expected. These findings suggest that in response to CNS injury, KC mediated mechanisms are responsible for increasing neutrophil entry to the site of CNS injury, but that neutrophil mobilisation is dependent on other non-KC mediated events. However, the suppression of KC activity may prevent secondary damage after acute brain injury. PMID- 18674547 TI - Post-hypoxic changes in rat cortical neuron GABA A receptor function require L type voltage-gated calcium channel activation. AB - Hypoxia modifies GABA(A) receptor (GABA(A)R) function and can cause seizures, encephalopathy or myoclonus. To characterize the effects of hypoxia on neuronal GABA(A)Rs, we subjected rat cortical neurons to 1% O2 for 2, 4 or 8h, followed by recovery times of 0-96h, and used whole-cell and perforated patch-clamp recording to assess GABA(A)R currents and pharmacology. Hypoxic exposure for 4h caused downregulation of maximal GABA current immediately following hypoxia and after 48h recovery without changing the EC50 for GABA. Two- and eight-hour hypoxic exposures had inconsistent effects on GABA(A)R currents. Maximal diazepam potentiation was increased immediately following 4h hypoxia, while potentiation by zolpidem was increased after 48h recovery. Pentobarbital enhancement and zinc inhibition of GABA currents were unchanged. Hypoxia also caused a depolarizing shift in the reversal potential of GABA-induced Cl(-) currents after 24h recovery. The L-type voltage-gated calcium channel (L-VGCC) blocker, nitrendipine, during hypoxia or control treatment prevented the reduction in GABA(A)R currents, and increased control currents over baseline. Nitrendipine also prevented the increase in zolpidem potentiation 48h after hypoxia, and blocked the depolarizing shift in Cl(-) reversal potential 24h after hypoxia. The effects of hypoxia on maximal GABA(A)R currents, zolpidem pharmacology and Cl(-) reversal potential thus require depolarization-induced calcium entry via L-VGCCs, and constitutive L-VGCC activity appears to reduce maximal GABA(A)R currents in control neurons via a calcium-dependent process. Calcium-dependent modulation of GABA(A)R currents via L-VGCCs may be a fundamental regulatory mechanism for GABA receptor function. PMID- 18674549 TI - The novel selective PDE9 inhibitor BAY 73-6691 improves learning and memory in rodents. AB - The present study investigated the putative pro-cognitive effects of the novel selective PDE9 inhibitor BAY 73-6691. The effects on basal synaptic transmission and long-term potentiation (LTP) were investigated in rat hippocampal slices. Pro cognitive effects were assessed in a series of learning and memory tasks using rodents as subjects. BAY 73-6691 had no effect on basal synaptic transmission in hippocampal slices prepared from young adult (7- to 8-week-old) Wistar rats. A dose of 10 microM, but not 30 microM, BAY 73-6691 enhanced early LTP after weak tetanic stimulation. The dose effective in young adult Wistar rats did not affect LTP in hippocampal slices prepared from young (7- to 8-week-old) Fischer 344 X Brown Norway (FBNF1) rats, probably reflecting strain differences. However, it increased basal synaptic transmission and enhanced early LTP after weak tetanic stimulation in hippocampal slices prepared from very old (31- to 35-month-old) FBNF1 rats. BAY 73-6691 enhanced acquisition, consolidation, and retention of long-term memory (LTM) in a social recognition task and tended to enhance LTM in an object recognition task. Bay 73-6691 attenuated the scoplamine-induced retention deficit in a passive avoidance task, and the MK-801-induced short-term memory deficits in a T-maze alternation task. The mechanism of action, possibly through modulation of the NO/cGMP-PKG/CREB pathway, is discussed. Our findings support the notion that PDE9 inhibition may be a novel target for treating memory deficits that are associated with aging and neurodegenerative disorders such as Alzheimer's disease. PMID- 18674550 TI - (+/-)-3,4-Methylenedioxymethamphetamine treatment in adult rats impairs path integration learning: a comparison of single vs once per week treatment for 5 weeks. AB - 3,4-Methlylenedioxymethamphetamine (MDMA) administration (4 x 15 mg/kg) on a single day has been shown to cause path integration deficits in rats. While most animal experiments focus on single binge-type models of MDMA use, many MDMA users take the drug on a recurring basis. The purpose of this study was to compare the effects of repeated single-day treatments with MDMA (4 x 15 mg/kg) once weekly for 5 weeks to animals that only received MDMA on week 5 and saline on weeks 1-4. In animals treated with MDMA for 5 weeks, there was an increase in time spent in the open area of the elevated zero maze suggesting a decrease in anxiety or increase in impulsivity compared to the animals given MDMA for 1 week and saline treated controls. Regardless of dosing regimen, MDMA treatment produced path integration deficits as evidenced by an increase in latency to find the goal in the Cincinnati water maze. Animals treated with MDMA also showed a transient hypoactivity that was not present when the animals were re-tested at the end of cognitive testing. In addition, both MDMA-treated groups showed comparable hyperactive responses to a later methamphetamine challenge. No differences were observed in spatial learning in the Morris water maze during acquisition or reversal but MDMA-related deficits were seen on reduced platform-size trials. Taken together, the data show that a single-day regimen of MDMA induces deficits similar to that of multiple weekly treatments. PMID- 18674551 TI - Don't look now or look away: two sources of saccadic disinhibition in Parkinson's disease? AB - Recent studies found evidence of impaired inhibition of saccades (fast eye movements) in non-demented people with PD. It has been suggested that impaired eye movement control reflects a general deficit of automatic response inhibition associated with impaired cognitive function in Parkinson's disease (PD). This study investigated the nature and source of saccadic disinhibition in PD. Eighteen non-demented PD patients and 18 control subjects completed prosaccade ('look towards'), delayed ('wait for cue') and antisaccade tasks ('look away') and a number of neuropsychological tests. There was evidence of saccadic disinhibition and cognitive impairment in the PD group. In the eye movement tasks the PD group made more express saccades (very fast reflexive responses) in the prosaccade task with a gap, more timing errors in the delayed response task and more directional errors in the antisaccade task than the control group. On average, neuropsychological test scores for the PD group were lower than for the control group. Subjects in the PD group who made a large number of directional errors in the antisaccade task did not necessarily also make a large number of timing errors in the delayed response task. Timing error rates, but not directional error rates, were negatively associated with neuropsychological test scores. Higher directional error rates in the antisaccade tasks were associated with higher proportions of express saccades in prosaccade tasks. This pattern of results suggests that there is more than one source of saccadic disinhibition in PD. We conclude that evidence of saccadic disinhibition may not necessarily reflect a general deficit of automatic response inhibition and cognitive impairment in PD. PMID- 18674552 TI - Progesterone suppresses an oxytocin-stimulated signal pathway in COS-7 cells transfected with the oxytocin receptor. AB - The present study was conducted to determine if progesterone (P4) would inhibit oxytocin-stimulated phosphoinositide hydrolysis in COS-7 cells expressing transfected ovine oxytocin receptor (OTR) with little or no nuclear P4 receptor (nPR) protein present. The relative absence of nPR in these cells was confirmed by immunocytochemistry and RT-PCR. To investigate the effects of P4 on oxytocin (OT) signaling, cells were transiently transfected with the ovine OTR. Radioreceptor assay for [(3)H]-OT binding confirmed the presence of a high affinity binding site for OT in transfected cells, while treatment with P4 and GTPgammaS (which uncouples the OTR from the heterotrimeric G-protein) increased the K(d) for OT binding slightly. Cells were then assayed for inositol phosphate hydrolysis 48 h post-transfection. Pre-treatment of cells with P4 for 10 min significantly interfered with rapid (20 min) OT-stimulated inositol trisphosphate (IP(3)) production. This inhibition was specific to P4, because pre-treatment of cells with promegestone (R5020), testosterone, mifepristone (RU 486), or cortisol did not decrease OT-stimulated IP(3) levels. By radioreceptor assay for PR, no measurable specific binding of R5020 was observed for either transfected or non transfected cells. We conclude that P4 can inhibit OTR-mediated phosphoinositide hydrolysis in COS-7 cells that express little or no nPR protein. These data support a role for a non-genomic action for P4 in OTR signaling via some mechanism other than by binding to a membrane progestin receptor in an immortalized, transfected cell. PMID- 18674553 TI - 7alpha- and 12alpha-Hydroxysteroid dehydrogenases from Acinetobacter calcoaceticus lwoffii: a new integrated chemo-enzymatic route to ursodeoxycholic acid. AB - We report the very efficient biotransformation of cholic acid to 7-keto- and 7,12 diketocholic acids with Acinetobacter calcoaceticus lwoffii. The enzymes responsible of the biotransformation (i.e. 7alpha- and 12alpha-hydroxysteroid dehydrogenases) are partially purified and employed in a new chemo-enzymatic synthesis of ursodeoxycholic acid starting from cholic acid. The first step is the 12alpha-HSDH-mediated total oxidation of sodium cholate followed by the Wolf Kishner reduction of the carbonyl group to chenodeoxycholic acid. This acid is then quantitatively oxidized with 7alpha-HSDH to 7-ketochenodeoxycholic acid, that was chemically reduced to ursodeoxycholic acid (70% overall yield). PMID- 18674554 TI - Particles from wood smoke and traffic induce differential pro-inflammatory response patterns in co-cultures. AB - The inflammatory potential of particles from wood smoke and traffic has not been well elucidated. In this study, a contact co-culture of monocytes and pneumocytes was exposed to 10-40 microg/cm(2) of particles from wood smoke and traffic for 12, 40 and 64 h to determine their influence on pro-inflammatory cytokine release (TNF-alpha, IL-1, IL-6, IL-8) and viability. To investigate the role of organic constituents in cytokine release the response to particles, their organic extracts and the washed particles were compared. Antagonists were used to investigate source-dependent differences in intercellular signalling (TNF-alpha, IL-1). The cytotoxicity was low after exposure to particles from both sources. However, wood smoke, and to a lesser degree traffic-derived particles, induced a reduction in cell number, which was associated with the organic fraction. The release of pro-inflammatory cytokines was similar for both sources after 12 h, but traffic induced a greater release than wood smoke particles with increasing exposure time. The organic fraction accounted for the majority of the cytokine release induced by wood smoke, whereas the washed traffic particles induced a stronger response than the corresponding organic extract. TNF-alpha and IL-1 antagonists reduced the release of IL-8 induced by particles from both sources. In contrast, the IL-6 release was only reduced by the IL-1 antagonist during exposure to traffic-derived particles. In summary, particles from wood smoke and traffic induced differential pro-inflammatory response patterns with respect to cytokine release and cell number. Moreover, the influence of the organic particle fraction and intercellular signalling on the pro-inflammatory response seemed to be source-dependent. PMID- 18674555 TI - Metabolomic profiling of a modified alcohol liquid diet model for liver injury in the mouse uncovers new markers of disease. AB - Metabolomic evaluation of urine and liver was conducted to assess the biochemical changes that occur as a result of alcohol-induced liver injury. Male C57BL/6J mice were fed an isocaloric control- or alcohol-containing liquid diet with 35% of calories from corn oil, 18% protein and 47% carbohydrate/alcohol for up to 36 days ad libitum. Alcohol treatment was initiated at 7 g/kg/day and gradually reached a final dose of 21 g/kg/day. Urine samples were collected at 22, 30 and 36 days and, in additional treatment groups, liver and serum samples were harvested at 28 days. Steatohepatitis was induced in the alcohol-fed group since a 5-fold increase in serum alanine aminotransferase activity, a 6-fold increase in liver injury score (necrosis, inflammation and steatosis) and an increase in lipid peroxidation in liver were observed. Liver and urine samples were analyzed by nuclear magnetic resonance spectroscopy and electrospray infusion/Fourier transform ion cyclotron resonance-mass spectrometry. In livers of alcohol-treated mice the following changes were noted. Hypoxia and glycolysis were activated as evidenced by elevated levels of alanine and lactate. Tyrosine, which is required for l-DOPA and dopamine as well as thyroid hormones, was elevated possibly reflecting alterations of basal metabolism by alcohol. A 4-fold increase in the prostacyclin inhibitor 7,10,13,16-docosatetraenoic acid, a molecule important for regulation of platelet formation and blood clotting, may explain why chronic drinking causes serious bleeding problems. Metabolomic analysis of the urine revealed that alcohol treatment leads to decreased excretion of taurine, a metabolite of glutathione, and an increase in lactate, n-acetylglutamine and n acetylglycine. Changes in the latter two metabolites suggest an inhibition of the kidney enzyme aminoacylase I and may be useful as markers for alcohol consumption. PMID- 18674557 TI - Role of water molecules and ion pairs in Dps and related ferritin-like structures. AB - A comparative study of water molecules and ion pairs in 11 Dps protein structures has been carried out. The invariant and common water molecules, the conserved residues interacting with them and the conserved ion pairs have been analyzed. Certain water molecules found on the interfaces between subunits are highly conserved and may be implicated in flexibility or continuing association of the subunits of the structure. It is possible that the water molecules, ion pairs and the special case of a water mediated charged network through a single water molecule are involved in maintaining the stability of the protein. PMID- 18674556 TI - Cocaine cue versus cocaine dosing in humans: evidence for distinct neurophysiological response profiles. AB - Subjective, physiological and electroencephalographic (EEG) profiles were studied in cocaine dependent study participants in response to cocaine cue exposure or a dose of smoked cocaine. Both stimuli increased subjective ratings of cocaine high and craving, enhanced negative affect, and boosted plasma ACTH and skin conductance levels. However, cocaine dose produced a greater increase in high and a more prolonged increase in plasma ACTH, while cocaine cue produced a decline in skin temperature. Both stimuli produced increases in absolute theta, alpha and beta EEG power over the prefrontal cortex. However, interhemispheric EEG coherence over the prefrontal cortex decreased during cocaine cue exposure but increased following cocaine dose. Moreover, correlation analysis of subjective, physiological and EEG responding to cocaine cue and dose revealed distinct profiles. Delta and theta activity were associated with negative affect during cocaine cue exposure, but were associated with cocaine craving and reward following cocaine dosing. In both conditions, alpha activity was marker for anxiousness but not high. These data demonstrate similar subjective, physiological responding in clinical laboratory states of cocaine craving and reward. However, differences in EEG response profiles, and their relationship to function, indicate distinct neurophysiological mediators of cocaine craving and reward within the prefrontal cortex. PMID- 18674558 TI - Olfaction and defense. PMID- 18674559 TI - Evaluation of neurite outgrowth anisotropy using a novel application of circular analysis. AB - Precise axon growth is required for making proper connections in development and after injury. One method of studying axon guidance and growth is through in vitro outgrowth assays that present controlled microenvironments. In this study, we applied circular statistical methods to evaluate directional neurite response. Visualization of data on a circular scale allows more accurate representation of the data, as neurite angles are inherently expressed on a circle. Here, the direction of neurite outgrowth from dorsal root ganglia derived neurons on different substrate types was quantitatively measured. Further, simulations of datasets with known circular parameters reflecting expected neurite angle distributions from different substrate types were also generated. Circular statistical methods were utilized and compared to linear statistical models widely used in the neuroscience literature. For small samples, Rao's spacing test showed the smallest occurrence of Type I errors (false positives) when tested against simulated uniform distributions. V-test and Rayleigh's test showed highest statistical power when tested against a unimodal distribution with known and unknown mean direction, respectively. For bimodal samples, Watson's U(2)-test showed the highest statistical power. Overall, circular statistical uniformity tests showed higher statistical power than linear non-parametric tests, particularly for small samples (n=5). Circular analysis methods represent a useful tool for evaluation of directionality of neurite outgrowth with applications including: (1) assessment of neurite outgrowth potential; (2) determination of isotropy of cellular responses to single and multiple cues and (3) determination of the relative strengths of cues present in a complex environment. PMID- 18674560 TI - A statistical method for quantifying songbird phonology and syntax. AB - Songbirds are the preeminent animal model for understanding how the brain encodes and produces learned vocalizations. Here, we report a new statistical method, the Kullback-Leibler (K-L) distance, for analyzing vocal change over time. First, we use a computerized recording system to capture all song syllables produced by birds each day. Sound Analysis Pro software [Tchernichovski O, Nottebohm F, Ho CE, Pesaran B, Mitra PP. A procedure for an automated measurement of song similarity. Anim Behav 2000;59:1167-76] is then used to measure the duration of each syllable as well as four spectral features: pitch, entropy, frequency modulation, and pitch goodness. Next, two-dimensional scatter plots of each day of singing are created where syllable duration is on the x-axis and each of the spectral features is represented separately on the y-axis. Each point in the scatter plots represents one syllable and we regard these plots as random samples from a probability distribution. We then apply the standard information-theoretic quantity K-L distance to measure dissimilarity in phonology across days of singing. A variant of this procedure can also be used to analyze differences in syllable syntax. PMID- 18674561 TI - Impact of microdialysis probes on vasculature and dopamine in the rat striatum: a combined fluorescence and voltammetric study. AB - Measuring extracellular dopamine in the brain of living animals by means of microdialysis and/or voltammetry is a route towards understanding both normal brain function and pathology. Previous reports, however, suggest that the tissue response to implantation of devices may affect the outcome of the measurements. To address the source of the tissue response and its impact on striatal dopamine systems microdialysis probes were placed in the striatum of anesthetized rats. Images obtained by dual-label fluorescence microscopy show signs of ischemia and opening of the blood-brain barrier near the probe tracks. Opening of the blood brain barrier was further examined by determining dialysate concentrations of carbi-DOPA, a drug that normally does not penetrate the brain. Although carbi DOPA was recovered in brain dialysate, it did not alter dialysate dopamine levels or evoked dopamine release as measured by voltammetry near the probes. Microdialysis probes also significantly diminished the effect of intrastriatal infusion of kynurenate on extracellular dopamine levels as measured by voltammetry near the probes. PMID- 18674562 TI - Testing for nested oscillation. AB - Nested oscillation occurs when the amplitude of a faster rhythm is coupled to the phase of a slower rhythm. It has been proposed to underlie the discrete nature of perception and the capacity of working memory and is a phenomenon observable in human brain imaging data. This paper compares three published methods for detecting nested oscillation and a fourth method proposed in this paper. These are: (i) the modulation index, (ii) the phase-locking value (PLV), (iii) the envelope-to-signal correlation (ESC) and (iv) a general linear model (GLM) measure derived from ESC. We applied the methods to electrocorticographic (ECoG) data recorded during a working-memory task and to data from a simulated hippocampal interneuron network. Further simulations were then made to address the dependence of each measure on signal to noise level, coupling phase, epoch length, sample rate, signal nonstationarity, and multi-phasic coupling. Our overall conclusion is that the GLM measure is the best all-round approach for detecting nested oscillation. PMID- 18674564 TI - A portable telemetry system for brain stimulation and neuronal activity recording in freely behaving small animals. AB - A portable multi-channel telemetry system which can be used for brain stimulation and neuronal activity recording in freely behaving small animals is described here. This system consists of three major components of headstage, backpack and portable Personal Digital Assistant (PDA). The headstage contains high precision instrument amplifiers with high input impedance. The backpack is comprised of two parts: (1) a main board (size: 36 mm x 22 mm x 3.5 mm and weight: 40 g with batteries, 20 g without), with current/voltage stimulator and special circuit suitable for neuronal activity recording and (2) and a bluetooth transceiver, with a high data transmission rate up to 70 kb/s, suitable for downloading stimulation commands and uploading acquired data. We recorded neuronal activities of the primary motor area of a freely behaving rat with 12-bit resolution at 12 k samples/s. The recorded data and analysis results showed that the system was successful by comparing with the commercial equipment Cerebus 128-Channel Data Acquisition System (Cyberkinetics Inc.). Using the PDA, we can control stimulation and recording. It provides a flexible method to do some research work in the circumstances where other approaches would be difficult or impossible. PMID- 18674563 TI - Fluorescent double-labeling with carbocyanine neuronal tracing and immunohistochemistry using a cholesterol-specific detergent digitonin. AB - The fluorescent carbocyanine dye DiI (1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate) has been widely used for tracing of neuronal pathways. To examine identities of the DiI-labeled neuronal pathways, it is desirable to combine DiI labeling with immunofluorescent staining. However, DiI labeling and immunofluorescent staining are not well compatible, mainly because treatment of DiI-labeled neurons with detergents, which are commonly used for immunohistochemistry, results in high levels of diffusion of the DiI label. In this study, we searched for detergents that are compatible with DiI labeling, and found that a cholesterol-specific detergent digitonin is useful for fluorescent double-labeling with DiI tracing and immunohistochemistry. We show that digitonin treatment, in contrast to Triton X-100, methanol and Nonidet P-40 treatment, preserves DiI labeling, even at higher concentrations. We also show that digitonin also preserves the signal of a DiI derivative CM-DiI. Moreover, we demonstrate that digitonin efficiently increases antibody penetration into brain sections. As a result, immunohistochemical images obtained with digitonin treatment are as good as those obtained with Triton X-100 treatment. In addition, we also try another cholesterol-specific detergent quillaja saponin, but find that it degrades the DiI label. Our simple double-labeling protocol using digitonin should prove useful in enabling detailed examination of the neuronal circuitry of the nervous system. PMID- 18674565 TI - The role of CD11b in phagocytosis and dendritic cell development. AB - Activation of resting T cells is highly dependent on dendritic cells (DCs), which take up antigens and present antigenic peptides to T cells in the context of the major histocompatibility complex (MHC). In this study, we generated a monoclonal antibody, which we call 1C4 that recognizes integrin alpha(M)beta(2) (CD11b/CD18) on the surface of conventional DCs (cDCs) and is internalized after binding. Addition of 1C4 inhibited the ability of immature DCs to phagocytose apoptotic cells. 1C4 treatment also partially inhibited the generation of cDCs from bone marrow in the presence of granulocyte macrophage colony-stimulating factor (GM CSF). Our findings suggest that not only CD11b is involved in the phagocytosis of apoptotic cells, but also that mAb such as 1C4 may be a useful tool for the delivery of specific proteins into the cytoplasm of immature DCs. PMID- 18674566 TI - In situ hybridization assays for localization of the chronic bee paralysis virus in the honey bee (Apis mellifera) brain. AB - Chronic bee paralysis virus (CBPV) is a common single-stranded RNA virus which may cause significant losses in honey bee colonies. As this virus seems to exhibit neurotropism, an in situ hybridization based method was developed to localize the genomic and antigenomic CBPV RNAs in infected honey bee brains. Double-stranded cDNA probes as well as genomic and antigenomic-specific single stranded cDNA probes were prepared, using the polymerase chain reaction in presence of labelled d-UTP with non-radioactive digoxigenin. Both genomic and antigenomic RNAs were detected the brain of honey bee infected naturally or artificially. Hybridization signals were obtained in some somata and neuropile regions of the brain. In particular, high signals were observed at the level of the mushroom bodies and central complex, regions that are known to be engaged in higher neuronal functions and in the optic and antennal lobes that are sensorial neuropiles. Thus, the presence of virus at these levels may explain the nervous symptoms observed in infected bees. The in situ hybridization procedure proved to be a useful tool to localize specifically CBPV and may be helpful for understanding the observed symptoms. PMID- 18674568 TI - A role of nitric oxide mechanism involved in the protective effects of venlafaxine in sleep deprivation. AB - The present study was designed to explore the possible nitric oxide mechanism in protective effect of venlafaxine in sleep deprivation in mice. Laca mice were sleep deprived for period of 72 h using grid suspended over water method. Venlafaxine (2.5, 5 and 10mg/kg, ip), l-arginine (50mg/kg, ip), l-NAME (10mg/kg, ip) and methylene blue (10mg/kg, ip) were administered for 5 days, starting 2 days before 72-h sleep deprivation. Various behavioral tests (plus maze, zero maze, mirror chamber tests for anxiety, and actophotometer test) followed by oxidative stress parameters (malondialdehyde level, glutathione, catalase, nitrite and protein) were assessed. The present study showed that venlafaxine (5 and 10mg/kg, ip) drug treatment significantly reversed 72-h sleep deprivation caused anxiety like behavior, impairment in locomotor activity and oxidative damage (increased lipid peroxidation and nitrite levels and depleted reduced glutathione and catalase activity) as compared to control. l-NAME (10mg/kg) and methylene blue (10mg/kg) pretreatment with lower dose of venlafaxine (5mg/kg) potentiated the protective effect of venlafaxine (5mg/kg). However, l-arginine (50mg/kg) pretreatment with venlafaxine (5mg/kg) reversed the protective effect of venlafaxine. Results of present study suggest that nitric oxide mechanism is involved in the protective effect of venlafaxine against sleep-deprivation induced behavior alteration and oxidative damage in mice. PMID- 18674567 TI - Towards the design of antiviral inhibitors against flaviviruses: the case for the multifunctional NS3 protein from Dengue virus as a target. AB - New treatments are urgently needed to combat the increasing number of dengue fever cases in endemic countries as well as amongst a large number of travellers from non-endemic countries. Of the 10 virus encoded proteins, NS3 (non-structural 3) and NS5 carry out all the enzymatic activities needed for polyprotein processing and genome replication, and are considered to be amenable to antiviral inhibition by analogy with successes for similar targets in human immunodeficiency virus and hepatitis C virus. The multifunctional NS3 protein of flavivirus forms a non-covalent complex with the NS2B cofactor and contains the serine-protease activity domain at its N-terminus that is responsible for proteolytic processing of the viral polyprotein and a ATPase/helicase and RNA triphosphatase at its C-terminal end that are essential for RNA replication. In addition, NS3 seems to be also involved in virus assembly. This review covers the recent biochemical and structural advances on the NS2B-NS3 protease-helicase and presents an outlook for the development of small molecules as antiviral drugs targeting this fascinating multifunctional protein. PMID- 18674569 TI - Two distinct arsenite-resistant variants of Leishmania amazonensis take different routes to achieve resistance as revealed by comparative transcriptomics. AB - Genome-wide search for the genes involved in arsenite resistance in two distinct variants A and A' of Leishmania amazonensis revealed that the two variants used two different mechanisms to achieve resistance, even though these two variants were derived from the same clone and selected against arsenite under the same conditions. In variant A, the variant with DNA amplification, the biochemical pathways for detoxification of oxidative stress, the energy generation system to support the biochemical and physiological needs of the variant for DNA and protein synthesis and the arsenite translocating system to dispose arsenite are among the primary biochemical events that are upregulated under the arsenite stress to gain resistance. In variant A', the variant without DNA amplification, the upregulation of aquaglyceroporin (AQP) gene and the high level of resistance to arsenate point to the direction that the resistance gained by the variant is due to arsenate which is probably oxidized from arsenite in the arsenite solution used for selection and the maintenance of the cell culture. As a result of the AQP upregulation for arsenite disposal, a different set of biochemical pathways for detoxification of oxidative stress, energy generation and cellular signaling are upregulated to sustain the growth of the variant to gain resistance to arsenate. From current evidences, reactive oxygen species (ROS) overproduced by the parasite soon after exposure to arsenite appear to play an instrumental role in both variants to initiate the subsequent biochemical events that allow the same clone of L. amazonensis to take two totally different routes to diverge into two different variants. PMID- 18674570 TI - Flow cytometric detection of progastrin interaction with gastrointestinal cells. AB - The unprocessed gastrin precursor, progastrin (PG), is often overexpressed in colon cancer and other malignancies where it appears to stimulate colonic growth. Overexpression of progastrin also stimulates proliferation of normal colonic mucosa, but the receptors mediating these effects have not been identified. Here we report the development of a non-radioactive assay for assessment of PG binding to normal and transformed cells. Progastrin was labeled using biotinylation, and binding of biotinylated PG to cells was assessed using flow cytometry. Using this approach, we show strong and specific binding of PG to some cell lines (IEC-6, IEC-18, HT-29, COLO320) and minimal binding to others (HeLa, DC2.4, Jurkat). We also found PG binding to several non-gut epithelial lines, such as CHO-K1, COS-6 and HEK293 cells. The specificity of binding was confirmed by competition with cold, unlabeled PG but not with glycine-extended gastrin or amidated gastrin-17. Binding was not influenced by the presence of the classical CCK-2 receptor, but was partially dependent on the charged glycosaminoglycans (GAG). The analysis of primary colonic tissues isolated from wild type C57BL/6 mouse, revealed a small epithelial subpopulation of non-hematopoietic (CD45-negative) cells that strongly interacted with PG. Surprisingly, this population was greatly expanded in gastrin knockout mice. This non-radioactive, FACS-based assay should prove useful for further characterization of cells expressing the progastrin receptor. PMID- 18674571 TI - Leukocyte Elastase Inhibitor, the precursor of L-DNase II, inhibits apoptosis by interfering with caspase-8 activation. AB - LEI (Leukocyte Elastase Inhibitor), the precursor of the pro-apoptotic molecule L DNase II, belongs to the ovalbumin subgroup of serpins. Several serpins can inhibit apoptosis: the viral serpin Crm A inhibits Fas or TNFalpha-induced apoptosis, and overexpression of PAI-2 or PI-9 protects cells from TNFalpha or granzyme B induced apoptosis. We have previously shown that LEI overexpression protects cells from etoposide-induced apoptosis. The molecular reason of this anti-apoptotic activity is now investigated. We show that, in BHK-21 and HeLa cells, LEI anti-protease activity is essential for its anti-apoptotic effect. The protease inhibited is cathepsin D, released from the lysosome during etoposide treatment. Cathepsin D enhances caspase activity in the cell by cleaving procaspase-8 and LEI overexpression slows down this cleavage, protecting cells from apoptosis. This let us presume that high expression of LEI in tumor cells may reduce the efficiency of etoposide as a chemotherapeutic agent. PMID- 18674572 TI - Evaluation of isolation methods and RNA integrity for bacterial RNA quantitation. AB - RNA integrity is critical for successful RNA quantitation for mammalian tissues, but the level of integrity required differs among tissues. The level of integrity required for quantitation has not been determined for bacterial RNA. Three RNA isolation methods were evaluated for their ability to produce high quality RNA from Dickeya dadantii, a bacterium refractory to RNA isolation. Bacterial lysis with Trizol using standard protocols consistently gave low RNA yields with this organism. Higher yields due to improved bacterial cells lysis was achieved with an added hot SDS incubation step, but RNA quality was low as determined by the RNA Integrity Number (RIN). Contaminating DNA remained a problem with the hot SDS Trizol method; RNA samples required repeated, rigorous DNase treatments to reduce DNA contamination to levels sufficient for successful real-time qRT-PCR. A hot SDS-hot phenol RNA method gave the highest RNA quality and required only two DNase treatments to remove DNA. The assessment of RNA integrity using the Agilent 2100 BioAnalyzer was critical for obtaining meaningful gene expression data. RIN values below 7.0 resulted in high variation and loss of statistical significance when gene expression was analyzed by real-time qRT-PCR. We found that RNA preparations of different quality yielded drastic differences in relative gene expression ratios and led to major errors in the quantification of transcript levels. This work provides guidelines for RNA isolation and quality assessment that will be valuable for gene expression studies in a wide range of bacteria. PMID- 18674573 TI - Separating deceptive and orienting components in a Concealed Information Test. AB - The Concealed Information Test (CIT) requires the examinee to deceptively deny recognition of known stimuli and to truthfully deny recognition of unknown stimuli. Because deception and orienting are typically coupled, it is unclear how exactly these sub-processes affect the physiological responses measured in the CIT. The present study aimed at separating the effects of deception from those of orienting. In a mock-crime study, using a modified CIT, thirty-six of seventy-two subjects answered truthfully ('truth group'), whereas the other thirty-six concealed their knowledge ('lie group'). Answering was delayed for 4 s after item presentation. Electrodermal activity (EDA), respiration (RLL), and phasic heart rate (HR) were recorded. A decomposition of EDA responses revealed two response components; the response in the first interval was expected to indicate orienting, stimulus evaluation, and answer preparation, whereas the response in the second interval was assumed to reflect answer-related processes. Inconclusively, both EDA components differentiated between 'probe' and 'irrelevant' items in both groups. Phasic HR and RLL differed between item classes only in the 'lie' group, thus reflecting answer-related processes, possibly deception, rather than merely orienting responses. The findings further support the notion that psychophysiological measures elicited by a modified CIT may reflect different mental processes involved in orienting and deception. PMID- 18674574 TI - Sapovirus-like particles derived from polyprotein. AB - We expressed full-length sapovirus genome constructs in insect cells and analyzed their products. The capsid protein was cleaved from the ORF1 polyprotein from a native-like genome construct and two full-length genome constructs with mutations in an active polymerase motif, whereas the capsid protein was not cleaved from a full-length genome construct with a mutation in an active protease motif. Our results showed that the sapovirus protease-polymerase precursor protein cleaved the capsid protein from the polyprotein at the putative conserved capsid start. Importantly, the cleaved capsid protein formed empty virus-like particles that were morphologically and antigenically similar to native sapovirus. PMID- 18674575 TI - Emerging G9 rotavirus strains in the northwest of China. AB - Although G9 rotaviruses have become one of the important rotavirus genotypes worldwide, they have been uncommon in China. Recently, we reported G9 rotaviruses as a highly prevalent genotype in Xinjiang, the northwest part of China [Yang, X., Matthijnssens, J., Sun, H., Muhamaiti, J., Zhang, B., Nahar, S., Van Ranst, M., Rahman, M., 2008. Temporal changes of rotavirus strain distribution in a northwest city of China, 1996-2005. Int. J. Infect. Dis., June (Epub ahead of print)]. Here we report the genetic variations of the Xinjiang-G9 rotaviruses isolated between 1999 and 2005. Sequence analysis of the VP7 genes of Xinjiang-G9 strains indicated that they were more closely related to the contemporary global G9 strains than to the prototype Chinese G9 strains. However, their VP4 genes were most similar to those from the locally circulating G1P[8], G2P[4], G3P[6] and G3P[8] strains. This indicates that reassortment rather than antigenic drift might be the preferred evolutionary mechanism for the emergence of the G9 rotaviruses in Xinjiang. These findings will be of major significance for understanding the emergence of newly introduced rotavirus strains. PMID- 18674576 TI - Isolation of a type 3 vaccine-derived poliovirus (VDPV) from an Iranian child with X-linked agammaglobulinemia. AB - Type 3 immunodeficiency-associated vaccine-derived polioviruses (iVDPVs) were isolated from a 15-month-old Iranian boy with acute flaccid paralysis (AFP) who was subsequently diagnosed with X-linked agammaglobulinemia (XLA). VP1 nucleotide sequences of the two isolates differed from Sabin 3 by 2.0% and 2.1% and from each other by 0.6%. Although the key determinant of attenuation and temperature sensitivity in the 5'-untranslated region (U(472)-->C) had reverted, a second capsid-region determinant (VP3:Phe(091)) was unchanged, but a presumptive suppressor (VP1:Ala(054)-->Val) was found. The isolates were Sabin 3/Sabin 1 recombinants, sharing a single recombination breakpoint in the 2C region. Although the two isolates were antigenically distinct from Sabin 3, only one amino acid replacement was found in the neutralizing antigenic sites (VP3:Ser(059)-->Asn in site 3). The patient was placed on intravenous immunoglobulin (IVIG) therapy within 9 days of onset of AFP, and iVDPV excretion ceased thereafter, but the patient remained severely paralyzed until his death approximately 11 months after paralysis. No secondary AFP cases were found, and none of the seven tested contacts of the patient were found to be infected with poliovirus. PMID- 18674577 TI - Molecular characterization of VP4, VP6 and VP7 genes of a rare G8P[14] rotavirus strain detected in an infant with gastroenteritis in Italy. AB - In this study, the molecular characterization of a rare G8P[14] group A rotavirus (GARV) strain detected in Northern Italy during the 2004-2005 epidemiological rotavirus season is described. Two hundred and seventy three rotavirus-like particle positive stools out of 856 stools from children (31.9%) hospitalized with gastroenteritis were analyzed using polyacrilamide gel electrophoresis and 271 GARVs were genotyped by VP7 and VP4 specific RT-PCRs. One strain (PR/1300/04) with a long electropherotype (e-type) displayed the G8 specificity and was VP4 un typeable. The P and the subgroup (SG) specificities were determined by sequencing the VP4 and the VP6 gene, respectively. The PR/1300/04 strain exhibited P[14] and SGI specificities. By sequence and phylogenetic analyses of the VP4, VP6 and VP7 amplicons, the PR/1300/04 VP4 and VP6 genes were demonstrated to be of human rotavirus origin, with the VP4 gene closely related to the human Italian PA169 strain (G6P[14]), while the VP7 gene was of animal origin (bovine). These data suggest that the Italian PR/1300/04 strain could be a reassortant between a PA169 like Italian strain with P[14] specificity, long e-type and SGI, and a G8 animal strain. The increasing number of reports of atypical GARVs in humans suggests that interspecies transmission of genes greatly contributes to the GARV genetic evolution. PMID- 18674578 TI - An efficient and low immunostimulatory nanoparticle formulation for systemic siRNA delivery to the tumor. AB - We have developed a nanoparticle formulation [liposomes-protamine-hyaluronic acid nanoparticle (LPH-NP)] for systemically delivering siRNA into the tumor. The LPH NP was prepared in a self-assembling process. Briefly, protamine and a mixture of siRNA and hyaluronic acid were mixed to prepare a negatively charged complex. Then, cationic liposomes were added to coat the complex with lipids via charge charge interaction to prepare the LPH-NP. The LPH-NP was further modified by DSPE PEG or DSPE-PEG-anisamide by the post-insertion method. Anisamide is a targeting ligand for the sigma receptor over-expressed in the B16F10 melanoma cells. The particle size, zeta potential and siRNA encapsulation efficiency of the formulation were approximately 115 nm, +25 mV and 90%, respectively. Luciferase siRNA was used to evaluate the gene silencing activity in the B16F10 cells, which were stably transduced with a luciferase gene. The targeted LPH-NP (PEGylated with ligand) silenced 80% of luciferase activity in the metastatic B16F10 tumor in the lung after a single i.v. injection (0.15 mg siRNA/kg). The targeted LPH-NP also showed very little immunotoxicity in a wide dose range (0.15-1.2 mg siRNA/kg), while the previously published formulation, LPD-NP (liposome-protamine DNA nanoparticle), had a much narrow therapeutic window (0.15-0.45 mg/kg). PMID- 18674579 TI - Sustained GM-CSF and PEI condensed pDNA presentation increases the level and duration of gene expression in dendritic cells. AB - Current techniques to educate dendritic cells (DCs) ex vivo for immunotherapy are plagued by inefficient protocols and DC modifications are often transient and lost upon transplantation. This study investigated the role of sustained presentation of GM-CSF and PEI condensed pDNA (PEI-DNA) on gene transfer and long term gene expression. Appropriate GM-CSF signaling during DC transfection promoted PEI-DNA uptake, although high cytokine concentrations induced intercellular DNA degradation, indicating the need for controlled presentation. Poly(lactide-co-glycolide) scaffolds that continuously stimulated DCs with both GM-CSF and PEI-DNA led to a 20-fold increase in gene expression, and high levels of expression persisted for a period of 10 days, in vitro. These results encourage the exploitation of biomaterials and GM-CSF to develop novel delivery vectors for genetically modified DCs or to genetically program host DCs in situ for vaccination and the treatment of autoimmunity. PMID- 18674580 TI - High-affinity choline uptake and acetylcholine-metabolizing enzymes in CNS white matter. A quantitative study. AB - The presence of nicotinic and muscarinic receptors suggests the occurrence of cholinergic neurotransmission in white matter; however no quantitative information exists on acetylcholine formation and breakdown in white matter. We compared white structures of pig brain (fimbria, corpus callosum, pyramidal tracts, and occipital white matter) to gray structures (temporal, parietal and cerebellar cortices, hippocampus, and caudate) and found that sodium-dependent, high-affinity choline uptake in white structures was 25-31% of that in hippocampus. White matter choline acetyltransferase activity was 10-50% of the hippocampal value; the highest activity was found in fimbria. Acetylcholine esterase activity in white structures was 20-25% of that in hippocampus. The caudate, which is rich in cholinergic interneurons, gave values for all three parameters that were 2.8-4 times higher than in hippocampus. The results suggest a certain capacity for cholinergic neurotransmission in central nervous white matter. The white matter activity of pyruvate dehydrogenase, which provides acetyl-CoA for acetylcholine synthesis, ranged between 33 and 50% of the hippocampal activity; the activity in the caudate was similar to that in hippocampus and the other gray structures, which was true also for other enzymes of glucose metabolism: hexokinase, phosphoglucomutase, and glucose-6-phosphate dehydrogenase. Acetylcholine esterase activity in white matter was inhibited by the nerve agent soman, which may help explain the reported deleterious effect of soman on white matter. Further, this finding suggests that acetylcholine esterase inhibitors used in Alzheimer's disease may have an effect in white matter. PMID- 18674581 TI - Attitudes towards vaccination among chiropractic and naturopathic students. AB - We have investigated the attitudes towards vaccination of undergraduate chiropractic and naturopathic students in the two major complementary and alternative medicine colleges in Canada. While the majority of the students were not averse to vaccination, we found in both colleges that anti-vaccination attitudes were more prevalent in the later years of the programs. Reasons for this are discussed, and we provide suggestions for strategies to address the situation. PMID- 18674582 TI - Cost-effectiveness analysis of universal childhood vaccination against varicella in Brazil. AB - This study conducts a cost-effectiveness analysis of a childhood universal varicella vaccination program in Brazil. An age and time-dependent dynamic model was developed to estimate the incidence of varicella for 30 years. Assuming a single-dose schedule; vaccine efficacy of 85% and coverage of 80%, the program could prevent 74,422,058 cases and 2905 deaths. It would cost R$ 3,178,396,110 and save R$ 660,076,410 to the society and R$ 365,602,305 to the healthcare system. The program is cost-effective (R$ 14,749 and R$ 16,582 per life-year saved under the societal and the healthcare system's perspective, respectively). The program's cost-effectiveness is highly sensitive to the vaccine price and number of doses. PMID- 18674583 TI - New vaccine approaches for seasonal and pandemic influenza. AB - Inactivated influenza vaccines have been available since the late 1940s for the prevention of influenza disease. Based on the available scientific evidence, many public health authorities, including the World Health Organization, recommend annual use of these vaccines for specific populations, including the elderly. Despite these recommendations, actual vaccination uptake rates are very limited in many countries. Influenza vaccine research is confounded by the variable nature of the influenza viruses and annual influenza epidemics and by non specific clinical diagnostic criteria. These confounding factors complicate evaluation not only of overall vaccine effectiveness, but also of the relative efficacy and effectiveness of different vaccine formulations. This paper summarizes recent advances in the development of seasonal and (pre-)pandemic vaccines, discusses the methodologic constraints on influenza vaccine research, and proposes measures to reduce the level of potential bias and confounding in influenza vaccine research. PMID- 18674584 TI - Addiction to topically used cyclopentolate hydrochloride: a case report. PMID- 18674585 TI - Can systems toxicology identify common biomarkers of non-genotoxic carcinogenesis? AB - For the rapid development of safe, efficacious chemicals it is important that any potential liabilities are identified as early as possible in the discovery/development pipeline. Once identified it is then possible to make rational decisions on whether to progress a chemical and/or series further; one such liability is chemical carcinogenesis, a highly undesirable characteristic in a novel chemical entity. Chemical carcinogens may be roughly divided into two classes, those that elicit their actions through direct damage to DNA (genotoxic carcinogens) and those that cause carcinogenesis through mechanisms that involve direct damage of the DNA by the agent (non-genotoxic carcinogens). Whereas the former group can be identified by in vitro screens to a good degree of accuracy, the latter group are far more problematic due to their diverse modes of action. This review will focus on the latter class of chemical carcinogens, examining how modern '-omic' technologies have begun to identify signatures that may represent sensitive, early markers for these processes. In addition to their use in signature generation the role of -omic level approaches to delineating molecular mechanisms of action will also be discussed. PMID- 18674588 TI - The impact of semantic-free second-language training on ERPs during case processing. AB - Some language-related ERP responses are only observed in high proficiency L2 speakers. It is unknown, however, how these ERP patterns are influenced by language training. We tested the effect of semantic-free training on ERPs related to syntactic processing in auditory sentence comprehension in German participants learning a miniature version of Japanese. When presented with correct sentences and sentences containing a case violation, the learners showed an N400-like negativity and a P600-like positivity resembling the ERP pattern reported for Japanese natives. They contrasted with a previously tested group of learners who, though they had been provided with full language training including semantic information, had only shown the P600. The results suggest that the absence of semantic input facilitates the development of some aspects of native-like language processing operations in L2 learners. PMID- 18674586 TI - Endocrine function in naturally long-living small mammals. AB - The complex, highly integrative endocrine system regulates all aspects of somatic maintenance and reproduction and has been widely implicated as an important determinant of longevity in short-lived traditional model organisms of aging research. Genetic or experimental manipulation of hormone profiles in mice has been proven to definitively alter longevity. These hormonally induced lifespan extension mechanisms may not necessarily be relevant to humans and other long lived organisms that naturally show successful slow aging. Long-lived species may have evolved novel anti-aging defenses germane to naturally retarding the aging process. Here, we examine the available endocrine data associated with the vitamin D, insulin, glucocorticoid and thyroid endocrine systems of naturally long-living small mammals. Generally, long-living rodents and bats maintain tightly regulated lower basal levels of these key pleiotropic hormones than shorter lived rodents. Similarities with genetically manipulated long-lived rodent models of aging suggest that evolutionary well-conserved hormonal mechanisms are integrally involved in lifespan determination. PMID- 18674587 TI - Hormonal control of aging in rodents: the somatotropic axis. AB - There is a growing body of literature focusing on the somatotropic axis and regulation of aging and longevity. Many of these reports derive data from multiple endocrine mutants, those that exhibit both elevated growth hormone (GH) and insulin-like growth factor I (IGF-1) or deficiencies in one or both of these hormones. In general, both spontaneous and genetically engineered GH and IGF-1 deficiencies have lead to small body size, delayed development of sexual maturation and age-related pathology, and life span extension. In contrast, characteristics of high circulating GH included larger body sizes, early puberty and reproductive senescence, increased cancer incidence and reduced life span when compared to wild-type animals with normal plasma hormone concentrations. This information, along with that found in multiple other species, implicates this anabolic pathway as the major regulator of longevity in animals. PMID- 18674589 TI - Emotional experience modulates brain activity during fixation periods between tasks. AB - Functional imaging studies have begun to identify a set of brain regions whose brain activity is greater during 'rest' (e.g., fixation) states than during cognitive tasks. It has been posited that these regions constitute a network that supports the brain's default mode, which is temporarily suspended during specific goal-directed behaviors. Exogenous tasks that require cognitive effort are thought to command reallocation of resources away from the brain's default state. However, it remains unknown if brain activity during fixation periods between active task periods is influenced by previous task-related emotional content. We examined brain activity during periods of FIXATION (viewing and rating gray-scale images) interspersed among periods of viewing and rating complex images ('PICTURE') with positive, negative, and neutral affective content. We show that a selected group of brain regions (PCC, precuneus, IPL, vACC) do exhibit activity that is greater during FIXATION (>PICTURE); these regions have previously been implicated in the "default brain network". In addition, we report that activity within precuneus and IPL in the FIXATION period is attenuated by the precedent processing of images with positive and negative emotional content, relative to non-emotional content. These data suggest that the activity within regions implicated in the default network is modulated by the presence of environmental stimuli with motivational salience and, thus, adds to our understanding of the brain function during periods of low cognitive, emotional, or sensory demand. PMID- 18674590 TI - Salicylaldehyde derivatives as new protein kinase CK2 inhibitors. AB - Protein kinase CK2 is a Ser/Thr kinase, with a constitutive activity, that is considered as a promising target for cancer therapy. The currently available CK2 inhibitors lack the potency and the pharmacological properties necessary to be suitable and successful in clinical settings. We report the development of new potent CK2 inhibitors from salicylaldehyde derivatives identified by automated screening of a proprietary small-molecule library. Docking simulations and analysis of the structure-activity relationship for the hits allowed to determine their binding modes on CK2, and to carry out the optimization of their structures. This strategy led to the discovery of potent CK2 inhibitors with novel structures, one of which was able to inhibit CK2 activity in living cells and promote tumor cell death. The essential features required for potent CK2 inhibitory activity of this class of compounds are discussed. PMID- 18674591 TI - Orexin peptides enhance median preoptic nucleus neuronal excitability via postsynaptic membrane depolarization and enhancement of glutamatergic afferents. AB - Subpopulations of neurons in the median preoptic nucleus (MnPO) located within the lamina terminalis contribute to thermoregulatory, cardiovascular and hydromineral homeostasis, and sleep-promotion. MnPO is innervated by lateral hypothalamic neurons that synthesize and secrete the arousal-promoting and excitatory orexin (hypocretin) neuropeptides. To evaluate the hypothesis that orexins modulate the excitability of MnPO neurons, we used patch-clamp recording techniques applied in rat brain slice preparations to assess the effects of exogenously applied orexin A and orexin B peptides on their intrinsic and synaptic properties. Whole cell recordings under current-clamp mode revealed that 11/15 tested MnPO neurons responded similarly to either orexin A or B (500-1000 nM) with a slowly rising, prolonged (10-15 min) and reversible membrane depolarization. Under voltage-clamp mode, orexin applications induced a tetrodotoxin-resistant inward current of -7.2+/-1.6 pA, indicating a direct (postsynaptic) activation, with a time course similar to the observed membrane depolarization. The orexin-induced responses in 4/7 neurons were associated with a significant decrease in membrane conductance and the net orexin-induced current that reversed at -99+/-5 mV, suggesting closure of potassium channels. Orexins did not attenuate the properties of excitatory (n=4) or inhibitory (n=7) postsynaptic currents evoked by subfornical organ stimulation. By contrast, orexins applications induce a significant increase in both frequency and amplitude of spontaneous glutamatergic postsynaptic currents (5/7 cells) but had no influence on spontaneous GABAergic currents (6/6 cells). Thus, in addition to a direct postsynaptic receptor-mediated excitation, orexins can also increase the excitability of MnPO neurons via increasing their excitatory inputs, presumably through an orexin receptor-mediated excitation of local glutamatergic neurons whose axons project to MnPO neurons. PMID- 18674592 TI - Error-specific medial cortical and subcortical activity during the stop signal task: a functional magnetic resonance imaging study. AB - The ability to detect errors and adjust behavior accordingly is essential for maneuvering in an uncertain environment. Errors are particularly prone to occur when multiple, conflicting responses are registered in a situation that requires flexible behavioral outputs. Previous studies have provided evidence indicating the importance of the medial cortical brain regions including the cingulate cortex in processing conflicting information. However, conflicting situations can be successfully resolved, or lead to errors, prompting a behavioral change in the observers. In particular, how does the brain use error signals specifically to adjust behavior on the fly? Here we employ a stop signal task (SST) to elicit errors approximately half of the time in high-conflict trials despite constant behavioral adjustment of the observers. Using functional magnetic resonance imaging, we show greater and, sequentially, less activation in the medial cortical regions when observers made an error, compared with when they successfully resolved high-conflict responses. Errors also evoked greater activity in the cuneus, retrosplenial cortex, insula, and subcortical structures including the thalamus and the region of the epithalamus (the habenula). We further showed that the error-related medial cortical activities are not correlated with post-error behavioral adjustment, as indexed by post-error slowing (PES) in go trial reaction time. These results delineate an error specific pattern of brain activation during the SST. The results also suggest that the relationship between error-related activity and post-error behavioral adjustment may be more complicated than has been conceptualized by the conflict monitoring hypothesis. PMID- 18674594 TI - The rat temporal association cortical area 2 (Te2) comprises two subdivisions that are visually responsive and develop independently. AB - In this study, we have used the expression of non-phosphorylated neurofilament (NNF), a protein that exhibits differential areal and laminar neuronal patterning, to assess the chemoarchitectural organization of the rat temporal association cortex (Te). Since expression of NNF is associated with the latter stages of neuronal development, this enabled us to profile the hierarchical development of this region of the cortex. We also examined the expression of the protein Fos, the product of the immediate-early gene cFos, as a neuronal activity marker to determine which areas within this region are visually responsive. Our findings reveal the existence of two previously undescribed subdivisions within the dorsal and ventral domains of the rat temporal association cortical area 2 (Te2) which we have termed Te2d and Te2v, respectively. We also demonstrated the early maturation of the caudal region of Te2d while preceding the primary visual cortex. Within this region of the cortex, the Fos protein indicates that both subdivisions are visually responsive. PMID- 18674593 TI - Mechanisms and implications of adaptive immune responses after traumatic spinal cord injury. AB - Traumatic spinal cord injury (SCI) in mammals causes widespread glial activation and recruitment to the CNS of innate (e.g. neutrophils, monocytes) and adaptive (e.g. T and B lymphocytes) immune cells. To date, most studies have sought to understand or manipulate the post-traumatic functions of astrocytes, microglia, neutrophils or monocytes. Significantly less is known about the consequences of SCI-induced lymphocyte activation. Yet, emerging data suggest that T and B cells are activated by SCI and play significant roles in shaping post-traumatic inflammation and downstream cascades of neurodegeneration and repair. Here, we provide neurobiologists with a timely review of the mechanisms and implications of SCI-induced lymphocyte activation, including a discussion of different experimental strategies that have been designed to manipulate lymphocyte function for therapeutic gain. PMID- 18674595 TI - The suprachiasmatic nuclei are involved in determining circadian rhythms during restricted feeding. AB - The circadian clock in the suprachiasmatic nuclei (SCN) responds to light and regulates peripheral circadian rhythms. Feeding regimens also reset the clock, so that time-restricted feeding (RF) dictates rhythms in peripheral tissues, whereas calorie restriction (CR) affects the SCN clock. To better understand the influence of RF vs. CR on circadian rhythms, we took advantage of the transgenic alphaMUPA mice that exhibit spontaneously reduced eating, and can serve as a model for CR under ad libitum feeding, and a model for temporal CR under RF compared with wild type (WT) mice. Our results show that RF advanced and generally increased the amplitude of clock gene expression in the liver under LD in both mouse types. However, under disruptive light conditions, RF resulted in a different clock gene phase in WT mice compared with alphaMUPA mice, suggesting a role for the reduced calories in resetting the SCN that led to the change of phase in alphaMUPA mice. Comparison of the RF regimen in the two lighting conditions in WT mice revealed that mPer1, mClock, and mBmal1 increased, whereas mPer2 decreased in amplitude under ultradian light in WT mice, suggesting a role for the SCN in determining clock gene expression in the periphery during RF. In summary, herein we reinforce a role for calorie restriction in resetting the SCN clock, and unravel a role for the SCN in determining peripheral rhythms under RF. PMID- 18674596 TI - Heme oxygenase 2 deficiency increases brain swelling and inflammation after intracerebral hemorrhage. AB - Intracerebral hemorrhage (ICH) remains a major medical problem and currently has no effective treatment. Hemorrhaged blood is highly toxic to the brain, and catabolism of the pro-oxidant heme, mainly released from hemoglobin, is critical for the resolution of hematoma after ICH. The degradation of the pro-oxidant heme is controlled by heme oxygenase (HO). We have previously reported a neuroprotective role for HO2 in early brain injury after ICH; however, in vivo data that specifically address the role of HO2 in brain edema and neuroinflammation after ICH are absent. Here, we tested the hypothesis that HO2 deletion would exacerbate ICH-induced brain edema, neuroinflammation, and oxidative damage. We subjected wild-type (WT) and HO2 knockout ((-/-)) mice to the collagenase-induced ICH model. Interestingly, HO2(-/-) mice had enhanced brain swelling and neuronal death, although HO2 deletion did not increase collagenase-induced bleeding; the exacerbation of brain injury in HO2(-/-) mice was also associated with increases in neutrophil infiltration, microglial/macrophage and astrocyte activation, DNA damage, peroxynitrite production, and cytochrome c immunoreactivity. In addition, we found that hemispheric enlargement was more sensitive than brain water content in the detection of subtle changes in brain edema formation in this model. Combined, these novel findings extend our previous observations and demonstrate that HO2 deficiency increases brain swelling, neuroinflammation, and oxidative damage. The results provide additional evidence that HO2 plays a critical protective role against ICH-induced early brain injury. PMID- 18674597 TI - Phenotypic characterization of cognition and social behavior in mice with heterozygous versus homozygous deletion of catechol-O-methyltransferase. AB - Catechol-O-methyltransferase is an important enzyme in the metabolism of dopamine and an important regulator of aspects of dopamine-dependent working memory in prefrontal cortex that are disturbed in schizophrenia. This study investigated the phenotype of mice with heterozygous deletion vs. homozygous knockout of the catechol-O-methyltransferase gene across paradigms that access processes relevant for psychotic illness. Homozygotes evidenced improved performance in spontaneous alternation, an index of immediate spatial working memory; this effect appeared more substantive in males and was reflected in performance in aspects of the Barnes maze, an index of spatial learning/memory. Heterozygotes evidenced impaired performance in object recognition, an index of recognition memory; this effect was evident for both sexes at a retention interval of 5 min but appeared more enduring in males. There were no material effects for either genotype in relation to sociability or social novelty preference. While homozygous catechol-O methyltransferase deletion results in improvement in spatial learning/working memory with little effect on social behavior, heterozygous deletion results in impairment of recognition memory. We have reported recently, using similar methods, that mice with deletion of the schizophrenia risk gene neuregulin-1 evidence disruption to social behavior, with little effect on spatial learning/working memory. The data suggest that catechol-O-methyltransferase and neuregulin-1 may influence, respectively, primarily cognitive and social endophenotypes of the overall schizophrenia syndrome. PMID- 18674598 TI - Response preparation and inhibition: the role of the cortical sensorimotor beta rhythm. AB - Paradigms requiring either a GO or a NO-GO response are often used to study the neural mechanisms of response inhibition. Here this issue is examined from the perspective of event-related beta (14-30 Hz) oscillatory activity. Two macaque monkeys performed a task that began with a self-initiated lever depression and maintenance (sustained motor output) and required a visual pattern discrimination followed by either a lever release (GO) or continued lever-holding (NO-GO) response. Analyzing simultaneous local field potentials (LFPs) from primary somatosensory, frontal motor, and posterior parietal cortices, we report two results. First, beta oscillation desynchronized shortly after stimulus presentation, the onset of which was approximately the same for both the GO and NO-GO conditions ( approximately 110 ms). Since it is well known that beta desynchronization is a reliable indicator of movement preparation, this result suggests that early motor preparation took place in both conditions. Second, following the GO/NO-GO decision ( approximately 190 ms), beta activity rebounded significantly ( approximately 300 ms) only in the NO-GO condition. Coherence and Granger causality measures revealed that the dynamical organization of the rebounded beta network was similar to that existing during the sustained motor output prior to stimulus onset. This finding suggests that response inhibition led to the restoration of the sensorimotor network to its prestimulus state. PMID- 18674599 TI - Anticipatory activity in the human thalamus is predictive of reaction times. AB - Responding to environmental stimuli in a fast manner is a fundamental behavioral capacity. The pace at which one responds is known to be predetermined by cortical areas, but it remains to be shown if subcortical structures also take part in defining motor swiftness. As the thalamus has previously been implicated in behavioral control, we tested if neuronal activity at this level could also predict the reaction time of upcoming movements. To this end we simultaneously recorded electrical brain activity from the scalp and the ventral intermediate nucleus (VIM) of the thalamus in patients undergoing thalamic deep brain stimulation. Based on trial-to-trial analysis of a Go/NoGo task, we demonstrate that both cortical and thalamic neuronal activity prior to the delivery of upcoming Go stimulus correlates with the reaction time. This result goes beyond the demonstration of thalamic activity being associated with but potentially staying invariant to motor performance. In contrast, it indicates that the latencies at which we respond to environmental stimuli are not exclusively related to cortical pre-movement states but are also correlated with anticipatory thalamic activity. PMID- 18674600 TI - Cholinergic neurons of mouse intrinsic cardiac ganglia contain noradrenergic enzymes, norepinephrine transporters, and the neurotrophin receptors tropomyosin related kinase A and p75. AB - Half of the cholinergic neurons of human and primate intrinsic cardiac ganglia (ICG) have a dual cholinergic/noradrenergic phenotype. Likewise, a large subpopulation of cholinergic neurons of the mouse heart expresses enzymes needed for synthesis of norepinephrine (NE), but they lack the vesicular monoamine transporter type 2 (VMAT2) required for catecholamine storage. In the present study, we determined the full scope of noradrenergic properties (i.e. synthetic enzymes and transporters) expressed by cholinergic neurons of mouse ICG, estimated the relative abundance of neurons expressing different elements of the noradrenergic phenotype, and evaluated the colocalization of cholinergic and noradrenergic markers in atrial nerve fibers. Stellate ganglia were used as a positive control for noradrenergic markers. Using fluorescence immunohistochemistry and confocal microscopy, we found that about 30% of cholinergic cell bodies contained tyrosine hydroxylase (TH), including the activated form that is phosphorylated at Ser-40 (pSer40 TH). Dopamine beta hydroxylase (DBH) and norepinephrine transporter (NET) were present in all cholinergic somata, indicating a wider capability for dopamine metabolism and catecholamine uptake. Yet, cholinergic somata lacked VMAT2, precluding the potential for NE storage and vesicular release. In contrast to cholinergic somata, cardiac nerve fibers rarely showed colocalization of cholinergic and noradrenergic markers. Instead, these labels were closely apposed but clearly distinct from each other. Since cholinergic somata expressed several noradrenergic proteins, we questioned whether these neurons might also contain trophic factor receptors typical of noradrenergic neurons. Indeed, we found that all cholinergic cell bodies of mouse ICG, like noradrenergic cell bodies of the stellate ganglia, contained both tropomyosin-related kinase A (TrkA) and p75 neurotrophin receptors. Collectively, these findings demonstrate that mouse intrinsic cardiac neurons (ICNs), like those of humans, have a complex neurochemical phenotype that goes beyond the classical view of cardiac parasympathetic neurons. They also suggest that neurotrophins and local NE synthesis might have important effects on neurons of the mouse ICG. PMID- 18674603 TI - Investigation of Alternanthera repens and Bidens odorata on gastrointestinal disease. AB - Alternanthera repens and Bidens odorata are used as medication for gastrointestinal diseases today, mainly in relation to diarrhea; therefore, pharmacological tests with aqueous and ethanol extracts of both species were carried out in mice. Using charcoal meal, the activity of the four extracts on intestinal motility was determined, finding that they inhibit the advance of the gastrointestinal content. Also, the lethal media doses were estimated in order to examine the plants' safety. The data confirmed the validity of the medicinal use for both plant species, contributing to explain the use of these plants as antidiarrheal agents in Mexican traditional medicine. PMID- 18674601 TI - Timing in the absence of supraspinal input I: variable, but not fixed, spaced stimulation of the sciatic nerve undermines spinally-mediated instrumental learning. AB - Rats with complete spinal transections are capable of acquiring a simple instrumentally trained response. If rats receive shock to one hind limb when the limb is extended (controllable shock), the spinal cord will learn to hold the leg in a flexed position that minimizes shock exposure. If shock is delivered irrespective of leg position, subjects do not exhibit an increase in flexion duration and subsequently fail to learn when tested with controllable shock (learning deficit). Just 6 min of variable intermittent shock produces a learning deficit that lasts 24 h. Evidence suggests that the neural mechanisms underlying the learning deficit may be related to those involved in other instances of spinal plasticity (e.g. windup, long-term potentiation). The present paper begins to explore these relations by demonstrating that direct stimulation of the sciatic nerve also impairs instrumental learning. Six minutes of electrical stimulation (mono- or biphasic direct current [DC]) of the sciatic nerve in spinally transected rats produced a voltage-dependent learning deficit that persisted for 24 h (experiments 1-2) and was dependent on C-fiber activation (experiment 7). Exposure to continuous stimulation did not produce a deficit, but intermittent burst or single pulse (as short as 0.1 ms) stimulation (delivered at a frequency of 0.5 Hz) did, irrespective of the pattern (fixed or variable) of stimulus delivery (experiments 3-6, 8). When the duration of stimulation was extended from 6 to 30 min, a surprising result emerged; shocks applied in a random (variable) fashion impaired subsequent learning whereas shocks given in a regular pattern (fixed spacing) did not (experiments 9-10). The results imply that spinal neurons are sensitive to temporal relations and that stimulation at regular intervals can have a restorative effect. PMID- 18674602 TI - Acute citalopram has different effects on regional 5-HT synthesis in FSL, FRL, and SDP rats: an autoradiographic evaluation. AB - In this study, we measured the effect of an acute treatment of citalopram on 5 hydroxytryptamine (5-HT) synthesis in a genetic rat model of depression, the Flinders Sensitive Line (FSL) rats, their counterparts, the Flinders Resistant Line (FRL) rats, and outbred Sprague-Dawley (SPD) rats, using the alpha [(14)C]methyl-l-tryptophan (alpha-MTrp) autoradiographic method. A comparison of 5-HT synthesis in the FSL rats treated with citalopram (FSL-CTP) and those treated with saline (FSL-SAL) indicate that citalopram reduces global 5-HT synthesis in the FSL rats, as well as in all the brain areas investigated. The reduced synthesis was also observed in the dorsal raphe (DR) nucleus and the median raphe (MR) nucleus. The comparison of the synthesis between the citalopram treated SPD rats (SPD-CTP) and the saline-treated SPD rats (SPD-SAL) revealed a global increase of 5-HT synthesis in the SPD-CTP group, as well as an increase in some terminal areas, but a reduction in the DR and the MR. In contrast to the reduction throughout the brain in the FSL rats, the FRL rats treated with citalopram (FRL-CTP), when compared to the saline group (FRL-SAL), showed a global increase of 5-HT synthesis, as well as in most of the terminal areas and in the DR and the MR. The reduction of 5-HT synthesis throughout the brain in the FSL rats is likely, in part, a result of reported supersensitivity of the 5 HT(1A) receptors. Comparing changes in the SPD, FRL, and FSL rats treated with citalopram to their respective controls (saline-treated rats), the FSL rats treated acutely with citalopram were the only rats that exhibited lower 5-HT synthesis rates in all of the limbic areas, the basal ganglia, and the neocortices. This may be related to the pathophysiological basis of depressive characteristics in FSL rats. The citalopram treatment produced unexpected results in the FRL rats: 5-HT synthesis was elevated not only in most of the terminal areas, but also in the cell body areas, the DR and MR. The increase of 5-HT synthesis throughout the brain in the FRL rats is likely, in part, a result of the reported subsensitivity of the 5-HT(1A) receptors, and possibly other sites through which 5-HT synthesis could be controlled (e.g., 5-HT(1B)). In addition differences in intracellular signaling could be at least in part responsible for these differences. PMID- 18674604 TI - Preferential learning from non-affiliated individuals in jackdaws (Corvus monedula). AB - It has been suggested that affiliated social relations may facilitate information transfer between individuals. We here tested this rarely examined hypothesis with juvenile and adult jackdaws (Corvus monedula) in three stimulus enhancement tasks, both in a non-food context (experiment 1) and in a food context (experiments 2 and 3). We first show that siblings and pair partners maintain stronger bonded social relations than do non-siblings and non-pair partners. We therefore tested individuals in sibling and non-sibling dyads and, later in ontogeny, in pair and non-pair dyads. Jackdaws either did not learn from any other conspecific (experiment 1), or they learned from non-affiliated individuals (non-siblings, non-pair partners in experiments 2 and 3). This may be related to two main characteristics of jackdaws' affiliated relationships. First, affiliates share food at a high rate and may rely on their knowledgeable partners to secure food rather than learning from them. Second, affiliates spend most time in close spatial proximity to each other which increases the probability that they simultaneously experience occurrences in their environment. Hence, spatially more distant individuals, which are more likely to be non-affiliated, face different foraging situations and may therefore provide more relevant information which may lead to selective social learning. PMID- 18674605 TI - Evolution and functional divergence of monocarboxylate transporter genes in vertebrates. AB - Monocarboxylate transporters (MCTs) form a gene family with an ancient past. The identification of MCTs (MCHs) from bacteria, protozoa, fungi, invertebrates, as well as vertebrates, but not from plants and virus, allowed illuminating the phylogenetic and evolutionary history of this gene family. The significant expansion of vertebrate MCT genes should have primarily occurred after the divergence of vertebrates and invertebrates, but before the divergence time between ray-finned fish and mammals. The divergence of insect MCTs should have at least occurred in the common ancestor of fruit fly, beetle, and honeybee. Fungi monocarboxylate transporter homologues (MCHs) might evolve independently from an ancient ancestor. The results of functional divergence analysis provided statistical evidences for shifted evolutionary rate and/or changes of amino acid property after gene duplication. The sliding window analysis of the d(N)/d(S) ratio values showed that strong functional constraints must impose on the N- and C-terminal domains of vertebrate MCTs. These corresponding regions may play crucial roles for functionality of MCT proteins. PMID- 18674606 TI - The Korean traditional medicine Gyeongshingangjeehwan inhibits obesity through the regulation of leptin and PPARalpha action in OLETF rats. AB - Gyeongshingangjeehwan (GGEx), which comprises Liriope platyphylla F.T. Wang & T. Tang (Liliaceae), Platycodongrandiflorum A. DC. (Campanulaceae), Schisandrachinensis K. Koch (Magnoliaceae), and Ephedra sinica Stapf (Ephedraceae), has traditionally been used as an anti-obesity drug in Korean local clinics, although there is no evidence concerning the scientific analyses of its effects and mechanism(s) of action. Thus, we investigated the effects of GGEx on obesity, as well as the mechanism by which GGEx functions, in Otsuka Long Evans Tokushima Fatty (OLETF) male rats. Compared with obese OLETF control rats, administration of GGEx for 8 weeks significantly decreased food intake and plasma leptin levels as well as body weight gain and abdominal fat in OLETF rats. GGEx treatment not only decreased circulating triglycerides, but also inhibited lipid accumulation in the liver. GGEx increased the hepatic mRNA levels of PPARalpha target genes responsible for fatty acid beta-oxidation. Consistent with the in vivo data, GGEx elevated PPARalpha reporter gene expression in NMu2Li liver cells. These results suggest that GGEx may effectively prevent obesity and hypertriglyceridemia in part through the inhibition of feeding and the activation of hepatic PPARalpha. PMID- 18674607 TI - Frazzled cytoplasmic P-motifs are differentially required for axon pathway formation in the Drosophila embryonic CNS. AB - Frazzled is a Netrin-dependent chemoattractive receptor required for axon pathway formation in the developing Drosophila embryonic CNS. The cytoplasmic domain is important and contains three conserved P-motifs (P1, P2, and P3) thought to initiate intracellular signaling cascades and to crosstalk with other receptors during axon pathway formation. Here, we rescue homozygous frazzled embryos by pan neurally expressing a series of mutants lacking either the cytoplasmic domain or one of the conserved P-motifs and assess the ability of these mutants to rescue frazzled defects in commissural, longitudinal and motor axon pathways. Surprisingly, while the cytoplasmic domain is required, removal of an individual P-motif does not prevent gross formation of commissures. However, removal of P3 from Fra does prevent eagle-expressing commissural axons from crossing the midline in the posterior commissure suggesting that some neurons have a stronger requirement for P3-dependent signaling. Indeed, axons within the longitudinal connective as well as a small subset of motor neurons within the ISNb pathway also specifically require P3 to project to their targets correctly. In these latter axon projections, deleting the P1-motif appears to de-regulate the receptor's activity, actually increasing the frequency of motor neuron projection errors and inducing ectopic midline crossing errors. Collectively, these data demonstrate the critical nature of both the P1 and the P3-motifs to Frazzled function in vivo during axon pathway formation. PMID- 18674608 TI - Enteroaggregative Escherichia coli (EAEC) strains enter and survive within cultured intestinal epithelial cells. AB - Enteroaggregative Escherichia coli (EAEC) is an emerging pathogen associated to cases of acute or persistent diarrhea in children and adults from developed and developing countries. These microorganisms also have been isolated from human immunodeficiency virus-infected patients. EAEC exhibits aggregative adherence (AA) in HEp-2 cells. This pattern is characterized by the production of bacteria aggregates adhered to monolayer cultured cells with a "stacked brick" phenotype. The AA pattern is related to the presence of a 60MDa plasmid (pAA). In the present study, we evaluated the adherence, invasion and persistent survival of five EAEC strains with Caco-2 and T84 cells, by a bacteria invasion assay and transmission electron microscopy. EAEC isolated from cases of acute infantile diarrhea can be internalized by intestinal epithelial cells cultivated in vitro, suggesting that these strains may employ a mechanism of host cell invasion to colonize the intestinal mucosa. Results showed that EAEC strains could survive intracellularly up to 72h. Our data support evidence that EAEC is able to invade, persist and replicate within intestinal cells for extended time. This strategy may be advantageous to EAEC in colonization and survival, favoring the exploitation of an intracellular niche where these strains are protected against host clearance mechanisms, immune system and antibiotic treatment. Intracellular persistence of EAEC may be associated with development of persistent diarrhea associated to these microorganisms. To our knowledge, this is the first report of EAEC intracellular survival in cultured intestinal epithelial cells. PMID- 18674609 TI - Inhibition of lymphocyte CD3 expression by Chlamydophila pneumoniae infection. AB - Since lymphocytes are a major immune cell besides macrophages in the development of atherosclerosis, interaction between lymphocytes and Chlamydophila pneumoniae may contribute to the pathogenesis of chronic inflammatory diseases associated with C. pneumoniae. In this regard, we examined a possible alteration of CD3 expression of human lymphocyte Molt-4 cells by C. pneumoniae infection. The expression levels of CD3 molecules of lymphocyte Molt-4 cells were significantly decreased by C. pneumoniae infection. In contrast, heat-killed C. pneumoniae as well as mock (cell lysates) did not cause any alteration of CD3 expression of the cells. Treatment of the infected cells with NS-398 (cyclo-oxyganase-2 inhibitor) or AH-23848 (EP(4) prostanoid receptor antagonist) abolished the inhibition of CD3 expression. The enhanced prostaglandin E(2) (PGE(2)) productions in the culture supernatants of infected cells were confirmed by competitive enzyme immunosorbent assay (ELISA). C. pneumoniae infection of enriched lymphocytes from human peripheral blood mononuclear cells also induced a decrease of CD3 expression. Thus, C. pneumoniae infection of lymphocytes induces a decrease of CD3 expression mediated by possibly PGE(2) production. PMID- 18674610 TI - Gene discovery and comparative analysis of X-degenerate genes from the domestic cat Y chromosome. AB - Mammalian sex chromosomes are the remnants of an ancient autosomal pair present in the ancestral mammalian karyotype. As a consequence of random decay and chromosome rearrangements over evolutionary time, Y chromosome gene repertoires differ between eutherian lineages. To investigate the gene repertoire and transcriptional analysis of the domestic cat Y chromosome, and their potential roles in spermatogenesis, we obtained full-length cDNA sequences for all known Y genes and their X chromosome gametologues and used those sequences to create a BAC-based physical map of the X-degenerate region. Our results indicate the domestic cat Y chromosome has retained most X-degenerate genes that were present on the ancestral eutherian Y chromosome. Transcriptional analysis revealed that most feline X-degenerate genes have retained housekeeping functions shared by their X chromosome partners and have not been specialized for testis-specific functions. Physical mapping data indicate that the cat SRY gene is present as multiple functional copies and that very little of the felid Y chromosome may be single copy. X-Y gene divergence time estimates obtained using Bayesian methods confirm an early origin of Stratum 1 genes prior to the origin of therian mammals. We observed no statistical difference in the ages of Stratum 2 and Stratum 3 gene pairs, suggesting that eutherian and marsupial Stratum 2 genes may have been independently retained in each lineage. PMID- 18674611 TI - Immunohistochemical localization of thyrotropin-releasing hormone in the brain of reptiles. AB - To gain insight into the evolution of the thyrotropin-releasing hormone (TRH) system in the brain of vertebrates we have conducted a comparative analysis of the distribution of TRH immunoreactive cell bodies and fibers in two reptiles, the turtle Pseudemys scripta elegans and the lizard Gekko gecko. Double labeling for TRH and tyrosine hydroxylase, the main catecholamine marker, was made to help the correct localization of the labeled structures and to evaluate the possible interaction of these two systems. Cells containing TRH were found in the hypothalamic paraventricular and periventricular nuclei and their projections to the median eminence were corroborated. In addition, with some species variation, extrahypothalamic cells were present in the medial amygdala, mesencephalic tegmentum, descending vestibular nucleus and in the retina. Fiber labeling was observed in all main brain subdivisions but was more abundant in regions such as the striatum, septum, amygdaloid complex, dorsal thalamus and tegmento-reticular zones. Actual colocalization of TRH and catecholamines in the same neurons was not observed but the codistribution of cells and fibers labeled for TRH and tyrosine hydroxylase strongly suggests that they can interact in diverse regions, not only in the hypothalamo-hypophysial system. The comparison of the distribution of TRH immunoreactive neurons and fibers found in reptiles with those reported for other vertebrates reveals a strong resemblance but also notable variations, not only across vertebrate classes but also within the same class. PMID- 18674612 TI - TRP-2 expression protects HEK cells from dopamine- and hydroquinone-induced toxicity. AB - We previously reported that melanogenic enzyme TRP-2 (or DCT for DOPAchrome tautomerase) expression in WM35 melanoma cells resulted in increased intracellular GSH levels, reduction in DNA damage induced by free radicals, and decreased cell sensitivity to oxidative stress. These effects seemed to depend on a particular cellular context, because none of them were found to occur in HEK epithelial cells. We postulated that the TRP-2 beneficial effect observed in WM35 cells in the oxidative stress situation may relate to quinone metabolization and, more precisely, to the ability of TRP-2 to clear off related toxic metabolites, resulting in a global redox status modification. Here, a comparative protein expression profiling of catecholamine biosynthesis enzymes and detoxification enzymes was conducted in WM35 melanoma cells and in HEK epithelial cells, in comparison with normal human melanocytes. Results showed that WM35 cells, but not HEK cells, expressed enzymes involved in catecholamine biosynthesis, suggesting that their quinone-related toxic metabolites were present in WM35 cells but not in HEK cells. To address the issue of a possible TRP-2 beneficial effect toward quinone toxicity, cell survival experiments were then conducted in HEK cells using dopamine and hydroquinone at toxic concentrations. We showed that TRP-2 expression significantly reduced HEK cell sensitivity to both compounds. This beneficial property of TRP-2 was likely to depend on the integrity of its DOPAchrome tautomerase catalytic site, because both TRP-2(R194Q) and TRP 2(H189G), which have lost their DOPAchrome tautomerase activity, failed to modify the HEK cell response to dopamine and hydroquinone. These results suggest that TRP-2 acts on quinone metabolites other than DOPAchrome, e.g., in the catecholamine pathway, and limits their deleterious effects. PMID- 18674613 TI - Neonatal exposure to chlorpyrifos affects maternal responses and maternal aggression of female mice in adulthood. AB - CD-1 mice were exposed to the organophosphate pesticide chlorpyrifos (CPF) throughout postnatal days (PND) 11-14 at the subtoxic dose of 3 mg/kg. At adolescent age, females and males underwent a sociability test in which level of sociability and social preference were measured. At adulthood only females' behavior was analyzed. Maternal behavior of CPF-exposed females was assessed on postpartum day 1 after removal of the pups for 1 h, while anxiety levels were measured in a 5 min dark-light test on postpartum day 2. Nest defense response to an unfamiliar male intruder was assessed on postpartum day 7. In addition, from birth to postpartum day 7 a detailed analysis of nest building activity was carried out. Neonatal CPF exposure does not interfere with social behavior and social preferences at adolescence, whereas at adulthood it induces significant behavioral alterations in lactating females. Motivation to build and defend the nest was decreased in CPF females that were also less anxious than controls in the dark-light paradigm. These results confirm that developmental exposure to CPF induces long-lasting alterations in selected sexual-dimorphic responses of the adult social repertoire, and suggest that early exposure to CPF might interfere with hypothalamic neuroendocrine mechanisms regulating social responses. PMID- 18674614 TI - KU70/80, DNA-PKcs, and Artemis are essential for the rapid induction of apoptosis after massive DSB formation. AB - KU70(-/-) and DNA-PKcs(-/-/-)chicken DT40 cells are reportedly highly sensitive to the DNA topoisomerase II inhibitor etoposide. Here we report that KU70 and DNA PKcs unexpectedly function together during the induction of apoptosis after exposure to high levels of etoposide. In the presence of 100 microM etoposide, apoptosis was induced within 1 h in wild type DT40 cells but not in KU70(-/-) and DNA-PKcs(-/-/-) cells. In addition, the DNA-PK inhibitors NU7026 and wortmannin, as well as the caspase inhibitor Z-VAD-FMK, inhibited etoposide-induced apoptosis in wild type cells. Although Artemis(-/-) cells also showed defects in the etoposide-induced apoptosis, the other mutants defective in nonhomologous end joining (NHEJ), LIG4(-/-), XRCC4(-), and XLF(-/-) cells were capable to induce apoptosis. When cells were treated with high doses of etoposide, the chromatin binding of DNA-PKcs was impaired by deletion of KU70 but not of Artemis, suggesting that KU70 acts upstream of DNA-PKcs and Artemis acts downstream of DNA PKcs in the apoptotic pathway like the NHEJ pathway. These results suggest that the proteins involved in the early stage of NHEJ pathway including Artemis but not the downstream factors decide the cell fate by selecting apoptosis or DNA repair according to the degree of DNA damage. PMID- 18674615 TI - Optimal HLA matching in hematopoietic cell transplantation. AB - Only 30% of patients in need of a hematopoietic cell transplant will have an HLA identical sibling to serve as the donor. Advances in the field of immunogenetics together with the growth of donor registries and cord blood banks worldwide have provided many of these patients the opportunity for a life saving transplant. Current data demonstrate the importance of matching the unrelated donor for HLA alleles and antigens. When a matched volunteer donor is not available, use of mismatched donors may be considered. New concepts in the selection of mismatched donors include consideration for the locus, the number of mismatches, differences between alleles and antigens, the location and nature of amino acid mismatches that define class I epitopes, and the presence of haplotype mismatching. When cord blood transplantation is an option, both cell dose and HLA matching are important variables. Optimizing the overall outcome of hematopoietic cell transplantation requires an appreciation for the relative importance of HLA factors with respect to other non-genetic factors that also influence transplant outcome. PMID- 18674616 TI - Genetic causality in schizophrenia and bipolar disorder: out with the old and in with the new. AB - Schizophrenia (SZ) and bipolar disorder are highly heritable forms of major mental illness. Great hopes were held out for a major breakthrough with the advent of genome wide association studies, but to date there have been no obvious low hanging fruit. A richer harvest of strong candidates has however emerged from molecular cytogenetics and copy number variant mapping strategies promise to add more. Both strategies identify gene mutations, paving a direct path to biological studies and tests of causality. Risk genes and convergent biological pathways common to both disorders are being revealed, offering hope for a better understanding of aetiological factors and for the development of biomarkers to aid in diagnosis and prognosis. PMID- 18674617 TI - New insights on the subcortical representation of reward. AB - Reward information is represented by many subcortical areas and neuron types, which constitute a complex network. Its output is usually mediated by the basal ganglia where behaviors leading to rewards are disinhibited and behaviors leading to no reward are suppressed. Midbrain dopamine neurons modulate these basal ganglia neurons differentially using signals related to reward-prediction error. Recent studies suggest that other types of subcortical neurons assist, instruct, or work in parallel with dopamine neurons. Such reward-related neurons are found in the areas which have been associated with stress, pain, mood, emotion, memory, and arousal. These results suggest that reward needs to be understood in a larger framework of animal behavior. PMID- 18674618 TI - Overcoming the challenges of membrane protein crystallography. AB - Membrane protein structural biology is still a largely unconquered area, given that approximately 25% of all proteins are membrane proteins and yet less than 150 unique structures are available. Membrane proteins have proven to be difficult to study owing to their partially hydrophobic surfaces, flexibility and lack of stability. The field is now taking advantage of the high-throughput revolution in structural biology and methods are emerging for effective expression, solubilisation, purification and crystallisation of membrane proteins. These technical advances will lead to a rapid increase in the rate at which membrane protein structures are solved in the near future. PMID- 18674620 TI - The involvement of cellular prion protein in the autophagy pathway in neuronal cells. AB - Apoptosis and autophagy are main mechanisms of neuronal death involved in prion diseases. Serum deprivation can induce both pathways to cell death in various types of cells. To investigate whether PrP(C) is involved in autophagy pathway, we analyzed the level of microtubule-associated protein 1 light chain 3 (LC3), an autophagy marker, by monitoring the conversion from LC3-I into LC3-II in Zurich I Prnp(-/-) hippocampal neuronal cells. We found that the expression level of LC3 II was increased in Prnp(-/-) compared to wild-type cells under serum deprivation. In electron microscopy, increased accumulation of autophagosomes in Prnp(-/-) cells was correlated with the increase in levels of LC3-II. Interestingly, this up-regulated autophagic activity was retarded by the introduction of PrP(C) into Prnp(-/-) cells but not by the introduction of PrP(C) lacking octapeptide repeat region. Thus, the octapeptide repeat region of PrP(C) may play a pivotal role in the control of autophagy exhibited by PrP(C) in neuronal cells. PMID- 18674619 TI - Decreased glutamate transport enhances excitability in a rat model of cortical dysplasia. AB - Glutamate transporters function to maintain low levels of extracellular glutamate and play an important role in synaptic transmission at many synapses. Disruption of glutamate transporter function or expression can result in increased extracellular glutamate levels. Alterations in glutamate transporter expression have been reported in human epilepsy and animal seizure models. Functional electrophysiological changes that occur when transporter expression is disrupted in chronic epilepsy models have not been examined. Here, we used a freeze-induced model of cortical dysplasia to test the role of glutamate transporters in synaptic hyperexcitability. We report that inhibiting glutamate transporters with the non-selective antagonist, DL-threo-beta-benzylozyaspartic acid (TBOA) preferentially prolongs postsynaptic currents (PSCs) and decreases the threshold for evoking epileptiform activity in lesioned compared to control cortex. The effect of inhibiting uptake is mediated primarily by the glia glutamate transporter (GLT-1) since the selective antagonist dihydrokainate (DHK) mimicked the effects of TBOA. The effect of uptake inhibition is mediated by activation of N-methyl-D-aspartate (NMDA) receptors since D-(-)-2-amino-5-phosphonovaleric acid (APV) prevents TBOA-induced effects. Neurons in lesioned cortex also have a larger tonic NMDA current. These results indicate that chronic changes in glutamate transporters and NMDA receptors contribute to hyperexcitability in cortical dysplasia. PMID- 18674621 TI - Positron emission tomography ligand activation studies in the sports sciences: measuring neurochemistry in vivo. AB - Functional neuroimaging with magnetic resonance imaging (fMRI) or positron emission tomography (PET) provides the methodology to unravel some of the fascinating, but hitherto largely unresolved interactions between physical exercise and brain function. Phenomena such as raised mood, pain modulation, and sport addiction associated with physical exercise are highly interesting psychophysical models that require further in depth understanding at the neurotransmitter level. PET ligand displacement studies allow in vivo monitoring of endogenous transmitter trafficking in the entire brain and, thereby, to identify the link between exercise-induced behavioral measures and the endogenous neurotransmitter release. This review focuses on the methodology of ligand displacement in the opioidergic system, which together with the dopaminergic system has been considered as a central neurotransmitter system underlying diverse sport-induced psychophysical effects. Understanding the basic principles of exercise-induced transmitter release in the brain will potentially aid clinical applications of endurance training, both as a preventative or therapeutic intervention. PMID- 18674622 TI - Immobilization of the first dimension in 2D blue native/SDS-PAGE allows the relative quantification of membrane proteomes. AB - In biological membranes many proteins are organized in complexes. The method of choice for the global analysis of the subunits of these complexes is two dimensional blue native (2D BN)/SDS-PAGE. In the 1st dimension complexes are separated by BN-PAGE, and in the 2nd dimension their subunits are resolved by SDS PAGE. In the currently available protocols the 1st dimension BN gel lanes get distorted during their transfer to the 2nd dimension separation gels. This leads to low reproducibility and high variation of 2D BN/SDS-gels, rendering them unsuitable for comparative analysis. We have developed a 2D BN/SDS-PAGE protocol where the 1st dimension BN gel is cast on a GelBond PAG film. Immobilization prevents distortion of BN gel lanes, which lowers variation and greatly improves reproducibility of 2D BN/SDS-gels. 2D BN/SDS-PAGE with an immobilized 1st dimension was used for the comparative analysis of the cytoplasmic membrane proteomes of Escherichia coli cells overexpressing a membrane protein and to create a 2D BN/SDS-PAGE reference map of the E. coli cytoplasmic membrane proteome with 143 identified proteins from 165 different protein spots. PMID- 18674623 TI - Effects of diazinon and diazoxon on the lymphoproliferation rate of splenocytes from Nile tilapia (Oreochromis niloticus): the immunosuppresive effect could involve an increase in acetylcholine levels. AB - The lymphoproliferation rate of spleen cells from Nile tilapia (Oreochromis niloticus) exposed to the organophosphorus pesticide diazinon, to its metabolite diazoxon and to the neurotransmitter acetylcholine, was evaluated in order to explore the immunotoxic mechanism of action of this widely used insecticide. The lymphoproliferative response of spleen cells to mitogenic stimulus was not affected by either diazinon or diazoxon, indicating that these xenobiotic substances do not have direct immunotoxic properties. Conversely, ex vivo assays showed that spleen from fish exposed to diazinon presented a lower acetylcholinesterase activity and a higher acetylcholine concentration than non exposed controls. Lymphoproliferation assays also indicated that pre-exposure to acetylcholine depleted the proliferative function of spleen cells. Thus the combined information from in vitro and ex vivo experiments suggest that the immunotoxic properties of diazinon in Nile tilapia are indirect and could involve the cholinergic system of lymphocytes. PMID- 18674624 TI - The manganese superoxide dismutase gene in bay scallop Argopecten irradians: cloning, 3D modelling and mRNA expression. AB - A novel manganese superoxide dismutase (MnSOD) was cloned from bay scallop Argopecten irradians by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA of MnSOD was of 1207 bp with a 678 bp open reading frame encoding 226 amino acids. The deduced amino acid sequence contained a putative signal peptide of 26 amino acids. Sequence comparison showed that the MnSOD of A. irradians shared high identity with MnSOD in invertebrates and vertebrates, such as MnSOD from abalone Haliotis discus discus (ABG88843) and frog Xenopus laevis (AAQ63483). Furthermore, the 3D structure of bay scallop MnSOD was predicted by SWISS-MODEL Protein Modelling Server and compared with those of other MnSODs. The overall structure of bay scallop MnSOD was similar to those of zebrafish Danio rerio, fruit fly Drosophila melanogaster, Chinese shrimp Fenneropenaeus chinensis, human Homo sapiens, and had the highest similarity to scallop Mizuhopecten yessoensis and abalone H. discus discus. A quantitative real-time PCR (qRT-PCR) assay was developed to detect the mRNA expression of MnSOD in different tissues and the temporal expression in haemocytes following challenge with the bacterium Vibrio anguillarum. A higher-level of mRNA expression of MnSOD was detected in gill and mantle. The expression of MnSOD reached the highest level at 3 h post-injection with V. anguillarum and then slightly recovered from 6 to 48 h. The results indicated that bay scallop MnSOD was a constitutive and inducible protein and thus could play an important role in the immune responses against V. anguillarum infection. PMID- 18674625 TI - Visual mental imagery and perception produce opposite adaptation effects on early brain potentials. AB - Event-related potentials (ERPs) were recorded during a rapid adaptation paradigm to determine whether visual perception and visual mental imagery of faces recruit the same early perceptual processes. The early effect of face and object adaptors, either perceived or visualized, on test stimuli, was assessed by measuring the amplitude of the N170/VPP complex, typically much larger for faces than for other object categories. Faces elicited a robust N170/VPP complex, localized to posterior ventrolateral occipitotemporal cortex. Both visualized and perceived adaptors affected the N170/VPP complex to test faces from 120 ms post stimulus, reflecting effects on neural populations supporting early perceptual face categorization. Critically, while perceived adaptors suppressed the amplitude of the N170/VPP, visualized adaptors enhanced it. We suggest that perceived adaptors affect neural populations in the neocortex supporting early perceptual processing of faces via bottom-up mechanisms, whereas visualized adaptors affect them via top-down mechanisms. Similar enhancement effects were found on the N170/VPP complex to non-face objects, suggesting such effects are a general consequence of visual imagery on processing of faces and other object categories. These findings support image-percept equivalence theories and may explain, in part, why visual percepts and visual mental images are not routinely confused, even though both engage similar neural populations in the visual system. PMID- 18674626 TI - Morphological priming in overt language production: electrophysiological evidence from Dutch. AB - The present study investigated morphological priming in Dutch and its time course in overt speech production using a long-lag priming paradigm. Prime words were compounds that were morphologically related to a picture name (e.g. the word jaszak, 'coat pocket' was used for a picture of a coat; Dutch jas) or form related monomorphemic words (e.g. jasmijn, 'jasmine'). The morphologically related compounds could be semantically transparent (e.g. eksternest, 'magpie nest') or opaque (e.g. eksteroog, lit. 'magpie eye', 'corn', for a picture of a magpie, Dutch ekster). Behavioral and event-related potential (ERP) data were collected in two sessions. The production of morphologically related and complex words facilitated subsequent picture naming and elicited a reduced N400 compared with unrelated prime words. The effects did not differ for transparent and opaque relations. Mere form overlap between a prime word and a target picture name did not affect picture naming. These results extend previous findings from German to another language and demonstrate the feasibility of measuring cognitive ERP components during overt speech. Furthermore, the results suggest that morphological priming in language production cannot be reduced to semantic and phonological processing. The time course of these priming effects as reflected in the ERP measure is in accordance with a meta-analytic temporal estimate of morphological encoding in speaking [Indefrey, P., & Levelt, W.J.M. (2004). The spatial and temporal signatures of word production components. Cognition, 92, 101 144.] suggesting that morphological relations are encoded at the word form level. PMID- 18674627 TI - Asymmetric control mechanisms of bimanual coordination: an application of directed connectivity analysis to kinematic and functional MRI data. AB - Mirror-symmetrical bimanual movement is more stable than parallel bimanual movement. This is well established at the kinematic level. We used functional MRI (fMRI) to evaluate the neural substrates of the stability of mirror-symmetrical bimanual movement. Right-handed participants (n=17) rotated disks with their index fingers bimanually, both in mirror-symmetrical and asymmetrical parallel modes. We applied the Akaike causality model to both kinematic and fMRI time series data. We hypothesized that kinematic stability is represented by the extent of neural "cross-talk": as the fraction of signals that are common to controlling both hands increases, the stability also increases. The standard deviation of the phase difference for the mirror mode was significantly smaller than that for the parallel mode, confirming that the former was more stable. We used the noise-contribution ratio (NCR), which was computed using a multivariate autoregressive model with latent variables, as a direct measure of the cross-talk between both the two hands and the bilateral primary motor cortices (M1s). The mode-by-direction interaction of the NCR was significant in both the kinematic and fMRI data. Furthermore, in both sets of data, the NCR from the right hand (left M1) to the left (right M1) was more prominent than vice versa during the mirror-symmetrical mode, whereas no difference was observed during parallel movement or rest. The asymmetric interhemispheric interaction from the left M1 to the right M1 during symmetric bimanual movement might represent cortical-level cross-talk, which contributes to the stability of symmetric bimanual movements. PMID- 18674628 TI - Sensitization and habituation regulate reinforcer effectiveness. AB - We argue that sensitization and habituation occur to the sensory properties of reinforcers when those reinforcers are presented repeatedly or for a prolonged time. Sensitization increases, and habituation decreases, the ability of a reinforcer to control behavior. Supporting this argument, the rate of operant responding changes systematically within experimental sessions even when the programmed rate of reinforcement is held constant across the session. These within-session changes in operant responding are produced by repeated delivery of the reinforcer, and their empirical characteristics correspond to the characteristics of behavior undergoing sensitization and habituation. Two characteristics of habituation (dishabituation, stimulus specificity) are particularly useful in separating habituation from alternative explanations. Arguing that habituation occurs to reinforcers expands the domain of habituation. The argument implies that habituation occurs to biologically important, not just to neutral, stimuli. The argument also implies that habituation may be observed in "voluntary" (operant), not just in reflexive, behavior. Expanding the domain of habituation has important implications for understanding operant and classical conditioning. Habituation may also contribute to the regulation of motivated behaviors. Habituation provides a more accurate and a less cumbersome explanation for motivated behaviors than homeostasis. Habituation also has some surprising, and easily testable, implications for the control of motivated behaviors. PMID- 18674629 TI - Spa2 is required for morphogenesis but it is dispensable for pathogenicity in the phytopathogenic fungus Ustilago maydis. AB - The increasing evidence linking regulation of polar growth and pathogenicity in fungi has elicited a significant effort devoted to produce a better understanding of mechanisms determining polarization in pathogenic fungi. Here we characterize in the phytopathogenic basidiomycete Ustilago maydis, the Spa2 protein, a well known component of polarisome, firstly described in Saccharomyces cerevisiae. U. maydis display a dimorphic switch between budding growth of hapoid cells and filamentous growth of the dikaryon. During yeast growth, a GFP-tagged Spa2 protein localized to distinct growth sites in a cell cycle-specific manner, while during hyphal growth is persistently located to hyphal tips. Deletion of spa2 gene produces rounder budding cells and thicker filaments than wild-type cells, suggesting a role of Spa2 for the determination of the growth area in U. maydis. We also address the connections between Spa2 and the actin- and microtubule cytoskeleton. We found that the absence of Spa2 does not affect cytoskeleton organization and strikingly, interference with actin filament or microtubule formation does not affect the polar localization of Spa2. In contrast, defects in the small GTPase Rac1 seems to affect the ability of Spa2 to locate to precise sites at the tip cell. Finally, to our surprise, we found that cells defectives in Spa2 function were as pathogenic as wild-type cells. PMID- 18674630 TI - Molecular cloning, tissue distribution and enzymatic characterization of cathepsin X from olive flounder (Paralichthys olivaceus). AB - In this study, we have cloned a cDNA encoding for cathepsin X (PoCtX) from the olive flounder, Paralichthys olivaceus. The presence of an HIP motif, which is conserved in the unique cathepsin X family, PoCtX, clearly shows its relation to the cathepsin X group, apart from the cathepsin L or B subfamily. The results of RT-PCR and real-time PCR analyses revealed ubiquitous PoCtX expression in normal and LPS-stimulated tissues. The cDNA encoding for the proenzyme of PoCtX (proPoCtX) was expressed in Escherichia coli as a 57 kDa fusion protein with glutathione S-transferase. Its activity was quantified via the cleavage of the synthetic fluorogenic peptide substrate Z-Phe-Arg-AMC, and the optimal pH for the protease activity was 5. The recombinant proPoCtX was inhibited by antipain and leupeptin. The PoCtX protein from P. olivaceus muscle extracts was purified 9.48 fold via a one-step purification process using a DEAE-Sephagel high performance liquid chromatography (HPLC) column. Western blotting and ELISA were conducted in order to evaluate the reaction ability and detection-specificity of the anti proPoCtX polyclonal antibody to native PoCtX and recombinant proPoCtX proteins. Our findings indicate that the P. olivaceus cathepsin X is highly conserved within the cathepsin X subfamily in terms of its amino acid sequence, tissue expression, and biochemical activity. PMID- 18674632 TI - Improving production of bioactive secondary metabolites in actinomycetes by metabolic engineering. AB - Production of secondary metabolites is a process influenced by several physico chemical factors including nutrient supply, oxygenation, temperature and pH. These factors have been traditionally controlled and optimized in industrial fermentations in order to enhance metabolite production. In addition, traditional mutagenesis programs have been used by the pharmaceutical industry for strain and production yield improvement. In the last years, the development of recombinant DNA technology has provided new tools for approaching yields improvement by means of genetic manipulation of biosynthetic pathways. These efforts are usually focused in redirecting precursor metabolic fluxes, deregulation of biosynthetic pathways and overexpression of specific enzymes involved in metabolic bottlenecks. In addition, efforts have been made for the heterologous expression of biosynthetic gene clusters in other organisms, looking not only for an increase of production levels but also to speed the process by using rapidly growing and easy to manipulate organisms compared to the producing organism. In this review, we will focus on these genetic approaches as applied to bioactive secondary metabolites produced by actinomycetes. PMID- 18674631 TI - Temporal and spatial expression pattern of the myostatin gene during larval and juvenile stages of the Chilean flounder (Paralichthys adspersus). AB - The full length cDNA sequence of the myostatin gene was cloned from a teleostean fish, the Chilean flounder (Paralichthys adspersus) through RT-PCR amplification coupled with the RACE approach to complete the 5'- and 3'-region. The deduced amino acid sequence encodes a protein of 377 amino acid residues, including the structural domains responsible for its biological activity. Amino acid sequence comparison revealed high sequence conservation, and confirmed that the isolated sequence corresponds to the MSTN1 gene. Gene expression analysis showed that cfMSTN mRNA is present in a wide variety of tissues in juvenile fish. In addition, we assessed the spatial expression pattern of the MSTN mRNA during embryos and larval stages through whole mount in situ hybridization. No expression was observed in embryos, whereas in larvae of 8 and 9 days post fertilization, the notochord, somites, intestine and some discrete territories in the head, such as brain and eye, were positive for MSTN mRNA. Our results contribute to the knowledge of the MSTN system in larval and juvenile stages; in particular the strong expression observed in the notochord suggests that MSTN, in synchronization with positive growth signals, may play an important role in the control of the development of larvae somites. PMID- 18674633 TI - Liver invariant NKT cells and listeriosis. AB - The invariant (i) NKT cells represent unique T lymphocytes expressing TCRValpha14. Although iNKT cells have been regarded as T lymphocytes expressing NK1.1, they do not consistently express this marker. NK1.1 allows recognition of "missing-self" and thus controls inhibition/activation of iNKT cells. It is thus tempting to assume that iNKT cells participate in the regulation of host immune responses during microbial infection by controlling NK1.1 expression. These findings shed light on the unique role of iNKT cells in microbial infection and provide an evidence for unique aspects of the NK1.1 on these cells as a regulatory molecule. PMID- 18674634 TI - The Plasmodium falciparum heat shock protein 40, Pfj4, associates with heat shock protein 70 and shows similar heat induction and localisation patterns. AB - Human cerebral malaria is caused by the protozoan parasite Plasmodium falciparum, which establishes itself within erythrocytes. The normal body temperature in the human host could constitute a possible source of heat stress to the parasite. Molecular chaperones belonging to the heat shock protein (Hsp) class are thought to be important for parasite subsistence in the host cell, as the expression of some members of this family has been reported to increase upon heat shock. In this paper we investigated the possible functions of the P. falciparum heat shock protein DnaJ homologue Pfj4, a type II Hsp40 protein. We analysed the ability of Pfj4 to functionally replace Escherichia coli Hsp40 proteins in a dnaJ cbpA mutant strain. Western analysis on cellular fractions of P. falciparum-infected erythrocytes revealed that Pfj4 expression increased upon heat shock. Localisation studies using immunofluorescence and immuno-electron microscopy suggested that Pfj4 and P. falciparum Hsp70, PfHsp70-1, were both localised to the parasites nucleus and cytoplasm. In some cases, Pfj4 was also detected in the erythrocyte cytoplasm of infected erythrocytes. Immunoprecipitation studies and size exclusion chromatography indicated that Pfj4 and PfHsp70-1 may directly or indirectly interact. Our results suggest a possible involvement of Pfj4 together with PfHsp70-1 in cytoprotection, and therefore, parasite survival inside the erythrocyte. PMID- 18674635 TI - A genotoxicity test system based on p53R2 gene expression in human cells: assessment of its reactivity to various classes of genotoxic chemicals. AB - The tumor suppressor, p53, plays an important role in DNA damage repair, by regulating the expression of target genes. One p53-target gene, p53R2, which encodes a subunit of ribonucleotide reductase, is activated by DNA damage. We have previously developed a genotoxicity test system, using human cell lines and a p53R2-dependent luciferase reporter gene assay. 80 chemicals have been examined with this system and 40 of 43 Ames-positive chemicals induced luciferase activity. Eight Ames-negative genotoxic chemicals also induced luciferase activity. Although this assay system could, potentially, be applied to the rapid screening of chemicals that are potentially genotoxic to humans, the ability of the assay to detect genotoxic effects was unclear. In this study, to evaluate the performance of this assay system, several different types of DNA damaging agents were screened. 27 chemicals, whose genotoxic mechanisms are well known, were screened. All genotoxic compounds, except for anti-metabolites and histone deacetylase HDAC inhibitors, showed significant luciferase activity with the following rank order of potency: topoisomerase II inhibitors, intercalaters>bleomycin>topoisomerase I inhibitors>alkylating agents=DNA cross linking agents=polycyclic aromatic hydrocarbons>spindle poisons. This assay showed greater response to those genotoxic agents that induce DNA double strand break damage compared to those agents that cause other forms of DNA damage. DNA double strand breakage initiates genomic instability, a feature of carcinogenicity. These results indicate that this assay system could be a helpful tool for predicting chemical genotoxicity and carcinogenicity in humans. PMID- 18674636 TI - Formaldehyde exposure and leukemia: a new meta-analysis and potential mechanisms. AB - Formaldehyde is an economically important chemical, to which more than 2 million U.S. workers are occupationally exposed. Substantially more people are exposed to formaldehyde environmentally, as it is generated by automobile engines, is a component of tobacco smoke and is released from household products, including furniture, particleboard, plywood, and carpeting. The International Agency for Research on Cancer (IARC) recently classified formaldehyde as a human carcinogen that causes nasopharyngeal cancer and also concluded that there is "strong but not sufficient evidence for a causal association between leukemia and occupational exposure to formaldehyde". Here, we review the epidemiological studies published to date on formaldehyde-exposed workers and professionals in relation to lymphohematopoietic malignances. In a new meta-analysis of these studies, focusing on occupations known to have high formaldehyde exposure, we show that summary relative risks (RRs) were elevated in 15 studies of leukemia (RR=1.54; confidence interval (CI), 1.18-2.00) with the highest relative risks seen in the six studies of myeloid leukemia (RR=1.90; 95% CI, 1.31-2.76). The biological plausibility of this observed association is discussed and potential mechanisms proposed. We hypothesize that formaldehyde may act on bone marrow directly or, alternatively, may cause leukemia by damaging the hematopoietic stem or early progenitor cells that are located in the circulating blood or nasal passages, which then travel to the bone marrow and become leukemic stem cells. To test these hypotheses, we recommend that future studies apply biomarkers validated for other chemical leukemogens to the study of formaldehyde. PMID- 18674637 TI - The vascular S1P gradient-cellular sources and biological significance. AB - Sphingosine 1-phosphate (S1P), a product of sphingomyelin metabolism, is enriched in the circulatory system whereas it is estimated to be much lower in interstitial fluids of tissues. This concentration gradient, termed the vascular S1P gradient appears to form as a result of substrate availability and the action of metabolic enzymes. S1P levels in blood and lymph are estimated to be in the muM range. In the immune system, the S1P gradient is needed as a spatial cue for lymphocyte and hematopoietic cell trafficking. During inflammatory reactions in which enhanced vascular permeability occurs, a burst of S1P becomes available to its receptors in the extravascular compartment, which likely contributes to the tissue reactions. Thus, the presence of the vascular S1P gradient is thought to contribute to physiological and pathological conditions. From an evolutionary perspective, S1P receptors may have co-evolved with the advent of a closed vascular system and the trafficking paradigms for hematopoietic cells to navigate in and out of the vascular system. PMID- 18674638 TI - Targeting insulin and insulin-like growth factor signalling in oncology. AB - The family of insulin/insulin-like growth factor (IGF) receptors regulates many crucial aspects of cellular and whole-organism physiology. Evidence that targeting these receptors may be useful in cancer treatment was first recognized more than 20 years ago. Drug development began relatively recently, justified both by laboratory studies and by circumstantial clinical evidence that this receptor family is involved in the molecular pathophysiology of neoplasia. Pharmacologic targeting strategies include both small molecule receptor tyrosine kinase inhibitors and anti-receptor antibodies. More than a dozen drug candidates have been studied preclinically, and several are now being evaluated in clinical trials. These trials have provided evidence suggesting safety of the anti-IGF-I receptor antibodies, a few anecdotes of impressive single-agent activity, and early evidence for a significant improvement in response rate to chemotherapy for lung cancer with co-administration of an anti-IGF-I receptor antibody. This experience has justified expanded clinical trials programs to evaluate several of the IGF-I receptor targeting agents in many different areas of clinical need. Most of these trials will involve assessing activity of rational combinations of IGF-I receptor targeting agents with currently approved drugs. PMID- 18674640 TI - Molecular epidemiology of dengue virus type 3 in Northern South America: 2000- 2005. AB - A phylogenetic approach was used to identify genetic variants of DENV-3 subtype III that may have emerged during or after its expansion throughout South America. We sequenced the capsid, premembrane/membrane and envelope genes from 22 DENV-3 strains isolated from Venezuela, Bolivia, Ecuador and Peru between 2000 and 2005. Phylogenetic analysis showed that the isolates sequenced in this study formed three clades within subtype III: one with the isolates from Venezuela, one with the Bolivian isolates and one with the isolates from Ecuador and Peru. PMID- 18674639 TI - Targeting the androgen receptor pathway in prostate cancer. AB - When prostate cancers progress following androgen depletion therapy, there are currently few treatment options with only one, docetaxel, that has been shown to prolong life. Recent work has shown that castration-resistant prostate cancers (CRPCs) continue to depend on androgen receptor (AR) signaling which is reactivated despite low serum androgen levels. Currently available AR-targeted therapy, including GnRH agonists and antiandrogens, cannot completely shut down AR signaling. Several mechanisms that enhance AR signaling in an androgen depleted environment have been elucidated. These include AR mutations that allow activation by low androgen levels or by other endogenous steroids, AR overexpression, increased local intracrine synthesis of androgens, and upregulation of tyrosine kinase pathways. This has led to the development of a number of novel agents targeting the AR signaling pathway, including more effective antiandrogens, inhibitors of CYP17, an enzyme required for androgen synthesis, inhibitors of 5alpha-reductase, inhibitors of HSP90 which protects AR from degradation, inhibitors of histone deacetylases which is required for optimal AR-mediated transcription, as well as inhibitors of tyrosine kinase inhibitors. Many of these strategies are currently being tested in clinical trials in CRPC. PMID- 18674641 TI - Diterpenes drive Th1 polarization depending on IL-12. AB - Sandaracopimaric acid and Sandaracopimaradiene-3beta-ol are diterpenes isolated from the heatwood of Cryptomeria japonica and are pharmacologically active substances. Dendritic cells (DC) are key antigen presenting cells (APC), which link innate and adaptive immunity, ultimately activating antigen-specific T cells. We demonstrate that Sandaracopimaric acid and Sandaracopimaradiene-3beta ol activate humans DC as documented by phenotypic and functional maturation and altered cytokine production. The expression of the co-stimulatory molecules such as CD83, CD86 and HLA-DR on Sandaracopimaric acid- and Sandaracopimaradiene-3beta ol-primed DC was enhanced. Sandaracopimaric acid- and Sandaracopimaradiene-3beta ol-primed DC also enhanced the T cell stimulatory capacity in an allo MLR. Naive T cells co-cultured with Sandaracopimaric acid- or Sandaracopimaradiene-3beta-ol primed DC turned into typical Th1 cells, which produced large quantities of IFN gamma and released small amounts of IL-4 depending on IL-12 secretion. Sandaracopimaric acid- or Sandaracopimaradiene-3beta-ol-primed DC had a high migration to macrophage inflammatory protein (MIP)-3beta. These results suggest that some diterpenes modulate human DC function in a fashion that favors Th1 cell polarization and may be used on DC-based vaccines for cancer immunotherapy. PMID- 18674642 TI - Implication of receptor for advanced glycation end product (RAGE) in pulmonary health and pathophysiology. AB - Receptor for advanced glycation end products (RAGE) is a membrane bound receptor and member of the immunoglobulin super family and is normally present in a highly abundant basal level expression in lung. This high expression of RAGE in lung alveolar epithelial type I (ATI) cells is presumably involved in the proliferation and differentiation of pulmonary epithelial cells. However, typically higher than basal level expression of RAGE may indicate the existence of severe pathophysiological condition in lung, e.g. acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). During pulmonary tissue injury an endogenous secretory isoform of RAGE called EsRAGE is noticed at high levels in broncho-alveolar lavage (BAL) and plasma. Recently, a soluble form of RAGE (sRAGE) produced by recombinant gene technology was shown to exhibit a therapeutic potential in experimental animal models. Detailed study of RAGE in the pulmonary tissues will facilitate the understanding of the importance of RAGE signaling in the pulmonary health and pathophysiology. PMID- 18674643 TI - Modeling airflow and particle transport/deposition in pulmonary airways. AB - A review of research papers is presented, pertinent to computer modeling of airflow as well as nano- and micron-size particle deposition in pulmonary airway replicas. The key modeling steps are outlined, including construction of suitable airway geometries, mathematical description of the air-particle transport phenomena and computer simulation of micron and nanoparticle depositions. Specifically, diffusion-dominated nanomaterial deposits on airway surfaces much more uniformly than micron particles of the same material. This may imply different toxicity effects. Due to impaction and secondary flows, micron particles tend to accumulate around the carinal ridges and to form "hot spots", i.e., locally high concentrations which may lead to tumor developments. Inhaled particles in the size range of 20nm< or =dp< or =3microm may readily reach the deeper lung region. Concerning inhaled therapeutic particles, optimal parameters for mechanical drug-aerosol targeting of predetermined lung areas can be computed, given representative pulmonary airways. PMID- 18674644 TI - Functional characterization of pathogen-responsive protein AtDabb1 with an antifungal activity from Arabidopsis thaliana. AB - A plant antifungal protein was purified from Arabidopsis thaliana leaves by using a typical procedure consisting of anion exchange chromatography and high performance liquid chromatography. We determined the amino acid sequence of the purified protein using MALDI-TOF/MS analysis, and found that the sequence matched that of a hypothetical Arabidopsis protein in GenBank (accession number NP_175547). We designated the protein as AtDabb1. After the cDNA encoding the AtDabb1 gene was cloned from an Arabidopsis leaf cDNA library, the recombinant protein was expressed in Escherichia coli and found to significantly inhibit cell growth of various pathogenic fungal strains. mRNA expression of the AtDabb1 gene was induced by pathogen-related signaling molecules including salicylic acid and jasmonic acid. These results suggest that AtDabb1 may contribute to the induced plant defense mechanism against diverse pathogenic fungi. PMID- 18674645 TI - M-banding characterization of a 16p11.2p13.1 tandem duplication in a child with autism, neurodevelopmental delay and dysmorphism. AB - We describe a partial duplication of the chromosome 16 short arm [46,XY,dup(16)(p11.2p13.1)] in an Iranian girl with autism, neurodevelopmental delay, mental retardation, very poor memory, and dysmorphism including sparse hair, upslanting palpebral fissures, long philtrum, micrognathia, hypotonia, small feet and hands, syndactyly of the fingers, and hypoplastic thumbs. The patient now four years old, has a normal twin sister, and the parents are unrelated. The abnormal 16p was originally detected by banding cytogenetic techniques, and was characterized by multicolour banding fluorescence in situ hybridization (MCB). The MCB pattern on the derivative chromosome 16 indicated a direct duplication of the region 16p11.2 to 16p13.1. PMID- 18674646 TI - Cytogenetic and molecular studies of an X;21 translocation previously diagnosed as complete monosomy 21. AB - We studied a child with apparent monosomy of chromosome 21. Cytogenetic, FISH and microsatellite analyses revealed a 45,X,-21,+der(X)t(X;21)(q25;q21.1) karyotype resulting from a de novo, unbalanced, X;21 non-reciprocal translocation of paternal origin, with partial monosomy of chromosomes 21 and X. An extreme, skewed X-inactivation pattern of the der(X) chromosome was demonstrated. Skewed inactivation probably accounted for a mild phenotype with respect to Xq25-->qter deletion while propagation of inactivation to the adjacent 21q region may account for mild clinical features associated to distal 21q monosomy. PMID- 18674647 TI - A cryptic unbalanced translocation t(2;9)(p25.2;q34.3) causes the phenotype of 9q subtelomeric deletion syndrome and additional exophthalmos and joint contractures. AB - We report on a 26-year-old woman with microcephaly, typical facial features of 9q subtelomeric deletion syndrome, exophthalmos, contractures of elbow and knee joints, severe muscular hypotonia, no ability to walk, and no speech development. Array CGH revealed a cryptic 9q34.3 deletion and 2p25.2-p25.3 duplication transmitted by her mother, who was carrying a balanced translocation of chromosomes 2p and 9q. There are about 50 reported cases of deletions of the subtelomeric part of chromosome 9q, however, duplications of only the terminal part of chromosome 2p are rare. Neuroblastoma, diaphragmatic hernia, neural tube defects, broncho-pulmonary abnormalities, and congenital heart defects are conditions associated with partial trisomy of larger fragments of 2p. To our knowledge there is only one case described with an isolated duplication as distal as in the patient reported here. Joint contractures and exophthalmos observed in this patient are also seen in our patient. These features are not allegeable by the deletion 9q34.3 identified in the patient reported here and may be a hint that terminal duplication of 2p could be associated with exophthalmos and contractures. PMID- 18674648 TI - Monosomy 19pter and trisomy 19q13-qter in two siblings arising from a maternal pericentric inversion: clinical data and molecular characterization. AB - Pericentric inversions of chromosome 19 are very rare rearrangements. Only one case was shown to have resulted in duplication deficiency in the offspring. We report a familial case of pericentric inversion of chromosome 19 not detectable by standard karyotype and usual subtelomeric FISH probes. This inversion was transmitted in its balanced and in its recombinant form to the offspring. The two children carrying the recombinant chromosome 19 presented with growth and mental retardation, microcephaly, mild facial dysmorphism and clinodactyly. The recombinant chromosome 19 was characterized by FISH and array CGH. It consisted of a 400kb 19pter deletion and a 6.9Mb (19q13.33-qter) duplication. This observation supports the recombination risk of pericentric inversion of chromosome 19 and emphasizes the role of molecular cytogenetics techniques in the characterization of chromosome 19 rearrangements. PMID- 18674650 TI - High molecular weight tropomyosins regulate osteoclast cytoskeletal morphology. AB - Tropomyosins are coiled-coil dimers that bind to the major groove of F-actin and regulate its accessibility to actin-modifying proteins. Although approximately 40 tropomyosin isoforms have been identified in mammals, they can broadly be classified into two groups based on protein size, that is, high molecular weight and low molecular weight isoforms. Osteoclasts, which undergo rounds of polarization and depolarization as they progress through the resorptive cycle, possess an unusual and highly dynamic actin cytoskeleton. To further define some of the actin regulatory proteins involved in osteoclast activity, we previously performed a survey of tropomyosin isoforms in resting and resorbing osteoclasts. Osteoclasts were found to express two closely related tropomyosins of the high molecular weight type, which are not expressed in monocytic and macrophage precursors. These isoforms, Tm-2 and Tm-3, are not strongly associated with actin rich adhesion structures, but are instead distributed diffusely throughout the cell. In this study, we found that Tm-2/3 expression occurs late in osteoclastogenesis and continues to increase as cells mature. Knockdown of these isoforms via RNA interference results in flattening and increased spreading of osteoclasts, accompanied by diminished motility and altered resorptive capacity. In contrast, overexpression of Tm-2, but not Tm-3, caused morphological changes that include decreased spreading of the cells and induction of actin patches or stress fiber-like actin filaments, also with effects on motility and resorption. Suppression of Tm-2/3 or overexpression of Tm-2 resulted in altered distribution of gelsolin and microfilament barbed ends. These data suggest that high molecular weight tropomyosins are expressed in fusing osteoclasts to regulate the cytoskeletal scaffolding of these large cells, due at least in part by moderating accessibility of gelsolin to these microfilaments. PMID- 18674649 TI - Transcriptional regulation of neuronal differentiation: the epigenetic layer of complexity. AB - The transcriptional programs of neural progenitor cells change dynamically during neurogenesis, a process regulated by both intrinsic and extrinsic factors. Although many of the transcription factors required for neuronal differentiation have long been identified, we are only at the brink of understanding how epigenetic mechanisms influence transcriptional activity and the accessibility of transcription factors to bind consensus cis-elements. Herein, we delineate the chief epigenetic modifications and the machinery responsible for these alterations. Further, we review the epigenetic modifications presently known to participate in the maintenance of the neural progenitor cell state and in the regulation of neuronal differentiation. PMID- 18674651 TI - Discussion: 'Reproductive factors associated with nocturia and urgency' by Tikkinen et al. AB - In the roundtable discussion that follows, clinicians discuss a study that is published in this issue of the Journal in light of its methods, relevance to practice, and implications for future research. Article discussed: Tikkinen KAO, Auvinen A, Tiitinen A, Valpas A, Johnson TM 2nd, Tammela TLJ. Reproductive factors associated with nocturia and urinary urgency in women: a population-based study in Finland. Am J Obstet Gynecol 2008;199:153.e1-153.e12. PMID- 18674652 TI - Steering Committee of the Eunice Kennedy Shriver National Institute of Child Health and Human Development's (NICHD) Maternal-Fetal Medicine Units Network. PMID- 18674653 TI - Group A Streptococcus vulvovaginitis in breastfeeding women. AB - Group A beta-hemolytic streptococcus-associated vulvovaginitis is uncommon in adult women. Clinicians should include group A beta-hemolytic streptococcus as a possible cause of vulvovaginal symptoms in breastfeeding women. Along with appropriate antibiotic therapy, vaginal estrogen therapy may be considered to diminish susceptibility to recurrent infection in women with vaginal atrophy. PMID- 18674654 TI - A call for an international consortium on the genetics of preterm birth. PMID- 18674655 TI - Toward gene therapy of endometriosis: transductional and transcriptional targeting of adenoviral vectors to endometriosis cells. AB - OBJECTIVE: The purpose of this study was to screen a panel of targeted adenoviruses as vectors for endometriosis gene therapy. STUDY DESIGN: Endometriotic cells were obtained from subjects with ovarian endometriomas. Liver tissues were taken from donors during hepatic transplantation surgery. Human endometriotic cells and liver tissues were transfected by targeted adenoviruses expressing luciferase reporter gene. Luciferase activity that was mediated by each virus was expressed as a percentage of adenovirus serotype 5 (Ad5-CMV-luc) activity. The 2-tailed Studentt test was used to compare the adenovirus data. RESULTS: In endometriotic cells, the adenovirus-RGD (Ad-RGD-luc), adenovirus under secretory leukocyte protease inhibitor promoter (Ad-SLPI-luc), and adenovirus under heparanase promoter (Ad-heparanase-luc) showed significantly higher activity, compared with the adenovirus serotype 5. In liver tissues, adenovirus-survivin (Ad-survivin-luc) and Ad-heparanase-luc had significantly lower activity, compared with adenovirus serotype 5. CONCLUSION: Ad-heparanase luc showed "endometriosis on, liver off" phenotype and is a promising vector for endometriosis gene therapy. PMID- 18674656 TI - Gynecologic malignancies in Ashkenazi families with the MSH2 A636P founder mutation. AB - OBJECTIVE: A founder mutation A636P in the MSH2 gene was found to be related to hereditary nonpolyposis colorectal cancer in Ashkenazi Jews. Although the incidence of colorectal cancer in carriers is relatively well established, the frequency of other tumors is less clear. STUDY DESIGN: We studied a consecutive series of 19 carrier families that were cared for by the Clalit Health Studies National Familial Cancer Consultation Service, most of whom were identified through a population-based case-control study of colorectal cancer in northern Israel. RESULTS: Gynecologic cancers, 88% of which (28 cases) were endometrial cancers, were diagnosed in 78.9% of the carrier families and in 26.2% of the women who were at risk, with a mean age at diagnosis of 51.2 years. Forty-six percent of the women with endometrial cancer reported at least 1 other primary tumor. CONCLUSION: Genetic counseling and testing for the MSH2 A636P mutation is indicated for Ashkenazi Jewish women with an endometrial cancer, especially if the cancer is detected before the age of 70 years in women with a personal or family history of colorectal cancer. PMID- 18674657 TI - Cervical biopsy and excision procedure specimens lack sufficient predictive value for lymph-vascular space invasion seen at hysterectomy for cervical cancer. AB - OBJECTIVE: The purpose of this study was to determine whether lymph-vascular space invasion (LVSI) that is discovered in cervical biopsy and excision specimens is associated with LVSI in the hysterectomy specimen of patients with cervical cancer. STUDY DESIGN: A retrospective pathologic review to determine the presence of LVSI in cervical biopsy specimens, cold-knife cone biopsy (CKC biopsy), and loop electrical excision procedure (LEEP) specimens that contained cervical cancer was performed if subsequent hysterectomy results were available for review. Data were analyzed with chi-square analysis testing. RESULTS: One hundred six patients were identified. The negative predictive value of the biopsy is lower at 0.45 than either the CKC biopsy (0.83) or LEEP (0.57); however, the positive predictive value (0.83) is higher than either CKC biopsy (0.50) or LEEP (0.75). LVSI, when present in cervical biopsy (odds ratio, 4.13; 95% CI, 0.414 98.446), CKC biopsy (odds ratio, 4.8; 95% CI, 0.542-46.280), and LEEP (odds ratio, 4.0; 95% CI, 0.439-43.793) specimens, is associated with a statistically insignificant increased risk of LVSI in the hysterectomy specimen. CONCLUSION: Cervical biopsy and excision specimens lack sufficient negative predictive value for the detection of LVSI in the hysterectomy specimen. PMID- 18674658 TI - Genetic regulation of cervical antiinflammatory cytokine concentrations during pregnancy. AB - OBJECTIVE: The purpose of this study was to evaluate the contribution of maternal genotype to the concentration of interleukin (IL)-4, -10, and -13 in the cervix during the first trimester in the context of bacterial vaginosis and cigarette smoking. STUDY DESIGN: In this prospective cohort of 116 pregnant women, we examined 72 single nucleotide polymorphisms (SNPs) in 6 genes: IL-4, IL-4 receptor, IL-10, IL-10 receptor alpha, IL-10 receptor beta, and IL-13. RESULTS: Among black women, an IL-10 receptor beta SNP (rs6517158) demonstrated significant association with IL-10 concentration (P = .04), adjusted for bacterial vaginosis. Among white women, there were 4 SNPs in IL-10 receptor alpha with a highly significant association with IL-10 concentration, adjusted for bacterial vaginosis and smoking. CONCLUSION: Our data demonstrate a significant relation between cervical concentrations of IL-10 and SNPs in the IL-10 receptor alpha and beta genes. The contribution of genotype to IL-10 concentration is significant, even in the consideration of bacterial vaginosis and smoking. PMID- 18674659 TI - Transplacental transfer and metabolism of 17-alpha-hydroxyprogesterone caproate. AB - OBJECTIVE: Determine transplacental transfer and metabolism of 17-alpha hydroxyprogesterone caproate and its distribution between the tissue and the maternal and fetal circuits of the dually perfused placental lobule. STUDY DESIGN: 17-alpha-Hydroxyprogesterone caproate (21 ng/mL) and its dual-labeled isotope, 17-alpha-hydroxy-[(3)H] progesterone [(14)C] caproate were added to the maternal circuit. The concentrations of the drug and its metabolite in trophoblast tissue and both circuits were determined by high performance liquid chromatography and liquid scintillation spectrometry. RESULTS: 17-alpha Hydroxyprogesterone caproate was transferred from the maternal to fetal circuit. After a 4-hour perfusion period, a metabolite of 17-alpha-hydroxyprogesterone caproate that retained both progesterone and caproate moieties was identified in the tissue and the maternal and fetal circuits. Neither 17-alpha hydroxyprogesterone caproate nor its metabolite, at the concentrations tested, had adverse effect on determined viability and functional parameters of placental tissue. CONCLUSION: 17-alpha-Hydroxyprogesterone caproate was metabolized by term placental lobule during its perfusion and both parent compound and its metabolite(s) transferred to the fetal circuit. PMID- 18674660 TI - Women in obstetrics and gynecology: appreciating the past, looking to the future. PMID- 18674661 TI - Do mechanical methods of cervical ripening increase infectious morbidity? A systematic review. AB - The purpose of this study was to review systematically randomized controlled trials that were associated with cervical ripening. We identified randomized controlled trials that compared the use of Foley catheter, with or without extraamniotic saline solution infusion, Laminaria, or hygroscopic dilators for cervical ripening or induction with pharmacologic agents or placebo. Randomized controlled trials that evaluated maternal or neonatal infection were selected. The outcomes that were assessed were maternal and neonatal infection, chorioamnionitis, and endomyometritis. Thirty studies met inclusion criteria. Compared with the use of pharmacologic methods alone, patients who underwent cervical ripening with mechanical agents had a significantly higher rate of maternal infection rates. Similar results were noted for patients who underwent ripening with Foley catheter alone in comparison with pharmacologic agents. No difference was noted in maternal infection rates for patients who underwent ripening with extraamniotic saline solution infusion, Laminaria, or hygroscopic dilators. Compared with the use of pharmacologic agents alone, maternal and neonatal infectious morbidity appears to be increased when mechanical agents are used for cervical ripening. PMID- 18674662 TI - Pelvic organ prolapse: is there a difference in POPQ exam results based on time of day, morning or afternoon? AB - OBJECTIVE: The purpose of this study was to determine if there is any difference between AM and PM POPQ exam results. STUDY DESIGN: Prospective, IRB approved study of women presenting with pelvic organ prolapse symptoms. Initial POPQ exams were performed in the morning or afternoon; a second exam was performed within 4 weeks at the opposite time of day. Statistical analysis included Wilcoxon signed rank test, paired t test, Spearman correlation, and Stuart-Maxwell test. RESULTS: The study consisted of 32 subjects. Mean age was 58.8 years, mean BMI 28.6, median parity 2.0, 75% white, 22% black, and 3% Hispanic. The POPQ exam stages were 47% stage II, 50% stage III, and 3.0% stage IV. There was no significant difference for the 9 POPQ exam measures between morning and afternoon exams. There was good agreement between AM and PM POPQ ordinal stages (Stewart Maxwell P = 1.0, kappa 0.76). CONCLUSION: There is no difference between AM and PM POPQ exam results. PMID- 18674663 TI - Assessment of ovarian reserve with anti-Mullerian hormone: a comparison of the predictive value of anti-Mullerian hormone, follicle-stimulating hormone, inhibin B, and age. AB - OBJECTIVE: The objective of this study was to evaluate basal anti-Mullerian hormone as a marker for ovarian responsiveness to fertility treatment. STUDY DESIGN: Frozen basal menstrual cycle day 3 serum samples were evaluated retrospectively for anti-Mullerian hormone, inhibin B, and follicle-stimulating hormone levels in 123 in vitro fertilization cycles (93 patients) and compared with in vitro fertilization records. RESULTS: Anti-Mullerian hormone values correlated the best with the number of retrieved oocytes (r = 0.539; P < .001) relative to age (r = -0.323; P < .01), follicle-stimulating hormone (r = -0.317; P < .01), inhibin B (P > .05), luteinizing hormone (P > .05), and estradiol (r = 0.190; P < .05). Receiver operating characteristic curve analysis demonstrated that, for the prediction of <4 oocytes retrieved, anti-Mullerian hormone had the largest area under the curve (AUC = 0.81; P = .0001) relative to age (r = 0.74; P = .005), follicle-stimulating hormone (0.71; P = .02), inhibin B (0.66; P = .03), and estradiol (0.54; P > .05). Similarly, for the prediction of >or=15 retrieved oocytes, anti-Mullerian hormone had the largest area under the curve (0.80; P = .0001) relative to age (0.63; P = .02), follicle-stimulating hormone (0.64; P = .005), inhibin B (r = 0.57; P > .05), and estradiol (0.58; P > .05). CONCLUSION: Anti-Mullerian hormone correlates better than age, follicle-stimulating hormone, luteinizing hormone, inhibin B, and estradiol with the number of retrieved oocytes. Receiver operating characteristic curves estimated that anti-Mullerian hormone accurately predicts ovarian responsiveness to controlled ovarian stimulation with high sensitivity and specificity. PMID- 18674664 TI - Reproductive factors associated with nocturia and urgency: Tikkinen et al. AB - The following article summarizes a roundtable discussion of a study that is published in this issue of the Journal in light of its methods, relevance to practice, and implications for future research. Article discussed: Tikkinen KAO, Auvinen A, Tiitinen A, Valpas A, Johnson TM, 2nd, Tammela TLJ. Reproductive factors associated with nocturia and urinary urgency in women: a population-based study in Finland. Am J Obstet Gynecol 2008;199:153.e1-153.e12. The full discussion appears at www.AJOG.org, page e1. PMID- 18674665 TI - Gather ye buds: fungus formation of the bladder after complicated cesarean section. PMID- 18674666 TI - Vulnerable populations. Preface. PMID- 18674667 TI - Collaboration for prevention of chronic disease in Kentucky: the Health Education Through Extension Leaders (HEEL) program. AB - Health Education Through Extension Leaders (HEEL) is one of the solutions the University of Kentucky College of Agriculture has created to address the problem of chronic disease in Kentucky. Building on the land grant model for education, outreach, and prevention, HEEL collaborates and partners with the academic health centers, area health education centers, the Center for Rural Health, the Kentucky Cancer Program, the Markey Cancer Center, the University of Kansas Wellness Program, and the Kentucky Cabinet for Health and Family Services to implement research-based preventive programs to the county extension agents across Kentucky. Extension agents are an instrumental bridge between the communities across Kentucky and the educational resources provided by the HEEL program. PMID- 18674668 TI - The Good Samaritan Nursing Center: a Commonwealth Collaborative. AB - The Good Samaritan Nursing Center (GSNC) is an integrated nurse-managed center that serves vulnerable populations in the community. Across its 10 clinics, the GSNC helps to improve access to health care for people of all ages. The purpose of this article is to (1) describe the services and goals of the GSNC, a Commonwealth Collaborative; (2) discuss selected outputs/outcomes from the GSNC clinics; and (3) propose recommendations for research related to the outcomes of this nurse-managed center. PMID- 18674669 TI - Determining standards of care for substance abuse and alcohol use in long-haul truck drivers. AB - The trucking industry employs approximately 9 million workers, with approximately 3 million being long-haul truck drivers. Truck drivers are exposed to a variety of stressful situations, such as working long hours, no sleep, inadequate rest and relaxation, being away from home and support systems, and driving in hazardous conditions. These risk factors place the long-haul truck driver at an increased risk for possible use or abuse of alcohol and drugs. Identification of those at risk and those who are abusing alcohol and drugs is vitally important for the health of these truckers. PMID- 18674670 TI - Promoting oral health among the inner city homeless: a community-academic partnership. AB - Oral health care resources for the homeless are scarce, underfunded, and generally inadequate to meet the oral health needs of this population. The purpose of this program was to improve oral health among the urban homeless in a faith-based inner city mission through education, screening, and improved access to oral health care. The program provided for expanded delivery of oral health care services to the homeless while preparing students in the health professions for community-based practice with at-risk and vulnerable populations. By proactively addressing oral health needs through prevention and earlier diagnosis and treatment, morbidity, quality of life, and cost can be positively affected. Innovative, cross-disciplinary, community delivery models that involve key stakeholders at all levels are needed to address the oral health needs of the homeless and underserved adequately. PMID- 18674671 TI - Transformation for health: a framework for conceptualizing health behaviors in vulnerable populations. AB - Shedding light on the factors and circumstances that operate to bring about marginalization of groups can facilitate appropriate responses to the issue of health disparities among vulnerable groups in society. This is showing to be a seemingly intractable problem; however, it may well be that the approaches currently used to respond to the issues are not appropriate because we overlook the "realties" that really matter: those emanating from the people being visited by these circumstances themselves. Under normal conditions, human behavior can only be controlled by the individual. Facilitating an environment in which an individual can comprehend his or her internal and external realities is the first step toward transformative behavior. PMID- 18674672 TI - Translating research on healthy lifestyles for children: meeting the needs of diverse populations. AB - This article provides two examples of approaches nursing can take to reach diverse populations of children and their families to enhance health lifestyles. First, a descriptive summary of a brief after-school intervention program aimed at influencing 8- and 9-year-old children's media habits and the prevention of negative health behaviors is presented. Design consideration for translating health lifestyles research findings into a nurse-managed inner city primary care practice is reviewed in the second example. PMID- 18674673 TI - Facilitators and barriers for implementing home visit interventions to address intimate partner violence: town and gown partnerships. AB - Pregnant women involved in violent relationships represent a population that is vulnerable for poor pregnancy and infant outcomes on several levels. This article describes the development of a "town and gown" partnership to assist pregnant women in violent relationships. Barriers and facilitating factors for research and home visitor (HV) nurse partnerships working with this vulnerable population were identified by HV participants in a qualitative focus group session. Methods used to develop and maintain the reciprocal relationship between the community (town) and academic researchers (gown) are described. PMID- 18674674 TI - African Americans with memory loss: findings from a community clinic in Lexington, Kentucky. AB - Kentucky's African-American Dementia Outreach Partnership (AADOP) has shown that African-American patients seek dementia care if a clinic is conveniently located and families are educated about the distinction between normal aging and signs of disease. The early identification of dementia allows African Americans access to pharmaceutic treatments that work best early in the course of the disease and provides the opportunity for the patient to plan future care. In the AADOP model, a conveniently located clinic and access to the patient's home were first steps in achieving equality of care. The trust that was built in the community through collaboration with African-American churches has allowed patients and their families to receive help with memory problems and to feel comfortable in seeking help for other medical problems. Maintaining this involvement and responsiveness to the community over the long term is the next challenge for the program. PMID- 18674675 TI - Maternal and newborn care during disasters: thinking outside the hospital paradigm. AB - Emergencies that challenge the infrastructure of the current health care system require a shift in the standard of usual practice. Pregnant women and their newborns are intimately linked special populations that require continued care despite the community circumstances. Pre-event planning with community partners can generate a safer alternative for providing care during a public health emergency. Lessons learned from international and United States public health emergencies have resulted in a better understanding of the essentials of care and the development of resources to guide planning for these populations. PMID- 18674676 TI - From ethics to palliative care: a community hospital experience. AB - One of the most vulnerable and voiceless groups of patients within American hospitals and institutions today are those who are dying. Health care institutions struggle with the challenge of providing excellent palliative and end-of-life care to patients while providing curative therapies at the same time. This article describes the efforts and accomplishments of the ethics committee of a community hospital system to provide for the palliative and end-of-life needs of its patients. PMID- 18674677 TI - Vulnerable populations: drug court program clients. AB - Substance abuse and addiction are chronic conditions characterized by an inability to control one's urge to use mood- or mind-altering drugs. Recognition of the association between addictions and crime to support the addiction, along with the relapsing nature of addictions, presents treatment and management challenges for clinicians and frustration for patients and their families. Pressures to reduce the burgeoning jail population have resulted in collaboration between the treatment community and the court--a diversion program called drug court. This article reviews the drug court programs, the clients, and the processes of accountability that direct the progress toward sobriety in the drug court clients. It also argues that the drug court clients have unique health needs requiring interventions best suited for the recovering addict enrolled in a diversion program within the criminal justice system. Nurses have the ability to influence these systems and provide safety-net clinics to drug court clients through outreach, case finding, and culturally and linguistically appropriate care that can ultimately help this population to reach a higher level of wellness. PMID- 18674678 TI - What is this thing called preventive medicine (III)? PMID- 18674679 TI - Do advanced glycation end products contribute to the development of long-term diabetic complications? PMID- 18674680 TI - Effect of contrast media on megavoltage photon beam dosimetry. AB - The purpose of this study was to quantify changes in photon beam dosimetry caused by using contrast media during computed tomography (CT) simulation and determine if the resulting changes are clinically significant. The effect of contrast on dosimetry was first examined for a single 6-MV photon beam incident on a plane phantom with a structure of varying electron densities (rho(e)) and thickness. Patient studies were then undertaken in which CT data sets were collected with and without contrast for 6 typical patients. Three patients received IV contrast (Optiray-240) only and 3 received IV plus oral (Gastrograffin) contrast. Each patient was planned using conformal multifield techniques in accordance with the department standards. Two methods were used to compare the effect of contrast on dosimetry for each patient. The phantom analysis showed that the change in dose at the isocenter for a single 10 x 10 cm2 6-MV photon beam traversing 10 cm of a contrast-enhanced structure with rho(e) 1.22 was 7.0% (1.22 was the highest average rho(e) observed in the patient data). As a result of using contrast, increases in rho(e) were observed in structures for the 6 patients studied. Consequently, when using contrast-enhanced CT data for multifield planning, increases in dose at the isocenter and in critical structures were observed up to 2.1% and 2.5%, respectively. Planning on contrast-enhanced CT images may result in an increase in dose of up to 2.1% at the isocenter, which would generally be regarded as clinically insignificant. If, however, a critical organ is in close proximity to the planning target volume (PTV) and is planned to receive its maximum allowable dose, planning on contrast-enhanced CT images may result in that organ receiving dose beyond the recommended tolerance. In these instances, pre-contrast CT data should be used for dosimetry. PMID- 18674681 TI - Split-volume treatment planning of multiple consecutive vertebral body metastases for cyberknife image-guided robotic radiosurgery. AB - Cyberknife treatment planning of multiple consecutive vertebral body metastases is challenging due to large target volumes adjacent to critical normal tissues. A split-volume treatment planning technique was developed to improve the treatment plan quality of such lesions. Treatment plans were generated for 1 to 5 consecutive thoracic vertebral bodies (CVBM) prescribing a total dose of 24 Gy in 3 fractions. The planning target volume (PTV) consisted of the entire vertebral body(ies). Treatment plans were generated considering both the de novo clinical scenario (no prior radiation), imposing a dose limit of 8 Gy to 1 cc of spinal cord, and the retreatment scenario (prior radiation) with a dose limit of 3 Gy to 1 cc of spinal cord. The split-volume planning technique was compared with the standard full-volume technique only for targets ranging from 2 to 5 CVBM in length. The primary endpoint was to obtain best PTV coverage by the 24 Gy prescription isodose line. A total of 18 treatment plans were generated (10 standard and 8 split-volume). PTV coverage by the 24-Gy isodose line worsened consistently as the number of CVBM increased for both the de novo and retreatment scenario. Split-volume planning was achieved by introducing a 0.5-cm gap, splitting the standard full-volume PTV into 2 equal length PTVs. In every case, split-volume planning resulted in improved PTV coverage by the 24-Gy isodose line ranging from 4% to 12% for the de novo scenario and, 8% to 17% for the retreatment scenario. We did not observe a significant trend for increased monitor units required, or higher doses to spinal cord or esophagus, with split volume planning. Split-volume treatment planning significantly improves Cyberknife treatment plan quality for CVBM, as compared to the standard technique. This technique may be of particular importance in clinical situations where stringent spinal cord dose limits are required. PMID- 18674682 TI - Feasibility of IMRT to cover pelvic nodes while escalating the dose to the prostate gland: dosimetric data on 35 consecutive patients. AB - Utilizing available dosimetric and acute toxicity data, we confirm the feasibility of intensity modulated radiotherapy (IMRT) to include treatment of the pelvic nodes (PN) while escalating the dose to the prostate. Data were obtained from 35 consecutive patients with prostate cancer with > or =15% risk of PN involvement. Patients received an initial boost to the prostate, delivering 16 Gy over 8 fractions using a 6-field conformal technique, followed by an 8-field coplanar inverse planning IMRT technique delivering an additional 60 Gy over 30 fractions to the prostate (76 Gy total) and 54 Gy over 30 fractions to the seminal vesicles (SV) and PN. Dose-volume histogram analysis was performed for planning target volumes and organs at risk. Acute toxicity (RTOG/EORTC scale) was prospectively and independently scored weekly for each patient. The maximum, mean, minimum dose, and D95 to each planning target volume is provided: prostate (82.2, 78.2, 72.6, 75.2 Gy), SV (79.0, 72.5, 56.9, 61.1 Gy), and PN (80.4, 59.7, 46.5, 53.3 Gy), respectively. The percent volume receiving a dose at or above "x" Gy (Vx) was recorded for V75, V70, V65, V60, and V50 as: bladder (14%, 24%, 32%, 39%, and 54%) and rectum (3%, 18%, 26%, 34%, and 51%), respectively. Acute toxicity was as follows: 54% grade 2+ GI (n = 19), 25% grade 2+ GU (n = 9). IMRT enables treatment of pelvic nodes while escalating dose to the prostate and is clinically feasible with acute toxicity within expected ranges. PMID- 18674683 TI - Evaluation of a single-isocenter technique for axillary radiotherapy in breast cancer. AB - The aim of this study was to develop a technique for axillary radiotherapy that minimizes the risk of radiation-induced damage to the surrounding normal tissue (i.e., arm, shoulder, lung, esophagus, and spinal cord) while keeping the risk of a nodal recurrence to a minimum. A planning study was performed in 20 breast cancer patients. The target volume of the axillary treatment encompassed the periclavicular and axillary lymph node areas. The 3-dimensional (3D) computed tomography (CT) information in this study was used to outline the lymph node areas and the organs at risk (i.e., the esophagus, spinal cord, brachial plexus, and lung). A conventional AP-PA technique (with a transmission plate placed in the AP beam) was evaluated. In addition, a new single-isocenter technique consisting of AP/PA fields using a gantry rotation of +/-20 degrees and a medial AP segment was developed. Both techniques were compared by evaluation of the calculated dose distributions and the dose-volume histograms of the target volume and surrounding organs at risk. The field borders and humeral shielding were redefined based on the 3D anatomical references. Adapting the humeral shielding reduced the irradiated volume by 19% and might contribute to a reduction of the incidence of arm edema and impairment of shoulder function. The maximum radiation dose in the esophagus and spinal cord was reduced by more than 50% using the single-isocenter technique. The difference between both techniques with respect to the mean doses in the target volume and lung, and the maximum dose in brachial plexus, was not statistically significant. Moreover, the single-isocenter technique allowed a fast and easy treatment preparation and reduced the execution time considerably (with approximately 10 minutes per fraction). PMID- 18674684 TI - A treatment planning investigation into the dosimetric effects of systematic prostate patient rotational set-up errors. AB - The purpose of this study was to investigate the potential dosimetric effects of systematic rotational setup errors on prostate patients planned according to the RTOG P-0126 protocol, and to identify rotational tolerances about either the anterior-posterior (AP) or left-right (LR) axis, under which no correction in setup is required. Eight 3-dimensional conformal radiation therapy (3D-CRT) treatment plans were included in the study, half planned to give 7020 cGy in 39 fractions (P-0126 Arm 1) and the other half planned to give 7920 cGy in 44 fractions (P-0126 Arm 2). Systematic rotations of the pelvic anatomy were simulated in a commercial treatment planning system by rotating opposing apertures in the opposite direction to the simulated anatomy rotation. Rotations were incremented in steps of 2.5 degrees to a maximum of +/-5.0 degrees and +/ 10.0 degrees about the AP and LR axis respectively. Dose distributions were evaluated with respect to the planning objectives set out in the P-0126 protocol. For patients on Arm 2 of the study, maintaining the prescribed dose to 98% of the PTV was found to be problematic for superior-end-posterior rotations beyond 5.0 degrees . The results also show that maintaining a rectal dose less than 7500 cGy to 15% of the volume can become problematic for cases of small rectal volume and large superior-end-anterior rotations. We found that setting rotational tolerances will depend on which Arm of the protocol the patient is, and how well the initial plan meets the protocol objectives. In general, we conclude that for rotations about the AP axis, no tolerance level is required; however, cases presenting extreme rotations should be investigated as routine practice. For rotations about the LR axis, we conclude that a tolerance level for patients on Arm 2 of the protocol should be set at +/-5.0 degrees . This tolerance represents the systematic setup error which would require correction if a variation to the initial plan was deemed unacceptable. PMID- 18674685 TI - Influence of Pro-Qura-generated plans on postimplant dosimetric quality: a review of a multi-institutional database. AB - The influence of Pro-Qura-generated plans vs. community-generated plans on postprostate brachytherapy dosimetric quality was compared. In the Pro-Qura database, 2933 postplans were evaluated from 57 institutions. A total of 1803 plans were generated by Pro-Qura and 1130 by community institutions. Iodine-125 (125I) plans outnumbered Palladium 103 (103Pd) plans by a ratio of 3:1. Postimplant dosimetry was performed in a standardized fashion by overlapping the preimplant ultrasound and the postimplant computed tomography (CT). In this analysis, adequacy was defined as a V100 > 80% and a D90 of 90% to 140% for both isotopes along with a V150 < 60% for 125I and < 75% for 103Pd. The mean postimplant V100 and D90 were 88.6% and 101.6% vs. 89.3% and 102.3% for Pro-Qura and community plans, respectively. When analyzed in terms of the first 8 sequence groups (10 patients/sequence group) for each institution, Pro-Qura planning resulted in less postimplant variability for V100 (86.2-89.5%) and for D90 (97.4 103.2%) while community-generated plans had greater V100 (85.3-91.2%) and D90 (95.9-105.2%) ranges. In terms of sequence groups, postimplant dosimetry was deemed "too cool" in 11% to 30% of cases and "too hot" in 12% to 27%. On average, no clinically significant postimplant dosimetric differences were discerned between Pro-Qura and community-based planning. However, substantially greater variability was identified in the community-based plan cohort. It is possible that the Pro-Qura plan and/or the routine postimplant dosimetric evaluation may have influenced dosimetric outcomes at community-based centers. PMID- 18674686 TI - Comparison of IMRT treatment plans between linac and helical tomotherapy based on integral dose and inhomogeneity index. AB - Intensity modulated radiotherapy (IMRT) is an advanced treatment technology for radiation therapy. There are several treatment planning systems (TPS) that can generate IMRT plans. These plans may show different inhomogeneity indices to the planning target volume (PTV) and integral dose to organs at risk (OAR). In this study, we compared clinical cases covering different anatomical treatment sites, including head and neck, brain, lung, prostate, pelvis, and cranio-spinal axis. Two treatment plans were developed for each case using Pinnacle(3) and helical tomotherapy (HT) TPS. The inhomogeneity index of the PTV and the non-tumor integral dose (NTID) were calculated and compared for each case. Despite the difference in the number of effective beams, in several cases, NTID did not increase from HT as compared to the step-and-shoot delivery method. Six helical tomotherapy treatment plans for different treatment sites have been analyzed and compared against corresponding step-and-shoot plans generated with the Pinnacle(3) planning system. Results show that HT may produce plans with smaller integral doses to healthy organs, and fairly homogeneous doses to the target as compared to linac-based step-and-shoot IMRT planning in special treatment site such as cranio-spinal. PMID- 18674687 TI - Case report and dosimetric analysis of an axillary recurrence after partial breast irradiation with mammosite catheter. AB - Partial breast irradiation (PBI) was designed in part to decrease overall treatment times associated with whole breast radiation therapy (WBRT). WBRT treats the entire breast and usually portions of the axilla. The goal of PBI is to treat a smaller volume of breast tissue in less time, focusing the dose around the lumpectomy cavity. The following is a case of a 64-year-old woman with early stage breast cancer treated with PBI who failed regionally in the ipsilateral axilla. With our dosimetric analysis, we found that the entire area of this axillary failure would have likely received at least 45 Gy if WBRT had been used, enough to sterilize microscopic disease. With PBI, this area received a mean dose of only 2.8 Gy, which raises the possibility that this regional failure may have been prevented had WBRT been used instead of PBI. PMID- 18674688 TI - The detectability and localization accuracy of implanted fiducial markers determined on in-room computerized tomography (CT) and electronic portal images (EPI). AB - Many different methods of image guidance are available for radiotherapy treatment (IGRT). The aims of the study were (1) to determine the optimal diameter of gold markers for IGRT to the prostate; (2) to compare, using the Siemens Primatom, the relative merits of in-room computerized tomography (CT) and electronic portal image (EPI) for locating the marker seeds. Gold markers of differing widths were embedded in 2 phantoms (perspex slabs and anthropomorphic). Images were acquired with an amorphous silicon flat panel detector (Siemens Optivue 500) and with the in-room CT scanner (Siemens Somatom Balance). The EPIs were reviewed independently by 6 operators to determine which diameter marker could be best visualized. The optimal marker technique was determined by comparing the investigators' observed marker co-ordinates with the known locations within the phantom. The visibility of all markers on anterior-posterior EPIs was 100%. On the lateral EPI, of a possible 180 visualizations of 1.2-, 1.0-, and 0.8-mm diameter markers, 176 (97.8%), 151 (83.9%), and 132 (73.3%), respectively, were successful. On EPI, the average deviation of fiducial markers from the known position was less than 0.5 mm in any direction. On CT, the largest deviation (2.17 mm) of markers from the known coordinate position was in the superior inferior direction, reflecting the 3.0-mm slice thickness used. EPI accurately located internal markers in all dimensions. The availability of "gold standard" CT imagery at the treatment unit does not improve how accurately the position of markers in a phantom can be defined compared with EPI. However, CT imagery does provide important soft tissue information, the benefits of which are being investigated further. PMID- 18674689 TI - A Bland-Altman analysis of the bias between computed tomography and ultrasound prostate volume measurements. AB - This study assesses the agreement between computed tomography (CT) measurements of prostate volume and those obtained by ultrasound (US), a well-established non invasive technique. Twenty-six patients aged between 58 and 74 years were evaluated for prostate seed implant therapy using both CT and US measurements. The level of agreement between these 2 methods, which were strongly correlated (R2 = 0.828; p < 0.0001), was determined through Bland-Altman analysis. The mean prostate volume (+/- one standard deviation) of the sample was 31.8 +/- 10.5 cc for the CT method and 27.0 +/- 8.2 cc for the US method. The prostate volumes obtained by CT were, on average, 17% larger than the corresponding volumes determined by US. The average bias between the 2 imaging methods is 4.80 cc or 15%, which is significantly larger than the clinically acceptable margin of 10%. PMID- 18674690 TI - Optimization of stereotactic radiotherapy treatment delivery technique for base of-skull meningiomas. AB - This study compares static conformal field (CF), intensity modulated radiotherapy (IMRT), and dynamic arcs (DA) for the stereotactic radiotherapy of base-of-skull meningiomas. Twenty-one cases of base-of-skull meningioma (median planning target volume [PTV] = 21.3 cm3) previously treated with stereotactic radiotherapy were replanned with each technique. The plans were compared for Radiation Therapy Oncology Group conformity index (CI) and homogeneity index (HI), and doses to normal structures at 6 dose values from 50.4 Gy to 5.6 Gy. The mean CI was 1.75 (CF), 1.75 (DA), and 1.66 (IMRT) (p < 0.05 when comparing IMRT to either CF or DA plans). The CI (IMRT) was inversely proportional to the size of the PTV (Spearman's rho = -0.53, p = 0.01) and at PTV sizes above 25 cm3, the CI (IMRT) was always superior to CI (DA) and CI (CF). At PTV sizes below 25 cm3, there was no significant difference in CI between each technique. There was no significant difference in HI between plans. The total volume of normal tissue receiving 50.4, 44.8, and 5.6 Gy was significantly lower when comparing IMRT to CF and DA plans (p < 0.05). There was significantly improved dose sparing for the brain stem and ipsilateral temporal lobe with IMRT but no significant difference for the optic chiasm or pituitary gland. These results demonstrate that stereotactic IMRT should be considered to treat base-of-skull meningiomas with a PTV larger than 25 cm3, due to improved conformity and normal tissue sparing, in particular for the brain stem and ipsilateral temporal lobe. PMID- 18674692 TI - Preface. Gastrointestinal endoscopy has come a long way. PMID- 18674693 TI - The molecular biology of gastrointestinal cancer: implications for diagnosis and therapy. AB - Cancers are caused by the sequential accumulation of alterations in genes that control the growth, differentiation, and other behaviors of cells. It has long been recognized that cancers are very heterogeneous pathologically, which is a reflection of the variable genetic lesions that give rise to the variety of lesions present in the gastrointestinal tract. Despite this complexity, certain types of genetic alterations are linked to specific pathologic lesions. This review summarizes the current understanding of the molecular pathogenesis of gastrointestinal neoplasia and provides explanations for some of the pathologic variability of lesions encountered by the endoscopist. PMID- 18674694 TI - Clinical application of magnification endoscopy and narrow-band imaging in the upper gastrointestinal tract: new imaging techniques for detecting and characterizing gastrointestinal neoplasia. AB - This article introduces one of the most advanced endoscopy imaging techniques, magnification endoscopy with narrow-band imaging. This technique can clearly visualize the microvascular (MV) architecture and microsurface (MS) structure. The application of this technique is quite useful for characterizing the mucosal neoplasia in the hypopharynx, oropharynx, esophagus, and stomach. The key characteristic findings for early carcinomatous lesions are an irregular MV pattern or irregular MS pattern as visualized by this technique. Such a diagnostic system could be applied to the early detection of mucosal neoplasia throughout the upper gastrointestinal tract. PMID- 18674695 TI - Endoscopic resection for premalignant and malignant lesions of the gastrointestinal tract from the esophagus to the colon. AB - The endoscopic mucosal resection and endoscopic submucosal dissection techniques, if performed with the right indications and with expertise, should be considered even in the West as elective treatment modalities for early gastrointestinal neoplasia. Because the experience is still limited, more should be done to strengthen the performance capacity and foster cooperation among skilled endoscopists. PMID- 18674696 TI - Confocal laser endomicroscopy for gastrointestinal diseases. AB - Confocal laser endomicroscopy enables in vivo microscopy of the mucosal layer of the gastrointestinal tract with subcellular resolution during ongoing endoscopy. Endomicroscopy opens the door to immediate tissue and vessel analysis. Different types of diseases can be diagnosed with optical surface and subsurface analysis. Analysis of the in vivo microarchitecture can be used for targeting biopsies to relevant areas, and subsurface imaging can unmask microscopic diseases or bacterial infection. Molecular imaging is becoming feasible, which will enable new indications in gastrointestinal endoscopy. This article reviews the current and rapidly expanding clinical data on endomicroscopy and gives a look into future research. PMID- 18674697 TI - Scanning single fiber endoscopy: a new platform technology for integrated laser imaging, diagnosis, and future therapies. AB - Remote optical imaging of human tissue in vivo has been the foundation for the growth of minimally invasive medicine. This article describes a new type of endoscopic imaging that has been developed and applied to the human esophagus, pig bile duct, and mouse colon. The technology is based on a single optical fiber that is scanned at the distal tip of an ultrathin and flexible shaft that projects red, green, and blue laser light onto tissue in a spiral pattern. The resulting images are high-quality color video that is expected to produce future endoscopes that are thinner, longer, more flexible, and able to directly integrate the many recent advances of laser diagnostics and therapies. PMID- 18674698 TI - Positron emission tomography for the diagnosis and management of patients with gastrointestinal malignancies. AB - F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) can play an important role in evaluating patients who have locally advanced diseases and in recurrence detection and restaging in patients who have gastrointestinal tract malignancies. Introduction of an integrated PET/CT system enabled the precise co evaluation of function and morphology and improved the diagnostic ability of FDG PET. Application of FDG-PET for treatment response evaluation and prognosis prediction is becoming important. Development of novel PET probes is expected to improve the characterization of individual cancer and to contribute to individualized patient management. PMID- 18674699 TI - Barrett's esophagus: pathogenesis, treatment, and prevention. AB - Esophageal adenocarcinoma is the most common type of esophageal cancer seen in the United States and Western Europe. Barrett's esophagus (BE) is a well-known risk factor for esophageal adenocarcinoma and is believed to be found in 6% to 12% of patients undergoing endoscopy for gastroesophageal reflux disease and in more than 1% of all patients undergoing endoscopy. This article focuses on the pathogenesis, treatment, and prevention of BE. PMID- 18674700 TI - Gastric cancer: pathogenesis, screening, and treatment. AB - Early gastric cancer is a curable disease regardless of its location, histologic type, genetic changes, or the ethnicity of the patient. To improve the detection rate of early gastric cancer, intensive training of endoscopists and the use of novel endoscopic techniques have been introduced into routine examinations in Japan. In the United States, where most gastric cancer is found in advanced stages, a similar approach should be advocated. Endoscopic resection of high grade dysplasia is also encouraged in the United States not only for proper diagnosis but also for achieving cure without surgical intervention. PMID- 18674701 TI - Intraductal papillary mucinous neoplasms. AB - Intraductal papillary mucinous neoplasm (IPMN) is characterized by enhanced mucus secretion. It is a benign or low-grade neoplasm associated with a dilated main pancreatic duct, patulous ampullary orifice, and abundant mucus secretion. Foci of aggressive cancer may arise and become invasive. Surgery is the only treatment that can cure IPMN, but the extent of pancreatic resection and the intraoperative margins remain areas of controversy. The risks of total pancreatectomy must be weighed against the risk for developing cancer in the residual pancreas. Risks must be factored against the natural course of the disease and the likelihood of malignancy developing over the life expectancy. PMID- 18674702 TI - Screening and surveillance approaches in familial pancreatic cancer. AB - Screening and surveillance for pancreatic cancer and its precursors is a relatively new indication for endoscopic ultrasound. It provides an alternative approach to the ineffective treatment of mostly incurable symptomatic pancreatic cancer. It is currently reserved for individuals with an increased risk for pancreatic ductal adenocarcinoma, such as those who have inherited genetic syndromes (eg, patients who have Peutz-Jeghers syndrome or hereditary pancreatitis, germline mutation carriers of p16 and BRCA2) and at-risk relatives of patients who have familial pancreatic cancer. This article discusses the rationale for performing screening and surveillance, the types of patients who are eligible for screening, the diagnostic modalities and technique for screening, the diagnostic yield of screening, and the ongoing research. PMID- 18674703 TI - Early detection of pancreatic cancer: the Japanese approach. AB - Pancreatic cancer is the fifth leading cause of cancer-related deaths in Japan. Small pancreatic cancers have some abnormal findings on ultrasonography, and diagnosis with endosonography is useful. Positron emission tomography and contrast-enhanced ultrasonography are expected to serve as new modalities for the early detection of pancreatic cancer. The identification of high-risk individuals is necessary to perform efficient screening. Intraductal papillary mucinous neoplasms and chronic pancreatitis are important risk factors for pancreatic cancer. PMID- 18674704 TI - Endoscopic management of biliary malignancies. AB - The current status of endoscopic diagnosis and treatment of biliary malignancies is discussed. Three ways are now available to approach the bile duct endoscopically. Endoscopic diagnostic modalities are quite useful for the assessment of local changes because of their high spatial resolution and ability in tissue and bile acquisition for histologic, cytologic, and molecular biologic evaluations. Endoscopic biliary drainage is less invasive and beneficial for patients with obstructive jaundice compared with surgical or percutaneous transhepatic drainage. Continuous evolution should expand its indications. PMID- 18674705 TI - Flat and depressed types of early colorectal cancers: from East to West. AB - Recent advances in colonoscopy have enabled us to diagnose early-stage colorectal tumors. Magnifying colonoscopy is useful for diagnosing histologic types by assessing the microstructure of the mucosal surface in detail, which also helps to predict the depth. This article describes clinicopathologic features and endoscopic treatment of flat and depressed types of early colorectal carcinoma. PMID- 18674706 TI - Screening, surveillance, and prevention of colorectal cancer. AB - Colon cancer screening can be effective, but only with a high-quality program that assures adherence to all elements of the program. There is evidence in the United States of greater acceptance of screening and decreased incidence and mortality of colorectal cancer. Patient education is a key element of any effective screening program. It is hoped that future screening will develop better risk-stratification tools and enable targeting of screening to high-risk individuals. PMID- 18674707 TI - Not your father's colonoscopy: a high-tech future for screening and surveillance of colorectal cancer. AB - The 20-year technology slump in endoscopic innovation is finally giving way to a flurry of technologies, of which many are directed specifically at improving or even replacing traditional colonoscopy. These technologies include "smart" overtubes, electronically mapped and driven instruments, and completely self propelled devices. In addition to nonendoscopic technologies such as CT, these innovations may dramatically alter the practice of colorectal cancer screening, the "bread and butter" of gastroenterologists in this country. There are multiple and complex forces driving these changes, including a mismatch between the supply and demand in colonoscopy, patient convenience and comfort, costs, and more recently, a growing concern about the miss rate of conventional colonoscopy. PMID- 18674708 TI - Hepatic cysts detected on Tc-99m sestamibi gated cardiac SPECT images. PMID- 18674709 TI - Sinus bradycardia during exercise: unusual manifestation of ischemia in the territory of the left anterior descending artery. PMID- 18674710 TI - Angioedema after injection of Tc-99m sestamibi tracer during adenosine nuclear stress testing. PMID- 18674711 TI - The rising cost of health care in the United States: is it making the United States globally noncompetitive? PMID- 18674712 TI - Intersecting techniques: the evaluation of left ventricular function with cardiac computed tomography and myocardial perfusion imaging. PMID- 18674713 TI - Detecting the impact of emerging cardiovascular risk factors: the role of positron emission tomography. PMID- 18674714 TI - Mental stress, a powerful provocateur of myocardial ischemia: diagnostic, prognostic, and therapeutic implications. PMID- 18674715 TI - The revolution and evolution of appropriateness in cardiac imaging. PMID- 18674716 TI - Comparison of 64-slice cardiac computed tomography with myocardial perfusion scintigraphy for assessment of global and regional myocardial function and infarction in patients with low to intermediate likelihood of coronary artery disease. AB - BACKGROUND: Cardiac computed tomography (CCT) has the potential to assess both coronary anatomy and ventricular function in a single study. We examined the agreement between CCT and myocardial perfusion scintigraphy (MPS) for the assessment of global and regional ventricular function. METHODS AND RESULTS: Research CCT was performed in 52 patients with a low to intermediate likelihood of coronary artery disease referred for MPS. Left ventricular end-diastolic volume, left ventricular end-systolic volume, left ventricular ejection fraction (LVEF), and myocardial wall motion and thickening were compared between techniques. In addition, myocardial contrast attenuation on CCT was compared with radiotracer uptake on MPS. LVEF values agreed well (mean difference, 4.1%; SD, 15.13%), but CCT left ventricular end-diastolic volume was greater compared with MPS (mean difference, 46.0 mL; SD, 33.34 mL) (P < .001). There was moderate agreement for segmental myocardial motion and thickening, with kappa values of 0.57 (95% confidence interval, 0.51-0.63) and 0.47 (95% confidence interval, 0.41 0.53), respectively. Seventeen patients had hypoattenuation in at least 1 myocardial segment on CCT. Three of four patients with concomitant abnormalities of wall motion and thickening on CCT had infarction in the same territory on MPS. CONCLUSIONS: There was good agreement for LVEF between CCT and MPS but myocardial volumes differed, and these modalities cannot be used interchangeably. Mild abnormalities of regional function are detected more commonly by CCT than by MPS. Myocardial hypoattenuation on CCT is highly specific for myocardial infarction when associated with reduction of systolic wall thickening and regional wall motion abnormality. PMID- 18674717 TI - Different manifestations of coronary artery disease by stress SPECT myocardial perfusion imaging, coronary calcium scoring, and multislice CT coronary angiography in asymptomatic patients with type 2 diabetes mellitus. AB - BACKGROUND: We sought to assess prospectively the evidence for silent coronary artery disease (CAD) in asymptomatic patients with type 2 diabetes mellitus by stress single-photon emission computed tomography (SPECT) myocardial perfusion imaging, coronary artery calcium (CAC) scoring, and multislice computed tomographic (MSCT) coronary angiography. METHODS: One hundred asymptomatic patients (aged 30 to 72 years) with type 2 diabetes mellitus and one or more risk factors for CAD were prospectively recruited from an outpatient diabetes clinic. All patients underwent adenosine technetium-99m sestamibi SPECT imaging, CAC scoring, and 64-slice MSCT coronary angiography. RESULTS: Twenty-three patients (23%) had abnormal stress SPECT imaging, consistent with inducible myocardial ischemia, whereas 60 patients (60%) had positive CAC scoring (18 patients [18%] with significant CAC >401), and 70 patients (70%) had abnormal MSCT coronary angiography (24 patients [24%] with significant, >or=50% stenosis). Of 77 patients with normal SPECT, 44 had a positive CAC score (10 patients [13%] >401), and 54 showed CAD on MSCT angiography (16 patients [21%] with >or=50% stenosis). Of 23 patients with an abnormal SPECT, 16 patients had a positive CAC score (8 patients [35%] >401), and 16 patients had CAD on MSCT angiography (8 patients [35%] with >or=50% stenosis). Overall, 17 patients (17%) had more than 2 significantly abnormal diagnostic test results, and 5 patients had three tests with significantly abnormal results. CONCLUSIONS: In this cohort of asymptomatic patients with type 2 diabetes mellitus, different modalities visualized different aspects of silent coronary atherosclerosis. Anatomic evidence of coronary atherosclerosis (CAC and MSCT) occurred more frequently than functional evidence (stress SPECT). However, clinically significant manifestations of CAD were observed in about one-quarter to one-fifth of patients by each modality, either separately or combined. The relative prognostic value of each modality needs to be determined by a follow-up of this cohort. PMID- 18674718 TI - Cardiovascular risk, obesity, and myocardial blood flow in postmenopausal women. AB - BACKGROUND: This study was designed to determine whether overweight or obese status is independently associated with myocardial flow reserve (MFR), an established predictor of cardiovascular mortality, in a group of postmenopausal women with no previous cardiovascular disease. Postmenopausal women are the largest group of overweight and physically inactive individuals in the United States. Increased body mass index (BMI) is consistently associated with increased cardiovascular mortality in this population. Whether this is because of obesity itself or the accompanying increase in cardiovascular risk factors (CRFs) remains controversial. METHODS: We examined the relationship of myocardial blood flow (MBF), coronary vascular resistance, and MFR to BMI in 60 postmenopausal women with no coronary heart disease. Subjects underwent dynamic N-13 ammonia positron emission tomography for the measurement of MBF and MFR. Baseline demographics, CRF, and hemodynamic parameters were recorded for each subject. Datasets were divided into 3 groups according to BMI: normal (18 to 24), overweight (25 to 29), and obese (>or=30). RESULTS: The overweight and obese groups showed significantly higher resting MBF and lower MFR than the normal-weight group (both P < .001), even after adjusting for CRF. A further analysis of subjects without any CRF (n = 35) showed that the MFR remained significantly lower in the obese compared with normal-weight subjects (P = .05). Levels of known markers of vascular inflammation (high-sensitivity C-reactive protein and homocysteine) and high density lipoprotein cholesterol levels correlated with declining MFR. CONCLUSIONS: These findings provide a mechanistic link between obesity and coronary heart disease in this population. PMID- 18674719 TI - Variability of myocardial ischemic responses to mental versus exercise or adenosine stress in patients with coronary artery disease. AB - BACKGROUND: Mental stress precipitates myocardial ischemia in a significant percentage of coronary artery disease (CAD) patients. Exercise or adenosine stresses produce different physiologic responses and cause myocardial ischemia via different mechanisms. Little is known about the comparative severity and location of myocardial ischemia provoked by these different stressors. In this study we sought to compare the within-individual ischemic responses to mental versus exercise or adenosine stress in a cohort of CAD patients. METHODS AND RESULTS: All patients underwent mental stress and either exercise or adenosine testing within a 1-week period. Mental stress was induced via a public speaking task. Rest-stress myocardial perfusion imaging was used with all testing protocols. Participants were 187 patients (65 women [35%]) with a documented history of CAD and a mean age of 64 +/- 9 years. Mental stress-induced myocardial ischemia (MSIMI) was less prevalent and frequently of less magnitude than exercise- or adenosine-induced ischemia. Ischemia induced by exercise or adenosine testing did not accurately predict the development or the location of MSIMI. The overall concordance between these stressors for provoking ischemia was weak (percent agreement, 71%; kappa [+/- SE], 0.26 +/- 0.07). In a minority of patients (11%) mental stress provoked ischemia in the absence of exercise- or adenosine-induced ischemia. Moreover, in patients who had myocardial ischemia during both stressors, there were significant within-individual differences in the coronary artery distribution of the ischemic regions. MSIMI was more likely to occur in a single-vessel distribution (86%) compared with exercise- or adenosine-induced ischemia (54%). The stressors had moderate agreement if the ischemic region was in the right coronary artery territory (percent agreement, 76%; kappa, 0.52 +/- 0.19) or the left anterior descending coronary artery (percent agreement, 76%; kappa, 0.51 +/- 0.19) and significantly lower agreement in the left circumflex territory (percent agreement, 62%; kappa, 0.22 +/- 0.18). CONCLUSIONS: Our findings indicate that mental and exercise or adenosine stresses provoke different myocardial ischemic responses. These observations suggest that exercise or adenosine testing may not adequately assess the likelihood of occurrence or severity of MSIMI and that different mechanisms are operative in each condition. PMID- 18674720 TI - Dobutamine-induced chest pain does not predict ischemic findings on myocardial perfusion imaging. AB - BACKGROUND: Chest pain occurs frequently during dobutamine stress testing and is commonly attributed to ischemia. However, the pathophysiologic significance of dobutamine-induced chest pain is uncertain. The purpose of this study is to explore the correlation between dobutamine-induced chest pain and evidence of ischemia on myocardial perfusion imaging (MPI). METHODS AND RESULTS: This study included 1608 patients who underwent dobutamine stress MPI at the Mid America Heart Institute (Kansas City, Mo) and were analyzed retrospectively. Patients were divided into those with chest pain during dobutamine infusion versus those without it. Multivariate and chi(2) analyses were conducted to explore the relationship between chest pain and ischemic changes on MPI. Of 1608 patients, 208 (13%) had chest pain with dobutamine whereas 1400 (87%) did not. MPI ischemia was seen in 47% of patients with chest pain and 43% without chest pain (P = .28). Chest pain was not any more predictive of ischemia when analyzed separately by gender (P = .31). Multivariate analysis did not identify chest pain as a predictor of ischemia (P = .19). Significant predictors of scintigraphic ischemic changes were age (P = .001), gender (P < .0001), smoking (P = .0149), and known coronary artery disease (P < .0001). CONCLUSION: This large retrospective study suggests that chest pain during dobutamine stress testing is not a predictor of ischemia when analyzed against reversible perfusion defects on SPECT MPI. PMID- 18674721 TI - Clinical evaluation of iterative reconstruction (ordered-subset expectation maximization) in dynamic positron emission tomography: quantitative effects on kinetic modeling with N-13 ammonia in healthy subjects. AB - BACKGROUND: The purpose of this study was to investigate the quantitative properties of ordered-subset expectation maximization (OSEM) on kinetic modeling with nitrogen 13 ammonia compared with filtered backprojection (FBP) in healthy subjects. METHODS AND RESULTS: Cardiac N-13 ammonia positron emission tomography (PET) studies from 20 normal volunteers at rest and during dipyridamole stimulation were analyzed. Image data were reconstructed with either FBP or OSEM. FBP- and OSEM-derived input functions and tissue curves were compared together with the myocardial blood flow and spillover values. The late area under the OSEM input functions during dipyridamole is overestimated by 30% (P < .0001) relative to FBP. Conversely, the area under the late part of the OSEM tissue curves is underestimated by 20% (P < .0001) compared with FBP during both rest and dipyridamole. These differences in tissue and input functions cause the resting myocardial blood flow to be underestimated by 15% (P < .0001). During dipyridamole, the OSEM flow is underestimated by 25% (P < .0001) relative to FBP, causing the myocardial flow reserve to be underestimated by 10% (P < .0001). Large inter-regional differences in FBP and OSEM flow values were observed with a flow underestimation of 45% (rest/dipyridamole) in the septum and of 5% (rest) and 15% (dipyridamole) in the lateral myocardial wall. CONCLUSIONS: OSEM reconstruction of myocardial perfusion images with N-13 ammonia and PET produces high-quality images for visual interpretation. However, compared with FBP, OSEM is associated with substantial underestimation of perfusion on quantitative imaging. Our findings indicate that OSEM should be used with precaution in clinical PET studies. PMID- 18674722 TI - Tc-99m-N-MPO: novel cationic Tc-99m radiotracer for myocardial perfusion imaging. AB - BACKGROUND: Technetium 99m-N-MPO ([Tc-99m-N(mpo)(PNP5)](+)) is a cationic Tc-99m nitrido complex. The objective of this study is to evaluate its potential as a new radiotracer for myocardial perfusion imaging. METHODS AND RESULTS: Biodistribution studies were performed in Sprague-Dawley rats and guinea pigs to compare the myocardial uptake and excretion kinetics of Tc-99m-N-MPO from noncardiac organs, such as the liver and lungs, with those of the known cationic Tc-99m radiotracers: Tc-99m-N-DBODC5 and Tc-99m-sestamibi. Planar imaging was performed in Sprague-Dawley rats to evaluate the utility of Tc-99m-N-MPO as a myocardial perfusion imaging agent. Metabolism studies were carried out by use of both Sprague-Dawley rats and guinea pigs. In general, the heart uptake of Tc-99m N-MPO was between that of Tc-99m-sestamibi and Tc-99m-N-DBODC5 over the 2-hour study period. However, the heart-liver ratio of Tc-99m-N-MPO (12.75 +/- 3.34) at 30 minutes after injection was more than twice that of Tc-99m-N-DBODC5 (6.01 +/- 1.45) and approximately 4 times higher than that of Tc-99m-sestamibi (2.90 +/- 0.22). The heart uptake and heart-liver ratio of Tc-99m-N-MPO and Tc-99m sestamibi in guinea pigs were significantly lower than those obtained in Sprague Dawley rats. The metabolism studies demonstrated no detectable Tc-99m-N-MPO metabolites in the urine and feces samples of the Sprague-Dawley rats at 120 minutes after injection. In guinea pigs no Tc-99m-N-MPO metabolites were detected in the urine at 120 minutes, but only approximately 60% of Tc-99m-N-MPO remained intact in the feces samples. In contrast, there was no intact Tc-99m-sestamibi detected in urine samples, and less than 15% of Tc-99m-sestamibi remained intact in the feces samples. Planar imaging studies indicated that clinically useful images of the heart may be obtained as early as 15 minutes after injection of Tc 99m-N-MPO. CONCLUSION: The combination of favorable organ biodistribution and myocardial uptake with rapid liver clearance makes Tc-99m-N-MPO a very promising myocardial perfusion radiotracer worthy of further evaluation in various preclinical animal models. PMID- 18674723 TI - Ordered subset expectation maximization and wide beam reconstruction "half-time" gated myocardial perfusion SPECT functional imaging: a comparison to "full-time" filtered backprojection. AB - OBJECTIVES: Compared to filtered back projection (FBP), OSEM with resolution recovery (OSEM-RR) and wide beam reconstruction (WBR)(UltraSPECT Ltd.), which resolve resolution and suppress noise simultaneously during reconstruction, have been shown to maintain/improve myocardial perfusion SPECT quality, even with low count density half-time acquisitions. We postulated that their characteristics would be advantageous for gated SPECT, where each frame is only 1/8th the count density of the summed perfusion images. METHODS: An 9 mCi rest/32 mCi (333/1184 MBq) stress Tc(99m) sestamibi protocol was used. 15-min FBP, and additional 7-min OSEM-RR and WBR post-stress 8-frame/cardiac cycle SPECT scans were acquired with 90 degrees -angled dual-headed detectors equipped with high resolution collimators in 156 patients. In 82 patients (48F, 34M) (123-252 lbs) with perfusion defects gated image quality was graded visually: 1 (poor)-5 (excellent) Regional LV wall motion (WM) was scored: 0 (normal)-4 (dyskinesis) in a total of 50 vascular territories with defects. Using Myometrix software (GE Healthcare), post-stress EDV, ESV, and EF were calculated for each method. Additionally, for purposes of comparison, the FBP gated tomograms were processed with other commercially available packages, Emory Toolbox and Cedars QGS. RESULTS: Despite half-time acquisitions, compared to FBP, image quality increased marginally with OSEM-RR (P = .09) but very significantly with WBR (P = 1.9 x 10(-21)). The WM score was greater only for WBR (P = 4.8 x 10(-8)). Although quantitative parameters correlated well with those determined by FBP (all EF r's > 0.85; all volume r's > 0.93), EFs were significantly lower (P = .0001 for OSEM-RR, 3.4 x 10(-14) for WBR), primarily due to a decrease in EDV with OSEM-RR (P = 7.3 x 10( 13)) and an increase in ESV with WBR (P = 9.2 x 10(-5)). However, inter-method differences in these parameters were of similar magnitude to differences encountered among the commercially available software methods. CONCLUSIONS: Half time OSEM-RR and particularly WBR improve gated SPECT diagnostic quality compared to full-time FBP due to increased resolution and reduced noise. However, these attributes, which affect endocardial edge detection, result in a systematic offset in EDV, ESV, and EF. PMID- 18674724 TI - Acute chest pain imaging in the emergency department with cardiac computed tomography angiography. AB - In practice, the determination of ischemic chest pain in the emergency department (ED) population is difficult and errors are common. Cardiac computed tomography angiography has recently emerged for accurate noninvasive evaluation of coronary artery disease, and it may offer a promising new approach to improve the triage of patients presenting to the ED with acute chest pain, in particular in terms of a faster and accurate way to determine the diagnosis, which could effectively reduce hospital admissions and costs. The focus of this article is to review the current literature on the use of cardiac computed tomography angiography in the ED setting by providing pooled sensitivity, specificity, and positive and negative predictive values of the published literature to date. Moreover, different protocols for detection of patients with cardiac and other, noncardiac causes of chest pain (triple rule-out protocol) are discussed. PMID- 18674725 TI - Molecular imaging of cardiovascular disease using ultrasound. AB - Molecular imaging using probes that specifically home to function- or disease specific targets is a promising tool for both basic research investigations as well as clinical diagnostics. Ultrasound-based molecular imaging utilizes acoustically active particles (contrast agents) bearing targeting ligands that specifically bind to a molecule of interest. In the presence of an ultrasound field, the bound particles are detectable as a persistent contrast effect during ultrasound imaging. Different types of targeted contrast agents have been reported, most of which share in common the presence of a gas encapsulated by a shell of varying chemical formulation. These agents, or "microbubbles," are typically 2 to 4 mum in diameter, and have a natural resonance frequency that corresponds to the frequencies used in diagnostic echocardiography. This attribute makes it possible to induce microbubble resonance and non-linear oscillation at diagnostic ultrasound frequencies, leading to acoustic emissions from the microbubbles that can be detected as specific signals during two dimensional ultrasound imaging. Targeting ligands that have been attached to microbubbles include monoclonal antibodies, peptides, and the naturally occurring ligands for the receptor of interest, such as vascular endothelial growth factor. Because the contrast agents stay within the intravascular space, they are ideally suited for detection of endothelial epitopes, such as leukocyte adhesion molecules or angiogenesis receptors. Ultrasound molecular imaging with targeted contrast agents has been used to detect inflammation association with ischemia/reperfusion (ischemic memory), cardiac transplant rejection, early atherosclerosis, and angiogenesis. Application to tumor angiogenesis has also been reported using peptides that specifically bind to angiogenic tumor endothelium. Translation of ultrasound molecular imaging to the clinical arena will require optimization of contrast agent design to maximize specific binding, and customization of imaging systems to sensitively detect the binding events. PMID- 18674726 TI - Basic science curriculums in nuclear cardiology and cardiovascular imaging: evolving and emerging concepts. AB - The teaching of basic science with regard to physics, instrumentation, and radiation safety has been part of nuclear cardiology training since its inception. Although there are clear educational and quality rationale for such, regulations associated with the Nuclear Regulatory Commission Subpart J of old 10 CFR section 35 (Title 10, Code of Federal Regulations, Part 35) from the 1960s mandated such prescriptive instruction. Cardiovascular fellowship training programs now have a new opportunity to rethink their basic science imaging curriculums with the era of "revised 10 CFR section 35" and the growing implementation of multimodality imaging training and expertise. This review focuses on the history and the why, what, and how of such a curriculum arising in one city and suggests examples of future implementation in other locations. PMID- 18674727 TI - Cardiac sarcoidosis: a challenge to measure disease activity. PMID- 18674728 TI - Coronary artery vasospasm or true transient left ventricular apical ballooning? Differentiation by nuclear imaging. PMID- 18674729 TI - Re: Combined therapy with carvedilol and amiodarone is more effective in improving cardiac symptoms, function, and sympathetic nerve activity in patients with dilated cardiomyopathy: comparison with carvedilol therapy alone. PMID- 18674730 TI - Pancreaticopleural fistula visualized by multidetector row computed tomography. PMID- 18674731 TI - Appendiceal orifice inflammation and ulcerative proctitis. PMID- 18674733 TI - Imaging in intestinal ischemic disorders. AB - Intestinal ischemia and infarction are a heterogeneous group of diseases that have as their unifying theme hypoxia of the small bowel and/or colon. The incidence of bowel ischemia is increasing for a number of reasons: the aging of the population, the ability of intensive care units to salvage critically ill patients, and heightened clinical awareness of these disorders. Improvements in diagnostic imaging techniques have contributed greatly to the earlier diagnosis of intestinal ischemia, which can have a positive influence on patient outcomes. In this review, advances in multidetector computerized tomography, magnetic resonance, and ultrasound in the detection of intestinal ischemia are highlighted and placed in the context of expeditious patient management. PMID- 18674734 TI - Regression of hepatic fibrosis after intestinal transplantation in total parenteral nutrition liver disease. AB - BACKGROUND & AIMS: Hepatic fibrosis may occur in patients with intestinal failure requiring total parenteral nutrition, leading to liver dysfunction necessitating combined intestinal and liver transplantation. The decision to perform combined transplantation as opposed to an isolated intestinal transplant is based on the presence of hyperbilirubinemia, portal hypertension, and advanced hepatic fibrosis. METHODS: We identified 4 patients who underwent isolated intestinal transplantation having significant liver fibrosis. A novel image analysis technique was applied to serial liver biopsies to more precisely quantitate posttransplantation fibrosis regression separately within both portal and centrilobular areas. RESULTS: All patients were found to have significant portal and centrilobular fibrosis regression, which occurred more rapidly in the former. Two patients had improvement in fibrosis despite infections and continuation of total parenteral nutrition, suggesting that hepatic fibrosis associated with intestinal failure may in part be related to adequate anatomic and functional bowel length. CONCLUSIONS: Significant hepatic fibrosis and liver dysfunction may regress after intestinal transplantation and fibrosis regresses more rapidly in portal areas. This suggests that some patients with intestinal failure and associated liver disease may safely undergo isolated intestinal transplant without the need for concurrent liver transplantation and its attendant higher morbidity and mortality. PMID- 18674735 TI - Increased risk of primary sclerosing cholangitis and ulcerative colitis in first degree relatives of patients with primary sclerosing cholangitis. AB - BACKGROUND & AIMS: The importance of genetic factors for the development of primary sclerosing cholangitis (PSC) is incompletely understood. This study assessed the risk of PSC and inflammatory bowel disease (IBD) among first-degree relatives of patients with PSC, compared with the first-degree relatives of a cohort without PSC. METHODS: Subjects from the national Swedish cohort of PSC patients (n = 678) were matched for date of birth, sex, and region to up to 10 subjects without a diagnosis of PSC (n = 6347). Linkage through general population registers identified first-degree relatives of subjects in both the PSC and comparison cohorts (n = 34,092). Diagnoses among first-degree relatives were identified by using the Inpatient Register. RESULTS: The risk of cholangitis was statistically significantly increased in offspring, siblings, and parents of the PSC patient cohort, compared with relatives of the comparison cohort, with the hazard ratios and 95% confidence intervals, 11.5 (1.6-84.4), 11.1 (3.3-37.8), and 2.3 (0.9-6.1), respectively. The hazard ratios for ulcerative colitis (UC) among first-degree relatives of all PSC patients was 3.3 (2.3-4.9) and for Crohn's disease 1.4 (0.8-2.5). The risk of UC for relatives of PSC patients without IBD was also increased, 7.4 (2.9-18.9). CONCLUSIONS: First-degree relatives of patients with PSC run an increased risk of PSC, indicating the importance of genetic factors in the etiology of PSC. First-degree relatives of PSC patients without IBD are also at an increased risk of UC, which might indicate shared genetic susceptibility factors for PSC and UC. PMID- 18674737 TI - Risk of pancreatitis in patients with celiac disease: is autoimmune pancreatitis a biologically plausible mechanism? PMID- 18674738 TI - Understanding the molecular regulation of T-wave alternans. PMID- 18674739 TI - Effect of common KCNE1 and SCN5A ion channel gene variants on T-wave alternans, a marker of cardiac repolarization, during clinical exercise stress test: the Finnish Cardiovascular Study. AB - T-wave alternans (TWA) in electrocardiography (ECG) is a marker of cardiac repolarization, the molecular regulation of which is incompletely understood. High TWA and prolonged QT intervals are both associated with ventricular arrhythmias and sudden death. Therefore, we tested the hypothesis of whether the same mutations that influence the QT interval also affect TWA variation. We examined the effect of 3 ion channel gene single nucleotide polymorphisms (SNPs), rs1805127, rs727957 KCNE1, and rs1805124 SCN5A, on TWA during a clinical exercise test. A total of 2008 subjects from the Finnish Cardiovascular Study underwent an exercise test with online ECG recording. TWA was measured by using the time domain, modified moving average method. Maximum values at rest, during maximal exercise, and during recovery were used as outcome measures in statistical analysis. Moreover, 4-year survival data were collected and ion channel SNPs were determined. TWA was lowest in subjects with the TT genotype of rs1805127 during all phases of the exercise test (RANOVA main effect for genotype, P = 0.018). The result remained significant after adjustment for age, existing coronary heart disease, and beta-blocker medication status (RANCOVA, P = 0.035). Of the polymorphisms studied, only rs1805127 had a significant association with mortality (P = 0.047). The most common G-C haplotype, formed by rs727957 and rs1805127, was associated with TWA (RANOVA, P = 0.007) but not with mortality. The rs1805124 polymorphism was not associated with TWA. The common KCNE1 gene variant rs1805127 is associated with TWA during an exercise test in a Finnish population, which provides additional evidence that KCNE1 genetics may influence cardiac repolarization and cardiovascular mortality. PMID- 18674740 TI - In vitro studies of amikacin-loaded human carrier erythrocytes. AB - Erythrocyte-encapsulated antibiotics have the potential to provide an effective therapy against intracellular pathogens. The advantages over the administration of free antibiotics include a lower systemic dose, decreased toxicity, a sustained delivery of the antibiotic at higher concentrations to the intracellular site of pathogen replication, and increased efficacy. In this study, the encapsulation of amikacin by human carrier erythrocytes prepared using a hypo-osmotic dialysis was investigated. The effects of the initial amikacin dialysis concentration and hypo-osmotic dialysis time on the encapsulation efficiency of amikacin were determined, and the osmotic fragility and hematologic parameters of amikacin-loaded carrier erythrocytes were measured. The efficiency of amikacin entrapment by carrier erythrocytes was dependent on the initial dialysis concentration of the drug. Statistically significant differences in the osmotic fragility profiles between control and carrier erythrocytes were observed, which were dependent on the hypo-osmotic dialysis time and on the dialysis concentration of amikacin. Mean hematologic parameters were evaluated and compared with unloaded, native erythrocytes; the mean corpuscular volume (MCV) of amikacin-loaded carrier erythrocytes was statistically significant smaller. Amikacin demonstrated a sustained release from loaded erythrocytes over a 48-h period, which suggests a potential use of the erythrocyte as a slow systemic-release system for antibiotics. PMID- 18674741 TI - Zinc supplementation decreases oxidative stress, incidence of infection, and generation of inflammatory cytokines in sickle cell disease patients. AB - Zinc deficiency is common in adult sickle-cell disease (SCD) patients. We previously demonstrated that zinc supplementation to adult SCD patients decreased the incidences of infections and hospital admissions. We hypothesize that zinc supplementation improves T-helper cell function and decreases vascular endothelial cell activation, oxidative stress, and nuclear factor-kappa B (NF kappaB)-DNA binding in mononuclear cells (MNCs) in SCD patients. To test this hypothesis, 36 SCD patients were recruited and randomly divided into 2 groups. One group (n = 18) received 25-mg zinc orally thrice a day for 3 months. The other group (n = 18) received placebo. The results indicate that the zinc supplemented group had decreased incidence of infections compared with the placebo group. After zinc supplementation, red blood cell, hemoglobin (Hb), hematocrit, (Hct), plasma zinc, and antioxidant power increased; plasma nitrite and nitrate (NOx), lipid peroxidation products, DNA oxidation products, and soluble vascular cell adhesion molecule-1 decreased in the zinc-supplemented group, compared with the placebo group. Zinc-supplemented patients exhibited significant decreases in lipopolysaccharide-induced tumor necrosis factor-alpha (TNF-alpha) and IL-1beta mRNAs, and TNF-induced nuclear factor of kappaB-DNA binding in MNCs, compared with the placebo group. Ex vivo addition of zinc to MNCs isolated from the placebo subjects decreased TNF-alpha and IL-1beta mRNAs. Zinc supplementation also increased relative levels of IL-2 and IL-2Ralpha mRNAs in phytohemagglutinin-p-stimulated MNCs. These results suggest that zinc supplementation may be beneficial to SCD patients. PMID- 18674742 TI - Growth factors expression in patients with erosive esophagitis. AB - Although the pathogenesis and treatment of erosive esophagitis (EE) is well recognized, little is known about the cellular and molecular mechanisms of mucosal healing in EE patients. In this pilot study, we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa. Forty endoscopically proved EE patients were consecutively enrolled. Messenger RNA expressions, which includes keratinocyte growth factor (KGF) and its receptor (KGFR), epidermal growth factor (EGF) and its receptor (EGFR), hepatocyte growth factor (HGF) and its receptor (HGFR), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and cyclooxygenase (COX)-1 and COX-2, were measured using real-time polymerase chain reaction (PCR). Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE. The mRNA expressions of HGF, HGFR, EGF, VEGF, and COX-2, but not EGFR, KGF, KGFR, bFGF, and COX-1, were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa (P < 0.05). The study found that HGF, HGFR, EGF, VEGF, and, COX 2 are activated in the injured mucosa of EE patients; their activation might be involved in mucosal repair and ulcer healing of EE. PMID- 18674743 TI - Responses to tiotropium in African-American and Caucasian patients with chronic obstructive pulmonary disease. AB - Sparse information exists about chronic obstructive pulmonary disease (COPD) outcomes among different ethnic groups. To determine whether the effect of tiotropium on COPD exacerbation differs between African Americans and Caucasians, we performed a post hoc analysis of African-American (n = 150) and Caucasian (n = 1670) subgroups from a previously reported 6-month trial of tiotropium in patients with moderate-to-very-severe COPD. Compared with placebo, tiotropium reduced the likelihood of having at least 1 exacerbation in the entire group (RR, 0.81; 95% CI, 0.66-0.99, P = 0.037) with no statistically significant difference between African-American and Caucasian subgroups (P = 0.34). For African Americans, tiotropium significantly reduced the number of antibiotic days for COPD, hospitalizations for exacerbations, and hospitalization days for COPD. For Caucasians, tiotropium significantly reduced the number of exacerbations, exacerbation days, unscheduled clinic visits for COPD, and hospitalizations for exacerbations. Tiotropium reduced the frequencies of antibiotic days and of COPD hospital days to a significantly greater extent in African Americans compared with Caucasians (P = 0.027 and P = 0.025, respectively). No statistically significant ethnic-related differences were observed in the effect of tiotropium on the frequencies of exacerbations, exacerbation days, systemic corticosteroid days, unscheduled clinic visits, or COPD hospitalizations. Spirometry improved to a similar extent in both subgroups for the entire duration of the 6-month trial. African Americans used fewer respiratory medications than Caucasians in this study. We conclude that tiotropium reduces COPD exacerbations and associated health-care use to a similar extent in African Americans compared with Caucasians. PMID- 18674744 TI - Thrombospondin-1 and transforming growth factor beta are pro-inflammatory molecules in rheumatoid arthritis. AB - Thrombospondin-1 (TSP1/THBS1) plays a major role in the pathophysiology of rheumatoid arthritis (RA); however, its interface with the cytokine network involved in RA has not been delineated. Correlations were performed between plasma levels of TSP1 and selected cytokines from blood samples collected from 20 patients affected by RA and 13 healthy donors (control). Plasma levels of TSP1 and tissue growth factor beta (TGFbeta) were determined by standard enzyme-linked immunosorbent assay, and cytokines were measured by protein profiling rolling circle amplification (RCA). TSP1 circulating levels in plasma were found significantly increased in the RA patients when compared with control individuals (P = 0.039). The plasma levels of TGFbeta were also increased in the RA patients, which indicates a statistical trend. Cytokine levels of interleukin (IL)-4, IL-5, IL-12, chemokine CXC 10 (CXCL10/IP10), and chemokine CC 4 (CCL4)/MIP1beta were significantly increased in the RA patients when compared with the control group. In summary, this study demonstrates increased plasma levels of TSP1, which correlated with increased levels of proinflammatory cytokines in plasma of RA patients. More detailed research is required to explore the cytokine imprint yielded by this study and its interface with TSP1 and TGFbeta. PMID- 18674745 TI - Impairment of SLC17A8 encoding vesicular glutamate transporter-3, VGLUT3, underlies nonsyndromic deafness DFNA25 and inner hair cell dysfunction in null mice. AB - Autosomal-dominant sensorineural hearing loss is genetically heterogeneous, with a phenotype closely resembling presbycusis, the most common sensory defect associated with aging in humans. We have identified SLC17A8, which encodes the vesicular glutamate transporter-3 (VGLUT3), as the gene responsible for DFNA25, an autosomal-dominant form of progressive, high-frequency nonsyndromic deafness. In two unrelated families, a heterozygous missense mutation, c.632C-->T (p.A211V), was found to segregate with DFNA25 deafness and was not present in 267 controls. Linkage-disequilibrium analysis suggested that the families have a distant common ancestor. The A211 residue is conserved in VGLUT3 across species and in all human VGLUT subtypes (VGLUT1-3), suggesting an important functional role. In the cochlea, VGLUT3 accumulates glutamate in the synaptic vesicles of the sensory inner hair cells (IHCs) before releasing it onto receptors of auditory-nerve terminals. Null mice with a targeted deletion of Slc17a8 exon 2 lacked auditory-nerve responses to acoustic stimuli, although auditory brainstem responses could be elicited by electrical stimuli, and robust otoacoustic emissions were recorded. Ca(2+)-triggered synaptic-vesicle turnover was normal in IHCs of Slc17a8 null mice when probed by membrane capacitance measurements at 2 weeks of age. Later, the number of afferent synapses, spiral ganglion neurons, and lateral efferent endings below sensory IHCs declined. Ribbon synapses remaining by 3 months of age had a normal ultrastructural appearance. We conclude that deafness in Slc17a8-deficient mice is due to a specific defect of vesicular glutamate uptake and release and that VGLUT3 is essential for auditory coding at the IHC synapse. PMID- 18674746 TI - Class-specific correlations of gene expressions: identification and their effects on clustering analyses. AB - Current microarray studies primarily focus on identifying individual genes with differential expression levels across different conditions or classes. A potential problem is that they may disregard multidimensional information hidden in gene interactions. In this study, we propose an approach to detect gene interactions related to study phenotypes through identifying gene pairs with correlations that appear to be class or condition specific. In addition, we explore the effects of ignoring class-specific correlations (CSC) on correlation based gene-clustering analyses. Our simulation studies show that ignoring CSC can significantly decrease the accuracy of gene clustering and increase the dissimilarity within clusters. Our results from a DLBCL (distinct types of diffuse large B cell lymphoma) data set illustrate that CSC are clearly present and have great adverse effects on gene-clustering results if ignored. Meanwhile, interesting biological interpretations may be derived from studying gene pairs with CSC. This study demonstrates that our algorithm is simple and computationally efficient and has the ability to detect gene pairs with CSC that are informative for uncovering interesting regulation patterns. PMID- 18674747 TI - Pathogenic mitochondrial DNA mutations are common in the general population. AB - Mitochondrial DNA (mtDNA) mutations are a major cause of genetic disease, but their prevalence in the general population is not known. We determined the frequency of ten mitochondrial point mutations in 3168 neonatal-cord-blood samples from sequential live births, analyzing matched maternal-blood samples to estimate the de novo mutation rate. mtDNA mutations were detected in 15 offspring (0.54%, 95% CI = 0.30-0.89%). Of these live births, 0.00107% (95% CI = 0.00087 0.0127) harbored a mutation not detected in the mother's blood, providing an estimate of the de novo mutation rate. The most common mutation was m.3243A-->G. m.14484T-->C was only found on sub-branches of mtDNA haplogroup J. In conclusion, at least one in 200 healthy humans harbors a pathogenic mtDNA mutation that potentially causes disease in the offspring of female carriers. The exclusive detection of m.14484T-->C on haplogroup J implicates the background mtDNA haplotype in mutagenesis. These findings emphasize the importance of developing new approaches to prevent transmission. PMID- 18674748 TI - ATM gene mutations result in both recessive and dominant expression phenotypes of genes and microRNAs. AB - The defining characteristic of recessive disorders is the absence of disease in heterozygous carriers of the mutant alleles. However, it has been recognized that recessive carriers may differ from noncarriers in some phenotypes. Here, we studied ataxia telangiectasia (AT), a classical recessive disorder caused by mutations in the ataxia telangiectasia mutated (ATM) gene. We compared the gene and microRNA expression phenotypes of noncarriers, AT carriers who have one copy of the ATM mutations, and AT patients with two copies of ATM mutations. We found that some phenotypes are more similar between noncarriers and AT carriers compared to AT patients, as expected for a recessive disorder. However, for some expression phenotypes, AT carriers are more similar to the patients than to the noncarriers. Analysis of one of these expression phenotypes, TNFSF4 level, allowed us to uncover a regulatory pathway where ATM regulates TNFSF4 expression through MIRN125B (also known as miR-125b or miR125b) [corrected] In AT carriers and AT patients, this pathway is disrupted. As a result, the level of MIRN125B is lower and the level of its target gene, TNFSF4, is higher than in noncarriers. A decreased level of MIRN125B is associated with breast cancer, and an elevated level of TNFSF4 is associated with atherosclerosis. Thus, our findings provide a mechanistic suggestion for the increased risk of breast cancer and heart disease in AT carriers. By integrating molecular and computational analyses of gene and microRNA expression, we show the complex consequences of a human gene mutation. PMID- 18674749 TI - Extensive copy-number variation of the human olfactory receptor gene family. AB - As much as a quarter of the human genome has been reported to vary in copy number between individuals, including regions containing about half of the members of the olfactory receptor (OR) gene family. We have undertaken a detailed study of copy-number variation of ORs to elucidate the selective and mechanistic forces acting on this gene family and the true impact of copy-number variation on human OR repertoires. We argue that the properties of copy-number variants (CNVs) and other sets of large genomic regions violate the assumptions of statistical methods that are commonly used in the assessment of gene enrichment. Using more appropriate methods, we provide evidence that OR enrichment in CNVs is not due to positive selection but is because of OR preponderance in segmentally duplicated regions, which are known to be frequently copy-number variable, and because purifying selection against CNVs is lower in OR-containing regions than in regions containing essential genes. We also combine multiplex ligation-dependent probe amplification (MLPA) and PCR to assay the copy numbers of 37 candidate CNV ORs in a panel of approximately 50 human individuals. We confirm copy-number variation of 18 ORs but find no variation in this human-diversity panel for 16 other ORs, highlighting the caveat that reported intervals often overrepresent true CNVs. The copy-number variation we describe is likely to underpin significant variation in olfactory abilities among human individuals. Finally, we show that both homology-based and homology-independent processes have played a recent role in remodeling the OR family. PMID- 18674750 TI - WW-domain-containing oxidoreductase is associated with low plasma HDL-C levels. AB - Low serum HDL-cholesterol (HDL-C) is a major risk factor for coronary artery disease. We performed targeted genotyping of a 12.4 Mb linked region on 16q to test for association with low HDL-C by using a regional-tag SNP strategy. We identified one SNP, rs2548861, in the WW-domain-containing oxidoreductase (WWOX) gene with region-wide significance for low HDL-C in dyslipidemic families of Mexican and European descent and in low-HDL-C cases and controls of European descent (p = 6.9 x 10(-7)). We extended our investigation to the population level by using two independent unascertained population-based Finnish cohorts, the cross-sectional METSIM cohort of 4,463 males and the prospective Young Finns cohort of 2,265 subjects. The combined analysis provided p = 4 x 10(-4) to 2 x 10(-5). Importantly, in the prospective cohort, we observed a significant longitudinal association of rs2548861 with HDL-C levels obtained at four different time points over 21 years (p = 0.003), and the T risk allele explained 1.5% of the variance in HDL-C levels. The rs2548861 resides in a highly conserved region in intron 8 of WWOX. Results from our in vitro reporter assay and electrophoretic mobility-shift assay demonstrate that this region functions as a cis-regulatory element whose associated rs2548861 SNP has a specific allelic effect and that the region forms an allele-specific DNA-nuclear-factor complex. In conclusion, analyses of 9,798 subjects show significant association between HDL-C and a WWOX variant with an allele-specific cis-regulatory function. PMID- 18674751 TI - Mutations in the cilia gene ARL13B lead to the classical form of Joubert syndrome. AB - Joubert syndrome (JS) and related disorders are a group of autosomal-recessive conditions sharing the "molar tooth sign" on axial brain MRI, together with cerebellar vermis hypoplasia, ataxia, and psychomotor delay. JS is suggested to be a disorder of cilia function and is part of a spectrum of disorders involving retinal, renal, digital, oral, hepatic, and cerebral organs. We identified mutations in ARL13B in two families with the classical form of JS. ARL13B belongs to the Ras GTPase family, and in other species is required for ciliogenesis, body axis formation, and renal function. The encoded Arl13b protein was expressed in developing murine cerebellum and localized to the cilia in primary neurons. Overexpression of human wild-type but not patient mutant ARL13B rescued the Arl13b scorpion zebrafish mutant. Thus, ARL13B has an evolutionarily conserved role mediating cilia function in multiple organs. PMID- 18674752 TI - Diabetes mellitus and birth defects. AB - OBJECTIVE: The purpose of this study was to examine associations between diabetes mellitus and 39 birth defects. STUDY DESIGN: This was a multicenter case-control study of mothers of infants who were born with (n = 13,030) and without (n = 4895) birth defects in the National Birth Defects Prevention Study (1997-2003). RESULTS: Pregestational diabetes mellitus (PGDM) was associated significantly with noncardiac defects (isolated, 7/23 defects; multiples, 13/23 defects) and cardiac defects (isolated, 11/16 defects; multiples, 8/16 defects). Adjusted odds ratios for PGDM and all isolated and multiple defects were 3.17 (95% CI, 2.20 4.99) and 8.62 (95% CI, 5.27-14.10), respectively. Gestational diabetes mellitus (GDM) was associated with fewer noncardiac defects (isolated, 3/23 defects; multiples, 3/23 defects) and cardiac defects (isolated, 3/16 defects; multiples, 2/16 defects). Odds ratios between GDM and all isolated and multiple defects were 1.42 (95% CI, 1.17-1.73) and 1.50 (95% CI, 1.13-2.00), respectively. These associations were limited generally to offspring of women with prepregnancy body mass index > or =25 kg/m(2). CONCLUSION: PGDM was associated with a wide range of birth defects; GDM was associated with a limited group of birth defects. PMID- 18674753 TI - Accounting for measurement error: a critical but often overlooked process. AB - AIMS: Due to instrument imprecision and human inconsistencies, measurements are not free of error. Technical error of measurement (TEM) is the variability encountered between dimensions when the same specimens are measured at multiple sessions. A goal of a data collection regimen is to minimise TEM. The few studies that actually quantify TEM, regardless of discipline, report that it is substantial and can affect results and inferences. This paper reviews some statistical approaches for identifying and controlling TEM. Statistically, TEM is part of the residual ('unexplained') variance in a statistical test, so accounting for TEM, which requires repeated measurements, enhances the chances of finding a statistically significant difference if one exists. METHODS: The aim of this paper was to review and discuss common statistical designs relating to types of error and statistical approaches to error accountability. This paper addresses issues of landmark location, validity, technical and systematic error, analysis of variance, scaled measures and correlation coefficients in order to guide the reader towards correct identification of true experimental differences. CONCLUSIONS: Researchers commonly infer characteristics about populations from comparatively restricted study samples. Most inferences are statistical and, aside from concerns about adequate accounting for known sources of variation with the research design, an important source of variability is measurement error. Variability in locating landmarks that define variables is obvious in odontometrics, cephalometrics and anthropometry, but the same concerns about measurement accuracy and precision extend to all disciplines. With increasing accessibility to computer-assisted methods of data collection, the ease of incorporating repeated measures into statistical designs has improved. Accounting for this technical source of variation increases the chance of finding biologically true differences when they exist. PMID- 18674754 TI - Structure of the O-specific polysaccharide of the lipopolysaccharide of Rahnella aquatilis 95 U003. AB - The O-polysaccharide of Rahnella aquatilis 95 U003 was obtained by mild acid degradation of the lipopolysaccharide and studied by sugar and methylation analyses, Smith degradation and (1)H and (13)C NMR spectroscopy, including 2D (1)H,(1)H COSY, TOCSY, ROESY, H-detected (1)H,(13)C HSQC and HMQC-TOCSY experiments. The O-polysaccharide was found to have a branched hexasaccharide repeating unit of the following structure: [carbohydrate structure: see text] PMID- 18674755 TI - IgE peptide-specific CTL inhibit IgE production: a transient IgE suppression model in wild-type and HLA-A2.1 transgenic mice. AB - Effect of IgE peptide-specific CTL on IgE antibody production was studied in mouse models. CTL elicited in B6.A2Kb tg mice against a human IgE peptide nonamer, pWV, lysed human IgE-secreting U266 myeloma cells and inhibit IgE production by these cells. U266 transfected with mouse A2Kb transgene (U266-A2Kb) were optimally lysed by these CTL, because the alpha3 domain of A2Kb interacts well with the CD8 co-receptors. The CTL generated were more effective in inhibiting IgE production by U266-A2Kb cells than lysing these cells. IgE production by and progression of U266 myeloma were suppressed in B6.A2Kb tg mice rendered tolerant to these cells and vaccinated with pWV along with CpG. We also studied the CTL response elicited in wild-type mice by a mouse nonameric IgE peptide, PI-1, along with CpG. This treatment caused a transient suppression of the IgE response in mice previously sensitized to an antigen. In mice treated with this regimen repeatedly, the IgE response was fully recovered 20 days after each treatment. Notably, while IgE peptide/CpG-treated mice remained unresponsive to antigen challenge in vivo, antigen-specific IgE production can be elicited by antigen in cultured splenocytes from these mice. Moreover, IgE peptide/CpG also inhibited an on-going IgE response, including IgE production by bone marrow cells. Taken together, these observations indicate that a CTL-based IgE peptide vaccine targeting IgE-secreting B/plasma cells may be safely employed as a therapeutic approach for suppressing IgE production. PMID- 18674756 TI - Long-term effects of human butyrylcholinesterase pretreatment followed by acute soman challenge in cynomolgus monkeys. AB - Human serum butyrylcholinesterase (Hu BChE) was demonstrated previously to be an effective prophylaxis that can protect animals from organophosphate nerve agents. However, in most of those studies, the maximum dose used to challenge animals was low (<2x LD(50)), and the health of these animals was monitored for only up to 2 weeks. In this study, six cynomolgus monkeys received 75 mg of Hu BChE followed by sequential doses (1.5, 2.0, 2.0 x LD(50)) of soman 10h later for a total challenge of 5.5x LD(50). Four surviving animals that did not show any signs of soman intoxication were transferred to WRAIR for the continuous evaluation of long-term health effects for 14 months. Each month, blood was drawn from these monkeys and analyzed for serum chemistry and hematology parameters, blood acetylcholinesterase (AChE) and BChE levels. Based on the serum chemistry and hematology parameters measured, no toxic effects or any organ malfunctions were observed up to 14 months following Hu BuChE protection against exposure to 5.5x LD(50) of soman. In conclusion, Hu BChE pretreatment not only effectively protects monkeys from soman-induced toxicity of the immediate acute phase but also for a long-term outcome. PMID- 18674757 TI - Artificial cranial deformation in Kow Swamp 1 and 5: a response to Curnoe (2007). AB - Recently, Curnoe (2007) tested the predictions of competing models of modern human origins using three crania from Australia: Kow Swamp 1 and 5 and Keilor. The Kow Swamp specimens have long been suspected of having been altered through artificial deformation of the skull. Though Curnoe (2007) provided assurances that no evidence of deformation is present in those specimens, the current study retests the hypothesis that these Australian specimens are artificially deformed. The Australian crania are compared to known deformed individuals from New Britian through canonical variates analysis, and the resulting Mahalanobis distances are examined for statistical significance with random expectation statistics. The results show that Kow Swamp 1 and 5 have strong shape similarities to known deformed individuals, and both crania are very different in shape from Keilor. Keilor is statistically significantly different in shape from both Kow Swamp specimens and all of the known deformed specimens. These findings cast doubt on Curnoe's (2007) conclusions of a shared Australian cranial morphology as well as the retention of an archaic suite of morphologies in the Australians. PMID- 18674758 TI - Chronic maternal stress affects growth, behaviour and hypothalamo-pituitary adrenal function in juvenile offspring. AB - Maternal stress during pregnancy, particularly that combined with low socioeconomic status (SES), has been linked to an increased risk for impaired behavioural and emotional development and affective disorders in children. In animal models, acute periods of prenatal stress have profound effects on hypothalamo-pituitary-adrenal (HPA) function and behaviour. However, few studies have determined the impact of chronic exposure to stress in animal models. The objective of this study was to determine the effects of chronic maternal stress (CMS) during the 2nd half of pregnancy and nursing on growth, locomotor behaviour and HPA axis function in juvenile guinea pig offspring. Pregnant guinea pigs were exposed to a random combination of variable stressors every other day over the 2nd half of gestation and from postnatal day (pnd) 1 until weaning (pnd25). CMS mothers displayed increased basal salivary cortisol levels in the later stages of pregnancy compared to control mothers (p<0.05). The male offspring of CMS mothers had a lower bodyweight, which was maintained to weaning (p<0.01). In open-field testing, CMS male offspring showed a decrease in activity compared to controls (p<0.05). There was no effect of CMS on bodyweight or activity in female offspring. In contrast, both male and female offspring born to CMS mothers displayed increased (p<0.05) basal salivary cortisol at pnd25, but a blunted adrenocortical response to exposure to the novel open-field enclosure. In conclusion, CMS leads to modification of growth trajectory, locomotor activity and adrenocortical responses to stress in juvenile offspring. Further, males appear considerably more vulnerable to these effects than females. PMID- 18674759 TI - Is mechanism of injury alone a useful predictor of major trauma? AB - INTRODUCTION: The Review of Trauma and Emergency Services in Victoria -1999 left unresolved the predictive value of mechanism of injury in pre-hospital trauma triage guidelines. Ethics approval was granted. The objective of this study is to determine if mechanism of injury alone is a useful predictor of major trauma in pre-hospital trauma triage. METHODS: A retrospective cohort study was undertaken of all Victorian ambulance trauma Patient Care Records (PCRs) for 2002. PCRs where patients were physiologically stable, had no significant pattern of injury, but had a significant mechanism of injury were identified and compared with the State Trauma Registry to determine those patients who sustained hospital defined major trauma. RESULTS: There were 4571 incidents of mechanism of injury only, of which 62% were males, median age was 28 years. Two criteria had statistically significant results. A fall from greater than 5m (n=52) of whom 5 (RR 10.86, CI 4.47 to 26.42, P<0.0001) sustained major trauma and a patient trapped greater than 30 min (n=36) of whom 3 (RR 9.0, CI 2.92 to 27.70, P=0003) sustained major trauma. The overall results are not clinically significant. CONCLUSION: This study suggests that individual mechanism of injury criteria have no clinical or operational significance in pre-hospital trauma triage of patients who have an absence of physiological distress and no significant pattern of injury. These results add to the knowledge base of trauma presentation in the pre-hospital setting, especially in Australia, and are the baseline for further studies. PMID- 18674760 TI - The incision angle of mediolateral episiotomy before delivery and after repair. AB - OBJECTIVE: To assess the relevance of the suture angle when evaluating mediolateral episiotomy, and the safety of the accepted lower limit of 40 degrees for the incision angle. METHOD: The angles formed by the incision and suture lines with the midline were measured, and any perineal tear noted, in 50 consecutive women undergoing their first vaginal delivery. RESULTS: The angles were significantly different (40 degrees and 22.5 degrees , respectively; P<0.001). The only variable significantly linked to the difference was the timing of the episiotomy. CONCLUSION: The suture angle cannot currently be used to assess the adequacy of the incision angle; moreover, an incision angle of 40 degrees is probably too acute to prevent potential sphincter damage. PMID- 18674761 TI - Knowledge translation and implementation research in nursing. PMID- 18674763 TI - Plasma levels of beta(2)-microglobulin, a biomarker of peripheral arterial disease, are not affected by maximal leg exercise in patients with intermittent claudication. PMID- 18674762 TI - Gender differences in and factors related to self-care behaviors: a cross sectional, correlational study of patients with heart failure. AB - BACKGROUND: Although self-care may reduce exacerbations of heart failure, reported rates of effective self-care in patients with heart failure are low. Modifiable factors, including psychosocial status, knowledge, and physical factors, are thought to influence heart failure self-care, but little is known about their combined impact on self-care. OBJECTIVES: The objective of this study was to identify factors related to self-care behaviors in patients with heart failure. DESIGN: A cross-sectional, correlational study design was used. PARTICIPANTS AND SETTINGS: One hundred twenty-two patients (77 men and 45 women, mean age 60+/-12 years old, 66% New York Heart Association functional class III/IV) were recruited from the outpatient clinics of an academic medical center and two community hospitals. METHODS: Data on self-care behaviors (Self-Care of Heart Failure Index), depressive symptoms, perceived control, self-care confidence, knowledge, functional status, and social support were collected. Factors related to self-care were examined using hierarchical multiple regression. RESULTS: Mean self-care behavior scores were less than 70 indicating the majority of men and women with HF did not consistently engage in self-care behaviors. Higher self-care confidence and perceived control and better heart failure management knowledge were associated with better self-care (r2=.25, p<.001). Higher perceived control and better knowledge were related to better self-care behaviors in men (r2=.18, p=.001), while higher self-care confidence and poorer functional status were related to better self-care behaviors in women (r2=.35, p<.001). CONCLUSION: This study demonstrates the substantial impact of modifiable factors such as confidence in one's self-care abilities, perceived control, and knowledge on self-care behaviors. This study demonstrates that there are gender differences in factors affecting self-care, even though at baseline men and women have similar knowledge levels, physical, psychological, and behavioral status. Effective interventions focusing on modifiable factors and the unique characteristics of men and women should be provided to improve self-care behaviors in patients with heart failure. PMID- 18674764 TI - Investigation of hepatic gluconeogenesis pathway in non-diabetic Asian Indians with non-alcoholic fatty liver disease using in vivo ((31)P) phosphorus magnetic resonance spectroscopy. AB - OBJECTIVE: To study hepatic gluconeogenesis pathway in non-diabetic Asian Indian males having non-alcoholic fatty liver disease (NAFLD) using in vivo ((31)P) phosphorous magnetic resonance spectroscopy (MRS) and correlate these data with anthropometry and insulin resistance. RESEARCH DESIGN AND METHODS: Forty non diabetic patients with NAFLD and 20 healthy controls were divided into (i) obese with NAFLD (group I, n=20), (ii) non-obese with NAFLD (group II, n=20) and (iii) non-obese without NAFLD (group III, n=20). Anthropometric and biochemical profiles, short insulin tolerance test (SITT), liver ultrasound, and (31)P MRS (to determine hepatic gluconeogenesis metabolite; phosphomonoesters (PMEs), inorganic phosphate (Pi) and their ratios with respect to ATP) were done. RESULTS: Insulin resistance (Kitt value) was highest in group I (p<0.05; compared to other two groups), but was also higher in group II as compared to group III (p=ns). The values of PME/Pi, PME/gammaATP, PME/betaATP, PME/tATP ratios were higher (p<0.05) in group I compared to other two groups. Interestingly, non-obese subjects with NAFLD also showed more derangements of hepatic gluconeogenesis metabolites than non-obese subjects without NAFLD. Positive correlation was observed between PME and other ratios in relation to body mass index, waist circumference, body fat percentage and fasting serum insulin levels in all the three groups. CONCLUSIONS: Derangements in hepatic gluconeogenesis as assessed non-invasively using (31)P MRS, was observed in obese and non-obese, non-diabetic Asian Indians with NAFLD. Further research is warranted whether this investigation in NAFLD subjects could be developed as a non-invasive tool to assess those predisposed to develop hyperglycemia. PMID- 18674765 TI - Study of the collateral capacity of the circle of Willis of patients with severe carotid artery stenosis by 3D computational modeling. AB - This numerical study aims to investigate the capacity of the circle of Wills (CoW) to provide collateral blood supply for patients with unilateral carotid arterial stenosis. The basic 3D geometry of the CoW was reconstructed based on a magnetic resonance angiogram of a normal human subject. A total of 52 computational fluid dynamics simulations were performed for four geometry configurations of the CoW with an artificially inserted axisymmetric stenosis of different luminal area reductions in an internal carotid artery (ICA) under a variety of boundary conditions. The CoW geometric configurations included (a) a normal CoW with all communicating arteries; (b) as model (a) but with enlarged communicating arterial diameters; (c) as (a) but with the ipsilateral posterior communicating artery missing, and (d) as (c) but with enlarged communicating arteries. It is found that the blood perfusion pressure drop between the ipsilateral ICA and the middle cerebral artery (MCA) only becomes significant when the degree of stenosis is greater than 86%. The cerebral autoregulation range varied significantly between the different CoW configurations for the severe stenosis cases. Without causing the flow rates to decrease at the efferent arterial ends, the mean perfusion pressure in the ipsilateral ICA can drop from 100 to 73, 67, 92 and 84 mmHg for the CoW models (a)-(d) with 96% luminal area reduction stenosis, respectively. The additional pathways are able to raise the ipsilateral MCA pressure significantly without reducing the total flow perfusion. Cerebral autoregulation effects were not directly included in the study. Therefore, the findings in the study should be interpreted with cautions when comes to the biological and clinical significance. PMID- 18674766 TI - An approach to the mechanical constitutive modelling of arterial tissue based on homogenization and optimization. AB - This paper is concerned with characterizing the quasistatic mechanical behaviour of arterial tissue undergoing finite deformation through hyperelastic constitutive functions. Commonly the parameters of constitutive functions are established by a process of optimization based on experimental data. Instead we construct a finite element model of a representative volume element of the material and compute its homogenized response to a range of deformations. These data are then used to provide objective functions for optimizing the parameters of two analytical models from the literature. PMID- 18674767 TI - Determination of priority carcinogenic polycyclic aromatic hydrocarbons in wastewater and surface water by coacervative extraction and liquid chromatography fluorimetry. AB - The US Environmental Protection Agency (EPA) and the European Union (EU) have set restrictive limits for priority carcinogenic polycyclic aromatic hydrocarbons (CPAHs) in surface waters (EPA 3.8 ng L(-1) and EU 2-100 ng L(-1)) in order to protect aquatic life and human health. Currently, methods meeting these sensitivity criteria are not suitable for routine analysis of CPAHs. Here, we present a simple, rapid and low-cost method for the routine monitorization of these pollutants in aquatic environments based on their extraction with coacervates of decanoic acid reverse micelles in the nano- and microscale, and determination by liquid chromatography-fluorimetry (LC-FL). The method involves the stirring of filtered aqueous samples (36 mL) with 4 mL of THF containing 70 mg of decanoic acid for 5 min, its centrifugation for 10 min and the analysis of 20 microL of the resulting coacervate containing the CPAHs by LC/FL. The method is robust, the extractions being independent on salt concentration (up to 1 M), temperature (up to 60 degrees C) and pH (below 4). Besides, the coacervate prevents the CPAHs from adsorption onto the surface of containers during sample storage. No clean-up steps are necessary and the method is matrix-independent. The quantification and detection limits of the method ranged between 0.4 and 3.5 ng L(-1) and 0.1 and 1 ng L(-1), respectively, for the seven priority CPAHs. The method has been successfully applied to the determination of these pollutants in raw and treated sewage from three mechanical-biological treatment plants, two rivers and a reservoir with frequent motorized recreational craft activities, all of them located in the South of Spain. Recoveries for spiked samples in the range 2-30 ng L(-1) were between 88 and 95% with relative standard deviations from 1 to 7%. CPAHs were present in wastewater influents at concentrations in the range 3.9 37 ng L(-1), while the treatment at the WWTPs studied reduced their concentration in their respective effluents in a percentage near 100%. Three CPAHs were present at quantifiable levels in Guadajoz river (1.8-6.6 ng L(-1)) and six in La Brena reservoir (1.39-4.8 ng L(-1)). PMID- 18674768 TI - Combining high-performance liquid chromatography-positron detection and on-line microdialysis for animal metabolism study of positron emission tomography probes. AB - Automated ultra-performance liquid chromatography (LC) with a radiometric detection system has been developed to analyze compounds labeled with short-lived positron (beta(+))-emitting radionuclides in microdialysates. This system involves (1) on-line microdialysis sampling to submit dialysates directly into the LC, (2) ultra-performance LC for improving sensitivity, resolution and speed of analysis, and (3) tandem flow-through of beta(+) and electrochemical detectors for real-time monitoring of radioactive and endogenous compounds simultaneously. This system permitted ultrasensitive measurement of radioactive compounds at the lowest detection limit of about 1Bq with a high-temporal resolution of sampling and less operator attention. The developed system was used for the continuous monitoring of L-[beta-(11)C]-3,4-dihydroxyphenylalanine (L-[beta-(11)C]DOPA) metabolites in the rat brain. PMID- 18674769 TI - Simultaneous analysis of amino acids and amines as their o-phthalaldehyde ethanethiol-9-fluorenylmethyl chloroformate derivatives in cheese by high performance liquid chromatography. AB - A high-performance liquid chromatographic method (HPLC) with combined diode array and fluorescence detection of amino acids and amines in various cheese samples is described. The proposal is based on acidic deproteinization, derivatization and gradient optimization studies, resulting in the identification and quantification of 21 amino acids and 9 amines from a single solution, by one injection. The optimized, simple protocol consists of deproteinization (1M perchloric acid), centrifugation, filtration and the subsequent derivatization with the o phthalaldehyde-ethanethiol-9-fluorenylmethyl chloroformate (OPA-ET-FMOC) reagent. The method can be characterized with a linearity of wide concentration range (6.25-1000 pM/injection), a good chromatographic reproducibility (average: 2.69% RSD) and an excellent recovery (average: 100.2%; average 3.84% RSD). The developed method was successfully applied in the determination of the amino acid and amine contents of port salut cheese, blue cheese and smoked cheese samples. PMID- 18674770 TI - Development of a new analytical method for the determination of fumonisins B1 and B2 in food products based on high performance liquid chromatography and fluorimetric detection with post-column derivatization. AB - A sensitive and selective analytical method was developed for the quantitative determination of fumonisins B(1) and B(2) in maize-based foods for direct human consumption. The method, based on high-performance liquid chromatography and fluorescence detection, presents a rapid and automated on-line post-column derivatization, performed with o-phtalaldehyde and N,N-dimethyl-2 mercaptoethylamine. Several factors affecting the separation and detection of fumonisins were investigated, including mobile phase composition, column features, derivatization agent flow-rate and both the excitation and the emission wavelengths. Optimal fluorescence detection was obtained by using a lambda(exc) of 343 nm and a lambda(em) of 445 nm. Under the optimized experimental conditions, a complete separation of fumonisins was obtained in less than 13 min by using a C(18) column and a gradient elution at 0.8 mL/min with methanol and 0.1M phosphate buffer at pH 3.15. The limits of detection for FB(1) and FB(2) were 4 and 5 microg/L corresponding to 5 and 6 microg/kg in matrix. Each fumonisin was determined in the range 40-320 microg/L that correspond to 50-400 microg/kg in matrix. The necessary requirements for accuracy, reproducibility and sensitivity were fulfilled and recovery values ranged from 87 to 94% for FB(1) and from 70 to 75% for FB(2) in cornflake samples at three fortification levels in the range 100-300 microg/kg. The potential of this method, combined with a simple clean-up procedure, was assessed by the measurements of FB(1) and FB(2) in maize-based products, such as maize flour, "polenta", tortillas and cookies. PMID- 18674771 TI - Size-controllable synthesis of monodispersed colloidal silica nanoparticles via hydrolysis of elemental silicon. AB - A new method is presented for preparing monodisperse and uniform-size silica nanoparticles using a two-stage hydrolysis of silicon powder in aqueous medium. The influence of synthesis conditions including solution composition and temperature on the formation of silica nanoparticles were systematically investigated. The structure and morphology of the silica particles were characterized via transmission electron microscopy (TEM) and dynamic light scattering (DLS). Various-sized particles in the range 10-100 nm were synthesized. The size of the nanoparticles can be precisely controlled by using a facile regrowth procedure in the same reaction media. PMID- 18674772 TI - Mechanisms of equilibrium shape transitions of liquid droplets in electrowetting. AB - Liquid droplets bridging the gap between two dielectric-coated horizontal electrode plates suffer breakup instabilities when a voltage applied between the electrodes exceeds a threshold. Interestingly enough, broken liquid bridges (i.e. a pair of a sessile and a pendant drop) can spontaneously rejoin if the voltage is still applied to the electrodes. Here we study the electro-hydrostatics of the liquid bridges in the joined or broken state and we illuminate the mechanisms of the shape transitions that lead to bridge rupture or droplet joining. The governing equations of the capillary electro-hydrostatics form nonlinear and free boundary problems which are solved numerically by the Galerkin/finite element method. On one hand, we found that capillary bridges become unstable at a turning point bifurcation in their solution space. The solutions past the turning point are unstable and the instability signals the bridge rupture. On the other hand, the separate droplets approach each other as the applied voltage increases. However, solutions become unstable past a critical voltage at a turning point bifurcation and the droplets join. By studying the relative position of the turning points corresponding to bridge rupture and droplet joining, respectively, we define parameter regions where stable bridges or separate droplets or oscillations between them can be realized. PMID- 18674773 TI - Growth kinetics for AgI nanoparticles in AOT reverse micelles: effect of molecular length of hydrocarbon solvents. AB - The growth kinetics for AgI nanoparticles formed in the solutions of water/AOT reverse micelles in n-hexane, n-octane, n-decane, and n-dodecane were investigated. In small micelles, the rate of nanoparticles growth was found to be independent of the type of solvent, while in large micelles the growth rate grew with increasing length of solvent molecules. The effect was explained by a different amount of free water in the micelle pools of the same size. PMID- 18674775 TI - Surface charging of layered double hydroxides during dynamic interactions of anions at the interfaces. AB - In this research, we investigated the effect of dynamic anion adsorption/exchange on the surface charging property of Mg(2)AlClLDH and Mg(2)AlCO(3)LDH particles that show the average zeta potential of 41 and 34 mV in the as-prepared suspension, respectively. The addition of NaCl up to 3x10(-3) M in the suspension does not obviously affect the zeta potential of both LDHs, which can be attributed to the less affinity of Cl(-) to LDH. The introduction of Na(2)CO(3) severely reduces the zeta potential at the CO(3)(2-) concentration higher than 1x10(-4) M, and to the negative value in both LDH systems at ca. 2x10(-3) M, which is presumably resulted from the exchange and the re-orientation of CO(3)(2 ) in a tilt/vertical style on the surface. All four organic anions (dodecyl sulfate, folate, citrate and polyacrylate) also significantly affect the zeta potential of the LDH particles. At the lower concentrations of organic anionic groups (<1x10(-4) M), the zeta potential was slightly affected, i.e. limited exchange/adsorption. However, the concentration increasing to some point suddenly decreases and reverses the zeta potential of the LDH particles, which is presumably caused by the hydrophobic interactions that bind the hydrophobic hydrocarbon chains (especially in dodecyl sulfate) into the micelle-like bilayer bunches on the LDH surface. In addition, the effect of pH in 5.5-11.0 on the LDH particle surface charging is mainly reflected through the conversion of CO(3)(2-) to HCO(3)(-)/H(2)CO(3) when pH decreases from ca. 11 to 6, with limited contribution from protonation/deprotonation and exchange/adsorption. PMID- 18674774 TI - Surfactant dependent morphology of polymeric capsules of perfluorooctyl bromide: influence of polymer adsorption at the dichloromethane-water interface. AB - In a strategy to develop more stable ultrasound contrast agents (UCAs), we have designed a process to obtain nano/microcapsules with a single core of liquid perfluorocarbon within a biodegradable polymeric shell of homogeneous thickness. During the optimization of perfluorooctyl bromide (PFOB) encapsulation by solvent emulsion-evaporation, a marked influence of surfactants has been observed. While sodium cholate leads to spherical capsules of homogeneous thickness, sodium taurocholate induces to the formation of "acorn"-particles with one hemisphere of PFOB and another one of PLGA, and polyvinyl alcohol is responsible for the coexistence of both morphologies. Whereas the theoretical model proposed by Torza and Mason [J. Colloid Interface Sci. 33 (1970) 67] fails to predict the observed morphologies, microscopic observations of the evaporation and interfacial tension measurements provide an insight into the mechanism of formation of these structures. Most probably, there is a competition between PLGA and the surfactant stabilizing the emulsion at the dichloromethane-water interface. If PLGA is able to adsorb at the interface, the core-shell morphology is obtained, otherwise the acorn morphology is preferentially formed. When the surfactant rearrangement at the interface is long (>30 min), a coexistence of morphologies can be obtained. PMID- 18674776 TI - Structure and dynamical mechanical properties of suspensions of sodium caseinate. AB - Sodium caseinate is derived from casein which is the major milk protein and forms small star-like aggregates in aqueous solution. The dynamic mechanical properties of dense sodium caseinate suspensions were studied by measuring the shear viscosity and the frequency dependent loss and storage shear modulus. The viscosity is shown to increase strongly above a concentration, C(c), due to jamming of the aggregates. C(c) depends little on the pH or on the ionic strength. The repulsion between the aggregates increases with decreasing temperature leading to jamming at lower concentrations. As a consequence the viscosity decreases strongly with increasing temperature for concentrated suspensions. The frequency dependent shear modulus of concentrated suspensions shows a solid like behaviour at high frequencies. Shear thinning is observed at Peclet numbers larger than unity. PMID- 18674777 TI - Shear rheology and porous media flow of wormlike micelle solutions formed by mixtures of surfactants of opposite charge. AB - The rheology of solutions of wormlike micelles formed by oppositely charged surfactant mixtures (cationic cetyl trimethylammonium p-toluene sulfonate, CTAT, and anionic sodium dodecyl sulfate, SDS), in the dilute and semi-dilute regimes, were studied under simple shear and porous media flows. Aqueous mixtures of CTAT and SDS formed homogeneous solutions for SDS/CTAT molar ratios below 0.12. Solutions of mixtures exhibited a strong synergistic effect in shear viscosity, especially in the semi-dilute regime with respect to wormlike micelles, reaching a four order of magnitude increase in the zero-shear rate viscosity for solutions with 20 mM CTAT. Oscillatory shear results demonstrated that the microstructure of CTAT wormlike micelles is sensitive to SDS addition. The cross-over relaxation times of wormlike micelles of 20 mM CTAT solutions increased by three orders of magnitude with the addition of up to 2 mM of SDS, and the solutions became increasingly elastic. The shear thickening process observed in shear rheology became more pronounced in porous media flow due to the formation of stronger cooperative structures induced by the extensional component of the flow. PMID- 18674778 TI - New explanation of the high electron concentration at the surface of a carbon electrode. AB - It is pointed out that the high surface electron concentration in the carbon electrode gives rise to surface repulsion forces between ensembles of electrons and carbon atoms. PMID- 18674779 TI - Characterization of crystal structure in binary mixtures of latex globules. AB - Colloidal crystals formed by two types of polystyrene particles of different sizes (94 and 141 nm) at various number ratios (94:141 nm) are studied. Experiments showed that the formation time of crystals lengthens as the number ratio of the two components approaches 1:1. The dependence of the mean interparticle distance (D(0)), crystal structure and alloy structure on the number ratio of the two types of particles was studied by means of Kossel diffraction technique and reflection spectra. The results showed that as the number ratio decreased, the mean interparticle distance (D(0)) became larger. And the colloidal crystal in binary mixtures is more preferably to form the bcc structure. This study found that binary systems form the substitutional solid solution (sss)-type alloy structure in all cases except when the number ratio of two types of particles is 5:1, which results instead in the superlattice structure. PMID- 18674780 TI - Preparation of polyaniline-metal composite nanospheres by in situ microemulsion polymerization. AB - Nanosized metal and polyaniline (PANi) composite spheres have been prepared via the polymerization of aniline using PdCl(2) or HAuCl(4) as the oxidant in a microemulsion system. The oxidization of aniline and the reduction of metal ion happened together during the reaction, yielding PANi and elemental metal simultaneously. The results of FTIR spectra suggested that the oxidation degree of PANi was affected by the initial ratio of metal ions to monomer in the microemulsion system. The PANi-metal nanospheres were characterized using X-ray photoelectron spectroscopy and the conductivity of the composite nanospheres was measured by conventional four-probe method. Scanning and transmission electron microscopy were used to show the morphology of the composites. PMID- 18674781 TI - Id-1 induces proteasome-dependent degradation of the HBX protein. AB - Id-1 is a member of the HLH protein family that regulates a wide range of cellular processes such as cell proliferation, apoptosis, senescence and overexpression of Id-1 was recently suggested to play roles in the development and progression of different cancers. Previously, Id-1 was shown to physically interact with the viral protein E1A. Meanwhile, Id-1 expression was found to be regulated by several of the virus-encoded proteins, suggesting that Id-1 may be a common cellular target of the viral proteins. Here, we report that Id-1 interacts with the Hepatitis-B virus (HBV)-encoded protein HBX and regulates its stability in hepatocellular carcinoma (HCC) cells. We found that in HCC cells, ectopic Id-1 expression significantly decreased the half-life of the HBX protein, indicating that HBX is destabilized by Id-1. Meanwhile, the Id-1-induced HBX degradation was found to be inhibited by treatment with proteasome inhibitor, suggesting that this process is mediated through the proteasome pathway. Interestingly, while Id 1 did not induce HBX-ubiquitination, we found that removal of all the lysine residues of the HBX protein protects it from the effect of Id-1, indicating that ubiquitination is still required for the Id-1-mediated HBX degradation. Meanwhile, we found that Id-1 binds to the proteasome subunit C8 and facilitates its interaction with the HBX protein and disruption of this interaction completely abolishes the negative effect of Id-1 on HBX protein stability. Taken together, our results demonstrated a novel function of Id-1 in regulating HBX protein stability through interaction with the proteasome. PMID- 18674782 TI - DNA packaging motor assembly intermediate of bacteriophage phi29. AB - Unraveling the structure and assembly of the DNA packaging ATPases of the tailed double-stranded DNA bacteriophages is integral to understanding the mechanism of DNA translocation. Here, the bacteriophage phi29 packaging ATPase gene product 16 (gp16) was overexpressed in soluble form in Bacillus subtilis (pSAC), purified to near homogeneity, and assembled to the phi29 precursor capsid (prohead) to produce a packaging motor intermediate that was fully active in in vitro DNA packaging. The formation of higher oligomers of the gp16 from monomers was concentration dependent and was characterized by analytical ultracentrifugation, gel filtration, and electron microscopy. The binding of multiple copies of gp16 to the prohead was dependent on the presence of an oligomer of 174- or 120-base prohead RNA (pRNA) fixed to the head-tail connector at the unique portal vertex of the prohead. The use of mutant pRNAs demonstrated that gp16 bound specifically to the A-helix of pRNA, and ribonuclease footprinting of gp16 on pRNA showed that gp16 protected the CC residues of the CCA bulge (residues 18-20) of the A-helix. The binding of gp16 to the prohead/pRNA to constitute the complete and active packaging motor was confirmed by cryo-electron microscopy three-dimensional reconstruction of the prohead/pRNA/gp16 complex. The complex was capable of supercoiling DNA-gp3 as observed previously for gp16 alone; therefore, the binding of gp16 to the prohead, rather than first to DNA-gp3, represents an alternative packaging motor assembly pathway. PMID- 18674784 TI - Synthetic bioabsorbable stent material for duct-to-duct biliary reconstruction. AB - Biliary stricture remains a significant cause of morbidity after liver transplantation. We performed duct-to-duct biliary anastomosis by using an absorbable stent tube with a diameter equal to that of pig common bile duct as an internal stent. The stent tube was constructed using a synthetic biodegradable material-a lactic glycolic acid and epsilon-caprolactone copolymer. Three pigs were alive without cholestasis for 180 d; however, 1 pig died on the 65th postoperative d, and autopsy revealed no cholestasis or biliary sledge in the biliary stent tube. The 3 pigs were euthanized for histological examinations 180 d after surgery; the biliary stent tube was completely absorbed by this time. These experimental results showed the good patency of the absorbable biliary stent tube. In the future, the absorbable biliary stent tube is expected to be clinically developed as a biliary stent for biliary anastomosis, which may protect the biliary anastomotic stricture. PMID- 18674783 TI - Prepulse inhibition of startle in adults with ADHD. AB - Prepulse inhibition of startle (PPI) is a measure of sensorimotor gating, a pre conscious regulator of attention. PPI is impaired in adults with schizophrenia and several other neuropsychiatric disorders associated with attentional abnormalities. The core feature of ADHD involves deficits in attention and, like schizophrenia, ADHD is associated with dysregulation of cortical-striatal circuits and dopamine transmission. Therefore, PPI may be disrupted in ADHD. While ADHD persists into adulthood in approximately half the children with ADHD, there has not been any published report of PPI in ADHD adults. In this study, PPI was measured in a sample of ADHD adults and compared to a sample of healthy comparison (HC) subjects. Twenty unmedicated adults with ADHD (11 inattentive subtype, 9 combined subtype) and 17 HC subjects were administered an eyeblink startle PPI paradigm. The PPI of ADHD adults was not significantly different from that of HC subjects in any of the PPI conditions. There was no significant effect of ADHD subtype nor of gender. The lack of PPI deficits in ADHD adults has important implications and suggests that, despite the presence of PPI dysregulation in a large number of disparate neuropsychiatric disorders, it is not a general feature of all neuropsychiatric disorders with attention abnormalities. Furthermore, the attentional deficiency in ADHD may have a neurobiological substrate somewhat distinct from schizophrenia and other neuropsychiatric disorders that are associated with PPI deficits. This distinction may be related to a relative sparing of pre-conscious attentional functions in ADHD compared to other disorders with PPI impairment. PMID- 18674785 TI - General surgery versus specialty rotations: a new paradigm in surgery clerkships. AB - BACKGROUND: The most common 3rd-year surgery clerkship uses general surgery services while limiting the involvement of subspecialty services. A novel surgery clerkship in which students were assigned to either general surgery or subspecialty services for the entire clerkship was tried at a large Midwestern medical school. The purpose of this study was to investigate the outcomes of clerks from subspecialty services and compare them with clerks from general surgery services. METHODS: Outcome measures included scores on the National Board of Medical Examiners Surgery Content Examination, faculty evaluation scores, and residency match results. A multivariate analysis of covariance compared National Board of Medical Examiners test scores and faculty evaluation ratings across service groups. United States Medical Licensing Examination Step 1 scores served as a covariate. RESULTS: Results showed significant differences between groups on faculty evaluation scores (F = 28.03; P 0.45) can prevent or reduce short circuiting and dead zones in peatlands treating wastewater. PMID- 18674793 TI - Seasonal variation in accurate identification of Escherichia coli within a constructed wetland receiving tertiary-treated municipal effluent. AB - As the reuse of municipal wastewater escalates worldwide as a means to extend increasingly limited water supplies, accurate monitoring of water quality parameters, including Escherichia coli (E. coli), increases in importance. Chromogenic media are often used for detection of E. coli in environmental samples, but the presence of unique levels of organic and inorganic compounds alters reclaimed water chemistry, potentially hindering E. coli detection using enzyme-based chromogenic technology. Over seven months, we monitored E. coli levels using m-Coli Blue 24 broth in a constructed wetland filled with tertiary treated municipal effluent. No E. coli were isolated in the wetland source waters, but E. coli, total coliforms, and heterotrophic bacteria increased dramatically within the wetland on all sampling dates, most probably due to fecal inputs from resident wildlife populations. Confirmatory testing of isolates presumptive for E. coli revealed a 41% rate of false-positive identification using m-Coli Blue 24 broth over seven months. Seasonal differences were evident, as false-positive rates averaged 35% in summer, but rose sharply to 75% in the late fall and winter. Corrected E. coli levels were significantly correlated with electrical conductivity, indicating that water chemistry may be controlling bacterial survival within the wetland. This is the first study to report that accuracy of chromogenic media for microbial enumeration in reclaimed water may show strong seasonal differences, and highlights the importance of validation of microbiological results from chromogenic media for accurate analysis of reclaimed water quality. PMID- 18674794 TI - Determination of the persistence of pharmaceuticals in biosolids using liquid chromatography tandem mass spectrometry. AB - Sludge generated in waste water treatment process can be a major sink for some pharmaceutical and personal care products (PPCPs). The land application of sewage sludge (in the form of biosolids in the United States) can therefore potentially introduce PPCPs into the environment. After treatment, biosolids are often subjected to a storage period before land application. However, little information is available with regard to the fate of PPCPs in biosolids during the storage. In this work, the persistence of seven pharmaceuticals and one antibacterial was evaluated using ultrasonic extraction and liquid-chromatography tandem mass spectrometry (LC-MS/MS). The impacts of aeration and sunlight exposure were investigated. During the experiment, no elimination was observed for carbamazepine, triclosan, and ciprofloxacin while elimination was found for tetracycline, doxycycline, clindamycin, erythromycin, and clarithromycin. Using an availability-adjusted kinetic model, the 50% dissipation time was 37 to >77d for tetracycline, 53 to >77d for doxycycline, 1.0-1.6d for clindamycin, 1.1-1.9d for clarithromycin, and 7.0-17d for erythromycin. Those compounds were found more persistent under anaerobic conditions than aerobic condition with a longer 50% dissipation time by a factor of 1.5-2. However, minor impact was observed from sunlight irradiation. PMID- 18674795 TI - Improving ready biodegradability testing of fatty amine derivatives. AB - This study assesses the biodegradation potential of a number of fatty amine derivatives in tests following the OECD guidelines for ready biodegradability. A number of methods are used to reduce toxicity and improve the bioavailability of the fatty amine derivatives in these tests. Alkyl-1,3-diaminopropanes and octadecyltrimethylammonium chloride are toxic to microorganisms at concentrations used in OECD ready biodegradability tests. The concentration of these fatty amine derivatives in the aqueous phase can be reduced by reacting humic, or lignosulphonic acids with the derivatives or through the addition of silica gel to the test bottles. Using these non-biodegradable substances, ready biodegradability test results were obtained with tallow-1,3-diaminopropane and octadecyltrimethylammonium chloride. Demonstration of the ready biodegradability of the water-insoluble dioctadecylamine under the prescribed standard conditions is almost impossible due to the limited bioavailability of this compound. However, ready biodegradability results were achieved by using very low initial test substance concentrations and by introducing an organic phase. The contents of the bottles used to assess the biodegradability of dioctadecylamine were always mixed. False negative biodegradability results obtained with the fatty amine derivatives studied are the result of toxic effects and/or limited bioavailability. The aids investigated therefore improve ready biodegradability testing. PMID- 18674796 TI - Serum dioxin levels in Sydney Harbour commercial fishers and family members. AB - BACKGROUND: Commercial fishing in Sydney Harbour (SH) was banned in February 2006 as a result of high dioxin levels in some SH seafood. Because of concerns by some fishers about their own dioxin levels, testing of serum dioxin levels was offered to SH commercial fishers and their families. OBJECTIVES: Aims of this study were to describe blood dioxin levels of SH fishers and family members; compare these levels to background Australian levels and international data; analyze association between intake of SH seafood and dioxin levels; and assess whether blood dioxin levels were useful to inform ongoing care of SH fishers. METHODS: NSW Department of Health conducted clinics at which 112 fishers and family members gave blood for analysis of dioxin levels. Dioxin exposure was assessed through a questionnaire. Seafood dioxin levels were provided by the NSW Department of Environment and Climate Change. RESULTS: For the fishers (n=26), median TCDD and total TEQ levels were 27.3 and 62.4 pg g(-1) lipid, respectively. For the whole group (n=112), median TCDD and total TEQ levels were 9.3 and 26.1 pg g(-1) lipid, respectively. Age was the strongest predictor of increased dioxin levels (p<0.01), and consumption of SH seafood was also strongly associated (p=0.01). CONCLUSIONS: Serum dioxin levels in the SH fisher cohort were higher than Australian background levels, but comparable with other international high fish-eating communities, and lower than occupationally and accidentally exposed cohorts. High TCDD levels in commonly eaten seafood by the SH fishers confirms this as the likely exposure source. PMID- 18674797 TI - Sorption of copper, zinc and lead on soil mineral phases. AB - Soil mineral phases play a significant role in controlling heavy metal mobility in soils. The effective study of their relation needs the integrated use of several analytical methods. In this study, analytical electron microscopy analyses were combined with sequential chemical extractions on soils spiked with Cu, Zn and Pb. Our aims were to study the metal sorption capacity of soil mineral phases and the effect of presence of iron oxide and carbonate on this property of soil minerals. Copper and Pb were found to be characterized by higher and stronger sorption on the studied samples than Zn. Only the former two metals showed significant differences in their immobilized metal amounts on the studied samples and soil mineral particles. Highest metal amounts were sorbed on the swelling clay mineral particles (smectites and vermiculites), but iron-oxide phases may also have similar lead sorption capacity. Alkaline conditions due to the carbonate content of soils resulted both in increased sorption on the mineral particles for Cu and in enhanced role of precipitation for all the studied metals. On the other hand, the intimate association of phyllosilicates and iron resulted in significant increase in metal sorption capacity of the given particle. The results of sequential extractions could be successfully completed by the analytical electron microscopy analyses for studying the sorption capacity of discrete mineral particles. Their integrated use helps us in better understanding the heavy metal-mineral interactions in soils. PMID- 18674798 TI - Polychlorinated biphenyl residues in sandstorm depositions in Beijing, China. AB - Sandstorms, which distribute many particles, are a special atmospheric occurrence and are frequent in northern China. We conducted this study to determine, for the first time, the concentration of polychlorinated biphenyls (PCBs) in sandstorm depositions. We collected 13 samples from urban areas of Beijing, and we measured a total of 144 PCB congeners. Thirteen samples all contained PCB residues. The total PCB concentration ranged from 1.6 to 15.6ngg(-1) (median, 4.8ngg(-1), dry weight), with trichlorinated biphenyls as the predominant homologue (>50.4%). Furthermore, we observed increasing PCB contamination from northwest to east Beijing. We later explored possible factors affecting contamination of the sandstorm depositions, which revealed a significant correlation between SigmaPCBs and the minimum particle size of the sandstorm deposition samples. Principal component analysis revealed that the major source of PCBs in Beijing may be potentially associated with the number-one commercial PCB through the long-range transmission. In previous results, PCBs were not a severe component of contamination in sandstorm depositions of Beijing. However, this study suggested that sandstorm deposition may be a potential source of exposure to PCBs for the residents of Beijing, China. PMID- 18674799 TI - Stable isotope tracing of trout hatchery carbon to sediments and foodwebs of limestone spring creeks. AB - Limestone springs support productive ecosystems and fisheries, yet aquaculture may modify or impair these ecosystems. We determined trout hatchery organic contribution to spring creek sediments and foodwebs with natural abundance stable isotope methods. Hatchery feed, waste, and trout were significantly enriched in delta(13)C relative to autotrophs and wild fish. Spring creek sediments were enriched in delta(13)C toward the hatchery endmember relative to reference streams without hatcheries and relative to a larger larger-order, spring influenced stream. Contribution of hatchery C to spring creek sediments was greatest during March and associated with greatest sediment %C. Contribution of hatchery C to pollution-tolerant isopod diet was 39-51% in a stream receiving limestone spring water via hatchery effluent. Isopods of one spring creek also relied on hatchery-derived C within one month of hatchery closure. Four years later, less pollution pollution-tolerant amphipods dominated and consumed non vascular over vascular autotrophs (86%). Isopods of a second spring creek with an active hatchery did not appear to be using hatchery matter directly, but were enriched in delta(34)S relative to a spring creek tributary with no hatchery influence. Isopods in both of these streams were relatively enriched in delta(15)N, indicating general nutrient enrichment from surrounding agricultural land use. The contribution of hatchery vs. wild fish in diet of herons and egrets was traced with delta(13)C of guano. These birds were strongly dependent on stocked trout in a spring creek with a recently closed state trout hatchery, and also near another large, state-run hatchery. Heron dependence on hatchery fish in the spring creek decreased with time since hatchery closure. Use of stable isotope natural abundance techniques in karst spring creeks can reveal stream impairment due to aquaculture, specific C sources to bio-indicating consumers, losses of farmed fish to predation, and potential exposure of higher order consumers to contaminants associated with aquaculture. PMID- 18674800 TI - The effect of growth hormone on ovarian follicular response and oocyte nuclear maturation in young and aged mice. AB - The aim of this study was to investigate the possible effects of recombinant human growth hormone (rhGH) on the ovarian follicle numbers and the nuclear maturity status of ovulated oocytes. Aged and young Balb-C mice were randomly divided into three groups: (i) control, (ii) stimulated with follicle stimulating hormone (FSH) and (iii) stimulated with FSH+rhGH. Ovaries were collected for histological and morphometric investigations. For the determination of nuclear maturation of ovulated oocytes, additional aged and young mice were randomly divided into two groups: (i) stimulated with FSH and (ii) stimulated with FSH+rhGH. In young mice, developing ovarian follicle numbers were significantly higher with rhGH; while in aged mice, there was no statistically significant difference between groups. In young mice co-stimulated with FSH and rhGH, atretic follicle numbers were significantly higher. In aged mice, atretic follicle numbers significantly decreased with FSH while there was a slight decrease with rhGH and this was not significant. The percentage of metaphase-II oocytes were significantly higher in both young and aged mice groups co-stimulated with FSH and rhGH. In conclusion, co-treatment with FSH and rhGH for ovarian stimulation in older mice appears to cause a slight decrease in the number of atretic follicles and thus improves the collection of high numbers of mature, fertilizable oocytes. PMID- 18674801 TI - Laparoscopic ureterolysis and omental wrapping. AB - OBJECTIVES: To describe our laparoscopic technique of ureterolysis and omental wrapping using the LigaSure device for the treatment of idiopathic retroperitoneal fibrosis. METHODS: Four bilateral laparoscopic ureterolyses (LUs) and two unilateral LUs were performed in 6 male patients (mean age 47 years). Of the 6 patients, 4 underwent LU without having undergone medical therapy before surgery and 2 underwent LU after medical therapy failure. All had had ureteral stents placed before surgery. The ureters were completely freed from the fibrotic tissue using an Overholt laparoscopic forceps and 10-mm LigaSure atlas. An omental wrap was passed behind the colonic flexure, placed around the ureter, and fixed to the psoas muscle. RESULTS: The mean operating time was 80 minutes (range 75 and 85) for the unilateral LUs and 200 minutes (range 180-225) for the bilateral procedures. The mean blood loss was 75 mL (range 50 and 100) during LUs and 150 mL (range 80-220) during bilateral LUs. The mean hospital stay was 3.33 days (range 2-5). All indwelling ureteral stents were removed at 4 weeks postoperatively. At a mean follow-up of 37.5 months (range 23-59), all patients were free of symptom and all renal units were unobstructed. CONCLUSIONS: In our experience of LUs and omental wrapping, the reproduction of each step of open surgery seems to offer excellent midterm outcomes. The use of the LigaSure simplified the laparoscopic procedure and made it feasible and safe. We believe that the minimally invasive nature and high effectiveness of LU suggest consideration of this procedure as first-line treatment of idiopathic retroperitoneal fibrosis. PMID- 18674802 TI - Outcomes and cost analysis of pyeloplasty for antenatally diagnosed ureteropelvic junction obstruction using Markov models. AB - OBJECTIVES: The optimal timing of pyeloplasty for children diagnosed with ureteropelvic junction obstruction (UPJO) after workup for antenatal hydronephrosis is disputed. We sought to examine the potential costs and clinical outcomes of treatment protocols featuring different indications for pediatric pyeloplasty using Markov models. METHODS: Cost and outcomes analysis using Markov modeling was performed for three treatment algorithms: medical management, immediate pyeloplasty (during the first year of life), and pyeloplasty after no improvement on imaging. The costs were determined from the perspective of the medical institution. The variables tracked during Markov model simulation included age at resolution of UPJO, the proportion of patients with worsened hydronephrosis, the number of pyeloplasties, the number of pyelonephritis episodes, and costs. Sensitivity analyses were performed to determine which elements affected the model and to determine threshold values. RESULTS: Immediate pyeloplasty and pyeloplasty after no improvement on imaging resulted in rapid resolution of UPJO (mean age at resolution younger than 2 years) with lower rates of worsened hydronephrosis and pyelonephritis compared with observation alone. For the surgical protocols, the costs per resolved case of UPJO were greater than those for medical management alone at the probability values tested in the Markov models. The sensitivity analysis of all variables over realistic ranges demonstrated that the costs of surgery, annual antibiotics and imaging, and the rate of pyelonephritis were critical in determining the costs. CONCLUSIONS: Pediatric urologists should include practice-specific features such as the costs of surgery, annual antibiotics and imaging, and pyelonephritis rates when considering efficacious, yet less costly, treatment protocols for UPJO. PMID- 18674803 TI - Stone attenuation and skin-to-stone distance on computed tomography predicts for stone fragmentation by shock wave lithotripsy. AB - OBJECTIVES: To determine whether stone attenuation and the skin-to-stone distance (SSD) can predict for stone fragmentation by SWL independently. Identifying the factors predictive of shock wave lithotripsy (SWL) outcome would help streamline the care of patients with stones. METHODS: A retrospective review was performed of 111 patients undergoing initial SWL for a solitary, 5-20 mm, renal calculus. Stone size, location, attenuation value, and SSD were determined on pretreatment noncontrast computed tomography. The outcome was categorized as stone free, complete fragmentation <5 mm, and incomplete fragmentation >or=5 mm or unchanged at 2 weeks on kidney/ureter/bladder radiography. RESULTS: After SWL, 44 (40%) were stone free, 27 (24%) had complete fragmentation, and 40 (36%) of 111 patients had incomplete fragmentation. The stone attenuation of the successfully treated patients (stone free and complete fragmentation groups) was 837 +/- 277 Hounsfield units (HU) vs 1092 +/- 254 HU for those with treatment failure (incomplete fragmentation; P < .01). The mean SSD also differed: 9.6 cm +/- 2.0 vs 11.1 cm +/- 2.5 for the successful treatment group vs the treatment failure group, respectively (P = .01). On multivariate analysis, the factors that independently predicted the outcome were stone attenuation, SSD, and stone composition. When patients were stratified into 4 risk groups (stone <900 HU and SSD <9.0 cm, stone <900 HU and SSD >or=9.0 cm, stone >or=900 HU and SSD <9.0 cm, and stone >or=900 HU and SSD >or=9.0 cm), the SWL success rate was 91%, 79%, 58%, and 41%, respectively (odds ratio 7.1, 95% confidence interval 1.6-32 for <900 HU and SSD <9.0 cm group vs other 3 risk groups; P = .01). CONCLUSIONS: The results of our study have shown that a stone attenuation of <900 HU, SSD of <9 cm, and stone composition predict for SWL success, independent of stone size, location, and body mass index. These factors will be considered important in the prospective design of a SWL treatment nomogram at our center. PMID- 18674804 TI - Application of the Yang-Monti channel in adult continent cutaneous urinary diversion. AB - OBJECTIVES: To report 8 cases using the Yang-Monti channel with ileum for adult continent cutaneous urinary diversion. METHODS: From 2004 to 2006, 108 patients underwent urinary diversion, 8 of whom had a continent cutaneous reservoir using the Yang-Monti principle. Channels were created by transversely retubularizing 2 to 3 cm of ileum. Indications, technique, complications, and outcomes, such as continence and ease of channel catheterization, are reported. RESULTS: Five patients underwent a right colon reservoir urinary reconstruction after extirpative surgery. Orthotopic neobladders were not performed, owing to anatomic or oncologic concerns. Two additional patients underwent an augmentation, with 1 undergoing an Indiana pouch revision. All urinary reservoirs but 1 were connected to the umbilicus. Two strictures occurred, which were treated with in-office dilation. Incontinence was identified in 2 patients and successfully treated with dextran microspheres injections. All patients were continent at final follow-up. Seven patients continue to perform intermittent catheterization, and 1 patient is managed with an indwelling catheter, owing to comorbidities. CONCLUSIONS: The Yang-Monti channel using ileum is an alternative as a neo-appendicoumbilicostomy in construction of continent cutaneous urinary diversion. The channel is surgically versatile and can be used in the creation of a right colon pouch when the appendix is unavailable. PMID- 18674805 TI - Successful management of an "overlooked" ureteral stent in a transplant kidney. AB - The authors present a case of successful management of an encrusted ureteral stent in a transplant kidney using cystolitholapaxy and percutaneous nephrolithotomy with electromechanical lithotripsy. PMID- 18674806 TI - Re: Busby JE, Brown GA, Matin SF. Comparing lymphadenectomy during radical nephroureterectomy: open versus laparoscopic (Urology 71:413-416, 2008). PMID- 18674807 TI - Predictive significance of surgical margin status after prostatectomy for prostate cancer during PSA era. AB - OBJECTIVES: The presence of positive surgical margins (PSMs) after prostatectomy for prostate cancer has long been an indicator of poor survival outcomes. However, with the downstaging of cancer occurring in the prostate-specific antigen testing era, we sought to determine whether the risk associated with PSMs retains the same effect on prognosis as before the prostate-specific antigen testing era. METHODS: Of the 3460 patients in the Columbia University Urologic Oncology database, 2215 (64%) were identified who had undergone radical prostatectomy from 1991 to 2005 and had sufficient pathologic data to be analyzed and >or=1 year of follow-up. Three epochs were chosen: 1991-1995, 1996-2000, and 2001-2005. RESULTS: The median age, preoperative prostate-specific antigen, and Gleason score was 61.6 years, 6 ng/mL, and 7, respectively, and >50% of patients had pathologic Stage T2 disease. On multivariate analysis, PSMs were a risk factor for biochemical failure for each epoch (P < .01). The Wald's test indicated that the significance of PSMs had not changed over time (P = .8). The contribution of PSMs to the accuracy of predicting biochemical failure in a multivariate model was found only for the earliest epoch, because it improved the model by 0.15 (95% confidence interval 0.03-0.27). In the second epoch, it was 0.13 (95% confidence interval -0.01 to 0.27), and it was 0.13 for the third (95% confidence interval -0.06 to 0.32). CONCLUSIONS: The results of this study suggest that the predictive contribution of PSMs to the accuracy of a multivariate model or nomogram used to predict the outcomes after prostatectomy has decreased during the past 15 years. PMID- 18674808 TI - Assessing the effects of lymphadenectomy and radiation therapy in patients with uterine carcinosarcoma: a SEER analysis. AB - OBJECTIVE: The purpose of this analysis is to determine the pathologic prognostic factors and treatment outcome of patients with carcinosarcoma of the uterus. METHODS: A retrospective analysis of data from the Surveillance, Epidemiology, and End Results program of the National Cancer Institute between January 1, 1988 and November 1, 2003 was conducted. A total of 1855 with AJCC Stages I-III disease were identified who received primary surgical treatment. Overall survival curves were constructed using Kaplan-Meier curves. Cox proportional hazards model was used to identify factors predictive of overall survival. RESULTS: AJCC stage of all patients was as follows: 65% Stage I (n=1099), 14% Stage II (n=245), 21% Stage III (n=353). 57% (n=965) patients underwent LND. The median number of lymph nodes removed was 12 (SD=10.2); 119 (14%) patients had positive lymph nodes. Five year overall survival (OS), disease free survival, and median survival were significantly improved for patients receiving lymph node dissection (LND) as compared to patients that received no LND, irrespective of radiotherapy. Adjuvant radiation therapy had no improvement on overall survival regardless of LND. There was no overall survival benefit to the addition of radiotherapy regardless of whether patients underwent a lymph node dissection or not. Age, race, marital status, lymph node dissection and stage were predictive of survival on multivariate analysis. CONCLUSIONS: Lymphadenectomy is significantly associated with improved overall survival in patients with Stage I-III uterine carcinosarcoma compared to no lymphadenectomy. The use of adjuvant radiotherapy conferred no overall survival benefit. PMID- 18674810 TI - New developments in low-dose insemination technology. AB - New nonsurgical procedures for inseminating swine with a low number of spermatozoa have been developed and/or evaluated over the last few years. These procedures allow the deposition of the insemination dose into the uterine body (post-cervical insemination) or directly into the uterine horn (deep intrauterine insemination). With the use of the post-cervical insemination, a threefold reduction in the number of fresh sperm has been successfully used to achieve pregnancy. Using deep intrauterine insemination (DUI), up to a 20-fold reduction in the number of fresh spermatozoa or a sixfold reduction in the number of frozen/thawed spermatozoa can be achieved, with reproductive performance very similar to that obtained after standard AI. Complementing these nonsurgical insemination techniques, a new procedure for depositing spermatozoa into the oviduct by laparoscopy has been recently described. This laparoscopic technique has proven to be applicable to diluted and sex-sorted spermatozoa. The development of new insemination procedures will help achieve more efficient application of currently available sperm technologies. Using appropriate insemination procedures, it is now feasible to achieve high fertility rates with cooled, frozen-thawed, or sex-sorted semen. PMID- 18674809 TI - AMPK and PPARdelta agonists are exercise mimetics. AB - The benefits of endurance exercise on general health make it desirable to identify orally active agents that would mimic or potentiate the effects of exercise to treat metabolic diseases. Although certain natural compounds, such as reseveratrol, have endurance-enhancing activities, their exact metabolic targets remain elusive. We therefore tested the effect of pathway-specific drugs on endurance capacities of mice in a treadmill running test. We found that PPARbeta/delta agonist and exercise training synergistically increase oxidative myofibers and running endurance in adult mice. Because training activates AMPK and PGC1alpha, we then tested whether the orally active AMPK agonist AICAR might be sufficient to overcome the exercise requirement. Unexpectedly, even in sedentary mice, 4 weeks of AICAR treatment alone induced metabolic genes and enhanced running endurance by 44%. These results demonstrate that AMPK-PPARdelta pathway can be targeted by orally active drugs to enhance training adaptation or even to increase endurance without exercise. PMID- 18674811 TI - Influence of halogen irradiance on short- and long-term wear resistance of resin based composite materials. AB - OBJECTIVE: The Oregon Health Science University (OHSU) four-chamber oral wear simulator was used to examine the impact of halogen irradiance on the short- and long-term wear behavior of four-methacrylate resin-based composites (RBCs). The hypothesis proposed was that exacerbated wear would occur following the long-term wear of RBCs irradiated under non-optimized irradiance conditions. METHODS: Disc shaped specimens (12.0+/-0.1mm diameter and 2.0+/-0.1mm height) of each RBC were prepared at irradiances of 650+/-14, 530+/-8, 420+/-14, 350+/-8, 270+/-8, 230+/ 14, 190+/-8, and 150+/-8 mW/cm(2), respectively. Short-term three-body wear simulation was carried out for 50,000 cycles and long-term was performed up to 500,000 cycles. The wear facets were analyzed using a non-contact optical profilometer. One-, two-, and three-way analyses of variance (ANOVA) were made at P=0.05, guided as necessary by Tukeys correction in multiple partial analyses of the mean total volumetric wear and mean maximum wear depth. RESULTS: For the short-term wear resistance, the two-way ANOVA identified no significant difference in the mean total volumetric wear (P=0.188) although the mean total wear depth was identified to be significant (P=0.024). The long-term wear resistance for the mean total volumetric wear identified significant differences for each RBC at varying irradiances with the number of cycles but not for the mean maximum wear depth. SIGNIFICANCE: The increased number of ploughing actions of the antagonist on the RBC (long-term) results in increased friction which would be expected to play a major role in the wear process. The significant increase in the mean total volumetric wear, but not the mean maximum wear depth, observed over time which is exacerbated at reduced halogen irradiances emphasizes the arguments of DeLong for the employment of volume for evaluating material loss due to wear following in vitro wear simulation of RBCs. PMID- 18674812 TI - Effects of technology or maternal factors on perinatal outcome after assisted fertilisation: a population-based cohort study. AB - BACKGROUND: Research suggests that singleton births following assisted fertilisation are associated with adverse outcomes; however, these results might be confounded by factors that affect both fertility and pregnancy outcome. We therefore compared pregnancy outcomes in women who had singleton pregnancies conceived both spontaneously and after assisted fertilisation. METHODS: In a population-based cohort study, we assessed differences in birthweight, gestational age, and odds ratios (OR) of small for gestational age babies, premature births, and perinatal deaths in singletons (gestation >/=22 weeks or birthweight >/=500 g) born to 2546 Norwegian women (>20 years) who had conceived at least one child spontaneously and another after assisted fertilisation among 1 200 922 births after spontaneous conception and 8229 after assisted fertilisation. FINDINGS: In the whole study population, assisted-fertilisation conceptions were associated with lower mean birthweight (difference 25 g, 95% CI 14 to 35), shorter duration of gestation (2.0 days, 1.6 to 2.3) and increased risks of small for gestational age (OR 1.26, 1.10 to 1.44), and perinatal death (1.31, 1.05 to 1.65) than were spontaneous conceptions. In the sibling relationship comparisons, the spontaneous versus the assisted-fertilisation conceptions showed a difference of only 9 g (-18 to 36) in birthweight and 0.6 days (-0.5 to 1.7) in gestational age. For assisted fertilisation versus spontaneous conception in the sibling-relationship comparisons, the OR for small for gestational age was 0.99 (0.62 to 1.57) and that for perinatal mortality was 0.36 (0.20 to 0.67). INTERPRETATION: Birthweight, gestational age, and risks of small for gestational age babies, and preterm delivery did not differ among infants of women who had conceived both spontaneously and after assisted fertilisation. The adverse outcomes of assisted fertilisation that we noted compared with those in the general population could therefore be attributable to the factors leading to infertility, rather than to factors related to the reproductive technology. PMID- 18674813 TI - Neonatal outcome in singletons conceived after ART. PMID- 18674814 TI - Bacterial adhesion and growth on a polymer brush-coating. AB - Biomaterials-related infections pose serious problems in implant surgery, despite the development of non-adhesive coatings. Non-adhesive coatings, like polymer brush-coatings, have so far only been investigated with respect to preventing initial bacterial adhesion, but never with respect to effects on kinetics of bacterial growth. Here, we compare adhesion and 20 h growth of three bacterial strains (Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa) on pristine and brush-coated silicone rubber in a parallel plate flow chamber. Brush-coatings were made using a tri-block copolymer of polyethylene oxide (PEO) and polypropylene oxide (PPO). Brush-coatings prevented adhesion of staphylococci to below 5 x 10(5)cm(-2) after 30 min, which is a 10-fold reduction compared to pristine silicone rubber. Biofilms grew on both brush-coated and pristine silicone rubber, while the viability of biofilms on brush-coatings was higher than on pristine silicone rubber. However, biofilms on brush-coatings developed more slowly and detached almost fully by high fluid shear. Brush coating remained non-adhesive after S. epidermidis biofilm formation and subsequent removal whereas a part of its functionality was lost after removal of S. aureus biofilms. Adhesion, growth and detachment of P. aeruginosa were not significantly different on brush-coatings as compared with pristine silicone rubber, although here too the viability of biofilms on brush-coatings was higher. We conclude that polymer brush-coatings strongly reduce initial adhesion of staphylococci and delay their biofilm growth. In addition, biofilms on brush coatings are more viable and easily removed by the application of fluid shear. PMID- 18674815 TI - Modeling the variability of shapes of a human placenta. AB - BACKGROUND: Placentas are generally round/oval in shape, but "irregular" shapes are common. In the Collaborative Perinatal Project data, irregular shapes were associated with lower birth weight for placental weight, suggesting variably shaped placentas have altered function. METHODS: (I) Using a 3D one-parameter model of placental vascular growth based on Diffusion Limited Aggregation (an accepted model for generating highly branched fractals), models were run with a branching density growth parameter either fixed or perturbed at either 5-7% or 50% of model growth. (II) In a data set with detailed measures of 1207 placental perimeters, radial standard deviations of placental shapes were calculated from the umbilical cord insertion, and from the centroid of the shape (a biologically arbitrary point). These two were compared to the difference between the observed scaling exponent and the Kleiber scaling exponent (0.75), considered optimal for vascular fractal transport systems. Spearman's rank correlation considered p<0.05 significant. RESULTS: (I) Unperturbed, random values of the growth parameter created round/oval fractal shapes. Perturbation at 5-7% of model growth created multilobate shapes, while perturbation at 50% of model growth created "star shaped" fractals. (II) The radial standard deviation of the perimeter from the umbilical cord (but not from the centroid) was associated with differences from the Kleiber exponent (p=0.006). CONCLUSIONS: A dynamical DLA model recapitulates multilobate and "star" placental shapes via changing fractal branching density. We suggest that (1) irregular placental outlines reflect deformation of the underlying placental fractal vascular network, (2) such irregularities in placental outline indicate sub-optimal branching structure of the vascular tree, and (3) this accounts for the lower birth weight observed in non-round/oval placentas in the Collaborative Perinatal Project. PMID- 18674816 TI - Expression of pig7 gene in acute leukemia and its potential to modulate the chemosensitivity of leukemic cells. AB - We analyzed expression of p53-induced gene 7 (pig7), at the transcript level, in bone marrow samples from patients with de novo acute leukemia (AL) and normal controls by quantitative reverse transcription PCR (RT-PCR), and revealed a markedly decreased pig7 expression in the patient group, as well as in the relapsed/refractory patients compared with those at initial diagnosis. By endonuclease analysis, we detected only one form of pig7 transcript, i.e., small integral membrane protein of late endosome (simple), in AL patients. In addition, up-regulated pig7 expression could be detected in differentiated leukemic cells induced by drugs. Transient expression of pig7 in leukemic cells exhibited no evident effect on cell proliferation and differentiation, but could intensify inhibitory efficacy of etoposide (VP16) and daunorubicin (DNR). Conclusively, the present study provides the evidence that pig7 is a silenced gene affected by perturbed differentiation in acute leukemia and restoration of pig7 expression sensitizes leukemic cells to chemotherapeutic agents. PMID- 18674817 TI - CD38 as a prognostic factor in Chinese patients with chronic lymphocytic leukaemia. AB - B-cell chronic lymphocytic leukemia (CLL) is the most common type of adult leukemia in the Western countries, however, infrequent in the Eastern. It shows a remarkable heterogeneity, with some patients having an almost normal lifespan, others surviving only several years after diagnosis despite intensive therapy. To explore the prognostic significance of CD38 expression in Chinese patients with CLL, multi-parameter flow cytometry was used to detect the expression of CD38 on CD5(+)CD19(+) cells of 147 patients. CD38 expression and its association with some other prognostic factors such as Binet stage, lymphocyte count in peripheral blood, serum lactate dehydrogenase (LDH), beta2-microglobulin (beta2-MG), ZAP-70 expression and cytogenetic abnormalities were analyzed. The Kaplan-Meier method was used to construct survival curves, and results were compared using the log rank test. Univariate and multivariate Cox regression analyses were used to assess associations between survival time and potential risk factors. Out of the 147 CLL patients, positive expression of CD38 was found in 45 (30.6%) cases. CD38 positivity identified a subgroup of CLL patients with aggressive disease of Binet stage at the time of the test (P=0.036). Furthermore, the presence of higher serum LDH and beta2-MG levels at diagnosis was strongly correlated with CD38 positive (P=0.016 and 0.025, respectively). Prognostically unfavorable cytogenetic abnormalities, including 17p13 and 11q22 deletions, were significantly more frequent in CD38-positive patients than in CD38-negative ones (P=0.047 and 0.001, respectively). There was no significant difference between CD38-positive and CD38-negative groups in molecular cytogenetic aberrations of del(6q23), del(13q14), 14q32 translocation, or trisomy 12. In addition, in CD38 positive patients, the percentage of leukemic cells expressing ZAP-70 protein was not significantly higher than in CD38-negative ones (P=0.120). CD38 expression was associated with poor outcome. Patients with positive expression of CD38 had significantly shorter overall survival (mean, 81 months) than patients without CD38 expression (mean, 179 months) (P=0.015). Univariate analysis showed that serum levels of LDH and beta2-MG, del(17p13) and CD38 expression were the significant factors in determining overall survival (OS). Del(17p13) and CD38 expression were the variables strongly associated with OS by multivariate Cox regression analysis. It was showed that the patients with high level of CD38 expression had poorer outcome; CD38 was a good predictor of OS in Chinese patients with CLL. PMID- 18674819 TI - Enhanced DNA photocleavage properties of Ru(II) terpyridine complexes upon incorporation of methylphenyl substituted terpyridine and/or the polyazine bridging ligand dpp (2,3-bis(2-pyridyl)pyrazine). AB - The heteroleptic complexes, [(MePhtpy)RuCl(dpp)](PF(6)) and [(tpy)RuCl(dpp)](PF(6)), have been synthesized, characterized, and investigated with respect to their photophysical, redox, and DNA photocleavage properties (where MePhtpy=4'-(4-methylphenyl)-2,2':6',2''-terpyridine and dpp=2,3-bis(2 pyridyl)pyrazine, tpy=2,2':6',2''-terpyridine). The X-ray crystal structure confirms the identity of the new [(MePhtpy)RuCl(dpp)](PF(6)) complex. These heteroleptic complexes were found to photocleave DNA in the presence of oxygen, unlike the previously studied complex, [Ru(tpy)(2)](PF(6))(2). The photophysical, redox, and DNA photocleavage properties of the heteroleptic complexes were compared with those of the homoleptic complexes, [Ru(MePhtpy)(2)](PF(6))(2) and [Ru(tpy)(2)](PF(6))(2). The heteroleptic complexes showed intense metal to ligand charge transfer (MLCT) transition at lower energy ([(MePhtpy)RuCl(dpp)](PF(6)), 522nm; [(tpy)RuCl(dpp)](PF(6)), 516nm) and longer excited state lifetimes as compared to the homoleptic complexes. The [Ru(MePhtpy)(2)](2+) complex was found to photocleave DNA in contrast to [Ru(tpy)(2)](2+). The introduction of a methylphenyl group on the tepyridine ligand not only enhances the (3)MLCT excited state lifetime but also increases the lipophilicity and thereby the DNA binding ability of the molecule. An increase in lipophilicity upon addition of a methylphenyl group on the 2,2':6',2''-terpyridine ligand was confirmed by determination of the partition coefficient ([(MePhtpy)RuCl(dpp)](PF(6)), logP=+1.16; [(tpy)RuCl(dpp)](PF(6)), logP=-1.27). The heteroleptic complexes photocleave DNA more efficiently than the homoleptic complexes, with the greatest activity being observed for the newly prepared [(MePhtpy)RuCl(dpp)](PF(6)) complex. PMID- 18674820 TI - Selective low-dose valganciclovir for prevention of cytomegalovirus disease following kidney transplantation. AB - OBJECTIVES: To assess the safety of a cytomegalovirus prophylaxis regimen using selective low-dose valganciclovir. METHODS: Retrospective before-after comparison, including consecutive kidney transplant recipients between 2002 and 2004. Excluded were deaths or graft loss within one month after transplantation. During the first period, universal prophylaxis with oral ganciclovir was used. During the second period, selective prophylaxis with low-dose valganciclovir (450 mg/day) was given to patients at high-risk for CMV disease (D+/R- recipients and patients treated with anti-lymphocyte antibodies) and oral acyclovir to all other patients. We compared the incidence of CMV disease between the two periods. RESULTS: Two hundred and sixty-four patients underwent kidney transplantation of which 221 were included. The incidence of CMV disease was higher with selective low-dose valganciclovir compared to universal ganciclovir without statistical significance, by intention to treat (4.7% vs. 8.5%, respectively, OR 1.88, 95% CI 0.63-5.60) and among patients adhering to the prophylaxis protocol (3.1% vs. 6.8% respectively, OR 2.85, 95% CI 0.65-12.38). Only late-onset CMV was observed for compliant patients. On multivariate analysis, only recipients' serostatus remained significantly associated with CMV disease (OR 0.22, 95% CI 0.07-0.70). CONCLUSIONS: Selective low-dose valganciclovir may provide similar protection against CMV when compared to universal oral ganciclovir. Prolongation of prophylaxis beyond 100 days should be explored. PMID- 18674818 TI - Biological activities of C1 inhibitor. AB - Broadly speaking, C1 inhibitor plays important roles in the regulation of vascular permeability and in the suppression of inflammation. Vascular permeability control is exerted largely through inhibition of two of the proteases involved in the generation of bradykinin, factor XIIa and plasma kallikrein (the plasma kallikrein-kinin system). Anti-inflammatory functions, however, are exerted via several activities including inhibition of complement system proteases (C1r, C1s, MASP2) and the plasma kallikrein-kinin system proteases, in addition to interactions with a number of different proteins, cells and infectious agents. These more recently described, as yet incompletely characterized, activities serve several potential functions, including concentration of C1 inhibitor at sites of inflammation, inhibition of alternative complement pathway activation, inhibition of the biologic activities of gram negative endotoxin, enhancement of bacterial phagocytosis and killing, and suppression of the influx of leukocytes into a site of inflammation. C1 inhibitor has been shown to be therapeutically useful in a variety of animal models of inflammatory diseases, including gram negative bacterial sepsis and endotoxin shock, suppression of hyperacute transplant rejection, and treatment of a variety of ischemia-reperfusion injuries (heart, intestine, skeletal muscle, liver, brain). In humans, early data appear particularly promising in myocardial reperfusion injury. The mechanism (or mechanisms) of the effect of C1 inhibitor in these conditions is (are) not completely clear, but involve inhibition of complement and contact system activation, in addition to variable contributions from other C1 inhibitor activities that do not involve protease inhibition. PMID- 18674821 TI - In vivo characterization of primitive hematopoietic cells in clonal ginbuna crucian carp (Carassius auratus langsdorfii). AB - Primitive hematopoietic cells in mammalian bone marrow are purified by flow cytometry using Hoechst 33342 (Hoechst) and rhodamine-123 (Rho), because these dyes efflux activities of hematopoietic cells widely conserved in mammals. Hematopoietic stem cells (HSCs) are identified as side population (SP) cells, characterized by specific Hoechst efflux pattern in flow cytometric analysis. We previously demonstrated that SP cells from teleost body kidney (BK) had the HSC activity by a transplantation experiment using clonal ginbuna crucian carp (Carassius auratus langsdorfii). In the present study, to isolate HSCs and hematopoietic progenitor cells (HPCs) from teleosts using Hoechst and Rho, we compared the hematopoietic activity of Rho-negative (Rho(-)) cells with that of SP cells by ginbuna transplantation experiments. Rho(-) cells were clearly identified from ginbuna BK, and the majority of these cells (85%) showed a non-SP phenotype. Transplantation experiments showed that long-term repopulating activity (HSC activity) of Rho(-) cells was lower than that of SP cells, while Rho(-) cells had higher short-term repopulating activity (HPC activity) than SP cells. These results suggest that Rho(-) cells in ginbuna BK contain various stages of hematopoietic cells, while SP cells are highly enriched for HSCs, and that these dyes are useful for purification of HSCs and HPCs in teleosts. PMID- 18674822 TI - The frequency of auditory neuropathy detected by universal newborn hearing screening program. AB - OBJECTIVE: Auditory neuropathy/auditory dyssynchrony (AN/AD) has become a well accepted clinical entity. The combined use of oto-acoustic emissions (OAEs) and auditory brainstem response (ABR) testing in the universal newborn hearing screening (UNHS) has led to the easy recognition of this disorder. Although, we are now able to diagnose AN/AD reliably, little is known about its epidemiology, etiology, and especially the frequency of its occurrence. The primary goal of this study was to determine the frequency of AN/AD in the Western Anatolian region of Turkey. The secondary goal was to compare the detection rate of AN/AD before and after the implementation of the UNHS in the audiology department of Dokuz Eylul University Hospital. METHOD: Between 2005 and 2007, among the 23,786 newborns who were screened by automated click evoked oto-acoustic emissions (a CEOAE) and automated auditory brainstem responses (a-ABRs), 2236 were referred to our department. All necessary audiological tests were performed for all the referred newborns. Among them, babies with deficient or abnormal ABR in combination with normal OAEs were considered as having AN/AD. These babies were evaluated with additional diagnostic audiological tests. Furthermore, comparison of the incidence of children diagnosed with AN/AD before and after the implementation of UNHS in our audiology department was also performed. RESULTS: Among the referred newborns, 65 had abnormal or deficient ABR test results. Ten of these 65 newborn babies (mean diagnostic age: 5.7 months) with hearing impairment showed electrophysiological test results that were consistent with AN/AD. The frequency of AN/AD in these 65 children with hearing loss was 15.38%. Moreover, the frequency of AN/AD within UNHS was found to be 0.044%. Seven of the 10 babies with AN/AD had hyperbilirubinemia as a risk factor, which is a high rate to be emphasized. On the other hand, the retrospective investigation of children diagnosed with AN/AD in the same audiology department between 1999 and 2005 (i.e. before the implementation of UNHS) revealed only 7 children, with an average diagnostic age of 34 months. CONCLUSION: After implementing the UNHS, the incidence of AN/AD in the audiology department increased from 1.16 to 4.13. Furthermore, the age of diagnosis of AN/AD decreased from 34 months to 5.7 months. This study shows that AN/AD, when screened, is a comparatively common disorder in the population of hearing-impaired infants. While newborn hearing screening provides early detection of babies with hearing loss, it also helps to differentiate AN/AD cases when the screening is performed with both a-ABR and automated oto-acoustic emission (a-OAE) tests. Thus, the routine combined use of a-ABR and a-OAE tests in UNHS programs, especially for the high-risk infants, can provide better detection of newborns with AN/AD. Furthermore, hyperbilirubinemia is merely an association and maybe etiologically linked. PMID- 18674824 TI - Predictors of falls and hospitalization outcomes in elderly patients admitted to an acute geriatric unit. AB - We aimed to identify fall incidence, predictors and characteristics and to investigate hospitalization outcomes for elderly inpatients. In 340 men and 280 women consecutively admitted to a Acute Geriatric Ward of a University Hospital the following variables were evaluated: demographics, clinical history, main disease responsible for hospitalization, comorbidity (cumulative illness rating scale: CIRS 1 and 2) gait and balance deficit (Tinetti's scales), cognition/function (short portable mental status questionnaire: SPMSQ); activities of daily living: ADL; instrumental activities of daily living: IADL; delirium (confusion assessment method: CAM), drugs administered during hospitalization. Overall 80 falls occurred in 70 patients. The incidence rate of falls was of 6.0 per 1000 patient-days with 2.0 falls per bed/year. Age (relative risk=RR=1.050; 95% confidence interval=CI=1.013-1.087), delirium (RR=3.577; 95% CI 1.096-11.672), diabetes (RR=5.913; 95% CI 1.693-20.644), balance deficit (RR=0.914; 95% CI 0.861-0.970) and polypharmacy (RR=1.226; 95% CI 1.122-1.340) were independently predictive of falling. Fallers had a prolonged length of stay (LOS) (35.5+/-47.8 days vs. 23.2+/-27.2; p=0.01) and more frequent nursing home placements (12.9% vs.5.6%; p<0.005). The knowledge of falling predictors might help in planning specific preventive strategies to improve the patients' global health status and to reduce the costs of medical care. PMID- 18674825 TI - The evaluation of the presence of comorbidity by the Marigliano-Cacciafesta polypathology scale (MCPS) and the cumulative illness rating scale (CIRS) in elderly subjects with disability. AB - Multiple factors associated with the frailty syndrome may be involved in the appearance of disability, including the presence of comorbidity. The CIRS is commonly used for the evaluation of comorbidity, consisting of two parts: the comorbidity index (CI) and the severity index (SI). A multidimensional scale, the MCPS, has been recently developed, predicting the risk to develop disability. Fifty-nine subjects were examined by a structured multidimensional geriatric assessment. The MCPS and CIRS was significantly correlated (r=0.410; p<0.01 with the CI; and r=0.443, p<0.001 with the SI). The patients were divided in two groups, according to the MCPS score. The mean activities of daily living (ADL) and instrumental activities of daily living (IADL), as well as the corrected mini mental state examination (MMSE) score (+/-S.E.M.) were: 3.19+/-0.26; 0.28+/-0.04 and 24.00+/-1.14 in moderate-severe polypathology (n=21); 2.16+/-0.22; 0.13+/ 0.02 and 21.23+/-0.72 in severe polypathology (n=38) (p<0.001, p<0.01 and p<0.05), respectively. The MCPS score was correlated with the main indices of disability. In conclusion, we found that the MCPS is a useful tool in order to quantify and classify the presence of comorbidity, with results significantly related to that obtained with the CIRS. The MCPS offers an important stratification of the patients on the base of a well-established classification, not supplied by the CIRS. PMID- 18674826 TI - Emotional support from family members and subjective health in caregivers of the frail elderly at home in Japan. AB - This study sought to determine the factors associated with subjective health in caregivers of the frail elderly living at home, using multivariate regression analysis. Data were collected from 130 caregivers of frail elderly persons listed on a roster for utilization of day service or short stay service from two Home Visit Nursing Care Stations, using self-administered questionnaire from January to February of 2005. Family caregivers were defined as co-residents family members who provided a minimum of 1 h of daily care for at least 3 months. Multiple logistic regression analysis was performed to examine the factors associated with subjective health in caregivers of the frail elderly at home. Subjects with good health had less depressive symptom (p=0.001), much emotional support (p=0.003) and low caregiver burden (p=0.03). Multivariate logistic regression analysis showed good health had a significantly positive association with only much emotional support after adjustment for confounders. Of the total effect of emotional support on subjective health, the direct effect (84.7%) was much greater than the indirect effect (15.3%). Results indicate that much emotional support from family member for caregiver is essential for better subjective health in caregivers. PMID- 18674827 TI - Clinical importance of short-long-short sequences: analysing the mode of onset of ventricular tachycardias and atrial fibrillation. AB - Clinical importance of atrial and ventricular ectopic beats are investigated in some experimental and clinical trials. They are common and occur in a broad spectrum of the population. This includes patients without structural heart disease and those with any form of cardiac disease, independent of severity. Although we know something about the mechanisms and etiology of serious ventricular arrhythmias and atrial fibrillation, we don't know a lot about mode of onset of those arrhythmias. Can short-long-short sequences, related ectopic beats predict AF and ventricular tachycardias in some selected groups of patients? In this paper we will try to answer this important question. PMID- 18674828 TI - Inflammation in lone atrial fibrillation: new insights by coronary sinus thermography. AB - BACKGROUND: In the clinical setting there are conflicting results regarding the role of inflammatory activation in atrial fibrillation (AF). Coronary sinus (CS) thermography assesses myocardial heat production and is correlated with inflammatory states. We investigated in patients with AF whether 1) there is increased CS blood temperature and 2) the correlation of heat production with systemic inflammation. METHODS: We included patients with AF and subjects with sinus rhythm. C-reactive protein (CRP) levels were measured in all patients. CS and right atrium (RA) blood temperature measurements were performed by a dedicated 7F thermography catheter. DeltaT was calculated by subtracting RA from CS blood temperature. RESULTS: We included 47 patients with AF and 23 subjects with sinus rhythm. We stratified patients with AF into two groups: normotensive (AFN) and hypertensive (AFH). DeltaT was lower in the RA compared with the CS in AFH (37.27+/-0.52 degrees C vs 37.47+/-0.54 degrees C, p<0.01), in AFN (37.13+/ 0.53 degrees C vs 37.34+/-0.54 degrees C, p<0.01), and in controls (37.41+/-0.69 degrees C vs 37.55+/-0.68 degrees C, p<0.01). DeltaTau was greater in AFH, and AFN compared to controls (0.20+/-0.07 degrees C, 0.20+/-0.08 degrees C, vs 0.14+/ 0.06 degrees C, p<0.01). DeltaT was similar between AFH and AFN (p=0.95). CRP was higher in AFH and AFN compared to controls (1.72+/-0.85 mg/Dl, 1.69+/-0.94 mg/dL, 0.98+/-0.71 mg/dL, p<0.01). CRP was similar between AFH and AFN (p=0.87). A correlation between CRP with DeltaT was observed in AFH and AFN (R=0.58, p<0.01, R=0.44, p=0.02). CONCLUSIONS: Patients with AF have increased myocardial heat production, which is correlated to the systemic inflammation. CS blood temperature measurement may provide significant information for the pathogenesis of AF. PMID- 18674829 TI - Antinuclear antibodies in recurrent idiopathic pericarditis: prevalence and clinical significance. AB - BACKGROUND: A positive result for antinuclear antibodies (ANA), often as a fortuitous observation, may be cause for concern in idiopathic recurrent pericarditis (IRP), nevertheless data are lacking on their prevalence and clinical significance. This study is sought to investigate the prevalence and clinical significance of ANA in IRP. METHODS: ANA titres were assessed in consecutive patients with recurrent pericarditis, and matched healthy controls. Baseline and follow-up data were recorded and compared according to ANA results. RESULTS: A total of 145 consecutive patients with recurrent pericarditis were studied: 122 patients with IRP, 23 patients with pericarditis due to known etiologies (rheumatologic diagnoses and postpericardiotomy syndrome), and 122 healthy controls. ANA were detected in 53 of 122 (43.4%) patients with IRP, and in only 12 of 122 (9.8%) controls (p<0.001). Low titres (1/40-1/80) were found in the majority of cases, while moderate positivity (1/160-1/320) was more common in patients with a known rheumatic disease (26.7% vs. 5.7%; p=0.020). High concentrations of ANA (> or =1/640) were not recorded. Women were at increased risk for ANA (OR 2.22 95%CI 1.07-4.60; p=0.033). During a mean follow-up of 32 months, complications and new diagnoses were similar in patients with or without ANA positivity. CONCLUSIONS: Low-positive titres are more common in patients with IRP than in controls, suggesting a possible autoimmune pathogenesis. Nevertheless, they are often a clinically non-specific finding. Routine serologic testing for ANA suggests a source for recurrent pericarditis in less than 10% of cases, and in these cases other evidence typically suggests the underlying disease. PMID- 18674830 TI - Cardiac resynchronization and cardioverter defibrillation therapy in a patient with isolated noncompaction of the ventricular myocardium. AB - Isolated noncompaction of the ventricular myocardium (INVM) is an unclassified cardiomyopathy and is thought to be due to arrest of myocardial morphogenesis. Left ventricular failure and ventricular arrhythmias may occur in approximately half of the patients and account for half of the death in this disorder. In this report, we describe a patient with INVM in whom cardiac resynchronization and cardioverter defibrillation therapy was effective for the improvement of left ventricular function and for the prevention of ventricular arrhythmias. PMID- 18674831 TI - D-ribose aids advanced ischemic heart failure patients. AB - Patients with advanced heart failure are exercise intolerant. Low cellular energy levels in the failing heart have been proposed. Energy enhancing substrates have revealed mixed results. Ribose, a pentose monosaccharide, has shown to replenish low myocardial energy levels, improving cardiac dysfunction following ischemia, and improving ventilation efficiency in patients with heart failure. As current pharmaceuticals do not address cellular energy levels, this study was designed to investigate the role of ribose on ventilation at anaerobic threshold in congestive heart failure patients. d-ribose (5 gms/dose, tid) was assessed in 16 NYHA class III-IV, heart failure patients with VO(2), tidal volume/VCO(2), heart rate/tidal volume evaluated at 8 weeks. All patients had a significant improvement in ventilatory parameters at anaerobic threshold, along with a 44% Weber class improvement. Ribose improved the ventilatory exercise status in advanced heart failure patients. PMID- 18674832 TI - Aortic insufficiency and cardiac herniation associated with blunt chest trauma. AB - Aortic insufficiency and cardiac herniation due to pericardial rupture after blunt chest trauma are rare complications and are usually associated with high mortality. We report on a patient with simultaneous aortic insufficiency and cardiac herniation. He was involved in a falling accident, but initially presented no symptoms associated with the heart. At a later stage, he developed dyspnea on effort and a final diagnosis was completed. He was successfully treated surgically. PMID- 18674833 TI - Comparison of incidence and angiography patterns in definite thrombosis between drug-eluting and bare-metal stents. AB - Stent thrombosis is a feared complication of percutaneous coronary intervention due to its catastrophic consequences. We analysed the incidence of angiographically-confirmed thrombosis in 5011 consecutive patients treated with drug-eluting stents or bare-metal stents. Although the incidence of thrombosis was similar between the two groups (0.8%), angiographically-documented late thrombosis was a relatively unusual complication after implantation of either stent, though it was more common with drug-eluting stents than bare-metal stents (0.25 vs. 0.06%, P=.03). PMID- 18674834 TI - Left ventricular lipomatous metaplasia following myocardial infarction. AB - We present 3 cases of left ventricular lipomatous metaplasia after myocardial infarction evaluated with cardiovascular magnetic resonance (CMR). Delayed enhancement CMR alone cannot differentiate lipomatous metaplasia from scar. T1 weighted images with and without fat suppression are needed to identify this condition. The aetiology, pathophysiology, and possible clinical significance of lipomatous metaplasia in infarcted myocardium are still unknown. The multi parametric capabilities of CMR make it the ideal modality to identify non invasively, and without exposure to radiation, individuals with lipomatous metaplasia. PMID- 18674835 TI - Aneurysm of the membranous septum detected during left ventriculography and demonstrated by cardiac magnetic resonance imaging. PMID- 18674836 TI - To wait or to pay for medical treatment? Restraining ex-post moral hazard in health insurance. AB - We explore the hierarchy of two instruments, waiting time and coinsurance for medical treatment, for optimally solving the tradeoff between the economic gains from risk sharing and the losses from moral hazard. We show that the optimal waiting time is zero, given that the coinsurance rate is optimally set. PMID- 18674837 TI - A comparison of the mechanical effect of arm swing and countermovement on the lower extremities in vertical jumping. AB - The purposes of this study were to quantify and compare how arm swing and countermovement affect lower extremity torque and work during vertical jumping and to gain insight into the mechanisms that enable the arm swing and countermovement to increase jump height. Five participants maximally performed two types of vertical squat jumps with (SJA) and without (SJ) an arm swing and two types of countermovement vertical jumps with (CJA) and without (CJ) an arm swing. The participants jumped from a force platform and all performances were videotaped with a high-speed video camera (200 Hz). Jump heights, joint torques and work were calculated by combining kinematic and kinetic data. It was found that of the four jumping conditions, the participants jumped highest when they used an arm swing with countermovement (i.e., CJA). The increase of the countermovement jump height with an arm swing is the result of the increase of the lower extremity work. In the hip joint, the increase in torque caused by the countermovement predominantly occurred at the beginning of the propulsion phase, while the increase in torque caused by the arm swing occurred in the rest of the propulsion phase. A key finding of our study is that arm swing and countermovement have independent effects on lower extremity work, and their effects are additive in CJA to produce greater jump height. PMID- 18674838 TI - [Perineal electrical stimulation and rehabilitation in urinary incontinence and other symptoms of non-neurologic origin]. AB - A literature survey of 106 articles shows that standard electrostimulation is an effective treatment of urinary incontinence and urinary disorders with bladder instability. Bladder inhibition is obtained by applying an alternating current at a frequency of between 5 and 25Hz and with a pulse width of between 0.2 and 0.5ms. In 19 articles (including three randomized, placebo-controlled studies), good results were achieved in 60 to 90% of cases, depending on the exact method (i.e. chronic or acute stimulation). Standard electrostimulation is also efficient in stress urinary incontinence. Urethral closure is obtained by applying a 50Hz alternating current with, again, a pulse width of between 0.2 and 0.5ms. In 21 articles (including two randomized, placebo-controlled studies), good results were achieved in 47.5 to 77% of cases. Treatments combining perineal rehabilitation (behavioural education, muscle improvement and biofeed-back) and electrostimulation are reported by 10 authors, with good results in 70 to 80% of cases after 10 to 12 sessions. According to 14 studies, neuromodulation is also an efficient treatment for complex urinary disorders, urgency, pollakiuria and dysuria. The recommended stimulation parameters are a frequency of 10 to 15Hz and a pulse width of 210ms. Good results were found in 34 to 94% of cases (with between 60 and 75% in an international, multicenter study). The overall results different from one study to another because of the need to harmonize stimulation parameters, choice of the study population and treatment follow-up with self training programs and therapeutic education. PMID- 18674839 TI - Factors responsible for elevation of 1-h postchallenge plasma glucose levels in Japanese men. AB - The 1-h postchallenge plasma glucose (1-h PG) level is considered to be a good index of the development of glucose intolerance and type 2 diabetes as well as of diabetic complications. In some cases, in Japanese, 1-h PG is elevated despite normal fasting glucose during oral glucose tolerance test (OGTT), but the factors responsible remain unclear. In the present study, subjects with normal glucose tolerance (NGT), isolated impaired fasting glucose (IFG), and isolated impaired glucose tolerance (IGT) were divided into subgroups at 1-h PG of 10.0mM, and the four indices of insulin secretion and insulin sensitivity were compared. In all three categories, the insulinogenic index in subjects with elevated 1-h PG was remarkably lower than in those without elevated 1-h PG. In addition, the insulinogenic index was the strongest factor in elevated 1-h PG according to the multiple regression analysis. Interestingly, one third of the NGT subjects enrolled in this study had elevated 1-h PG. These subjects showed significantly elevated area under the curve of glucose (G-AUC) compared to NGT subjects without 1-h PG elevation. Thus, elevated 1-h PG in Japanese subjects indicates mildly impaired glucose tolerance due to decreased early-phase insulin secretion. PMID- 18674840 TI - The role of serial measurement of serum HBV DNA levels in patients with chronic HBeAg(-) hepatitis B infection: association with liver disease progression. A prospective cohort study. AB - BACKGROUND/AIMS: To evaluate the fluctuating course of serum HBV-DNA levels during the natural history of chronic HBV infection in the general population of North-Eastern Greece, in association with liver disease progression. METHODS: Two hundred and sixty-three adults with chronic HBV, median 34 years of age, were randomly selected and prospectively followed-up for a maximum period of 12 years. Viral markers, liver biochemistry and physical examination were performed every 6 months, and liver biopsy/abdominal ultrasound every 2-4 years. RESULTS: At entry, 195/263 (76%) were HBeAg (-)/anti-HBe (+) inactive carriers: (a) almost all 195 individuals with undetectable or HBV-DNA levels <2000IU/ml had no liver disease at entry and at follow-up period by imaging or liver histology evaluation (b) only 4/195 (2%) showed HBV reactivation with HBV-DNA >2000IU/ml. At entry, 48/263 (18%) patients were chronic HBeAg(-); (a) 1/3 patients had intermittently HBV-DNA <2000IU/ml for at least one occasion and were misclassified as inactive carriers (b) 22/48 (46%) had moderate/severe histology at entry and 5/48 (10%) showed liver disease progression during follow-up. Logistic regression analysis was used to derive OR (95%CI) for factors associated with liver disease progression. CONCLUSIONS: Close monitoring of serum HBV-DNA levels is useful in the management of chronic HBeAg(-) patients, as associated with liver disease progression. PMID- 18674842 TI - Doppler ultrasonography for the diagnosis of liver vascular malformations in hereditary hemorrhagic telangiectasia. PMID- 18674841 TI - Probiotics improve high fat diet-induced hepatic steatosis and insulin resistance by increasing hepatic NKT cells. AB - BACKGROUND/AIMS: Dietary factors and intestinal bacteria play an important role in the rapidly increasing incidence of obesity and its associated conditions, such as steatosis and insulin resistance. In the current study, we evaluated the effect of probiotics, and their mechanisms on diet-induced obesity, steatosis and insulin resistance. METHODS: Wild-type male C57BL6 mice were fed either normal or high fat diets. Some mice received VSL#3 probiotics. Animal weight, hepatic steatosis, insulin resistance, and their relationship to hepatic Natural Killer T cells (NKT) cell number and inflammatory signaling were evaluated. RESULTS: High fat diet induced a depletion of hepatic NKT cells thus leading to insulin resistance and steatosis. Oral probiotic treatment significantly improved the high fat diet-induced hepatic NKT cell depletion, insulin resistance and hepatic steatosis. This effect was NKT cell dependant, resulted from the attenuation of the tumor necrosis factor-alpha and IkappaB kinase inflammatory signaling, and led to an improved sensitivity in insulin signaling. CONCLUSIONS: Probiotics improve high fat diet-induced steatosis and insulin resistance. These effects of probiotics are likely due to increased hepatic NKT cell numbers and reduced inflammatory signaling. PMID- 18674843 TI - Micromanagers of malignancy: role of microRNAs in regulating metastasis. AB - It has become evident that cancer pathogenesis involves, among other macromolecules, a class of small regulatory RNAs named microRNAs, and that microRNA expression profiles are associated with prognosis and therapeutic outcome in several human cancers. Although the oncogenic or tumor-suppressing functions of several microRNAs have been characterized, the mechanistic roles played by microRNAs specifically in mediating metastasis have been addressed only recently. In this review, we focus on our emerging understanding of the contributions of microRNAs to malignant progression, specifically their functions in mediating tumor invasion and metastasis. These findings illuminate the molecular basis of metastasis and begin to connect small-RNA discoveries to the development of novel clinical biomarkers and therapeutic targets in neoplastic diseases. PMID- 18674844 TI - Use of tandem circulation wells to measure hydraulic conductivity without groundwater extraction. AB - Conventional methods to measure the hydraulic conductivity of an aquifer on a relatively large scale (10-100 m) require extraction of significant quantities of groundwater. This can be expensive, and otherwise problematic, when investigating a contaminated aquifer. In this study, innovative approaches that make use of tandem circulation wells to measure hydraulic conductivity are proposed. These approaches measure conductivity on a relatively large scale, but do not require extraction of groundwater. Two basic approaches for using circulation wells to measure hydraulic conductivity are presented; one approach is based upon the dipole-flow test method, while the other approach relies on a tracer test to measure the flow of water between two recirculating wells. The approaches are tested in a relatively homogeneous and isotropic artificial aquifer, where the conductivities measured by both approaches are compared to each other and to the previously measured hydraulic conductivity of the aquifer. It was shown that both approaches have the potential to accurately measure horizontal and vertical hydraulic conductivity for a relatively large subsurface volume without the need to pump groundwater to the surface. Future work is recommended to evaluate the ability of these tandem circulation wells to accurately measure hydraulic conductivity when anisotropy and heterogeneity are greater than in the artificial aquifer used for these studies. PMID- 18674845 TI - Prevalence and factors associated with hidradenitis suppurativa: results from two case-control studies. AB - BACKGROUND: Conflicting opinions have been reported regarding the epidemiology of hidradenitis suppurativa. OBJECTIVE: We sought to evaluate the prevalence of hidradenitis suppurativa and to identify associated factors. METHODOLOGY: Prevalence was evaluated using a representative sample of the French population (n=10,000). Associated risk factors were assessed using two case-control studies, one population-based with 67 self-reported patients and 200 control subjects, and the other clinic-based with 302 medically assessed patients and 906 control subjects. RESULTS: The prevalence was 1% of the French population. Multivariate analyses showed a strong association with current smoking in self-reported (odds ratio=4.16, 95% confidence interval [2.99-8.69]) and in medically assessed (odds ratio=12.55 [8.58-18.38]) populations. Association with body mass index was significant in medically assessed patients (odds ratio=1.12 [1.08-1.15]) for each increase of 1 U of BMI. LIMITATIONS: A causal relationship could not be established with such a cross-sectional study. CONCLUSION: Hidradenitis suppurativa is a common disease, frequently associated with smoking and being overweight. PMID- 18674846 TI - Aging affects the direction selectivity of MT cells in rhesus monkeys. AB - The ability to accurately perceive the direction and speed of moving objects declines during normal aging. This is likely due to functional degradation of cortical neurons. Most neurons in the primate middle temporal area (MT) are direction-selective and their activity is closely linked to the perception of coherent motion. We investigated the mechanisms that underlie this age-related decline by comparing the proportions of direction-selective MT cells in old and young macaque monkeys, using in vivo single-cell recording techniques. Our results showed that the proportion of such cells was lower in old than in young monkeys. Moreover, one type of direction-sensitive cells, pattern cells, was especially sensitive to aging and was affected more severely than another class, component cells. We also found that direction selectivity was affected more severely in MT than in V1 of senescent monkeys. Thus, the functional degradation of MT and V1 cells may mediate perceptual decline in visual motion tasks in old primates. PMID- 18674848 TI - Organically bound tritium (OBT) for various plants in the vicinity of a continuous atmospheric tritium release. AB - In order to quantify tritium impact on the environmental, we studied vegetation continuously exposed to a tritiated atmosphere. We chose lichens as bio indicators, trees for determination of past tritium releases of the Valduc Centre, and lettuce as edible vegetables for dose calculation regarding neighbourhood. The Pasquill and Doury models from the literature were tested to estimate tritium concentration in the air around vegetable for distance from the release point less than 500 m. The results in tree rings show that organically bound tritium (OBT) concentration was strongly correlated with tritium releases. Using the GASCON model, the modelled variation of OBT concentration with distance was correlated with the measurements. Although lichens are recognized as bio indicators, our experiments show that they were not convenient for environmental surveys because their age is not definitive. Thus, tritium integration time cannot be precisely determined. Furthermore, their biological metabolism is not well known and tritium concentration appears to be largely dependent on species. An average conversion rate of HTO to OBT was determined for lettuce of about 0.20 0.24% h(-1). Nevertheless, even if it is equivalent to values already published in the literature for other vegetation, we have shown that this conversion rate, established by weekly samples, varies by a factor of 10 during the different stages of lettuce development, and that its variation is linked to the biomass derivative. PMID- 18674849 TI - Patterns of emergency maxillofacial referrals and provision of services. AB - The rising number of maxillofacial injuries in the UK requires a reappraisal of emergency services within our specialty. Although the impact of a dedicated trauma list has been reported, it has not been widely embraced, and we know of few data that help to plan the number or timing of such lists. We designed a minimum dataset to collect information about referral, cause of emergency, assessment of patients, and outcome during 2003 and 2004. Theatre data were examined retrospectively to analyse operations during the same period. We conclude that to serve a population of 2.6 million a minimum of 8 emergency lists are required each week to deal with non-complex cases; this equates to 3 lists/million population. The lists should be concentrated at weekends and early in the week. PMID- 18674847 TI - Age-related alterations in default mode network: impact on working memory performance. AB - The default mode network (DMN) is a set of functionally connected brain regions which shows deactivation (task-induced deactivation, TID) during a cognitive task. Evidence shows an age-related decline in task-load-related modulation of the activity within the DMN during cognitive tasks. However, the effect of age on the functional coupling within the DMN and their relation to cognitive performance has hitherto been unexplored. Using functional magnetic resonance imaging, we investigated functional connectivity within the DMN in older and younger subjects during a working memory task with increasing task load. Older adults showed decreased connectivity and ability to suppress low frequency oscillations of the DMN. Additionally, the strength of the functional coupling of posterior cingulate (pCC) with medial prefrontal cortex (PFC) correlated positively with performance and was lower in older adults. pCC was also negatively coupled with task-related regions, namely the dorsolateral PFC and cingulate regions. Our results show that in addition to changes in canonical task related brain regions, normal aging is also associated with alterations in the activity and connectivity of brain regions within the DMN. These changes may be a reflection of a deficit in cognitive control associated with advancing age that results in deficient resource allocation to the task at hand. PMID- 18674850 TI - Effects of thermal desorption on the composition of two coking plant soils: impact on solvent extractable organic compounds and metal bioavailability. AB - To evaluate the efficiency and the influence of thermal desorption on the soil organic compartment, contaminated soils from coking plant sites (NM and H) were compared to their counterparts treated with thermodesorption. The extractable organic matter, and the metal content and distribution with soil compartments were studied. In both thermodesorbed soils, PAH (polycyclic aromatic hydrocarbon) degradation exceeded 90%. However, the thermal desorption led not only to a volatilization of the organic compounds but also to the condensation of extractable organic matter. The treatments only affected the Fe and Zn distribution within the more stable fractions, whereas the organic compound degradation did not affect their mobility and availability. PMID- 18674851 TI - Seasonal and spatial occurrence and distribution of atmospheric polycyclic aromatic hydrocarbons (PAHs) in rural and urban areas of the North Chinese Plain. AB - Passive air sampling (PAS) was employed to study the occurrence of gaseous and particle-bound PAHs in the North Chinese Plain. The averaged concentrations of gaseous and particle-bound PAHs were 485+/-209 ng/m(3) and 267+/-161 ng/m(3), respectively. The PAHs concentrations at urban sites were generally higher than those at rural ones with ratios <1.5 in spring, summer and fall, but differences between them were not significant for the wintertime and annually averaged concentrations. This urban-rural distribution pattern was related to the PAHs emission sources. PAHs spatial variation can be partially (49%) explained by emission with a simple linear regression method. Both the gaseous and particle bound PAHs were highest in winter and lowest in summer, with winter/summer ratios of 1.8 and 8, respectively. Emission strength was the most important factor for the seasonality. PMID- 18674852 TI - Hg concentrations and related risk assessment in coral reef crustaceans, molluscs and fish from New Caledonia. AB - There is a dramatic lack of data on Hg levels in marine organisms from tropical areas, and in particular from New Caledonia. For the first time, this study reports the total Hg concentrations in the tissues of several marine taxa from the New Caledonian lagoon. Seafood from both wild and farmed populations was considered. Hg concentrations varied over three orders of magnitudes according to factors including species, age (size/weight), trophic level, lifestyle and geographical origin. Taking into account the edible tissues, estimations of the amount of flesh that should be consumed by a 60-kg person to reach the Hg Provisional Tolerable Weekly Intake (PTWI) reveal acceptable risk for Human health in general. However, a risk was clearly identified in one site of the lagoon (i.e. Grande Rade) where high Hg concentrations were measured. These concentrations were higher than values reported in the current literature. PMID- 18674854 TI - Where there's a will, is there a way?: is New Zealand's publicly funded health sector able to steer towards population health? AB - Since 2000, the substantive focus of health policy in New Zealand has been closely aligned to the agendas of improving population health and reducing health inequalities. Health system restructuring, through the introduction of locally based and partially elected District Health Boards (DHBs), was the structural mechanism chosen for reorienting the health sector towards population health. Strategic planning at the DHB level was the key mechanism by which central government population health objectives would be translated into local action. This analysis of the early years of elected DHBs (2001-2005) sets out to answer the following broad questions: (i) did strategic planning by District Health Boards reflect an orientation to population health?; (ii) to what extent was strategic planning towards population health shaped by community participation and input?; (iii) to what extent did strategic planning lead to a re prioritisation of resources? These questions were explored as part of a larger research project investigating the introduction and implementation of the DHB system. Data were collected from over 350 interviews of local and national stakeholders, and two surveys of DHB Members between 2002 and 2004-2005. Overall, DHBs demonstrated the 'will' to engage in strategic decision-making processes to enhance population health but have difficulty in finding the 'way'. The priorities and requirements of central government and the weight of institutional history were found to be the most influential factors on DHB decision-making and practice, with flexibility and innovation only exercised at the margins. This raises the key question of whether there is the governmental capacity at the local level to adequately address nationally determined population health policy priorities. PMID- 18674853 TI - Regional trends in soil acidification and exchangeable metal concentrations in relation to acid deposition rates. AB - The deposition of high levels of reactive nitrogen (N) and sulphur (S), or the legacy of that deposition, remain among the world's most important environmental problems. Although regional impacts of acid deposition in aquatic ecosystems have been well documented, quantitative evidence of wide-scale impacts on terrestrial ecosystems is not common. In this study we analysed surface and subsoil chemistry of 68 acid grassland sites across the UK along a gradient of acid deposition, and statistically related the concentrations of exchangeable soil metals (1 M KCl extraction) to a range of potential drivers. The deposition of N, S or acid deposition was the primary correlate for 8 of 13 exchangeable metals measured in the topsoil and 5 of 14 exchangeable metals in the subsoil. In particular, exchangeable aluminium and lead both show increased levels above a soil pH threshold of about 4.5, strongly related to the deposition flux of acid compounds. PMID- 18674855 TI - Reutilization of the Prolift mark system for the simultaneous correction of prolapse and incontinence in patients with pelvic organ prolapse and stress urinary incontinence. AB - OBJECTIVE: The reutilization of the Prolift mark system with the simultaneous creation of a midurethral sling from the same set was described. STUDY DESIGN: The technique was applied in 23 patients with pelvic organ prolapse (POP) and clinically evident or occult stress urinary incontinence (SUI). RESULTS: Correction of POP was achieved in 21 out of 23 (91.3%) patients, and complete continence after the surgery in 20 out of 23 (86.9%). There was a significant improvement in voiding symptoms without deterioration of voiding function. Morbidity of the surgery was not different from that with Prolift mark alone. CONCLUSION: Modification of the method with creation of a suburethral sling from the same set is more economically viable without compromising either pelvic organ support or continence. PMID- 18674856 TI - Photocatalytic degradation of rhodamine B by Bi(2)WO(6) with electron accepting agent under microwave irradiation: mechanism and pathway. AB - Bi(2)WO(6) was successfully synthesized by a facile hydrothermal method, and characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD) and BET. As a result, Bi(2)WO(6) crystals displayed mainly square-plate-like morphologies with a short edge and the average crystalline size was in the range of 50-150 nm. Then microwave-assisted photocatalytic degradation of rhodamine B (RhB) using Bi(2)WO(6) was investigated. The results illustrated that RhB (10 mg/L) was bleached effectively and the removal efficiency was about 94% in 60 min. Effect of electron accepting agent (air, H(2)O(2)) on the degradation efficiency of RhB was also examined. Degradation intermediates of RhB in the presence of H(2)O(2) were identified by LC/MS/MS and GC/MS. All five N-de ethylated intermediates were monitored by LC/MS/MS easily, and seven organic acids such as succinic acid, benzoic acid, adipic acid, 3-hydroxybenzoic acid, phthalic acid, etc., were also detected by GC/MS. The possible degradation mechanism of RhB in the presence of H(2)O(2) included four processes: N-de ethylation, chromophore cleavage, opening-ring and mineralization, which coexisted in microwave-assisted photocatalytic system. PMID- 18674857 TI - Effect of quartz sand replacement by agate rejects in triaxial porcelain. AB - The ceramics industry, given the high volume of materials processed, stands as one of the largest consumers of natural raw materials but has also the capacity and potential to make significant contributions to solving environmental problems associated with other industries rejects. This work investigates the effects of quartz sand replacement by agate rejects (scrap) in a traditional triaxial porcelain composition. The study was carried out using the design of experiments (DoE) method. Characterization results were used to calculate statistically significant and valid regression equations, relating dried and fired body properties with clay, feldspar and agate scrap contents in the unfired mixture. The regression models were then discussed against X-ray diffraction and scanning electron microscopy results and used simultaneously to delimit the combinations of those three raw materials most adequate to produce a porcelainized stoneware floor tile with specified properties. Thus, an alternative use of an otherwise waste material is proposed, which can be translated into economic benefits and an important and welcome relief on environmental and waste disposal concerns. PMID- 18674858 TI - The pH as a key parameter in the choice between coagulation and electrocoagulation for the treatment of wastewaters. AB - In this work, it is studied the influence of the pH of the waste in the coagulation with aluminum by conventional and electrochemical dosing. To do that, the speciation of aluminum as a function of the pH has been characterized, and this information has been used to interpret the results obtained in the treatments (by both coagulation methods) of a synthetic oil-in-water emulsion and an actual effluent of a door-manufacturing factory. Results show that a simple change in the pH of the wastes can result in a significant change in the efficiency of the coagulation process, and that if the same pH conditions are found at the end of the treatment, the efficiencies of the solution-dosing and of the electrochemical dosing technologies are very similar. PMID- 18674859 TI - Sulphates for skin preservation--a novel approach to reduce tannery effluent salinity hazards. AB - In tanneries microorganisms are able to find environment suitable for their growth. Raw hide of buffalo and other animals like goat that are economically important, are an ideal source of nutrients for bacterial and fungal growth. In the past, preservatives like sodium chloride provided effective protection to fresh hides however the ill effect of their excessive use was not evaluated. But recently concern over potential ecological hazards has become more deliberate and sodium chloride features lot of disadvantages in agriculture as most of the tannery effluent is flown in agricultural fields in India. After rigorous laboratory experimentation on moisture content, SEM of hide, pure sodium sulphate as well as sodium sulphate in addition with sodium chloride (i.e. 10% w/w and 20% w/w) proved as most preferable option for curing of buffalo hide which gives effective preservation. Pollution load studies put forward sodium sulphate as an effective curing agent for buffalo hide to apply at industrial scale also. PMID- 18674860 TI - Solvothermal-induced phase transition and visible photocatalytic activity of nitrogen-doped titania. AB - Nitrogen-doped titania nanoparticles consisting of pure anatase, pure rutile and bicrystallites (anatase+rutile and anatase+brookite) have been prepared in TiCl(3)-HMT (hexamethylene tetramine)-alcohol solution under solvothermal process. The effect of the solvent type and amount of HMT as pH adjuster on the phase composition of titania and its visible photocatalytic activity for degradation to MO (methyl orange) was investigated. It is found that anatase gradually transferred to rutile with increase of carbon chain using methanol, ethanol, 1-propanol and 1-butanol as solvent. The pure anatase formed at the pH value of 1-2, while bicrystalline titania (anatase+rutile and anatase+brookite) at that of 7-10 in the presence of methanol. The bicrystalline (anatase+brookite) titania have the best visible photocatalytic activity among all the samples. The (NO) and -(NH) dopants with an N (1s) binding energy of 400 eV may have positive effects on the visible light photocatalytic activity. PMID- 18674861 TI - Preconcentration and separation of trace amount of heavy metal ions on bis(2 hydroxy acetophenone)ethylendiimine loaded on activated carbon. AB - A sensitive and simple method for simultaneous preconcentration of trace heavy metal ions in some food samples has been reported. The method is based on the adsorption of Cr(3+), Fe(3+), Cu(2+), Ni(2+), Co(2+) and Zn(2+) on bis(2-hydroxy acetophenone)ethylendiimine (BHAPED) loaded on activated carbon (AC). The adsorbed metals on activated carbon were eluted using 2 mol L(-1) nitric acid in acetone. The influences of the analytical parameters including pH and sample volume were investigated. The effects of matrix ions on the recoveries of analyte ions were also investigated. The recoveries of analytes were generally higher than 94%. The method has been successfully applied for analysis of the metal contents in real samples including natural water samples. PMID- 18674862 TI - The influence of fear of movement and pain catastrophizing on daily pain and disability in individuals with acute whiplash injury: a daily diary study. AB - Previous research supports the fear-avoidance model in explaining the transition from acute to chronic non-specific musculoskeletal pain. However, there is still little knowledge on when this vicious circle of pain, disability, pain catastrophizing and fear of movement starts. We performed a daily diary study in 42 patients with acute whiplash injury. Pain, disability, pain catastrophizing and fear of movement were measured on a daily basis with paper diaries for 21 consecutive days. Most participants showed a decline in pain and disability from day 1 to day 21 and this was paralleled by a decline in the fear of movement and pain catastrophizing. Multilevel analyses showed that both between and within persons, high levels of pain catastrophizing and fear of movement are associated with more pain and disability. Moreover, the fear of movement was also predictive of pain and disability on the following day. We also examined the reverse association, that is, whether the changes in pain predict changes in the next day's fear of movement and pain catastrophizing. Although for the fear of movement the model reached significance, the amount of explained variance was negligible. In conclusion, this study provides evidence that already in the early stages of whiplash-related complaints, significant associations between fear of movement and pain intensity and disability occur, and that this association may be predictive of the persistence of pain. PMID- 18674863 TI - Consequences of childhood burn: findings from the largest community-based injury survey in Bangladesh. AB - In terms of morbidity and disability, burn is a major public health problem throughout the world, especially in low-income countries. It causes long-term disability and remains as a health, social and economic burden. A population based survey was conducted in Bangladesh between January and December 2003. Nationally representative data were collected from 171,366 rural and urban households comprising of a total 819,429 population, which included 351,651 children under 18 years of age. Mothers/head of households were interviewed with a structured instrument. The objective of this paper is to determine the consequences of childhood burn at social and economic levels in Bangladesh. In the survey, 1013 children were found with different degrees of burn in the preceding 1 year. Among them 20 children were permanently disabled. The rate of permanent disability was found to be 5.7 per 100,000. The average loss of school days was found to be about 21 days. More than two-thirds of the burn victims required assistance in their daily activities for different durations of time. More than 7% of the children required hospitalisation for their burns. The rate of hospitalisation was 21.9 per 100,000; the average duration of hospital stay was 13.4 days. The highest duration (40 days) of hospital stay was found among girls 10-14 years old. The highest expenditure for the treatment was also found in this age group. The average direct expenditure incurred by a family for treatment of severe burn was determined to be $462. In this study it was found that more than 61% of the families earn less than $50 a month. Burn is a devastating injury among all childhood injuries with significant additional economic consequences beyond the medical, pain, and suffering issues. Developing a national prevention program should be an immediate public health priority. PMID- 18674864 TI - Corneal calcification after chemical eye burns caused by eye drops containing phosphate buffer. PMID- 18674865 TI - Chronic stress during paradoxical sleep deprivation increases paradoxical sleep rebound: association with prolactin plasma levels and brain serotonin content. AB - Previous studies suggest that stress associated to sleep deprivation methods can affect the expression of sleep rebound. In order to examine this association and possible mechanisms, rats were exposed to footshock stress during or immediately after a 96-h period of paradoxical sleep deprivation (PSD) and their sleep and heart rate were recorded. Control rats (maintained in individual home cages) and paradoxical sleep-deprived (PS-deprived) rats were distributed in three conditions (1) no footshock--NF; (2) single footshock--SFS: one single footshock session at the end of the PSD period (6-8 shocks per minute; 100 ms; 2 mA; for 40 min); and (3) multiple footshock--MFS: footshock sessions with the same characteristics as described above, twice a day throughout PSD (at 7:00 h and 19:00 h) and one extra session before the recovery period. After PSD, animals were allowed to sleep freely for 72 h. Additional groups were sacrificed at the end of the sleep deprivation period for blood sampling (ACTH, corticosterone, prolactin and catecholamine levels) and brain harvesting (monoamines and metabolites). Neither SFS nor MFS produced significant alterations in the sleep patterns of control rats. All PS-deprived groups exhibited increased heart rate which could be explained by increased dopaminergic activity in the medulla. As expected, PS deprivation induced rebound of paradoxical sleep in the first day of recovery; however, PSD+MFS group showed the highest rebound (327.3% above the baseline). This group also showed intermediate levels of corticosterone and the highest levels of prolactin, which were positively correlated with the length of PS episodes. Moreover, paradoxical sleep deprivation resulted in elevation of the serotonergic turnover in the hypothalamus, which partly explained the hormonal results, and in the hippocampus, which appears to be related to adaptive responses to stress. The data are discussed in the realm of a prospective importance of paradoxical sleep for processing of traumatic events. PMID- 18674866 TI - Relationship between alexithymia, alexithymia factors and salivary cortisol in men exposed to a social stress test. AB - BACKGROUND: The fact that alexithymia is associated with several medical and psychiatric disorders suggests that it may be a vulnerability factor for various diseases, possibly by enhancing stress responses. To test this "alexithymia stress hypothesis", we measured the influence of alexithymia and alexithymia subfactors on the cortisol response to an acute stressor. METHODS: Twenty-eight male students were exposed to the Trier Social Stress Test (TSST), during which saliva samples for cortisol determination were collected. RESULTS: Subjects reacted to the stressor with a significant cortisol response. Subjects scoring high on alexithymia evidenced an increased basal anticipatory cortisol level but their peak cortisol and area under the curve were similar to that of low scorers. Multiple regression analyses revealed that the increased cortisol in high scorers was due to only one subfactor of alexithymia, "the difficulty in describing feelings" factor (DDF). DDF high scorers reacted with a large increase in cortisol during anticipation but not during exposure to the stress test. CONCLUSION: The observation that alexithymia scores were associated with differences in cortisol levels before social stress exposure raises the possibility that alexithymia modulates cortisol levels, possibly by affecting the anticipatory cognitive appraisal of situations. This may be essentially attributed to the DDF factor. This observation sheds new light on the "alexithymia-stress hypothesis", which may be of importance to better understand the relationship between alexithymia and diseases. Further studies to address this issue should focus on the factorial structure of the construct and on the importance of anticipation. PMID- 18674867 TI - A new track for understanding the pathogenesis of multiple sclerosis: from the perspective of early developmental deficit caused by the potential 5-HT deficiency in individuals in high-latitude areas. AB - The association between the prevalence of multiple sclerosis (MS) and latitude gradient indicates the importance of environmental factors in MS susceptibility. Studies on immigrants have shown that the living environment in the first two decades of life determines MS risk, suggesting that early development may be critical for the occurrence of MS in adulthood. The level of 5-hydroxytryptamine (5-HT) in individuals living in high-latitude areas might be decreased because of limited levels of its metabolic precursor, the essential amino acid tryptophan, and superabundant levels of its metabolic product, melatonin, attributable to long duration of darkness in high-latitude areas. Considering the significant loss and damage of myelin observed in many psychiatric disorders with the etiology of 5-HT deficiency, we hypothesize that 5-HT deficiency due to superabundant synthesis of melatonin in individuals living in high-latitude areas may potentially cause the developmental myelin deficit early in life. This developmental deficit may play an important role in triggering MS in adulthood. This is the first proposal of the potential role of early development in the susceptibility to MS, and we suggest monitoring 5-HT levels in both patients with MS and in individuals with high environmental risk, especially children living in high-latitude areas. This will validate our hypothesis and contribute to designing specific preventive strategies that can be applied early in life. PMID- 18674868 TI - Single umbilical artery in fetopathological investigations. AB - Single umbilical artery (SUA) is a relatively common malformation that may call attention to the possibility of associated malformations (often chromosome aberrations). The current study aimed at surveying malformations associated with SUA on the basis of fetopathological investigations, analyzing the role of history, summarizing the clinically important factors emerging together with this malformation. In this study, we processed the details of 204 cases in which SUA was confirmed fetopathologically after miscarriage or induced abortion between 1990 and 2007. In our sample, SUA occurred in 7.38% of the cases. The history was positive in almost 30% of the cases. The majority of the cases had a positive obstetric and the minority of them a positive genetic history. The highest association of SUA with other malformations was found for craniospinal ones, but an association with cardiovascular malformations should also be mentioned. Regarding the individual types of malformation, SUA was most commonly associated with hydrocephalus, but Potter's sequence, trisomy 21, and atrioventricular septal defect also reached a higher rate in associated SUA. Previously published articles dealing with associated malformations found that urogenital malformations were most commonly associated with SUA. 'Itemizing' the different non-chromosomal malformations in association with SUA, we found that hydrocephalus, Potter's sequence, and atrioventricular septal defect were the most frequent malformations, while in earlier studies, the association with non chromosomal malformations such as vertebral malformations, imperforated anus, cheilognathopalatoschisis, and renal agenesis occurred more frequently than usual. PMID- 18674869 TI - E-cadherin, beta-catenin adhesion complex and relation to matrilysin expression in pT3 rectosigmoid cancers. AB - E-cadherin/beta-catenin complex has a critical role in cell-cell adhesion. beta Catenin is a critical component of the highly conserved Wnt signaling pathway that regulates cell proliferation and differentiation. Wnt signaling leads to the stabilization of cytosolic beta-catenin and to translocation to the nucleus, where it binds with T-cell factor and promotes the transcription and changes in target gene expression, including matrix metalloproteinases. In this study, we analyzed paraffin-embedded specimens from 42 patients with pT3 rectosigmoid cancer for E-cadherin, beta-catenin, and matrix metalloproteinase-7(MMP-7, matrilysin) expression using immunohistochemistry. Seventy-four and 79% of tumors expressed beta-catenin and E-cadherin, respectively. Nuclear expression of beta catenin was detected only in 26.1% of tumors. Forty-five percent of the rectosigmoid cancers showed strong expression of MMP-7. It was revealed that membranous or cytoplasmic beta-catenin expression was significantly related to E cadherin and MMP-7 expression. No significant association was seen between E cadherin, beta-catenin, or MMP-7 expression and some clinicopathologic features. Our results may contribute to the functional interaction between beta-catenin and MMP-7. Further studies on Wnt/beta-catenin and MMP-7 gene activity and protein expression are necessary to better understand the pathogenesis of colorectal carcinoma. PMID- 18674870 TI - Epithelioid angiomatous nodule of the nasal cavity. AB - Epithelioid angiomatous nodule (EAN) is a recently described cutaneous vascular lesion with epithelioid features. We report two additional cases occurring in an unusual location in the nasal cavity. The patients were two young adults (17 and 26 years) presenting with recurrent unilateral epistaxis. Computerized tomography scan and endoscopy revealed small tumors that had developed in the nasal septum and the turbinate, measuring 1 and 1.5 cm, respectively. Endoscopic resection was performed. No recurrence was observed in the follow-up. Pathological examination revealed a well-limited but unencapsulated nodule composed of large epithelioid cells with round nuclei. Some tumor cells contained intracytoplasmic vacuoles. Mitotic activity was low without necrotic areas. Tumor cells were strongly immunostained with antibodies to CD34 and CD31. The main differential diagnosis is an epithelioid hemangioma sharing many morphologic features with EAN. We report these cases to inform of a possibly unusual location of EAN. PMID- 18674871 TI - Low prefrontal perfusion linked to depression symptoms in methadone-maintained opiate-dependent patients. AB - BACKGROUND: Clinically depressed patients without substance use disorders, compared to controls, exhibit significantly lower resting regional cerebral blood flow (rCBF) in the prefrontal cortex (PFC). In this study, we examined the link between resting rCBF in the PFC and current depressive symptoms in methadone maintained opiate-dependent (MM) patients with or without major depression. METHODS: Arterial spin labeled perfusion fMRI at 3 Tesla was used to measure resting rCBF in 21 MM patients. Perfusion data were analyzed using SPM2. The relationship between Beck Depression Inventory (BDI) score and resting rCBF was examined in a single regression analysis. RESULTS: The BDI scores ranged between 0 and 18 (m=7.0, S.D.=4.8), and 30% of the sample had mild to moderate depression symptoms according to BDI scores. A negative correlation was observed between BDI scores and relative rCBF in the bilateral ventrolateral prefrontal cortex, and middle frontal gyri. CONCLUSIONS: The inverse relationship between prefrontal paralimbic rCBF and depression scores suggests a link between reduced fronto limbic activity and depressive symptoms in MM patients. A significant subgroup of opiate-dependent patients has clinical or sub-clinical depression that is often undetected; our data identify brain substrates of depression symptoms that may also be a potential marker of relapse in this population. Treatment strategies targeting these brain regions may improve outcomes in depressed substance abusers. PMID- 18674873 TI - [Acceptance of influenza vaccination by caregivers in a geriatric and long-term care institution]. AB - INTRODUCTION: Seasonal influenza is a viral transmissible infectious disease causing increased morbidity or mortality in frail subjects, especially those living in institutions. Measures to prevent the impact of infectious diseases were proposed based on the use of influenza vaccination among health-care professionals. We wanted to evaluate the acceptance of our institutional vaccination procedure initiated in 2005 and possible improvement. METHODS: A questionnaire was sent in May 2007 to all health-care professionals (n=730) to identify their current vaccine status in 2006 and their opinion concerning vaccination against influenza in 2007. RESULTS: Subsequently, 369 (50.2%) responses were obtained. Amongst those responding, 31.7% were vaccinated in 2006, 77.8% using the institutional procedure. Also, 221 (87.7%) nonvaccinated health care professionals indicated their position concerning influenza vaccination: 37% of them would accept the vaccination in 2007 (on the condition that our current institutional procedure be improved), 63% of them would refuse influenza vaccines in spite of any improvement. CONCLUSION: Our study emphasizes that the use of an adapted procedure for influenza vaccination among health-care professionals could improve vaccine coverage. It also emphasizes that a margin of those professionals are still reluctant to vaccination. PMID- 18674872 TI - Tri-city study of Ecstasy use problems: a latent class analysis. AB - This study used latent class analysis to examine distinctive subtypes of Ecstasy users based on 24 abuse and dependence symptoms underlying standard DSM-IV criteria. Data came from a three site, population-based, epidemiological study to examine diagnostic nosology for Ecstasy use. Subject inclusion criteria included lifetime Ecstasy use exceeding five times and once in the past year, with participants ranging in age between 16 and 47 years of age from St. Louis, Miami, U.S. and Sydney, Australia. A satisfactory model typified four latent classes representing clearly differentiated diagnostic clusters including: (1) a group of sub-threshold users endorsing few abuse and dependence symptoms (negatives), (2) a group of 'diagnostic orphans' who had characteristic features of dependence for a select group of symptoms (mild dependent), (3) a 'transitional group' mimicking the orphans with regard to their profile of dependence also but reporting some abuse symptoms (moderate dependent), and (4) a 'severe dependent' group with a distinct profile of abuse and dependence symptoms. A multinomial logistic regression model indicated that certain latent classes showed unique associations with external non-diagnostic markers. Controlling for demographic characteristics and lifetime quantity of Ecstasy pill use, criminal behavior and motivational cues for Ecstasy use were the most efficient predictors of cluster membership. This study reinforces the heuristic utility of DSM-IV criteria applied to Ecstasy but with a different collage of symptoms that produced four distinct classes of Ecstasy users. PMID- 18674874 TI - [Warning for error risk between isoprenaline and suxamethonium injectable vials]. PMID- 18674875 TI - [Evaluation of perioperative risk by anaesthesiologists]. AB - OBJECTIVE: The aim of this survey was to evaluate anaesthesiologists' ability to quantify a given risk of perioperative morbidity. STUDY DESIGN: Descriptive study. METHODS: Nine hypothetical patients with well-defined single-organ failure related to three validated perioperative risk indexes (cardiac risk, respiratory failure risk and cirrhosis decompensation) were described in vignettes and submitted to 30 anesthesiologists issued from two institutions and distributed in two groups (10 residents and 20 staff anesthesiologists). Physicians were asked to estimate the patient's risk using qualitative (scored using a categorical scale) and quantitative (scored using a number, i.e., the estimated frequency that a given complication would occur in the given case) evaluations of the considered risk for each case proposed. RESULTS: Both qualitative and quantitative evaluation of the most severe cases showed a tendency to underestimate respiratory and hepatic risks and a tendency to overestimate cardiac risk. No major difference in the accuracy of evaluations was found between trainees and staff anaesthesiologists. Both provided imprecise risk evaluation and had difficulty to define the expected level of risk, even for clear-cut clinical situations described in the vignettes. CONCLUSION: Both qualitative and quantitative evaluations were overall very imprecise. This survey showed that a rigorous evaluation of a patient's perioperative risk cannot be considered as intuitive and needs objective material supports to be performed. PMID- 18674876 TI - [Cardioprotection and volatile anaesthetics]. PMID- 18674877 TI - [Pulmonary oedema in scuba-diving: frequency and seriousness about a series of 19 cases]. AB - OBJECTIVES: Pulmonary oedema in self-contained underwater breathing apparatus diving is an accident whose risk factors, conditions of occurrence and incidence are not well-known. The aim of this study was to evaluate the frequency, the risk factors and the evolution of this accident. STUDY DESIGN: Retrospective case study and prospective frequency study. PATIENTS AND METHODS: Study covering the Brittany region and performed in two steps with distinct objectives: a review of cases diagnosed between 2002 and 2007, and a one-year study of cases reported by emergency physicians. Diagnosis was based on the history, a respiratory distress, auscultation and radiologic features. RESULTS: Nineteen cases were reported, of which one was recurrent. The mean age of patients was 49 years. Divers without heart disease were involved, as well as divers with hypertension (eight cases) or valve abnormalities (three cases). Stress and/or physical exertion were involved. Dyspnoea, cough and haemoptysis were the most common symptoms; in addition, two cases of cardiac arrest and three of loss of consciousness were observed. Chest radiography was unsensitive (normal in four cases), contrasting with abnormal thoracic CT scan in all cases. Symptoms resolved rapidly with oxygen, except for two divers who died. We identified five cases over one year, one of which lethal. CONCLUSION: This accident is not a rare event and may have serious consequences. Oldest divers submitted to stress and/or effort are at higher risk. PMID- 18674878 TI - Resveratrol protects primary rat hepatocytes against necrosis induced by reactive oxygen species. AB - Reactive oxygen species can be important mediators of damage to cell molecules and structures. Besides the endogen antioxidant defences, the antioxidant intake in the diet has an important role in the protection against the development of diseases produced by oxidative damage. Resveratrol is a naturally occurring compound present in many plants some of which are part of the human diet. This molecule has been thoroughly investigated because of its antioxidant and anticarcinogenic properties among others. We investigated whether resveratrol could provide protective antioxidant action in primary rat hepatocyte cultures. Primary rat hepatocytes cultures were exposed to 300 microM tert-butyl hydroperoxide; 25, 50 or 75 microM resveratrol or to 300 microM tert-butyl hydroperoxide plus 25, 50 or 75 microM resveratrol for different time periods. Necrosis was evaluated by lactate dehydrogenase liberation to the medium. Apoptosis was evaluated by caspase 3 activity measurement. Changes in cellular morphology after the different treatments were recorded using bright field microscopy. Inhibition of the reactive oxygen species by resveratrol was studied by confocal microscopy and spectrofluorimetrically. Resveratrol inhibited necrosis induced by tert-butyl hydroperoxide. No apoptosis was observed in any treatment. It also was effective in eliminating reactive oxygen species. At 75 microM, the highest concentration tested, resveratrol became slightly cytotoxic. Our results show that resveratrol protects primary rat hepatocytes in culture from oxidative stress induced cell death. These results suggest that resveratrol could enhance the antioxidant status of hepatic cells. PMID- 18674879 TI - Resveratrol inhibits hypoxia-induced metastasis potential enhancement by restricting hypoxia-induced factor-1 alpha expression in colon carcinoma cells. AB - Resveratrol has been shown recently to exhibit antimetastatic effect on various human solid tumors. However, the possible molecular mechanism for its antimetastatic action needs to be elucidated. In this study, we investigated the effect of resveratrol on metastasis potential of colon carcinoma cells under normoxia and hypoxia in vitro. These results showed that, resveratrol can restrict the migration, adhesion, invasion and MMP-9 and MMP-2 secretion in Lovo cells cultured under normoxia and hypoxia. Hypoxia and iron chelator 2,2' dipyridyl treatment can stimulate the invasion and migration enhancement of Lovo cells, while resveratrol exhibited substantial resistance on the metastasis potential stimulation by inhibiting the mRNA expression of VEGF and MMP-9 in colon carcinoma cells under normoxia and hypoxia, reducing HIF-1 alpha protein expression under hypoxia. Also, iron chelator 2,2'-dipyridyl treatment showed approximately the same effect on metastasis potential as Lovo cells cultured under hypoxia. These data demonstrated that, the antimetastatic effect of resveratrol under hypoxia were associated with the restriction of HIF-1 alpha protein expression and stabilization, which could be a promising drug target for resveratrol in the development of an effective chemopreventive and anticancer therapy in human tumors. PMID- 18674880 TI - Fulkerson procedure for chronic patella component dislocation after total knee arthroplasty. AB - Chronic patella component dislocation after primary knee arthroplasty can be a challenging problem. The purpose of this case series was to review our results of a chronic patella dislocation after total knee arthroplasty treated with a Fulkerson procedure (anteromedialization of the tibial tubercle). Five patients with an average age of 68.8 years (range, 60-76 years) underwent a Fulkerson procedure to restore extensor mechanism alignment. The tibial tubercle osteotomy was secured with lag screws with an average medialization of approximately 1.5 cm. The average preoperative Knee Society score was 70.5 points that improved to 85 points postoperatively. The average knee flexion was 93 degrees preoperatively that improved to 101 degrees postoperatively. Adequate patellofemoral alignment was achieved in all patients along with union of the tibial tubercle osteotomy site. The Fulkerson procedure appears to be an excellent treatment option in patients with a chronically dislocated patella component after total knee arthroplasty. PMID- 18674881 TI - IL-1beta signaling initiates inflammatory hypernociception. PMID- 18674882 TI - A study of voice therapy dropout. AB - Treatment dropout is a consistent problem among behavior change therapies. A recent study by the authors demonstrated that 38% of patients did not attend a voice evaluation after referral by the otolaryngologist. Further, 47% of patients who attended a voice evaluation did not return for therapy. No previous study has set out to document completion/dropout rates for voice therapy, but high rates of attrition are reported as problematic within voice therapy research studies. The purpose of this study was to quantify the problem of voice therapy dropout over the course of voice therapy and to analyze factors that may predict dropout using a retrospective chart review. Data were collected at two voice centers and included demographics (gender, age, and race/ethnicity), quality-of-life impact (Voice Handicap Index [VHI]), severity of dysphonia (Consensus Auditory Perceptual Evaluation of Voice [CAPE-V] overall severity score), diagnosis, and completion/dropout status. Results indicated a 65% voice therapy dropout rate in this study. There was no significant difference in dropout rates for gender, age, race, VHI, CAPE-V, or diagnosis. No factor studied was strongly associated with dropout. The 65% dropout rate in this study was consistent with literature published in other behavior change fields. The variables analyzed in this study were not predictive of dropout. Future research should examine methods to effect a reduction in dropout, from otolaryngologist referral through completion of therapy. PMID- 18674883 TI - Neural network based pattern matching and spike detection tools and services--in the CARMEN neuroinformatics project. AB - In the study of information flow in the nervous system, component processes can be investigated using a range of electrophysiological and imaging techniques. Although data is difficult and expensive to produce, it is rarely shared and collaboratively exploited. The Code Analysis, Repository and Modelling for e Neuroscience (CARMEN) project addresses this challenge through the provision of a virtual neuroscience laboratory: an infrastructure for sharing data, tools and services. Central to the CARMEN concept are federated CARMEN nodes, which provide: data and metadata storage, new, thirdparty and legacy services, and tools. In this paper, we describe the CARMEN project as well as the node infrastructure and an associated thick client tool for pattern visualisation and searching, the Signal Data Explorer (SDE). We also discuss new spike detection methods, which are central to the services provided by CARMEN. The SDE is a client application which can be used to explore data in the CARMEN repository, providing data visualization, signal processing and a pattern matching capability. It performs extremely fast pattern matching and can be used to search for complex conditions composed of many different patterns across the large datasets that are typical in neuroinformatics. Searches can also be constrained by specifying text based metadata filters. Spike detection services which use wavelet and morphology techniques are discussed, and have been shown to outperform traditional thresholding and template based systems. A number of different spike detection and sorting techniques will be deployed as services within the CARMEN infrastructure, to allow users to benchmark their performance against a wide range of reference datasets. PMID- 18674885 TI - Isoflurane exacerbates electrically evoked seizures in amygdala-kindled rats during recovery. AB - Neuroexcitatory effects of isoflurane during or following anesthesia are controversial, particularly in epileptic patients. In contrast, halothane is generally considered to be highly anticonvulsant. Kindling is an animal model of epilepsy suitable for studying the effects of anesthetic agents on the epileptic brain. Fully kindled, Sprague-Dawley rats were either untreated or received a 5 min exposure to isoflurane or halothane 30 min prior to a seizure and compared to seizures in the absence of prior anesthesia. Afterdischarge duration was assessed via electroencephalographs recorded from electrodes implanted in the basolateral amygdala and behavioral seizure stereotypy (stages I-V) was simultaneously recorded and analyzed using digital video for all seizures. Total seizure duration and clonus duration were significantly (P<0.05) increased 30 min after isoflurane but not halothane exposure relative to pre-treatment control. These results are the first to demonstrate that isoflurane exacerbates electrically evoked secondarily generalized seizures in fully kindled animals during recovery. These results also show that the kindling paradigm is useful for evaluating the mechanism of anesthetic agents that may be proconvulsant in epileptic subjects. PMID- 18674886 TI - Therapeutic effect of a novel anilidoquinoline derivative, 2-(2-methyl-quinoline 4ylamino)-N-(2-chlorophenyl)-acetamide, in Japanese encephalitis: correlation with in vitro neuroprotection. AB - 2-(2-Methyl-quinoline-4ylamino)-N-(2-chlorophenyl)-acetamide, a novel anilidoquinoline derivative, was synthesised and evaluated for its therapeutic efficacy in treating Japanese encephalitis. The compound showed significant antiviral and antiapoptotic effects in vitro. Significant decreases in viral load (P<0.01) combined with an increase in survival was observed in Japanese encephalitis virus-infected mice treated with 2-(2-methyl-quinoline-4ylamino)-N (2-chlorophenyl)-acetamide. PMID- 18674888 TI - Expression of NR1, NR2A and NR2B NMDA receptor subunits is not altered in the genetically-inbred Balb/c mouse strain with heightened behavioral sensitivity to MK-801, a noncompetitive NMDA receptor antagonist. AB - The genetically-inbred Balb/c mouse strain shows heightened sensitivity to the ability of MK-801 (dizocilpine), a noncompetitive NMDA receptor antagonist, to raise the threshold voltage necessary to precipitate tonic hindlimb extension and elicit irregular episodes of intense jumping behavior (referred to as "popping"), relative to other inbred mouse strains and the outbred NIH Swiss mouse. Moreover, an allosteric modulatory effect of sarcosine, a glycine reuptake inhibitor, on MK 801's antagonism of electrically precipitated seizures was detected 24 h after Balb/c mice were forced to swim in cold water for up to 10 min; this was not observed in unstressed Balb/c mice or stressed or unstressed NIH Swiss mice. Phencyclidine (PCP), a noncompetitive NMDA receptor antagonist that binds to the same hydrophobic channel domain as MK-801, precipitates a schizophreniform psychosis in susceptible individuals that shares descriptive similarities with schizophrenia. This observation has led to the hypothesis that NMDA receptor hypofunction (NRH) is involved in the pathophysiology of schizophrenia and the testing of pharmacotherapeutic strategies to facilitate NMDA receptor-mediated neurotransmission in patients with this disorder (e.g., glycine reuptake inhibitors). The heightened behavioral sensitivity of the Balb/c mouse to MK-801 suggests that this mouse strain may be a useful model to study "psychosis proneness" and screen for positive allosteric modulators of NMDA receptor mediated neurotransmission. Conceivably, strain differences in the pharmacology of the NMDA receptor are due to differences in the relative expression of individual NMDA receptor subunits to each other (i.e., combinatorial regulation). The current study compared the normal protein expression patterns of six of the eight identified splice variant isoforms of the NR1 NMDA receptor subunit, and NR2A and NR2B subunits in the hippocampus and cerebral cortex of Balb/c and NIH Swiss mice. The heightened behavioral sensitivity of the Balb/c genetically inbred mouse strain to MK-801, compared to the outbred NIH Swiss mouse strain, does not appear to result from relative alterations of expression of these NMDA receptor protein subunits that were examined. PMID- 18674887 TI - Dynamic changes of the endogenous cannabinoid and opioid mesocorticolimbic systems during adolescence: THC effects. AB - Adolescence is a critical phase of active brain development often characterized by the initiation of marijuana (Cannabis sativa) use. Limited information is known regarding the endogenous cannabinoid system of the adolescent brain as well as related neurotransmitters that appear sensitive to cannabis exposure. We recently observed that adult rats pre-exposed to Delta-9-tetrahydrocannabinol (THC) during adolescence self-administered higher amounts of heroin and had selective impairments of the enkephalin opioid system within the nucleus accumbens (NAc) implicated in reward-related behavior. To explore the ontogeny of the cannabinoid and opioid neuronal systems in association with adolescence THC exposure, rats were examined at different adolescent stages during an intermittent THC paradigm (1.5 mg/kg i.p. every third day) from postnatal days (PNDs) 28-49. Rat brains were examined 24 h after injection at PND 29 (early adolescence), PND 38 (mid adolescence) and PND 50 (late adolescence) and analyzed for endocannabinoids (anandamide and 2-arachidonoylglycerol), Met-enkephalin, cannabinoid CB(1) receptors and micro opioid receptors (microOR) in the NAc, caudate-putamen and prefrontal cortex (PFC). Of the markers studied, the endocannabinoid levels had the most robust alterations throughout adolescence and were specific to the PFC and NAc. Normal correlations between anandamide and 2 arachidonoylglycerol concentrations in the NAc (positive) and PFC (negative) were reversed by THC. Other significant THC-induced effects were confined to the NAc - increased anandamide, decreased Met-enkephalin and decreased microORs. These findings emphasize the dynamic nature of the mesocorticolimbic endocannabinoid system during adolescence and the selective mesocorticolimbic disturbance as a consequence of adolescent cannabis exposure. PMID- 18674889 TI - [Malignancy risk and Wiedemann-Beckwith syndrome: what follow-up to provide?]. AB - Wiedemann-Beckwith syndrome (WBS) is a syndrome of excessive growing with a high predisposition to developing embryologic tumours within the first years of life. This risk is evaluated between 7.5 and 10%; it varies with the mechanisms of mutations involved. These take place in two distinct domains of 11p15, which are under parental printing. Emerging techniques of cytogenetic and molecular biology now have shown correlations between genotypes and phenotypes, and can identify the 30% of WBS who are especially at risk of developing tumours. A specific follow-up, integrating the specificity of developing tumours of each 11p15 mutations involved, is now proposed to patients with WBS. PMID- 18674890 TI - Metabolic syndrome and left ventricular hypertrophy in the prediction of cardiovascular events: the Strong Heart Study. AB - BACKGROUND AND AIMS: Metabolic syndrome (MetS) is associated with increased prevalence of echocardiographic LV hypertrophy (LVH), a potent predictor of cardiovascular (CV) outcome. Whether MetS increases risk of CV events independently of presence of LVH has never been investigated. It is also unclear whether LVH predicts CV risk both in the presence and absence of MetS. METHODS AND RESULTS: Participants in the 2nd Strong Heart Study examination without prevalent coronary heart disease, congestive heart failure or renal insufficiency (plasma creatinine >2.5mg/dL) were studied (n=2758; 1746 women). MetS was defined by WHO criteria. Echocardiographic LV hypertrophy was defined using population specific cut-point value for LV mass index (>47.3g/m(2.7)). After controlling for age, sex, LDL-cholesterol, smoking, plasma creatinine, diabetes, hypertension and obesity, participants with MetS had greater probability of LVH than those without MetS (OR=1.55 [1.18-2.04], p<0.002). Adjusted hazard of composite fatal and non fatal CV events was greater when LVH was present, in participants without (HR=2.03 [1.33-3.08]) or with MetS (HR=1.64 [1.31-2.04], both p<0.0001), with similar adjusted population attributable risk (12% and 14%). After adjustment for LVH, risk of incident CV events remained 1.47-fold greater in MetS (p<0.003), an effect, however, that was not confirmed when diabetic participants were excluded. CONCLUSION: LVH is a strong predictor of composite 8-year fatal and non-fatal CV events either in the presence or in the absence of MetS and accounts for a substantial portion of the high CV risk associated with MetS. PMID- 18674892 TI - Waist circumference cut-off values to predict the incidence of hypertension: an estimation from a Brazilian population-based cohort. AB - BACKGROUND AND AIMS: Central obesity is a key component in the definition of the metabolic syndrome, but the cut-off values proposed to define abnormal values vary among different guidelines and are mostly based on cross-sectional studies. In this study, we identify the best cut-off values for waist circumference (WC) associated with the incidence of hypertension. METHODS AND RESULTS: Participants for this prospectively planned cohort study were 589 individuals who were free of hypertension and selected at random from the community of Porto Alegre, Brazil. Hypertension was defined by a blood pressure measurement >or= 140/90 mmHg or the use of blood pressure lowering drugs. A logistic regression model established the association between WC and the incidence of hypertension. A receiver operating characteristics (ROC) curve analysis was used to select the best WC cut-off point to predict the incidence of hypertension. During a mean follow-up of 5.5+/-0.9 years, 127 subjects developed hypertension. The hazard ratios for the development of hypertension, adjusted for age, baseline systolic blood pressure, alcohol consumption, gender and scholarship were 1.02 (95% CI; 1.00-1.04; P=0.02) for WC. The best cut-off WC values to predict hypertension were 87 cm in men and 80 cm in women, with an area under the curve of 0.56 (95% CI; 0.47-0.64; P=0.17) and 0.70 (95% CI; 0.63-0.77; P<0.001), respectively. CONCLUSION: Excess visceral adiposity is a major risk factor for hypertension in individuals living in communities in Brazil, and this risk begins at lower values of WC that those recommended by some guidelines. PMID- 18674891 TI - Short-term mortality risk in children and young adults with type 1 diabetes: the population-based Registry of the Province of Turin, Italy. AB - Short-term mortality risk in young diabetic people is an indicator of quality of care. We assessed this in the Italian incident population-based registry of Turin. The study base included 1210 incident cases (n=677 aged 0-14 years and n=533 aged 15-29 years) with diabetes, onset period 1974-2000 in the Province of Turin, Italy. The relevant timescale for analysis was the time since the onset of diabetes to death, or till 31 December 2003. Standardized mortality ratio (SMR) for all-cause mortality was computed using the Italian population as a standard, by 5 years, age group, sex, and calendar period. Mean attained age of the incident cohort was 29.7 years (range 5.2-49.7 years). During a mean follow-up period of 15.8 years (range 2.0-29.9 years), there were 19 deaths in 15,967. Nine person-years of observation (n=9.5 expected deaths), giving an all-cause mortality rate of 1.19/1000 person-years (95% CI 0.76-1.87) and an SMR of 1.96 (1.25-3.08). In no cases did death occur at the onset of diabetes or in childhood. Out of 19 deaths, 9 were diabetes related (n=6 coma and n=3 end-stage renal disease). In Cox regression analysis, the hazard ratio (HR) was higher in adult-onset than in childhood-onset diabetes (HR=3.90, 95% CI 1.14-13.39), independently of calendar period and gender. (1) Children and young adults with type 1 diabetes experienced a two-fold higher short-term mortality risk than Italian people of similar age and sex and (2) the risk was higher in adult-onset than in childhood-onset diabetes. The quality of diabetes care should be improved to prevent early deaths. PMID- 18674893 TI - Cranial morphology and bite force in Chamaeleolis lizards--adaptations to molluscivory? AB - Anolis lizards have become a model system for the study of adaptive radiations as species with similar morphologies occupying similar habitats have arisen independently on all the larger islands in the Caribbean. However, on both, Cuba and Hispaniola unique forms have evolved that seemingly have no counterparts on any of the other Caribbean islands. Anoles of the genus Chamaeleolis comprise such a unique form and have been termed 'twig giants' because of their cryptic life style, slow locomotor mode, and short limbs. However, some of the most unusual features of these lizards are their large heads and molluscivorous diet. Here, we compare head shape, bite force, and muscle structure among sexes and age classes of Chamaeleolis lizards with Anolis crown giants. Our data show that Chamaeleolis lizards have a dramatically different head shape characterized by tall heads with a pronounced temporal ridge and long snouts. Analyses of bite force, surprisingly, show no differences between adult Chamaeleolis and Anolis crown giants. Juveniles of Chamaeleolis, however, have very tall heads for their size and bite harder than Anolis juveniles do. This can be related to the propensity of juveniles of this genus to eat snails, food items for which high bite forces are crucial. This observation is corroborated by the presence of well developed jaw adductors in juveniles. Thus, our data suggest that the unusual phenotype of adults with large and tall heads may be due to selection on the juvenile life history stages. PMID- 18674894 TI - Repeatability of health economic data in COPD. AB - BACKGROUND: Little is known concerning the precision of self-reported health economic data. AIM: To investigate the repeatability of self-reported health economic cost and utilization data in subjects with chronic obstructive pulmonary disease (COPD). METHODS: Twelve weeks after inclusion, a physician and a nurse in random order interviewed 29 subjects with post-bronchodilator COPD included from a research registry of COPD patients. The interview recorded healthcare utilization and costs, sick leave, exacerbations and quality of life (QoL). Variation of individual agreement of observations was described estimating kappa statistics and 95% limits of agreement. RESULTS: Mean age was 63 years (standard deviation (SD) 10) and 17 participants were men. Average FEV(1)% predicted was 56% (SD 15). For sick leave, exacerbation, healthcare provider visit, change of medication, assistance and leisure time the kappa values were 1.00, 0.73, 0.73, 0.66, 0.63 and 0.54, respectively. The physician recorded fewer days of exacerbation and fewer contacts with healthcare providers than the nurse (p=0.01 and p=0.05, respectively). The 95% limits of agreement for costs of drugs were 690 to +710 Norwegian Kroner (NOK), -1200 to +899 NOK for costs of healthcare providers and -20 to +26 for QoL as measured by a visual analogue scale. CONCLUSION: Repeatability of economic data from COPD patients showed considerable variation. This issue should be addressed when analyzing cost data from interviews and when designing studies on health economy. PMID- 18674895 TI - Social inequality in incidence of and survival from cancer in a population-based study in Denmark, 1994-2003: Summary of findings. AB - The purpose of this nationwide, population register-based study was to describe variations in cancer incidence and survival by social position in a social welfare state, Denmark, on the basis of a range of socioeconomic, demographic and health-related indicators. Our study population comprised all 3.22 million Danish residents born in 1925-1973 and aged >or=30 years, who were followed up for cancer incidence in 1994-2003 and for survival in 1994-2006, yielding 147,973 cancers. The incidence increased with lower education and income, especially for tobacco- and other lifestyle-related cancers, although for cancers of the breast and prostate and malignant melanoma the association was inverse. Conversely there was a general increase in incidence among early retirement pensioners, persons living in rented housing and those living in the smallest dwellings. Also incidence rates were generally higher in persons living alone compared to those living with a partner and in the capital area compared to the rural areas. Social inequality in the prognosis of most cancers was observed, despite the equal access to health care in Denmark, with poorer relative survival related to fewer advantages, regardless of how they were measured, often most pronounced in the first year after diagnosis. Also living alone and having somatic or psychiatric comorbidity negatively impacted the relative survival after most cancers. Our study shows that inequalities in cancer incidence and survival must be addressed in all aspects of public health, with interventions both to reduce incidence and to prolong survival. PMID- 18674896 TI - Biocatalytic conversion of wheat bran hydrolysate using an immobilized GH43 beta xylosidase. AB - To investigate the concept of a xylosidase-based process for the continuous production of xylose from arabinoxylan-containing feedstocks, a beta-xylosidase from Bacillus halodurans C-125 was immobilized and deployed in packed bed reactor (PBR). Among the several immobilization methods tested, glutaraldehyde-mediated immobilization on chitosan was the best both in terms of immobilization and activity yields (91% and 72.9%, respectively). In batch experiments the immobilized enzyme hydrolyzed wheat bran hydrolysates quite efficiently, consuming nearly all xylobiose and xylotriose after 6h. Its reusability showed only a 50% decrease of its activity after 92h. Using the chitosan-immobilized beta-xylosidase in a PBR, xylose productivity was 7.2g xylose l(-1)h(-1) and the conversion factor was 0.55 (derived from initial xylose in the substrate). The operational stability of the PBR was good, because only 25% of productivity was lost after the treatment of three batches of substrate over a 72-h period. PMID- 18674897 TI - Formation of aerobic granules and their PHB production at various substrate and ammonium concentrations. AB - Aerobic granular sludge rich in polyhydroxybutyrate (PHB) was cultivated in a sequencing batch reactor (SBR) by seeding anaerobic granular sludge. The PHB content in aerobic granules was investigated and the experimental results reveal that both influent chemical oxygen demand (COD) and ammonium concentrations had a significant effect on the morphological characteristics and the PHB production of the aerobic granular sludge. At a COD and ammonium concentration of 750 mg/L and 8.5mg/L, respectively, the PHB content of the granules reached 44%, but their poor settling ability, as evidenced by a high sludge volume index, was observed. This was attributed to the outgrowth of filamentous bacteria on the granule surface. However, an increase in the ammonium concentration resulted in an elevated sludge concentration and a decrease in the PHB content in the granules. In this case, the aerobic granular sludge with a regular and compact structure was formed. The results suggest that, through controlling the COD and ammonium concentrations in the influent, the PHB-rich aerobic granular sludge with good settling ability could be cultivated. PMID- 18674898 TI - Aminomethylpiperazines as selective urotensin antagonists. AB - Aminomethylpiperazines, reported previously as being kappa-opioid receptor agonists, were identified as lead compounds in the development of selective urotensin receptor antagonists. Optimized substitution of the piperazine moiety has provided high affinity urotensin receptor antagonists with greater than 100 fold selectivity over the kappa-opioid receptor. Select compounds were found to inhibit urotensin-induced vasoconstriction in isolated rat aortic rings consistent with the hypothesis that an urotensin antagonist may be useful for the treatment of hypertension. PMID- 18674900 TI - Interaction of human serum albumin and its clinically relevant modification with oligoribonucleotides. AB - Human serum albumin (HSA) was shown to mediate oligoribonucleotide cleavage. Nonenzymatic glycation of HSA decreased the ribonuclease-like activity of the protein. According to (31)P NMR data, both native and glycated albumins induced hydrolysis of RNA molecule through 2',3'-cyclophosphate intermediates. A feasible mechanism of RNA hydrolysis by native albumin and its clinically relevant modification was discussed. PMID- 18674899 TI - Synthesis and antiviral properties of some polyphenols related to Salvia genus. AB - An efficient synthesis of the acid part of salvianolic acid E 2 is described. Compound 2 was obtained from vanillin in 10 steps and 21% overall yield. During the synthesis of 2 an unexpected 5-oxo-4b,9b-dihydroindano[1,2-b]benzofuran rac 12 was isolated. Both compounds together with the acid part of salvianolic acid D were active as HIV-1 integrase inhibitors at the submicromolar level. But they did not inhibit the replication of the virus on MT-4 cells. PMID- 18674901 TI - Synthesis and biological evaluation of N-mercaptoacylproline and N mercaptoacylthiazolidine-4-carboxylic acid derivatives as leukotriene A4 hydrolase inhibitors. AB - We studied the synthetic modification of structurally similar N-mercaptoacyl-L proline and (4R)-N-mercaptoacylthiazolidine-4-carboxylic acid to obtain potent leukotriene A(4) (LTA(4)) hydrolase inhibitors. An N-mercaptoacyl group, (2S)-3 mercapto-2-methylpropionyl group, was effective for both scaffolds. Additional introduction of a large substituent such as 4-isopropylbenzylthio (3f), 4-tert butylbenzylthio (3l) or 4-cyclohexylbenzylthio group (3m) with (S)-configuration at the C(4) position of proline yielded much more potent LTA(4) hydrolase inhibitors (IC(50); 52, 31, and 34 nM, respectively) than captopril (IC(50); 630,000 nM). PMID- 18674902 TI - Synthesis and pharmacological evaluation of hydrophobic esters and ethers of butorphanol at opioid receptors. AB - We synthesized several hydrophobic esters and ethers of butorphanol and assessed their affinities at opioid receptors in CHO cell membranes. Tested compounds displayed moderate to high affinities to the mu and kappa receptors. The findings accord with previous evidence of a lipophilic binding pocket in the opioid receptors that can be accessed to afford good binding affinity without the need for a phenolic hydrogen-bond donor group. The most potent (K(i)=61 pM at mu and 48 pM at kappa) novel agent was (-)-N-cyclobutylmethylmorphinan-3-yl-14-ol phenoxyacetate (4d). PMID- 18674903 TI - Discovery of dihydroquinoxalinone acetamides containing bicyclic amines as potent Bradykinin B1 receptor antagonists. AB - Replacement of the core beta-amino acid in our previously reported piperidine acetic acid and beta-phenylalanine-based Bradykinin B1 antagonists by dihydroquinoxalinone acetic acid increases the in vitro potency and metabolic stability. The most potent compounds from this series have IC(50)s<0.2 nM in a human B1 receptor functional assay. A molecular modeling study of the binding modes of key compounds, based on a B1 homology model, explains the structure activity relationship (SAR) for these analogs. PMID- 18674904 TI - Design and synthesis of a novel DNA-encoded chemical library using Diels-Alder cycloadditions. AB - DNA-encoded chemical libraries are increasingly being employed for the identification of binding molecules to protein targets of pharmaceutical relevance. Here, we describe the synthesis and characterization of a DNA-encoded chemical library, consisting of 4000 compounds generated by Diels-Alder cycloaddition reactions. The compounds were encoded with unique DNA fragments which were generated through a stepwise assembly process and serve as amplifiable bar codes for the identification and relative quantification of library members. PMID- 18674905 TI - Factor VIIa inhibitors: target hopping in the serine protease family using X-ray structure determination. AB - Selective factor VIIa-tissue factor complex (FVIIa/TF) inhibition is regarded as a promising target for developing new anticoagulant drugs. Compound 1 was discovered from focused screening of serine protease-directed compounds from our internal collection. Using parallel synthesis supported by structure-based drug design, we identified peptidemimetic FVIIa/TF inhibitors (compounds 4-11) containing L-Gln or L-Met as the P2 moiety. However, these compounds lacked the selectivity of other serine proteases in the coagulation cascade, especially thrombin. Further optimization of these compounds was carried out with a focus on the P4 moiety. Among the optimized compounds, 12b-f showed improved selectivity. PMID- 18674906 TI - A new multi-gram synthetic route to labeling precursors for the D(2/3) PET agent 18F-fallypride. AB - This Letter describes a new multi-gram synthetic protocol for the preparation of the classic tosylate labeling precursor for the D(2/3) PET agent [(18)F]fallypride. In the course of our studies, we also discovered two novel labeling precusors, the previously undescribed mesylate and chloro congeners of fallypride. PMID- 18674907 TI - Double dissociation of PKC and AC manipulations on operant and classical learning in Drosophila. AB - Learning about relationships between stimuli (i.e., classical conditioning [1]) and learning about consequences of one's own behavior (i.e., operant conditioning [2]) constitute the major part of our predictive understanding of the world. Since these forms of learning were recognized as two separate types 80 years ago [3], a recurrent concern has been the issue of whether one biological process can account for both of them [4, 5, 6, 7, 8, 9]. Today, we know the anatomical structures required for successful learning in several different paradigms, e.g., operant and classical processes can be localized to different brain regions in rodents [9] and an identified neuron in Aplysia shows opposite biophysical changes after operant and classical training, respectively [5]. We also know to some detail the molecular mechanisms underlying some forms of learning and memory consolidation. However, it is not known whether operant and classical learning can be distinguished at the molecular level. Therefore, we investigated whether genetic manipulations could differentiate between operant and classical learning in Drosophila. We found a double dissociation of protein kinase C and adenylyl cyclase on operant and classical learning. Moreover, the two learning systems interacted hierarchically such that classical predictors were learned preferentially over operant predictors. PMID- 18674908 TI - Experience shapes the utility of natural statistics for perceptual contour integration. AB - Segmenting meaningful targets from cluttered scenes is a fundamental function of the visual system. Evolution and development have been suggested to optimize the brain's solution to this computationally challenging task by tuning the visual system to features that co-occur frequently in natural scenes (e.g., collinear edges) [1, 2, 3]. However, the role of shorter-term experience in shaping the utility of scene statistics remains largely unknown. Here, we ask whether collinearity is a specialized case, or whether the brain can learn to recruit any image regularity for the purpose of target identification. Consistent with long term optimization for typical scene statistics, observers were better at detecting collinear contours than configurations of elements oriented at orthogonal or acute angles to the contour path. However, training resulted in improved detection of orthogonal contours that lasted for several months, suggesting retuning rather than transient changes of visual sensitivity. Improvement was also observed for acute contours but only after longer training. These results demonstrate that the brain flexibly exploits image regularities and learns to use discontinuities typically associated with surface boundaries (orthogonal, acute alignments) for contour linking and target identification. Thus, short-term experience in adulthood shapes the interpretation of scenes by assigning new statistical utility to image regularities. PMID- 18674909 TI - Sponge genes provide new insight into the evolutionary origin of the neurogenic circuit. AB - The nerve cell is a eumetazoan (cnidarians and bilaterians) synapomorphy [1]; this cell type is absent in sponges, a more ancient phyletic lineage. Here, we demonstrate that despite lacking neurons, the sponge Amphimedon queenslandica expresses the Notch-Delta signaling system and a proneural basic helix loop helix (bHLH) gene in a manner that resembles the conserved molecular mechanisms of primary neurogenesis in bilaterians. During Amphimedon development, a field of subepithelial cells expresses the Notch receptor, its ligand Delta, and a sponge bHLH gene, AmqbHLH1. Cells that migrate out of this field express AmqDelta1 and give rise to putative sensory cells that populate the larval epithelium. Phylogenetic analysis suggests that AmqbHLH1 is descendent from a single ancestral bHLH gene that later duplicated to produce the atonal/neurogenin related bHLH gene families, which include most bilaterian proneural genes [2]. By way of functional studies in Xenopus and Drosophila, we demonstrate that AmqbHLH1 has a strong proneural activity in both species with properties displayed by both neurogenin and atonal genes. From these results, we infer that the bilaterian neurogenic circuit, comprising proneural atonal-related bHLH genes coupled with Notch-Delta signaling, was functional in the very first metazoans and was used to generate an ancient sensory cell type. PMID- 18674910 TI - The survival advantage of olfaction in a competitive environment. AB - Olfaction is generally assumed to be critical for survival because this sense allows animals to detect food and pheromonal cues. Although the ability to sense sex pheromones [1, 2, 3] is likely to be important for insects, the contribution of general odor detection to survival is unknown. We investigated the extent to which the olfactory system confers a survival advantage on Drosophila larvae foraging for food under conditions of limited resources and competition from other larvae. PMID- 18674911 TI - Ancestral TSH mechanism signals summer in a photoperiodic mammal. AB - In mammals, day-length-sensitive (photoperiodic) seasonal breeding cycles depend on the pineal hormone melatonin, which modulates secretion of reproductive hormones by the anterior pituitary gland [1]. It is thought that melatonin acts in the hypothalamus to control reproduction through the release of neurosecretory signals into the pituitary portal blood supply, where they act on pituitary endocrine cells [2]. Contrastingly, we show here that during the reproductive response of Soay sheep exposed to summer day lengths, the reverse applies: Melatonin acts directly on anterior-pituitary cells, and these then relay the photoperiodic message back into the hypothalamus to control neuroendocrine output. The switch to long days causes melatonin-responsive cells in the pars tuberalis (PT) of the anterior pituitary to increase production of thyrotrophin (TSH). This acts locally on TSH-receptor-expressing cells in the adjacent mediobasal hypothalamus, leading to increased expression of type II thyroid hormone deiodinase (DIO2). DIO2 initiates the summer response by increasing hypothalamic tri-iodothyronine (T3) levels. These data and recent findings in quail [3] indicate that the TSH-expressing cells of the PT play an ancestral role in seasonal reproductive control in vertebrates. In mammals this provides the missing link between the pineal melatonin signal and thyroid-dependent seasonal biology. PMID- 18674912 TI - Male fish deceive competitors about mating preferences. AB - A fundamental question in animal communication is whether the information provided is honest or deceptive [1, 2]. This problem has received much attention, both in theoretical [1, 3] and experimental [4] work. Here we show that male Atlantic mollies (Poecilia mexicana), when observed during mate choice by another male, reduce their mating activity and no longer prefer mating with one of two females presented, which can be interpreted as an attempt to avoid unintended interception of information by the rival male. Most importantly, focal males directed their first sexual interaction (after they were presented with the rival male) toward the initially nonpreferred female, suggesting that males deceive other males about their mating preferences. Deception by the choosing male may be an adaptation to avoid sperm competition, because surrounding males may use public information and copy the focal male's mate choice. PMID- 18674913 TI - D1 receptor activation in the mushroom bodies rescues sleep-loss-induced learning impairments in Drosophila. AB - BACKGROUND: Extended wakefulness disrupts acquisition of short-term memories in mammals. However, the underlying molecular mechanisms triggered by extended waking and restored by sleep are unknown. Moreover, the neuronal circuits that depend on sleep for optimal learning remain unidentified. RESULTS: Learning was evaluated with aversive phototaxic suppression. In this task, flies learn to avoid light that is paired with an aversive stimulus (quinine-humidity). We demonstrate extensive homology in sleep-deprivation-induced learning impairment between flies and humans. Both 6 hr and 12 hr of sleep deprivation are sufficient to impair learning in Canton-S (Cs) flies. Moreover, learning is impaired at the end of the normal waking day in direct correlation with time spent awake. Mechanistic studies indicate that this task requires intact mushroom bodies (MBs) and requires the dopamine D1-like receptor (dDA1). Importantly, sleep-deprivation induced learning impairments could be rescued by targeted gene expression of the dDA1 receptor to the MBs. CONCLUSIONS: These data provide direct evidence that extended wakefulness disrupts learning in Drosophila. These results demonstrate that it is possible to prevent the effects of sleep deprivation by targeting a single neuronal structure and identify cellular and molecular targets adversely affected by extended waking in a genetically tractable model organism. PMID- 18674914 TI - C. elegans anaplastic lymphoma kinase ortholog SCD-2 controls dauer formation by modulating TGF-beta signaling. AB - BACKGROUND: Different environmental stimuli, including exposure to dauer pheromone, food deprivation, and high temperature, can induce C. elegans larvae to enter the dauer stage, a developmentally arrested diapause state. Although molecular and cellular pathways responsible for detecting dauer pheromone and temperature have been defined in part, other sensory inputs are poorly understood, as are the mechanisms by which these diverse sensory inputs are integrated to achieve a consistent developmental outcome. RESULTS: In this paper, we analyze a wild C. elegans strain isolated from a desert oasis. Unlike wild type laboratory strains, the desert strain fails to respond to dauer pheromone at 25 degrees C, but it does respond at higher temperatures, suggesting a unique adaptation to the hot desert environment. We map this defect in dauer response to a mutation in the scd-2 gene, which, we show, encodes the nematode anaplastic lymphoma kinase (ALK) homolog, a proto-oncogene receptor tyrosine kinase. scd-2 acts in a genetic pathway shown here to include the HEN-1 ligand, the RTK adaptor SOC-1, and the MAP kinase SMA-5. The SCD-2 pathway modulates TGF-beta signaling, which mediates the response to dauer pheromone, but SCD-2 might mediate a nonpheromone sensory input, such as food. CONCLUSIONS: Our studies identify a new sensory pathway controlling dauer formation and shed light on ALK signaling, integration of signaling pathways, and adaptation to extreme environmental conditions. PMID- 18674915 TI - Synthesis of stable and selective inhibitors of human galectins-1 and -3. AB - The syntheses of glycolytically stable galactosides and lactosides have been made toward the selective inhibition of human galectins-1 and -3. Transition metal catalyzed cross-coupling reactions were used to create carbon-carbon bond formation (Sonogashira, Suzuki, Heck, Glaser). Additionally, Hantzsch condensation was used to create novel 2-aminothiazoles which reacted with a panel of acylating and sulfonylating reagents. Moreover, dimeric galactosides and lactosides bearing triazoles, regiospecifically prepared using copper-catalyzed Huisgen azide-alkyne [1,3]-dipolar cycloaddition, provided efficient galectins-1 and -3 inhibitors. Best monovalent inhibitor among the tested series was (E) methyl 2-phenyl-4-(beta-D-galactopyranosyl)-but-2-enoate 15 with inhibitory potency of 313 microM against galectin-1 and best dimers were bis-lactoside 68 and 75 having both inhibitory properties of 160 microM against Galectin-3. PMID- 18674916 TI - 9-N-Substituted berberine derivatives: stabilization of G-quadruplex DNA and down regulation of oncogene c-myc. AB - A series of 9-N-substituted berberine derivatives (2a-j) were synthesized and evaluated as a new class of G-quadruplex binding ligands. G-quadruplex of DNA had been proven to be the transcription controller of human c-myc gene. The interaction of 9-N-substituted berberine derivatives with G-quadruplex DNA in c myc was examined via EMSA, CD spectroscopy, FRET-melting method, PCR-stop assay, competitive dialysis, cell proliferation assay, and RT-PCR assay. The experiment results indicated that these derivatives could selectively induce and stabilize the formation of intramolecular parallel G-quadruplex in c-myc, which led to down regulation of transcription of the c-myc in the HL60 lymphomas cell line. The related structure-activity relationships were also discussed. PMID- 18674917 TI - Synthesis, biological evaluation, and molecular modeling studies of methylene imidazole substituted biaryls as inhibitors of human 17alpha-hydroxylase-17,20 lyase (CYP17)--part II: Core rigidification and influence of substituents at the methylene bridge. AB - Thirty-five novel substituted imidazolyl methylene biphenyls have been synthesized as CYP17 inhibitors for the potential treatment of prostate cancer. Their activities have been tested with recombinant human CYP17 expressed in Escherichia coli. Promising compounds were tested for selectivity against CYP11B1, CYP11B2, and hepatic CYP enzymes 3A4, 1A2, 2B6 and 2D6. The core rigidified compounds (30-35) were the most active ones, being much more potent than Ketoconazole and reaching the activity of Abiraterone. However, they were not very selective. Another rather potent and more selective inhibitor (compound 23, IC(50)=345 nM) was further examined in rats regarding plasma testosterone levels and pharmacokinetic properties. Compared to the reference Abiraterone, 23 was more active in vivo, showed a longer plasma half-life (10h) and a higher bioavailability. Using our CYP17 homology protein model, docking studies with selected compounds were performed to study possible interactions between inhibitors and amino acid residues of the active site. PMID- 18674918 TI - Development of micromachining technology in ion microbeam system at TIARA, JAEA. AB - An ion-beam-lithography technique has been progressed in the microbeam systems at Japan Atomic Energy Agency (JAEA) Takasaki. In order to obtain a high-precision measure for microbeam size estimation with a high precision, we applied this technique combined with the electroplating process to make a Ni relief pattern as a resolution standard used in secondary electron imaging. As a result, the smallest beam size could be recorded. The scattering of ions in the materials influenced the spatial resolution and this is also discussed. PMID- 18674919 TI - Characteristics of focusing high-energy heavy ion microbeam system at the JAEA AVF cyclotron. AB - Ion optical analysis was made for a new focusing high-energy heavy ion microbeam system connected to the AVF cyclotron (K=110) at the accelerator facility, TIARA of JAEA Takasaki. The focusing performance of the microbeam system was estimated from both the calculation up to third-order term using TRANSPORT code and the measurement of beam resolution with the secondary electron imaging. As a result, a minimum beam size was evaluated at 0.56 and 0.62 microm in FWHM for the X and Y directions, respectively. The high-energy heavy ion microbeam system seemed to have been established as designed by the calculation with the TRANSPORT code, because it was confirmed that the calculation results was fairly reproduced by the measurement result. PMID- 18674920 TI - Investigation of the oxidation states of Pu isotopes in a hydrochloric acid solution. AB - The characteristics of the oxidation states of Pu in a hydrochloric acid solution were investigated and the results were applied to a separating of Pu isotopes from IAEA reference soils. The oxidation states of Pu(III) and Pu(IV) were prepared by adding hydroxylamine hydrochloride and sodium nitrite to a Pu stock solution, respectively. Also, the oxidation state of Pu(VI) was adjusted with concentrated HNO(3) and HClO(4). The stability of the various oxidation states of plutonium in a HCl solution with elapsed time after preparation were found to be in the following order: Pu(III) approximately Pu(VI)>Pu(IV)>Pu(V). The chemical recoveries of Pu(IV) in a 9M HCl solution with an anion exchange resin were similar to those of Pu(VI). This method for the determination of Pu isotopes with an anion exchange resin in a 9M HCl medium was applied to IAEA reference soils where the activity concentrations of (239,240)Pu and (238)Pu in IAEA-375 and IAEA 326 were consistent with the reference values reported by the IAEA. PMID- 18674921 TI - Complexation study on no-carrier-added astatine with insulin: a candidate radiopharmaceutical. AB - No-carrier-added astatine radionuclides produced in the (7)Li-irradiated lead matrix were separated from bulk lead nitrate target by complexing At with insulin, followed by dialysis. The method offers simultaneous separation of At from lead as well as its complexation with insulin. The At-insulin complex might be a potential radiopharmaceutical in the treatment of hepatocellular carcinoma. The stability of At-insulin complex was checked by dialysis against deionized water and Ringer lactate (RL) solution. It has been found that the half-life of At-insulin complex is about approximately 12h, when dialyzed against deionized water and is only 6h, when dialyzed against RL solution having the same composition as blood serum. The 6h half-life of this Insulin-At complex is perfect for killing cancer cells from external cell surfaces as the half-life of internalization of insulin molecule inside the cell is 7-12h. PMID- 18674922 TI - Plant pathogenesis-related (PR) proteins: a focus on PR peptides. AB - The novel classes of plant pathogenesis-related (PR) proteins identified during the last decade also include novel peptide families. This review specifically focuses on these pathogenesis-related peptides, including proteinase inhibitors (PR-6 family), plant defensins (PR-12 family), thionins (PR-13 family) and lipid transfer proteins (PR-14 family). For each family of PR peptides, the general features concerning occurrence, expression and possible functions of their members are described. Next, more specifically the occurrence of each PR peptide family in the model plant Arabidopsis thaliana is discussed. Single-gene studies performed on particular gene members of a PR peptide family are reported. In addition, expression data of yet undescribed gene members of that particular PR peptide family are presented by consultation of publicly available micro-array databases. Finally an update is provided on the potential role of these PR peptides in A. thaliana, with a focus on their possible involvement in plant defense. PMID- 18674923 TI - Ghost cell odontogenic carcinoma of the mandible: a case report demonstrating expression of tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor. AB - INTRODUCTION: Ghost cell odontogenic carcinoma is a rare neoplastic variant of calcifying odontogenic cyst, with aggressive growth characteristics. A painful swelling in the jaws with local paraesthesia is the most common symptom. Although it often causes irregular destruction of the adjacent bone, immunohistochemical expression of tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor has not previously been described in this carcinoma. CASE REPORT: This article describes a ghost cell odontogenic carcinoma affecting the mandible of a 55-year old man. The patient was treated by segmental mandibulectomy and there was no evidence of recurrence or metastasis for 1.8 years. Cytological features including the immunohistochemical expression of TRAP and vitronectin receptor were studied. CONCLUSION: Specimens revealed varying sized islands of anucleate cell clusters with homogenous, pale eosinophilic cytoplasm, so called ghost cells, admixed with sheets of tumour. TRAP and vitronectin receptor were detected in the ghost cells, but they were not expressed in the tumour cells. Our findings suggest that some of the cytokines produced by ghost cells may play important roles in causing extensive bone resorption in the ghost cell odontogenic carcinoma. PMID- 18674924 TI - Ewing's sarcoma of jaw bones in adult patients: 10-year experiences in a Paris university hospital. AB - INTRODUCTION: Despite using aggressive treatment, patients with Ewing's sarcoma (ES) always show a high recurrence and a low survival rate. Ki-67 has been used widely in surgical oncology. PATIENTS AND METHODS: This case report identified the Ki-67 expression in jaw bone ES from 4 adult patients operated upon between 1996 and 2005 in Pitie-Salpetriere University Hospital, Paris, France. The clinical data of each patient was also reviewed. RESULTS: Ki-67 reactivity was found in 3 cases. Two of 4 patients with 50% and 80% of Ki-67 positive tumour cells had local relapse at 5 years and 8 months after treatments, respectively. Furthermore, the patient with 80% Ki-67 expression exhibited resistance to chemotherapy and died a year after resection. The other 2 cases revealed no evidence of recurrence and metastasis to date. CONCLUSION: Ki-67 expression is likely to be associated with tumour recurrence and poor prognosis in jaw bone ES in adult patients. This marker probably helps surgeons to plan and employ appropriate treatment and/or surveillance for each patient; however, the number of cases in this series is very limited. A large-scale, prospective study is, therefore, required to confirm our suggestion. PMID- 18674925 TI - Effect of light on soluble guanylyl cyclase activity in Pharbitis nil seedlings. AB - Cyclic GMP acts as a chemical switch in plant cells to modulate cellular reactions. However, its metabolism has not been extensively explored and is still poorly understood. Previous experiments suggest that an endogenous cGMP system could participate in the mechanism of phytochrome controlled photoperiodic flower induction in Pharbitis nil. In order to gain further information on the role of cGMP, we have begun to study the enzyme of cGMP synthesis. In this article, the presence of the enzyme with guanylyl cyclase (GC) activity in soluble protein fractions of P. nil is reported. A large portion of the enzymatic activity is present in the cotyledons, where enzyme activity amounted to 0.45 pmol cGMP/min/mg protein. The enzyme exhibited a K(m) 0.5mM for GTP. A plot of 1/v versus 1/[GTP] was linear and V(max) was 0.74 pmol cGMP/min/mg protein. It was shown that the anti-sGC antibody recognise a 40 kDa protein. Moreover, the NO donor, sodium nitroprusside (SNP) and YC-1, as a NO-independent stimulator, enhanced enzyme activity. The NS 2028 (a potent GC inhibitor) treatments provoked a 3-fold reduction of the enzyme activity in comparison to the untreated fractions. Furthermore, the influence of light on GC activity was analysed. It was noted that cGMP level increased in cool white light, and darkness inhibited enzyme activity. Exposure to blue light acts to stimulate cGMP formation, whereas in red light a rapid decrease in GC activity was observed that returned to the high level when far-red light was applied after the red light treatment. The results presented in this work strongly argue that an enzyme with guanylyl cyclase activity is present in P. nil organs and its activity is controlled by light via the photoreceptors-dependent pathways. PMID- 18674926 TI - Advances in radiological imaging of gastrointestinal tumors. AB - This article summarizes the recent advances in radiological imaging in the hepatogastrointestinal field in terms of detection, characterization and staging of tumors. Introduction of multidetector row computed tomography technology has helped computed tomography to excel in its already established indications, and has expanded its capabilities by adding new clinical applications, such as computed tomography angiography, liver perfusion, computed tomography enterography or enteroclysis, and virtual colonoscopy. Contrast-enhanced ultrasound has dramatically changed the role of ultrasound in liver tumor characterization, detection, and other applications. Combining the advantages of unsurpassed soft tissue contrast and lack of ionizing radiation, magnetic resonance imaging of the gastrointestinal tract has become increasingly used clinically. Moreover, substantial progress in molecular and cellular magnetic resonance imaging has been achieved in the past few years. Particularly, the increasing number of available specific contrast agents dedicated to the evaluation of the liver and lymph nodes shows promises in the area of oncologic abdominal imaging. PMID- 18674927 TI - Imaging and cancer--I. PMID- 18674928 TI - Virus inactivation by solvent/detergent treatment using Triton X-100 in a high purity factor VIII. AB - Virus inactivation by solvent/detergent treatment using 0.3% tri-n-butyl phosphate and 1% Triton X-100 in the high purity factor VIII concentrate Replenate has been investigated. A wide range of model enveloped viruses were confirmed to be inactivated by >4 to >6 log after 30 min at 22 degrees C under standard conditions. Using Sindbis as a representative enveloped virus, the effect of various parameters on the inactivation process was tested. Virus inactivation was confirmed to be effective in different batches of product and was not influenced by changing the process conditions with regard to protein and salt concentration or pH. Virus inactivation was effective even at a temperature as low as 4-5 degrees C. Although solvent/detergent concentration was the most critical parameter, a concentration as low as 0.15% TnBP/0.5% Triton X-100 was still completely effective. At a lower concentration an extended incubation period was required. These studies demonstrate the robustness of this solvent/detergent procedure based on Triton X-100 and allow suitable process limits to be set for this manufacturing step. PMID- 18674929 TI - An overview of animal cell substrates for biological products. AB - The issue of which cells to use as substrates for the production of biological products, and especially vaccines, has been with us in one form or another ever since the development of cell cultures in the 1950s. The major cell substrate events that occurred over the past 50 years are reviewed briefly. Although numerous conferences were held during that period, incomplete resolution of some cell substrate issues has remained. Specifically, the potential oncogenicity of cellular DNA derived from continuous cell lines, and the tests that are used to rule out the presence of adventitious agents have been recognized as areas that could benefit greatly from studies using state-of-the-art techniques. A collaborative effort involving WHO, NIAID, and IABS resulted from consensus recommendations of a 2004 conference, and the prospects for revised guidance in the near future on the characterization and use of animal cell substrates are bright. PMID- 18674930 TI - Pattern of osteophyte distribution in primary osteoarthritis of the elbow. AB - The goal of this study was to look at the pattern of osteophyte distribution on a 3-dimensional computed tomography scan of patients with symptomatic osteoarthritis in the elbow. We recruited 22 consecutive patients with symptomatic osteoarthritis of the elbow for the study. Three-dimensional reconstructed anterior, posterior, medial, and lateral views of the elbow were reviewed. Ulnohumeral osteophytes were found in 21 patients (95%), and radiohumeral osteophytes were found in 13 (59%). Cadaveric and biomechanical studies suggest that the radiohumeral joint appears to be more prone to wear and stress than the ulnohumeral joint. Our study showed that the percentage of patients with ulnohumeral joint osteophytes (95%) was higher than that of radiohumeral joint (59%). Therefore, this study challenges the conventional belief that osteoarthritis starts from the radiohumeral joint and progress toward the ulnohumeral joint. PMID- 18674931 TI - Clinical review of chondroitin sulfate in osteoarthritis. AB - Symptomatic slow-acting drugs for the treatment of osteoarthritis (SYSADOA; OA) are compounds which are prescribed as drugs in European countries since many years, whereas they are sold as nutraceuticals in USA. In Europe, the publication of the EULAR Recommendations for the Treatment of Knee OA in 2003 has listed oral chondroitin sulfate (CS) as evidence 1A and strength of recommendation A which represents the highest level for a therapeutic strategy. Symptomatic slow-acting drugs are intended to be used as ground therapy for OA; these compounds are not rapidly acting agents such as Non Steroidal Anti-Inflammatory Drugs (NSAIDs), and their clinical efficacy on algo-functional symptoms can only be demonstrated after a couple of weeks of regular intake. Interestingly, once the administration is stopped, they do show carry-over effects of various durations, from about 3 months with the oral formulations to 6-9 months with intra-articular formulations. The main rationale behind the use of the SYSADOA therapeutic class is the reduction of NSAIDs in the overall drug management of OA disease and therefore consequently to limit the very significant risks of upper Gastro intestinal (GI) tract erosions, ulcers with bleeding and/or deleterious renal effects in elderly patients. The evidence for clinical efficacy of oral CS as a drug able to significantly improve the algo-functional symptoms of OA disease does come from a set of randomized clinical studies published a couple of years ago. Indeed, it was demonstrated that the drug was effective in knee and finger OA, whereas previous data suggested that hip OA patients could also benefit from it. In addition, oral CS supported the comparison with NSAIDs such as diclofenac sodium in a medium/long-term clinical study in patients with knee OA. A dose finding study in patients with knee OA did provide strong data supporting the administration of 800 mg of CS orally which had nearly the same effects as 1200 mg/day, whereas the use of a sequential 3 months administration mode, twice a year was also shown to provide the same results as a continuous treatment. The good tolerability and safety aspects of oral CS were largely documented in these CTs. Taking these important points into account, we definitively have enough clinical data available supporting the view that oral CS is a valuable and safe symptomatic treatment for OA disease. More recent data based on a couple of previous trials and two pivotal studies do provide further evidence that oral CS does also have structure-modifying effects in knee OA patients. A couple of other compounds such as hyaluronan, diacerein, avocado and soya unsaponifiables, doxycycline have also been tested with respect to their potential disease modifying effects. Additional compounds including receptor activator of NF-kappaB (RANK) ligand inhibitors, cathepsin K inhibitors, bisphosphonates are further assessed regarding their potential structure-modifying effect. PMID- 18674932 TI - Autologous conditioned serum (Orthokine) is an effective treatment for knee osteoarthritis. AB - OBJECTIVE: Osteoarthritis (OA) is prevalent and difficult to treat. Autologous conditioned serum (ACS), marketed under the trade name Orthokine, is a novel, injectable antiarthritic derived from the patient's own blood. The present study is the first time ACS has undergone a controlled clinical trial. METHOD: We investigated 376 patients with knee OA in a prospective, randomized, patient- and observer-blinded, placebo-controlled trial using an intention-to-treat analysis (ITT). The clinical effects of ACS were compared to hyaluronan (HA) and saline (placebo) as assessed by patient-administered outcome instruments (Western Ontario and McMaster Universities osteoarthritis index, global patient assessment, visual analog scale, Short-Form 8) after 7, 13 and 26 weeks. After 104 weeks an observer-blinded follow-up was carried out. Frequency and severity of adverse events were used as safety parameters. RESULTS: In all treatment groups, intra-articular injections produced a reduction in symptoms as well as an improvement in quality of life. However, the effects of ACS were significantly superior to those of HA and saline for all outcome measures and time points, and improvements were clinically relevant; there were no differences between the effects of HA and saline. The frequency of adverse events was comparable in the ACS and saline groups, but higher in the HA group. CONCLUSION: The data demonstrate that ACS injection considerably improves clinical signs and symptoms of OA. It remains to be determined whether ACS is disease-modifying, chondroprotective, or chondroregenerative. PMID- 18674933 TI - E2A proteins promote development of lymphoid-primed multipotent progenitors. AB - The first lymphoid-restricted progeny of hematopoietic stem cells (HSCs) are lymphoid-primed multipotent progenitors (LMPPs), which have little erythromyeloid potential but retain lymphoid, granulocyte, and macrophage differentiation capacity. Despite recent advances in the identification of LMPPs, the transcription factors essential for their generation remain to be identified. Here, we demonstrated that the E2A transcription factors were required for proper development of LMPPs. Within HSCs and LMPPs, E2A proteins primed expression of a subset of lymphoid-associated genes and prevented expression of genes that are not normally prevalent in these cells, including HSC-associated and nonlymphoid genes. E2A proteins also restricted proliferation of HSCs, MPPs, and LMPPs and antagonized differentiation of LMPPs toward the myeloid fate. Our results reveal that E2A proteins play a critical role in supporting lymphoid specification from HSCs and that the reduced generation of LMPPs underlies the severe lymphocyte deficiencies observed in E2A-deficient mice. PMID- 18674934 TI - A synaptic basis for paracrine interleukin-2 signaling during homotypic T cell interaction. AB - T cells slow their motility, increase adherence, and arrest after encounters with antigen-presenting cells (APCs) bearing peptide-MHC complexes. Here, we analyzed the cell-cell communication among activating T cells. In vivo and in vitro, activating T cells associated in large clusters that collectively persisted for >30 min, but they also engaged in more transient interactions, apparently distal to APCs. Homotypic aggregation was driven by LFA-1 integrin interactions. Ultrastructural analysis revealed that cell-cell contacts between activating T cells were organized as multifocal synapses, and T cells oriented both the microtubule-organizing complex and interleukin-2 (IL-2) secretion toward this synapse. T cells engaged in homotypic interactions more effectively captured IL-2 relative to free cells. T cells receiving paracrine synaptic IL-2 polarized their IL-2 signaling subunits into the synaptic region and more efficiently phosphorylated the transcription factor STAT5, likely through a synapse associated signaling complex. Thus, synapse-mediated cytokine delivery accelerates responses in activating T cells. PMID- 18674936 TI - Effect of presentation bias on selection of treatment option for failed endodontic therapy. AB - OBJECTIVE: The purpose of the study was to determine if treatment option presentation bias affects decision making by patients when they have failed endodontic therapy. STUDY DESIGN: First-year dental students simulated the role of patients. All students were given the same scenario of a symptomatic tooth with failed endodontic therapy and asked to select from between 2 treatment options: nonsurgical endodontic retreatment or extraction and implant placement. One half of the students had characterizations of the treatment options biased toward nonsurgical retreatment, and the other half had characterizations of the treatment options biased toward extraction and implant selection. Statistical analysis was performed with chi-squared test. RESULTS: Biased presentations significantly influenced the treatment selection by the students (P = .0006). CONCLUSION: If treatment options are presented in a biased manner to favor one option, the patient is more likely to select that treatment option. PMID- 18674935 TI - A bony fish immunological receptor of the NITR multigene family mediates allogeneic recognition. AB - Novel immune-type receptors (NITRs) comprise an exceptionally large, diversified family of activating and inhibitory receptors that has been identified in bony fish. Here, we characterized the structure of an activating NITR that is expressed by a cytotoxic natural killer (NK)-like cell line and that specifically binds an allogeneic B cell target. A single amino acid residue within the NITR immunoglobulin variable (V)-type domain accounts for specificity of the interaction. Structures solved by X-ray crystallography revealed that the V-type domains of NITRs form homodimers resembling rearranging antigen-binding receptor heterodimers. CDR1 elements of both subunits of NITR dimers form ligand-binding surfaces that determine specificity for the nonself target. In the evolution of immune function, it appears that a specific NK type of innate recognition may be mediated by a complex germline multigene family of V structures resembling those that are somatically diversified in adaptive immunological responses. PMID- 18674937 TI - Management considerations in reconstruction of postablative defects of the mandible: vertical distraction of a scapular bone flap and removable lip support: a case report. AB - OBJECTIVE: To describe vertical distraction osteogenesis of a scapular flap and removable lip support for oral rehabilitation after surgical creation of an ablative defect of the mandible. CASE REPORT: A 70-year-old man who was diagnosed with squamous cell carcinoma of the left lower gingiva underwent segmental mandibulectomy for tumor ablation and reconstruction with a scapular bone flap. To augment bone height of this flap, vertical distraction osteogenesis was performed. After denture fabrication, a removable lip support was placed between the implant-supported denture and the lower lip. RESULTS: The bone height of the scapular bone flap increased by 9 mm. Implants with adequate length could be placed in the distracted bone. The lip support was effective. Two years after masticatory loading, the implants remained stable. CONCLUSION: Vertical distraction osteogenesis of the scapular bone flap was suitable to facilitate postoperative functional and esthetic restoration after tumor resection. A removable lip support was also useful as a supplementary tool for oral rehabilitation. PMID- 18674938 TI - Veterinary science and the Internet: tangible help for a better understanding of equine atypical myopathy. PMID- 18674939 TI - Inherited disease in the horse: mapping complex disease variants is on the horizon. PMID- 18674940 TI - Frequently encountered cranial ultrasound features in the white matter of preterm infants: correlation with MRI. AB - BACKGROUND: Bilateral symmetrical echogenic and echolucent areas in the white matter are frequently seen on the cranial ultrasound scans of apparently well preterm infants without overt pathology. AIM: To determine whether these features reflect maturational processes as seen on MRI. METHODS: Preterm and term-born infants without overt pathology on contemporaneous brain ultrasound and MRI were studied. Ultrasound scans were compared with T(2)-weighted MRI to identify MR correlates for the bilateral and symmetrical echogenic and echolucent phenomena in the white matter seen on ultrasound. RESULTS: Forty-four sets of scans (26 preterm, 8 term-born infants) were assessed. Echogenic features were better and more frequently seen on early ultrasound as compared to nearer term age. Echogenic blushes in the white matter correlated well with high signal intensity areas and echogenic lines with low signal intensity lines on MRI. Echolucent areas correlated with the site of the internal capsule and the myelinated posterior pons. The subplate was not reliably identified. CONCLUSION: Many echogenic and echolucent features in the white matter of well preterm and some term-born infants correlated well with areas of differing signal intensity on MRI. They most likely reflect normal maturational processes but the echogenic hemispheric features may represent delayed or abnormal maturation. PMID- 18674941 TI - Relaxation-based distance measurements between a nitroxide and a lanthanide spin label. AB - Distance measurements by electron paramagnetic resonance techniques between labels attached to biomacromolecules provide structural information on systems that cannot be crystallized or are too large to be characterized by NMR methods. However, existing techniques are limited in their distance range and sensitivity. It is anticipated by theoretical considerations that these limits could be extended by measuring the enhancement of longitudinal relaxation of a nitroxide label due to a lanthanide complex label at cryogenic temperatures. The relaxivity of the dysprosium complex with the macrocyclic ligand DOTA can be determined without direct measurements of longitudinal relaxation rates of the lanthanide and without recourse to model compounds with well defined distance by analyzing the dependence of relaxation enhancement on either temperature or concentration in homogeneous glassy frozen solutions. Relaxivities determined by the two calibration techniques are in satisfying agreement with each other. Error sources for both techniques are examined. A distance of about 2.7 nm is measured in a model compound of the type nitroxide-spacer-lanthanide complex and is found in good agreement with the distance in a modeled structure. Theoretical considerations suggest that an increase of the upper distance limit requires measurements at lower fields and temperatures. PMID- 18674942 TI - Meningococcal meningitis in Mali: a long-term study of persistence and spread. AB - OBJECTIVES: Meningococcal meningitis (MM) is still a huge threat in the African meningitis belt. To fight against epidemics, a strengthened health information system, based upon weekly collected data, was set up in Mali. We aimed to study the spatio-temporal dynamics of MM in this country between 1992 and 2003. METHODS: We were first interested in the impact of population size on the disease persistence. We then used cross-correlation analysis to study the spread of the disease on three different spatial scales, i.e., inter-region (global) and inter district and intra-district (local) levels. RESULTS: We found no persistence of MM at district level in Mali during the whole of the study period. However, we found persistence on a nationwide scale after the 1997 big epidemics, as opposed to the 1992-1996 time periods. In terms of spread, two main regions seem to lead MM dynamics in Mali, even if on a local scale the 'cities-villages' diffusion pattern was not systematically observed. CONCLUSIONS: This study improves knowledge on the spread and persistence of MM in Mali in recent years. It constitutes a first spatial study describing persistence and spread of MM in an African meningitis belt country. The next step should be the integration of vaccination and genetic variability data to clarify the route of spread of the disease in the human population. PMID- 18674943 TI - Multiple septic complications of disseminated candidiasis following anorectal sepsis in a non-immunocompromised patient. PMID- 18674944 TI - Serum levels of MMP-3 and cathepsin K in patients with ankylosing spondylitis: effect of TNFalpha antagonist therapy. AB - OBJECTIVE: To measure serum levels of MMP-3 and cathepsin K in patients with ankylosing spondylitis (AS) and in controls and to look for changes in these variables during TNFalpha antagonist therapy. METHODS: We prospectively studied a group of patients who met New York criteria for AS and a group of healthy volunteers. We recorded age, disease duration, main features of the disease, BASDAI, erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP). Serum MMP-3 and cathepsin K were assayed in duplicate using ELISA kits (Quantikine MMP 3, R&D Systems; and Cathepsin K, Biomedica). We also assayed IL-17 (Quantikine IL 17, R&D Systems) and BMP-7 (human BMP-7 DuoSet, R&D Systems). In patients treated with TNFalpha antagonists, the assays were repeated 10 weeks after treatment initiation. The Mann-Whitney test was used for between-group comparisons and the Wilcoxon test for evaluations of changes under treatment. Correlation testing was performed. P-values less than 0.05 were considered significant. RESULTS: We studied 23 outpatients with AS and 21 controls, with mean age of 39.9 years and 41.2 years, respectively (NS). Disease duration was 10.1 years (1.3); most patients had axial disease (n=21) and carried HLA-B27 (n=19). At baseline, the mean BASDAI was 44.1 mm (4.1) and the mean CRP level was 22.3 mg/L (4.7). Serum MMP-3 levels were significantly higher in the patients than in the controls (4.71 vs. 2.79 ng/ml, P=0.04); levels were also higher for cathepsin K (6.4 vs. 3.6 pg/ml) and IL-17 (60.4 vs. 32 pg/ml), but the differences were not statistically significant. No difference was noted for BMP-7. The only positive correlation was between the ESR and the CRP level (P=0.0002). Thirteen patients were evaluated 10 weeks into TNFalpha antagonist therapy (adalimumab, n=7; etanercept, n=4; or infliximab, n=2). Serum MMP-3 decreased significantly (P=0.04); significant decreases were also noted for the ESR, CRP, and BASDAI. CONCLUSION: MMP-3 is significantly increased in patients with active AS but fails to correlate significantly with conventional variables used to assess disease activity. TNFalpha antagonist therapy induces a significant decrease in MMP-3 levels, together with decreases in conventional variables (ESR, CRP, and BASDAI). MMP-3 may be a biomarker for disease activity in AS but supplies no additional information to the clinician. PMID- 18674945 TI - Herpes zoster in patients taking TNFalpha antagonists for chronic inflammatory joint disease. AB - OBJECTIVE: To assess the rate of occurrence and outcomes of herpes zoster in patients taking TNFalpha antagonists. METHODS: Retrospective review of the medical records of 300 patients who received TNFalpha antagonists to treat chronic inflammatory joint disease. RESULTS: We identified 9 (9/300, 3%) patients who experienced herpes zoster, 6 women and 3 men, with rheumatoid arthritis (n=7) or ankylosing spondylitis (n=2). The drug was infliximab in 4 patients, adalimumab in 2 patients, and etanercept in 3 patients, including 2 patients with a prior history of infliximab therapy (for 12 and 36 months, respectively). Mean treatment duration at the occurrence of herpes zoster was 27 months (range, 6-42 months). DISCUSSION: Glucocorticoid therapy (n=7) and methotrexate therapy (n=6) were the only risk factors identified in our study. Mean follow-up was 26 months. All 9 patients achieved a full recovery with antiviral treatment and interruption of the TNFalpha antagonist. One patient experienced a recurrence after resuming TNFalpha antagonist therapy. CONCLUSION: The scant data in the literature suggest a higher risk of herpes zoster with anti-TNFalpha antibodies than with the soluble receptor. The role for concomitant treatments (glucocorticoids and methotrexate) should be taken into account. PMID- 18674946 TI - Efficacy of anakinra in a patient with refractory relapsing polychondritis. PMID- 18674947 TI - Mycobacterial coxitis masked by concomitant staphylococcal infection after infliximab for Crohn's disease. PMID- 18674948 TI - Ultrasonography for diagnosing atypical gout. Two case reports. AB - Gout is characterized by the deposition of monosodium urate crystals on the surface of the articular cartilage, within periarticular tissues, and within bone and skin. The diagnosis rests on identification of the crystals in joint fluid or a tophus. However, joint aspiration is not always feasible, and the presentation may be atypical. We describe two cases of chronic gouty arthritis misdiagnosed as psoriatic arthritis. Ultrasonography of the bone and joints disclosed two patterns recently described as highly suggestive of gout, namely, the double contour appearance of the cartilage and the snowstorm appearance of the synovial membrane. In addition, ultrasonography was useful for guided aspiration of joint fluid or other material containing monosodium urate crystals. Thus, ultrasonography may contribute to improve the diagnosis and treatment of gout. PMID- 18674949 TI - Teenage pregnancy-new tools to support local health campaigns. AB - Teenage pregnancy has remained high in many inner city areas despite several years of campaigns to reduce numbers and to support young people and their families tackle the problem. In this paper we propose new methods to focus local strategies on high-risk areas as well as ranking secondary schools and GP practices most likely to be in contact with young people at risk. The proposed methods proved successful in engaging local schools in a new campaign and have provided a framework for evaluation of local teenage pregnancy rates in years to come. PMID- 18674950 TI - Actinic cheilitis: epithelial expression of COX-2 and its association with mast cell tryptase and PAR-2. AB - Cyclooxygenase-2 (COX-2) is overexpressed in various types of human malignancies, including oral cancers. Recent studies have shown that mast cell-derived protease tryptase can induce COX-2 expression by the cleavage of proteinase-activated receptor-2 (PAR-2). Actinic cheilitis (AC) is a premalignant form of lip cancer characterized by an increased density of tryptase-positive mast cells. To investigate the possible contribution of tryptase to COX-2 overexpression during early lip carcinogenesis, normal lip (n=24) and AC (n=45) biopsies were processed for COX-2, PAR-2 and tryptase detection, using RT-PCR and immunohistochemistry. Expression scores were obtained for each marker and tested for statistical significance using Mann-Whitney and Spearmann's correlation tests as well as multivariate logistic regression analysis. Increased epithelial co-expression of COX-2 and PAR-2, as well as, elevated subepithelial density of tryptase-positive mast cells were found in AC as compared to normal lip (P<0.001). COX-2 overexpression was found to be a significant predictor of AC (P<0.034, forward stepwise, Wald), and to be correlated with both tryptase-positive mast cells and PAR-2 expression (P<0.01). The results suggest that epithelial COX-2 overexpression is a key event in AC, which is associated with increased tryptase positive mast cells and PAR-2. Therefore, tryptase may contribute to COX-2 up regulation by epithelial PAR-2 activation during early lip carcinogenesis. PMID- 18674951 TI - Detection of constituent layers of histological oral sub-mucous fibrosis: images using the hybrid segmentation algorithm. AB - This paper concentrates on the segmentation of histological images of oral sub mucous fibrosis (OSF) into its constituent layers. In this regard hybrid segmentation algorithm shows very interesting results. The segmentation results depict the superiority of hybrid segmentation algorithm (HSA) in comparison to region growing algorithm (RGA). In clinical sense, the presented method provides an automatic means for segmenting histological layers (reference class map provided by the expert). The method shows potential in mimicking clinical acumen to act as a support system to oncologist. PMID- 18674952 TI - Current concepts in management of oral cancer--surgery. AB - Oral cancer is the sixth most common cancer worldwide, with a high prevalence in South Asia. Tobacco and alcohol consumption remain the most dominant etiologic factors, however HPV has been recently implicated in oral cancer. Surgery is the most well established mode of initial definitive treatment for a majority of oral cancers. The factors that affect choice of treatment are related to the tumor and the patient. Primary site, location, size, proximity to bone, and depth of infiltration are factors which influence a particular surgical approach. Tumors that approach or involve the mandible require specific understanding of the mechanism of bone involvement. This facilitates the employment of mandible sparing approaches such as marginal mandibulectomy and mandibulotomy. Reconstruction of major surgical defects in the oral cavity requires use of a free flap. The radial forearm free flap provides excellent soft tissue and lining for soft tissue defects in the oral cavity. The fibula free flap remains the choice for mandibular reconstruction. Over the course of the past thirty years there has been improvement in the overall survival of patients with oral carcinoma largely due to the improved understanding of the biology of local progression, early identification and treatment of metastatic lymph nodes in the neck, and employment of adjuvant post-operative radiotherapy or chemoradiotherapy. The role of surgery in primary squamous cell carcinomas in other sites in the head and neck has evolved with integration of multidisciplinary treatment approaches employing chemotherapy and radiotherapy either sequentially or concurrently. Thus, larynx preservation with concurrent chemoradiotherapy has become the standard of care for locally advanced carcinomas of the larynx or pharynx requiring total laryngectomy. On the other hand, for early staged tumors of the larynx and pharynx, transoral laser microsurgery has become an effective means of local control of these lesions. Advances in skull base surgery have significantly improved the survivorship of patients with malignant tumors of the paranasal sinuses approaching or involving the skull base. Surgery thus remains the mainstay of management of a majority of neoplasms arising in the head and neck area. Similarly, the role of the surgeon is essential throughout the life history of a patient with a malignant neoplasm in the head and neck area, from initial diagnosis through definitive treatment, post treatment surveillance, management of complications, rehabilitation of the sequelae of treatment, and finally for palliation of symptoms. PMID- 18674953 TI - Gypenosides induced G0/G1 arrest via CHk2 and apoptosis through endoplasmic reticulum stress and mitochondria-dependent pathways in human tongue cancer SCC-4 cells. AB - Gypenosides (Gyp), a component of Gynostemma pentaphyllum Makino, was selected for examining the effects on the cell viability, cell cycle and induction of apoptosis in human tongue cancer SCC-4 cells. Gyp induced cytotoxicity (decreased the percentage of viable cells) in SCC-4 cells appeared to be associated with induction of cell cycle arrest (G0/G1 arrest), apoptotic cell death based on Gyp induced morphological changes and DNA fragmentation and increased the sub-G1 group in examined SCC-4 cells. The production of reactive oxygen species and Ca(2+) and the depolarization of mitochondrial membrane potential were observed, dose- and time-dependently, after treatment of SCC-4 cells with various concentrations of Gyp. Gyp inhibited the levels of the anti-apoptotic proteins Bcl-2 and Bcl-xl, but promoted the levels of the pro-apoptotic protein Bax. Western blotting showed the releases of cytochrome c and Endo G and both were also confirmed by confocal laser microscopic systems. The GADD153 moved to nuclei (nuclear translocation). In conclusion, Gyp induced ER stress and production of reactive oxygen species and Ca(2+), change the ratio of Bcl-2 and Bax, followed by the dysfunction of mitochondria, caused cytochrome c release, activation of caspase-3 before leading to apoptosis. These results provide information towards an understanding of the mechanisms by which Gyp induces cell cycle arrest and apoptosis in human tongue cancer cells. PMID- 18674954 TI - Potentially malignant disorders of the oral and oropharyngeal mucosa; terminology, classification and present concepts of management. AB - In a recently held WHO workshop it has been recommended to abandon the distinction between potentially malignant lesions and potentially malignant conditions and to use the term potentially malignant disorders instead. Of these disorders, leukoplakia and erythroplakia are the most common ones. These diagnoses are still defined by exclusion of other known white or red lesions. In spite of tremendous progress in the field of molecular biology there is yet no single marker that reliably enables to predict malignant transformation in an individual patient. The general advice is to excise or laser any oral of oropharyngeal leukoplakia/erythroplakia, if feasible, irrespective of the presence or absence of dysplasia. Nevertheless, it is actually unknown whether such removal truly prevents the possible development of a squamous cell carcinoma. At present, oral lichen planus seems to be accepted in the literature as being a potentially malignant disorder, although the risk of malignant transformation is lower than in leukoplakia. There are no means to prevent such event. The efficacy of follow-up of oral lichen planus is questionable. Finally, brief attention has been paid to oral submucous fibrosis, actinic cheilitis, some inherited cancer syndromes and immunodeficiency in relation to cancer predisposition. PMID- 18674955 TI - Gene polymorphisms related to angiogenesis, inflammation and thrombosis that influence risk for oral cancer. AB - Genetic association studies have implicated functional DNA polymorphisms in genes encoding factors related to angiogenesis, inflammation and thrombosis with increased risk for oral squamous cell carcinoma (OSCC). This study examines possible interactions between nine such genotype polymorphisms and their combinatory effect in assessing the OSCC risk in a European population. OSCC cases (N=162) and healthy controls (N=168) of comparable age, gender, and ethnicity (Greeks and Germans) were studied. Multivariate logistic regression models were constructed in order to assess the contribution of homozygous or heterozygous variant genotypes of polymorphisms MMP-1 (-1607 1G/2G), MMP-3 (-1171 5A/6A), MMP-9 (-1562C/T), TIMP-2 (-418C/G), VEGF (+936C/T), GPI-alpha (+807C/T), PAI-1 (4G/5G), ACE (intron 16D/I) and TAFI (+325C/T) upon overall, early and advanced stages of OSCC. Four out of nine polymorphisms affecting PAI-1, MMP-9, TIMP-2 and ACE expression contributed significantly in OSCC prediction in the various logistic regression models. Based on these findings and previous reports, possible interactions of the implicated factors leading to OSCC development, as well as an algorithm of risk estimation are discussed. PMID- 18674956 TI - Lifestyle risk factors for oral cancer. AB - The "style of life is the unique way in which individuals try to realize their fictional final goal and meet or avoid the three main tasks of life: work, community, love" (Alfred Adler, founder of the Individual Psychology). Lifestyle refers to the way individuals live their lives and how they handle problems and interpersonal relations. The lifestyle behaviours associated to oral cancer with convincing evidence are tobacco use, betel quid chewing, alcohol drinking, low fruit and vegetable consumption (the detrimental lifestyle is high fat and/or sugar intake, resulting in low fruit and/or vegetable intake). Worldwide, 25% of oral cancers are attributable to tobacco usage (smoking and/or chewing), 7-19% to alcohol drinking, 10-15% to micronutrient deficiency, more than 50% to betel quid chewing in areas of high chewing prevalence. Carcinogenicity is dose-dependent and magnified by multiple exposures. Conversely, low and single exposures do not significantly increase oral cancer risk. These behaviours have common characteristics: (i) they are widespread: one billion men, 250 million women smoke cigarettes, 600-1200 million people chew betel quid, two billion consume alcohol, unbalanced diet is common amongst developed and developing countries; (ii) they were already used by animals and human forerunners millions of years ago because they were essential to overcome conditions such as cold, hunger, famine; their use was seasonal and limited by low availability, in contrast with the pattern of consumption of the modern era, characterized by routine, heavy usage, for recreational activities and with multiple exposures; (iii) their consumption in small doses is not recognized as detrimental by the human body and activates the dopaminergic reward system of the brain, thus giving instant pleasure, "liking" (overconsumption) and "wanting" (craving). For these reasons, effective Public Health measures aimed at preventing oral cancer and other lifestyle-related conditions fail to realize their final goal to eradicate these lifestyles. Following Adler's theory and the principles of the "Ottawa Charter for Health Promotion", conditions such as education, sustainable resources, social justice, and equity must be satisfied before the implementation of physical health promotion campaigns. PMID- 18674957 TI - PTCH1 isoforms in odontogenic keratocysts. AB - Odontogenic keratocyst (OKC) is an aggressive benign odontogenic neoplasm associated with PTCH1 alteration. PTCH1 has several isoforms generated by use of different first exon (1b, 1d and 1e). These isoforms code for proteins with different functions, expression profiles and transcriptional regulation. The aim of the present study was to investigate the expression of PTCH1 first exons in OKC tumors to shed light on scenery whereby PTCH1 coordinates OKC tumorigenesis. Forty OKC, including 12 sporadic and 28 associated with Nevoid Basal Cell Carcinoma Syndrome (NBCCS), were included in the study. The variants 1b, 1d and 1e were investigated by RT-PCR. The exon 1b was detected in 90% of OKC and none of the dental follicle (control). Most of the OKC, sporadic and syndromic, and all of the samples of dental follicles demonstrated the expression of 1d mRNA. All primary OKC had 1b mRNA while 4 (24%) lesions marsupialized lost 1b expression. In addition, the pattern of exon 1 expression observed in oral mucosa adjacent to the OKC was similar to the OKC tumor. In conclusion, this report showed overactivity of Hedgehog (HH) pathway in OKC lesion and at adjacent oral mucosa. We also demonstrated that marsupialization could alter PTCH1 variants profiling in some OKC cases. PMID- 18674958 TI - Applying the molecular biology and epigenetics of head and neck cancer in everyday clinical practice. AB - During the past decade, there has been a significant increase in knowledge regarding the molecular biology and epigenetics of head and neck squamous cell carcinoma (HNSCC). This has been aided by the steady development of new technology and novel techniques aimed at elucidating additional aberrant molecular alterations characteristic of HNSCC, including the advent of high throughput assays and the development of more sophisticated bioinformatics tools. In addition, advancements in the field of cancer epigenetics and microRNA have increased the complexity of understanding HNSCC tumorigenesis. These advances have lead to an increasing number of translational studies in the diagnosis, prognosis, and treatment of head and neck cancer. The end result is that molecular biomarkers, gene detection panels and targeted therapeutics are becoming a reality for the care of patients with HNSCC. In this article, we will focus on the many implications of this research as it pertains to clinical practice and the treatment of HNSCC patients. PMID- 18674959 TI - Survival following primary surgery for oral cancer. AB - The main aims of this article are to report the overall and disease-specific survival of a consecutive series of patients presenting with oral cancer from 1992 to 2002 and to relate survival to clinical and pathological factors. The article uses population-based age-sex mortality rates in the North-West of England to highlight differences in overall and disease-specific survival. 541 patients with oral squamous cell carcinoma presented to the Regional Maxillofacial Unit from 1992 to 2002. Curative treatment favoured radical primary surgery, 10% (52) received primary radiotherapy. These patients were on average 8 years older with more advanced tumours and overall poorer survival at 5 years, 23% (SE 7%). The remainder of the results refer to 489 patients who had primary curative surgery, 40% (194) of whom received adjuvant radiotherapy. The overall survival (OS) was 56% (SE 2%) and the disease-specific survival (DSS) was 74% (SE 2%). There was a local recurrence rate of 10% (50) and the loco-regional recurrence rate was 21% (103). The second primary rate was 7% (35). Survival figures had improved over the 10-year period from 63% DSS for the first 4 years of the study (1992-1995) compared to 81% for the last 3 years (2000-2002). In stepwise Cox regression the two predictors selected for disease-specific survival were pN status and margins (both p<0.001). Age-sex mortality rates for the North West indicate that 15.0% of the 489 primary surgery patients might have been expected to die within 5 years if they were typical of the general population and the observed difference between all causes and oral-cancer specific survival was 18.3%. These data emphasise the value of disease-specific survival as an indicator of successful treatment in a cohort that tends to be elderly, from social deprived backgrounds, with life styles and comorbidity that influence overall survival. PMID- 18674961 TI - A non-invasive XRF study supported by multivariate statistical analysis and reflectance FTIR to assess the composition of modern painting materials. AB - The palette used in two paintings by Paul Cezanne, L'etang des soeurs dated c. 1875 and La route tournante, made in the last year of his life (1902), were analyzed using non-invasive spectroscopic methods. X-ray fluorescence combined with principal components analysis (PCA) and supported by reflectance near- and mid-FTIR was shown to be a powerful analytical tool to draw conclusions about the chemical identification of inorganic materials in paintings. Pigments and fillers such us Thenard's blue, Prussian blue, red ochre, kaolin, vermilion, lead white, zinc white and barium sulphate, were identified. Evidence for three different pigments, namely a copper arsenite pigment, chrome green (a mixture of chrome yellow and Prussian blue) and viridian has been obtained by the PCA analysis of elemental compositions of green hues. PMID- 18674960 TI - Molecular techniques and genetic alterations in head and neck cancer. AB - It is well known that cellular DNA alterations can lead to the formation of cancer, and there has been much discovery in the pathways involved in the development of head and neck squamous cell carcinoma (HNSCC). With novel genome wide molecular assays, our ability to detect these abnormalities has increased. We now have a better understanding of the molecular complexity of HNSCC, but there is still much research to be done. In this review, we discuss the well described genetic alterations and touch on the newer findings, as well as some of the future directions of head and neck cancer research. PMID- 18674962 TI - Determination of dextran sulfate sodium with crystal violet by triple-wavelength overlapping resonance Rayleigh scattering. AB - A triple-wavelength overlapping resonance Rayleigh scattering (TWO-RRS) method was developed to detect dextran sulfate sodium (DSS) with crystal violet (CV). At pH 10.0 Britton Robinson buffer solution medium, the interaction of CV with DSS occurred which greatly enhanced the RRS intensity with the new RRS peaks appearing at 340, 501 and 671 nm and all these three peaks enhanced with the increase of DSS concentration in the range of 0.04-2.5 microg ml(-1) and the detection limit for the three single peaks was 0.024, 0.027, and 0.027 microg ml( 1), respectively, whereas that of the TWO-RRS method was 0.013 microg ml(-1). The TWO-RRS method was found to have much better flexibility and high sensitivity than the single-wavelength method. In this paper, the interaction conditions were optimized. The affecting factors and characteristics of RRS for the interaction of DSS with CV were investigated and a sensitive method for the determination of trace amounts of DSS using the TWO-RRS method was developed. PMID- 18674963 TI - A cross-case analysis of technology-in-use practices: EPR-adaptation in Canada and Norway. AB - PURPOSE: To identify and characterize enabling factors that support a continuous adaptation of technology and work practices in the health care sector. METHODS: Cross-case analysis of two longitudinal ethnographic studies of managing the gradual adaptation of electronic patient records, one in Canada and one Norway. RESULTS: The cross-case analysis revealed that technology-in-use practices developed more rapidly in one of the cases, and one of the major driving forces was the establishment of a special committee and the associated project meetings. Based on the literature and grounded in the empirical observations, we complement and expand the notion of project meetings as composed of continuous reflection-on practice activities to construct technology-in-use practices. CONCLUSION: We characterize reflection-on-practice activities as frequent encounters of negotiations of work practices and technology use, providing internal actors a space for systematic evaluation of suggested changes. Further we argue that representatives of the affected professions should not only participate, but also have a mandate to make and evaluate decisions of the technology-in-use practices of the particular group. PMID- 18674965 TI - One-step RT-PCR for detection of Zika virus. AB - BACKGROUND: Zika virus (ZIKV) is an emerging mosquito-borne flavivirus circulating in Asia and Africa. Human infection induces an influenza-like syndrome that is associated with retro-orbital pain, oedema, lymphadenopathy, or diarrhea. Diagnosis of Zika fever requires virus isolation and serology, which are time consuming or cross-reactive. OBJECTIVE: To develop a one-step RT-PCR assay to detect ZIKV in human serum. STUDY DESIGN: An assay targeting the envelope protein coding region was designed and evaluated for its specificity, detection limit, repeatability, and capacity to detect ZIKV isolates collected over a 40-year period from various African countries and hosts. RESULTS: The assay's detection limit and repeatability were respectively 7.7pfu/reaction and 100% in serum and L-15 medium; none of 19 other flaviviruses tested were detected. CONCLUSIONS: The assay is rapid, sensitive, and specific to detect ZIKV in cell culture or serum, but needs to be validated for diagnosis using clinical samples. PMID- 18674964 TI - Multiplex MassTag-PCR for respiratory pathogens in pediatric nasopharyngeal washes negative by conventional diagnostic testing shows a high prevalence of viruses belonging to a newly recognized rhinovirus clade. AB - BACKGROUND: Respiratory infections are the most common infectious diseases in humans worldwide and are a leading cause of death in children less than 5 years of age. OBJECTIVES: Identify candidate pathogens in pediatric patients with unexplained respiratory disease. STUDY DESIGN: Forty-four nasopharyngeal washes collected during the 2004-2005 winter season from pediatric patients with respiratory illnesses that tested negative for 7 common respiratory pathogens by culture and direct immunofluorescence assays were analyzed by MassTag-PCR. To distinguish human enteroviruses (HEV) and rhinoviruses (HRV), samples positive for picornaviruses were further characterized by sequence analysis. RESULTS: Candidate pathogens were detected by MassTag PCR in 27 of the 44 (61%) specimens that previously were rated negative. Sixteen of these 27 specimens (59%) contained picornaviruses; of these 9 (57%) contained RNA of a recently discovered clade of rhinoviruses. Bocaviruses were detected in three patients by RT-PCR. CONCLUSIONS: Our study confirms that multiplex MassTag-PCR enhances the detection of pathogens in clinical specimens, and shows that previously unrecognized rhinoviruses, that potentially form a species HRV-C, may cause a significant amount of pediatric respiratory disease. PMID- 18674966 TI - CD4 memory T cells on trial: immunological memory without a memory T cell. AB - Immunological memory crucially depends on CD4 T cells. In contrast with B cells, we find no decisive evidence that CD4 T cells are permanently altered by antigen stimulation. We propose that the memory response is derived from an increase in frequency of resting naive-like CD4 T cells with a half-life of years (or months in rodents), rather than the currently proposed specialized T-cell types that have a known lifespan of days. In addition, residual antigen will significantly influence the longevity of a memory response. Our model offers a new insight into immunological memory that could assist the development of CD4 T cell-based vaccines. PMID- 18674967 TI - let-7 microRNAs in development, stem cells and cancer. AB - MicroRNAs (miRNAs) are small noncoding RNAs, approximately 22 nucleotides in length, that repress target messenger RNAs (mRNAs) through an antisense mechanism. The let-7 miRNA was originally discovered in the nematode Caenorhabditis elegans, where it regulates cell proliferation and differentiation, but subsequent work has shown that both its sequence and its function are highly conserved in mammals. Recent results have now linked decreased let-7 expression to increased tumorigenicity and poor patient prognosis. Moreover, during normal development, accumulation of let-7 can be prevented by LIN28, a promoter of pluripotency. Based on these findings, we propose that let-7 regulates 'stemness' by repressing self-renewal and promoting differentiation in both normal development and cancer. A more complete understanding of its function will thus provide insights into these processes and might yield diagnostic and therapeutic advances for cancer treatment. PMID- 18674968 TI - Schistosome monogamy: who, how, and why? AB - Schistosomes represent a unique animal model for comparative analyses of monogamy. Indeed, schistosomes are classified at the lowest taxonomical level of monogamous species and lack complex social interactions, which could alter our understanding of their unusual mating system. Elements discussed here include the fact that monogamy in schistosomes could be an ancestral state between hermaphroditism and polygyny or polygynandry and the occurrence of mate changes. In addition, hypotheses are proposed to explain monogamy in schistosomes (e.g. female dispersion, the need for paternal care, oviposition site limitation or aggressiveness, and mate guarding). We also propose future experimental and analytical approaches to improve our understanding of the schistosomes' mating system. PMID- 18674969 TI - Inhibition of M. tuberculosis in vitro in monocytes and in mice by aminomethylene pyrazinamide analogs. AB - Pyrazinamide is unusual among anti-tuberculous agents in its ability to promote a durable cure and shorten the duration of therapy. Yet the basis for this effect is not well understood. A particularly effective strategy for the development of new drugs can be to synthetically manipulate the well-established structures to improve either the spectrum of activity or some pharmacological properties. Similar to previously described aminomethylene amides such as morphazinamide, it was found that novel aminomethylene amides can have in vitro activity at higher than the very acidic pH conditions where pyrazinamide is inactive as well as retaining activity against pyrazinamide-resistant M. tuberculosis. These new compounds have shown an improved anti-tuberculous activity in infected human macrophages relative to pyrazinamide. Compound 1, in combination with rifamycin, was especially effective in both infected human macrophages and in a murine model of infection. The activity of these analogs against pyrazinamide-resistant strains suggests that the development of second generation pyrazinamide analogs may be especially fruitful. PMID- 18674971 TI - Gender and analgesic use in nursing home residents with dementia. PMID- 18674970 TI - Cytokine mRNA expression in cattle infected with different dosages of Mycobacterium bovis. AB - Cytokine mRNA expression of pro-inflammatory cytokines, i.e., interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and anti-inflammatory cytokines, i.e., interleukin-4 (IL-4), interleukin-10 (IL-10) was quantified using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) in cattle infected with different doses (1-1000 colony-forming units (cfu)) of Mycobacterium bovis. RNA was extracted from the Hepes glutamic acid buffer mediated organic solvent protection effect (HOPE) fixed lymph node tissues using Trizol method. The expression levels of all the four cytokines gradually increased in cattle infected with 1 cfu-1000 cfu. Statistical significance (P<0.05) was observed for the cytokines IFN-gamma, IL-4 and IL-10 between the cattle infected with 1 cfu and 1000 cfu. Though there was an increase in the expression levels of TNF-alpha from cattle infected with 1 cfu-1000 cfu, this difference in expression was not statistically significant (P>0.05). The increase in the levels of IFN-gamma indicates that the host may be responding to control the infection and the increased level of IL-4 and IL-10 which are anti inflammatory cytokines, suggests that these cytokines are trying to protect the host by reducing the inflammation and also by controlling the levels of TNF-alpha (the cytokine that may cause tissue damage). PMID- 18674972 TI - Hyphenated techniques for global metabolite profiling. PMID- 18674973 TI - Liquid chromatography-tandem mass spectrometric assay for diclofenac and three primary metabolites in mouse plasma. AB - The first liquid chromatography-tandem mass spectrometric assay for the simultaneous determination of diclofenac, 4'-hydroxy-diclofenac, 5-hydroxy diclofenac and diclofenac-acyl-glucuronide in mouse plasma, using a simple sample pre-treatment procedure, was developed and validated. Analytes in plasma were stabilized using acetic acid and ascorbic acid. After addition of the internal standard D(4)-diclofenac to a 10 microl sample volume and protein precipitation with acetonitrile, the supernatant was supplemented with an equal volume of water and injected into the chromatographic system. A polar embedded reversed-phase column with gradient elution using formic acid and ammonium acetate in water methanol were used. The eluate was totally transferred into an electrospray interface with positive ionization and the analytes were quantified using triple quadrupole mass spectrometry. The assay was validated in the ranges 10-5000 ng/ml for 4'-hydroxy-diclofenac and 20-10,000 ng/ml for the other analytes, the lowest levels of these ranges (10 or 20 ng/ml) being the lower limits of quantification. Within day precisions were < or = 10%, between day precisions < or = 13% and accuracies were between 90 and 108%. The analytes were chemically stable under all relevant conditions. The assay was successfully applied in a pharmacokinetic study with diclofenac in mice. PMID- 18674974 TI - Near-fatal gastrointestinal haemorrhage: an unrecognised culprit. PMID- 18674975 TI - [Delayed measurement of PTH in patients with CKD: storage of the primary tube in the dialysis unit, which temperature? Which kind of tube?]. AB - Parathyroid hormone (PTH) is measured in patients with chronic kidney disease (CKD) to evaluate the spread of secondary hyperparathyroidism and to identify renal osteodystrophy subtypes. An important intermethod variability that can significantly influence the clinical decision has been highlighted recently. Similarly, it is acknowledged that the preanalytical conditions are important to optimize the interpretation of a PTH level by comparison with the K/DOQI guidelines. Considering the frequent case of a dialysis patient in whom blood is handled in the evening and addressed to the clinical laboratory the next morning, we have evaluated the stability of the PTH concentration during a 18-hour period. We thus measured PTH with three automated assays in three kinds of tubes (plain tubes with a gel separator, EDTA tubes, EDTA+aprotinin tubes) which were either immediately centrifugated with a prompt freezing of the serum or plasma, or stored for 18hours at room temperature or at 4 degrees C. Our results demonstrate that, whatever the kind of tube, the PTH concentration is not altered by a 18 hour storage at 4 degrees C which is not the case at room temperature. Using a tube with a gel separator necessitates however to centrifugate the tube in the dialysis unit. On the other hand, the use of serum, by contrast with EDTA plasma, allows the measurement of other biological parameters including calcium, does not need that the tube is fully filled and, according to our results, reduces the intermethod variability. In conclusion, this study shows that the measurement of PTH may be delayed by 18hours if the primary tube is kept at 4 degrees C. Assuming that the primary tube is centrifugated but not opened in the dialysis unit, serum may be the sample of choice for the measurement of PTH in patients with CKD. PMID- 18674976 TI - [Pheochromocytoma in end-stage renal disease patient treated by peritoneal dialysis]. AB - Pheochromocytoma is a rare tumor responsible for paroxysmal hypertension which is difficult to control. Diagnosis is important because it represents a curable form of hypertension. Few cases of pheochromocytoma patients with end-stage renal failure were reported in the literature. These cases are specially responsible for diagnosis and therapeutic problems. We report here a case of an end-stage renal failure patient who has pheochromocytoma, he was treated by automated peritoneal dialysis. The patient is a 47-year-old man who has an IgA glomerulonephritis. On peritoneal dialysis, his blood pressure level remains high despite four antihypertensive drugs association and adequate dialysis. Furthermore, the patient suffered from headaches, sweats and palpitations. This leads to suspect pheochromocytoma. Thus, urinary excretion rates of metanephrines and normetanephrines were high. Radiographic diagnosis tests were negative but MIBG scintigraphy was able to localise the tumor in the left suprarenal gland. He had coelioscopic left adrenalectomy without complications, microscopic studies showed an hyperplasia of the adrenal medulla. Soon after surgery his blood pressure was well controlled by one antihypertensive drug. We conclude that refractory hypertension, as a possible diagnosis, is uncommon in peritoneal dialysis patients. Pheochromocytoma must be eliminated by careful evaluation. PMID- 18674977 TI - The effects of hip and ankle stretching on gait function of older people. AB - OBJECTIVE: To examine effects of hip and ankle stretching on gait function of older people. DESIGN: Randomized controlled trial. SETTING: Flexibility training was performed in participants' homes. Assessments were performed in a biomechanics laboratory. PARTICIPANTS: Forty healthy volunteers (mean age +/- SD, 72.1+/-4.7 y) randomized to 2 groups: intervention (n=20) and control (n=20). INTERVENTION: Intervention participants performed an 8-week stretching program, and control group participants maintained activity level for 8 weeks. One investigator made weekly visits to instruct and monitor participants. MAIN OUTCOME MEASURES: Primary outcome measures were passive joint motion for hip extension and ankle dorsiflexion and freely chosen gait speed. Secondary outcome measures were gait parameters during freely chosen gait speed and set gait speed walking (stride length, joint displacement). RESULTS: Compared with the control group, the intervention group had increased combined hip and knee motion (P=.023), ankle motion (P=.020), and freely chosen gait speed (P=.016). The intervention group showed statistically nonsignificant trends of increased stride length at freely chosen gait speed and set gait speed. CONCLUSIONS: Findings suggest joint motion is a modifiable impairment that is effectively targeted with flexibility training for older people. Participants in the intervention group had improvements in joint motion as well as increased freely chosen gait speed. Mechanisms responsible for changes in freely chosen gait speed warrant further investigation. PMID- 18674978 TI - Preventing recurrent pressure ulcers in veterans with spinal cord injury: impact of a structured education and follow-up intervention. AB - OBJECTIVE: To test the hypothesis that enhanced education and structured follow up after pressure ulcer surgery will result in fewer recurrences. DESIGN: Randomized controlled trial. SETTING: Veterans Affairs medical center. PARTICIPANTS: Forty-nine veteran men with spinal cord injury or dysfunction were approached on admission for pressure ulcer surgery. Five never had surgery, 2 refused to participate, and one withdrew. Forty-one were randomized into 3 groups. Three participants' ulcers did not heal, so follow-up could not begin. INTERVENTIONS: Group 1 received individualized pressure ulcer education and monthly structured telephone follow-up (n=20); group 2 received monthly mail or telephone follow-up without educational content (n=11); and group 3 received quarterly mail or telephone follow-up without educational content (n=10). Follow up continued until recurrence, death, or 24 months. MAIN OUTCOME MEASURE: Time to pressure ulcer recurrence. RESULTS: Group 1 had a longer average time to ulcer recurrence or end of study than groups 2 and 3 (19.6 mo, 10.1 mo, 10.3 mo; P=.002) and had a smaller rate of recurrence (33%, 60%, 90%; P=.007). Survival analysis confirmed these findings (P=.009). CONCLUSIONS: Individualized education and structured monthly contacts may be effective in reducing the frequency of or delaying pressure ulcer recurrence after surgical repair of an ulcer. PMID- 18674979 TI - Risk of stroke, heart attack, and diabetes complications among veterans with spinal cord injury. AB - OBJECTIVES: To compare the rates of diabetes and macrovascular conditions in veterans with spinal cord injury (SCI) and to examine variations by patient-level demographic, socioeconomic, access, and health status factors. DESIGN: A retrospective analysis. Diabetes status was classified by merging with diabetes epidemiology cohort using a validated algorithm. Chi-square tests and logistic regressions used to compare rates in macro- and microvascular conditions in veterans with and without diabetes. SETTING: Veteran Health Administration clinic users in fiscal year (FY) 1999 to FY 2001. PARTICIPANTS: SCI patients (N=8769) with diabetes (n=1333), in FY 2000, identified through the SCI registry. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Macrovascular and microvascular conditions in the next year (February 2001). Derived from International Statistical Classification of Diseases, 9th Revision, Clinical Modification, codes in the patient treatment files. RESULTS: Overall, 15% of SCI veterans were identified with diabetes but this was an underestimate due to high mortality (8%). Among SCI veterans with diabetes, 49% had at least one macrovascular condition and 54% had microvascular conditions compared with 24% and 25% of those without diabetes (P<.001). CONCLUSIONS: Our study highlights the highly significant relationship between diabetes and macro- and microvascular conditions in veterans with SCI. Neurologic deficit combined with increased insulin resistance has a greater macrovascular impact on SCI veterans than on those who do not have diabetes. Increasing age and physical comorbidities compound the problem. PMID- 18674980 TI - Feasibility of a classification system for physical therapy, occupational therapy, and sports therapy interventions for mobility and self-care in spinal cord injury rehabilitation. AB - OBJECTIVE: To test the feasibility of a classification system developed to record the contents of treatment sessions intended to improve mobility and self-care by persons with a spinal cord injury (SCI) in clinical rehabilitation. DESIGN: Descriptive study. SETTING: Three Dutch SCI facilities. PARTICIPANTS: Participants (N=36) as well as physical therapists (n=20), occupational therapists (n=14), and sports therapists (n=2). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Questionnaires to assess the clarity of the classification system, time needed to record 1 treatment session, and the distribution of categories and interventions. The classification system consisted of 28 categories at 3 levels of functioning: basic functions (eg, muscle power), basic activities (eg, transfers), and complex activities (eg, walking and moving around outside). RESULTS: Therapists used 1625 codes to record 856 treatment sessions of 142 patients. For 93% of the treatment sessions, the coding caused little or no doubt. The therapists were able to classify 86.3% of the treatment sessions within 3 minutes. The classification system was rated as useful and easy to use. CONCLUSIONS: The findings support the suitability of our classification system as a tool to record the contents of SCI treatment sessions in different settings and by different therapists. PMID- 18674981 TI - Disparities in antihypertensive medication adherence in persons with disabilities and without disabilities: results of a Korean population-based study. AB - OBJECTIVE: To determine disparities in antihypertensive medication adherence between persons with disabilities and those without disabilities in South Korea. DESIGN: The study compared antihypertensive medication adherence between persons with disabilities and those without disabilities using medical claims data of the National Health Insurance (NHI). SETTING: We obtained data from claims submitted to the NHI, which covers almost the entire Korean population. Persons who were prescribed antihypertensive medication during the calendar year 2004 were identified. PARTICIPANTS: The study comprised data from persons with disabilities (n=85,098) and persons without disabilities (n=2,368,636). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: A cumulative medication adherence (CMA) greater than or equal to 80% was defined as an appropriate medication adherence. Multiple logistic regression was used to identify differences in antihypertensive medication adherence between persons with disabilities and without disabilities. Estimates were adjusted for demographic characteristics (sex, age), type of medical insurance, insurance contribution a month as a proxy for household income, residential area, and clinical characteristics (medication duration, comorbid conditions). RESULTS: People with disabilities had lower CMAs than those without (median CMA, 83.6% vs 85.7%; appropriate medication adherence, 54.5% vs 57.5%). Results of the multiple logistic regression adjusting other factors indicated that people with disabilities had decreased probabilities of appropriate adherence. CONCLUSIONS: Medication adherence is reduced by various types of disability and impairment such as those involving mobility and communication. Much effort should be made to investigate how and why these disparities take place and develop health policies to remove these disparities if they exist. PMID- 18674982 TI - Objective measures of neighborhood environment and self-reported physical activity in spinal cord injured men. AB - OBJECTIVES: To assess the relationship between objective neighborhood environment and self-reported physical activity (PA) and between PA and obesity-related risk factors in people with spinal cord injury (SCI). DESIGN: A cross-sectional study. SETTING: Urban university. PARTICIPANTS: Men with SCI (N=131), 20 to 59 years old, at least 1 year postinjury and using wheelchair for mobility most of the time. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Metabolic syndrome (abdominal obesity, elevated blood pressure, glucose, triglycerides, and low-high density lipoprotein cholesterol) and high C-reactive protein (CRP), as well as total PA metabolic equivalent score. RESULTS: Lower PA was associated with higher prevalence rate for elevated triglycerides, metabolic syndrome, and high CRP. Compared with those in low PA tertile, those in high PA tertile had significantly lower odds for elevated triglycerides (odds ratio [OR]=.19; 95% confidence interval [CI], .04-.80), metabolic syndrome (OR=.15; 95% CI, .03-.66) and high CRP (OR=.17; 95% CI, .04-.71) while adjusting for relevant factors. In crude analysis, lower PA was associated with neighborhood environmental characteristics including shorter distance to nearest transit stops, smaller mean block area, greater number of transit stops, high vacant housing, and higher neighborhood crime rate. In multivariate analysis higher total crime was the only risk factor significantly associated with lower PA level. Those living in higher crime rate neighborhoods had 86% lower odds of having greater than median PA level (OR=.14; 95% CI, .04-.49) than their counterparts. CONCLUSIONS: In men with SCI, lower PA is independently associated with having elevated triglycerides, metabolic syndrome, and high CRP. Additionally, lower PA is associated with higher neighborhood crime rate. PMID- 18674983 TI - Earnings among people with spinal cord injury. AB - OBJECTIVE: To identify differences in conditional and unconditional earnings among participants with spinal cord injury (SCI) attributable to biographic, injury, educational, and employment factors by using a 2-part model (employment, earnings). DESIGN: A secondary analysis of cross-sectional survey data. SETTING: A Midwestern university hospital and a private hospital in the Southeastern United States. PARTICIPANTS: All participants (N=1296) were adults between the ages of 18 and 64 who had a traumatic SCI at least 1 year before study initiation. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Earnings were defined by earnings within the previous 12 months and were measured by a single categoric item. Conditional earnings reflect the earnings of employed participants, whereas unconditional earnings reflect all participants with $0 in earnings recorded for those unemployed. RESULTS: Sex and race were significantly related to conditional earnings, even after controlling for educational and vocational variables. Additionally, conditional earnings (employed participants only) were related to 16 or more years of education, number of years employed, the percentage of time after SCI spent employed, and working in either government or private industry (not self-employed or family business). There was a greater number of significant variables for unconditional earnings, largely reflective of the influence of the portion employed (those not working having $0 in earnings). CONCLUSIONS: Efforts to improve employment outcomes should focus on facilitating return to work immediately after injury, returning to preinjury job, maintaining regular employment, and working for placement in government or private industry. Special efforts may be needed to promote vocational outcomes among women and nonwhites. PMID- 18674984 TI - A prospective study of health and risk of mortality after spinal cord injury. AB - OBJECTIVE: To test hypothesized relationships between multiple health parameters and mortality among persons with spinal cord injury (SCI) while controlling for variations in biographical and injury characteristics. DESIGN: Prospective cohort study with health data collected in late 1997 and early 1998 and mortality status ascertained in December 2005. SETTING: A large rehabilitation hospital in the Southeastern United States. PARTICIPANTS: A total of 1389 adults with traumatic SCI, at least 1 year postinjury. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The primary outcome was time from survey to mortality (or time of censoring). Mortality status was determined using the National Death Index and the Social Security Death Index. There were 225 deaths (16.2%) by December 31, 2005. RESULTS: Cox proportional hazards modeling identified several significant health predictors of mortality status, while controlling for biographic and injury factors. Two sets of analyses were conducted--the first identifying the significance of a single variable of interest and the second analysis building a comprehensive model based on an optimal group of variables. Multiple types of health conditions were associated with mortality. The best set of health predictors included probable major depression, surgeries to repair pressure ulcers, fractures and/or amputations, symptoms of infections, and days hospitalized. Inclusion of these variables, along with a general health rating, improved prediction of survival compared with biographic and injury variables alone, because the pseudo R(2) increased from .12 to .18 and the concordance from .730 to .776. CONCLUSIONS: In addition to secondary conditions that have been the traditional focus of prevention efforts (eg, pressure ulcers, urinary tract infections), amputations, fractures, and depressive symptoms were associated with higher risk for mortality; however, further research is needed to identify the association of specific conditions with causes of death and to determine whether interventions can modify these conditions and ultimately improve survival. PMID- 18674985 TI - Computer and internet use by persons after traumatic spinal cord injury. AB - OBJECTIVE: To determine whether computer and internet use by persons post spinal cord injury (SCI) is sufficiently prevalent and broad-based to consider using this technology as a long-term treatment modality for patients who have sustained SCI. DESIGN: A multicenter cohort study. SETTING: Twenty-six past and current U.S. regional Model Spinal Cord Injury Systems. PARTICIPANTS: Patients with traumatic SCI (N=2926) with follow-up interviews between 2004 and 2006, conducted at 1 or 5 years postinjury. INTERVENTIONS: Not applicable. RESULTS: Results revealed that 69.2% of participants with SCI used a computer; 94.2% of computer users accessed the internet. Among computer users, 19.1% used assistive devices for computer access. Of the internet users, 68.6% went online 5 to 7 days a week. The most frequent use for internet was e-mail (90.5%) and shopping sites (65.8%), followed by health sites (61.1%). We found no statistically significant difference in computer use by sex or level of neurologic injury, and no difference in internet use by level of neurologic injury. Computer and internet access differed significantly by age, with use decreasing as age group increased. The highest computer and internet access rates were seen among participants injured before the age of 18. Computer and internet use varied by race: 76% of white compared with 46% of black subjects were computer users (P<.001), and 95.3% of white respondents who used computers used the internet, compared with 87.6% of black respondents (P<.001). Internet use increased with education level (P<.001): eighty-six percent of participants who did not graduate from high school or receive a degree used the internet, while over 97% of those with a college or associate's degree did. CONCLUSIONS: While the internet holds considerable potential as a long-term treatment modality after SCI, limited access to the internet by those who are black, those injured after age 18, and those with less education does reduce its usefulness in the short term for these subgroups. PMID- 18674987 TI - Subcortical vascular lesions predict falls at 12 months in elderly patients discharged from a rehabilitation ward. AB - OBJECTIVE: To test whether subcortical vascular lesions are associated with falls in elderly patients with gait disorder discharged from a rehabilitation ward. DESIGN: Secondary 12-month follow-up analysis of an observational survey focusing on the prevalence of subcortical vascular lesions in a population of elderly patients discharged from rehabilitation hospitals. SETTING: A rehabilitation and aged care unit. PARTICIPANTS: Consecutively admitted elderly patients (N=214) with gait disorder. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: On admission, all patients underwent comprehensive geriatric assessment including sociodemographics, cognitive and depressive symptoms, nutritional status, physical health, and functional status. Subcortical vascular lesions were assessed on computed tomography films with a validated rating scale. All patients received a standardized rehabilitative program. Twelve months after discharge, all patients were interviewed by telephone, mainly focusing on the occurrence of falls during the follow-up period. Potential predictors of falls were assessed in univariate and multivariate analyses. RESULTS: Univariate predictors of falls were age, sex, Mini-Mental State Examination, Barthel Index on admission, and subcortical vascular lesions. In multivariate analyses, subcortical vascular lesions were the only significant predictor of risk of falling; patients with moderate and severe subcortical vascular lesions scores had a greater risk of falling (odds ratio [OR]=3.0; 95% confidence interval [CI], 1.3-7.1; P=.012; OR=3.9; 95% CI, 1.6-9.2; P=.002, respectively) than those with no subcortical vascular lesions. CONCLUSIONS: Subcortical vascular lesions are associated with falls at 12 months in elderly patients with gait disorder discharged from a rehabilitative ward. Future research is needed to confirm our results. PMID- 18674986 TI - Determinants of lung volumes in chronic spinal cord injury. AB - OBJECTIVE: To characterize determinants of lung volumes in chronic spinal cord injury (SCI). DESIGN: Cross-sectional. SETTING: VA Boston Healthcare System. PARTICIPANTS: White men (N=330) with chronic SCI. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Questionnaire responses and measurements of lung volumes. RESULTS: Adjusted for SCI severity and stature, greater body mass index (BMI) was associated (all P<.05) with lower total lung capacity (TLC) (-38.7 mL x kg(-1) x m(-2)), functional residual capacity (FRC) (-73.9 mL x kg(-1) x m(-2)), residual volume (RV) (-40.4 mL x kg(-1) x m(-2)), and expiratory reserve volume (ERV) ( 32.2 mL x kg(-1) x m(-2)). The effect of BMI on RV was most pronounced in quadriplegia (-72 mL x kg(-1) x m(-2)). Lifetime smoking was associated with a greater FRC (5.3 mL/pack-year) and RV (3.1 mL/pack-years). The effects of lifetime smoking were also greatest in quadriplegia (11 mL/pack-year for FRC; 7.8 mL/pack-year for RV). Time since injury, independent of age, was associated with a decrease in TLC, FRC, ERV, and RV (P<.05). Age was not a predictor of TLC once time since injury was considered. CONCLUSIONS: Determinants of FRC, TLC, ERV, and RV in chronic SCI include factors related and unrelated to SCI. The mechanisms remain to be determined but likely involve the elastic properties and muscle function of the respiratory system and perhaps the effects of systemic inflammation related to adiposity. Addressing modifiable factors such as obesity, muscle stiffness, and smoking may improve respiratory morbidity and mortality in SCI by improving pulmonary function. PMID- 18674988 TI - Accuracy of partial weight bearing after autologous chondrocyte implantation. AB - OBJECTIVE: To determine whether patients can accurately replicate and retain weight-bearing restrictions in both stationary (static) and dynamic conditions after autologous chondrocyte implantation (ACI). DESIGN: Case series. SETTING: Rehabilitation clinic. PARTICIPANTS: A consecutive sample of patients (N=48) who had undergone ACI to a medial or lateral femoral condylar defect in the knee. INTERVENTIONS: Patients were trained to partially weight bear using bathroom scales and forearm crutches prior to assessment. MAIN OUTCOME MEASURES: A force platform was used to measure peak vertical ground reaction forces in patients during static and dynamic conditions immediately after weight-bearing instruction and training, and again during gait 7 days after training. RESULTS: Immediately after instruction and weight-bearing practice on a set of scales, patients exerted a mean of 15.8% body weight more than expected during walking for 20% weight-bearing trials, 8.3% more for the 40% trials, 11.9% more for the 60% trials, and 1.2% less for the prescribed 80% trials. Accuracy of weight-bearing replication improved across all weight-bearing levels when assessed 7 days later, when patients exerted a mean of 6.6% body weight more than expected during walking for 20% weight-bearing trials (9.2% body weight improvement), 4.2% more for the 40% trials (4.1% body weight improvement), 9.9% more for the 60% trials (2% body weight improvement), and 0.2% more for the 60% trials (1% body weight improvement). CONCLUSIONS: Patients were unable to follow weight-bearing restrictions after instruction and practice on a set of scales, and patients were unable to replicate weight-bearing levels in both static and dynamic conditions. PMID- 18674989 TI - Classifying subgroups of chronic low back pain patients based on lifting patterns. AB - OBJECTIVE: To compare self-reported measures of chronic lower back pain (CLBP) patients who were assigned to 2 subgroups based on their lifting patterns performed during a repetitive lifting task. DESIGN: Cross-sectional study. SETTING: Research laboratory PARTICIPANTS: CLBP subjects (n=81) and pain-free controls (n=53). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Measures of lifting patterns and self-reported disability, pain, and psychosocial aspects. RESULTS: Two CLBP subgroups were found: 1 group that lifts similarly to control subjects (n=35) and 1 group that lifts very differently from controls (n=46). The CLBP group that lifted differently than controls reported higher pain intensity (P=.005), higher pain severity (P=.025), and lower self-efficacy (P=.013) than the CLBP group that lifted similarly to controls. CONCLUSIONS: A classification system based on lifting patterns identified 2 CLBP subgroups that were significantly different on lifting and self-reported measures, indicating the importance of physical functioning measures in classification systems. PMID- 18674990 TI - Functional oral intake and time to reach unrestricted dieting for patients with traumatic brain injury. AB - OBJECTIVES: To investigate the status of functional oral intake for patients with severe traumatic brain injury (TBI) and time to return to unrestricted dieting; and to investigate whether severity of brain injury is a predictor for unrestricted dieting. DESIGN: Observational retrospective cohort study. SETTING: Subacute rehabilitation department, university hospital. PARTICIPANTS: Patients age 16 to 65 years (N=173) with severe TBI (posttraumatic amnesia from 7d to >6 mo) admitted over a 5-year period. Patients are transferred to the brain injury unit as soon as they ventilate spontaneously. INTERVENTION: Facial oral tract therapy. MAIN OUTCOME MEASURE: Unrestricted dieting assessed by the Functional Oral Intake Scale (FOIS). RESULTS: We found that 93% of all patients had problems with functional oral intake at admission. Within 126 days of rehabilitation, 64% recovered to unrestricted dieting before discharge. The chance of returning to total oral diet depends on the severity of the brain injury and can be predicted by Glasgow Coma Scale (GCS; measured the day after cessation of sedation; Wald chi(2)=42.78, P<.01), Rancho Los Amigos Scale (RLAS) level (Wald chi(2)=11.84, P=.01), FIM instrument (Wald chi(2)=44.40, P<.01), and FOIS score at admission (Wald chi(2)=82.93, P<.01). CONCLUSIONS: Impairment in functional oral intake was found to be very common for patients with severe TBI admitted to a subacute rehabilitation department. For those who recovered during hospital rehabilitation, return to unrestricted dieting happened within 126 days of rehabilitation. The chance of returning to unrestricted dieting depends on the severity of the brain injury and can be predicted by GCS score, RLAS level, FIM score, and functional oral intake at admission. These results are important when planning rehabilitation, giving information to patients and relatives, and designing efficacy studies of facial oral tract therapy, which are highly recommended. PMID- 18674991 TI - Longitudinal stability of the Fugl-Meyer Assessment of the upper extremity. AB - OBJECTIVE: To investigate the longitudinal stability of the Fugl-Meyer Assessment (FMA) of the upper-extremity item difficulties by using Rasch analysis. DESIGN: Secondary analysis of existing data from a cohort longitudinal study of stroke recovery. SETTING: University research center. PARTICIPANTS: A total of 377 people, ages 69.2+/-11.2 years, to whom the assessment was administered at 2 weeks and 6 months poststroke. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Differential item function analysis performed by using the Winsteps software program examined whether the item difficulty hierarchical order of a modified 30-item FMA for the upper extremity (reflex items removed) was invariant across 2 testing occasions. RESULTS: Only 2 items (shoulder flexion to 180 degrees, movement with normal speed) showed large differences in test-retest item difficulty calibration. Item instability had no practical consequences on the longitudinal measurement of person ability. CONCLUSIONS: The 30-item assessment shows a longitudinally stable item difficulty order and is valid for measuring volitional arm motor ability over time. PMID- 18674992 TI - Home-based motor imagery training for gait rehabilitation of people with chronic poststroke hemiparesis. AB - OBJECTIVE: To test the feasibility and efficacy of a home-based motor imagery gait training program to improve walking performance of individuals with chronic poststroke hemiparesis. DESIGN: Nonrandomized controlled trial. SETTING: Local facility. PARTICIPANTS: Participants (N=17) were community-dwelling volunteers with hemiparesis caused by a unilateral stroke that occurred at least 3 months before the study. INTERVENTION: Participants received 15 minutes of supervised imagery gait training in their homes 3 days a week for 6 weeks. The intervention addressed gait impairments of the affected lower limb and task-specific gait training. Walking ability was evaluated by kinematics and functional scales twice before the intervention, 3 and 6 weeks after the intervention began, and at the 3 week follow-up. MAIN OUTCOME MEASURES: Spatiotemporal, kinematic, and functional walking measurements. RESULTS: Walking speed increased significantly by 40% after training, and the gains were largely maintained at the 3-week follow-up. The effect size of the intervention on walking speed was moderate (.64). There were significant increases in stride length, cadence, and single-support time of the affected lower limb, whereas double-support time was decreased. Improvements were also noted on the gait scale of the Tinetti Performance-Oriented Mobility Assessment as well as in functional gait. Sixty-five percent of the participants advanced 1 walking category in the Modified Functional Walking Categories Index. CONCLUSIONS: Although further study is recommended, the findings support the feasibility and justify the incorporation of home-based motor imagery exercises to improve walking skills for poststroke hemiparesis. PMID- 18674993 TI - Potential role of optimal velocity as a qualitative factor of physical functional performance in women aged 72 to 96 years. AB - OBJECTIVE: To assess the relationship of maximal leg power and its corresponding determinants (eg, optimal velocity and optimal torque) measured during maximal voluntary knee extension to physical functional performance of older women. DESIGN: Descriptive. SETTING: Community retirement homes. PARTICIPANTS: Women (N=39) aged 72 to 96 years. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Volunteers performed in sitting position maximal knee extensions on an Ergopower dynamometer to calculate maximal leg power, optimal velocity, and optimal torque. Three standardized tests were also performed to evaluate physical performance: walking speed over 6m, time taken to rise 5 times from a chair, and time to climb 6 stairs. RESULTS: On multiple regression analysis, leg power (mean, 1.37+/-0.80 W/kg) significantly correlated with physical performance as measured by 6-m walking speed (mean, .85+/-.40 m/s), chair-stand time (mean, 16.3+/-7.7s), and stair-climb time (mean, 7+/-4s), describing 16% to 33% of the variance. Optimal velocity (mean, 1.79+/-1.20 rad/s) also significantly correlated with 6-m walking speed, chair-stand time, and stair-climb time, describing 46% to 89% of the variance. Optimal torque (50.8+/-16.9 Nm) did not correlate with physical performance. CONCLUSIONS: Maximal power and moreover optimal velocity were thus found to be determinants of physical performance, both appearing as significant mobility factors in older adults. This may provide more focus on velocity oriented training as a means of improving functional status. PMID- 18674994 TI - Digital ischemia after carpal tunnel injection: a case report. AB - A 51-year-old woman described paresthesias in her right thumb, index, and middle fingers consistent with carpal tunnel syndrome. Using a sterile technique, a 25 G, 1 1/4-in needle was introduced ulnar to the palmaris longus tendon, and a mixture of 1.5 mL of 40 mg/mL of triamcinolone acetonide and 1.5 mL of 1% lidocaine was injected. After the injection, the patient's hand exhibited signs of ischemia including coolness and discoloration. Rewarming of the hand with paraffin was performed immediately, and normative color returned. At follow-up visits, the patient described burning in the hand, and blotchiness of the digits was noted. A magnetic resonance imaging angiogram of the right wrist showed a single deep palmar arch. Electromyography and nerve conduction study weeks after the injection showed bilateral median neuropathies, moderately severe on the right and mild on the left. She underwent an open carpal tunnel release 6 weeks postinjection. The patient did well and returned to her job without restrictions. The exact etiology of the hand ischemia is unclear but may be related to vasospasm as has been described in the spine-injection literature. Regardless of the etiology, this case shows an uncommon adverse event in a commonly performed procedure and raises questions for further review. PMID- 18674995 TI - Efficacy and safety of a hip flexion assist orthosis in ambulatory multiple sclerosis patients. AB - OBJECTIVE: To evaluate the efficacy and safety of a hip flexion assist orthosis (HFAO) in ambulatory patients with multiple sclerosis (MS). DESIGN: Fourteen week pre- and postintervention uncontrolled trial. SETTING: Outpatient rehabilitation clinic within an MS center. PARTICIPANTS: Ambulatory MS patients (N=21) with unilateral (or unilateral predominant) hip flexor weakness. INTERVENTION: Subjects were fitted with the HFAO on the weaker side, trained to use the device, and given a wear schedule. Subjects completed 2 baseline evaluations and follow up testing at 8 and 12 weeks. MAIN OUTCOME MEASURES: Lower-extremity manual muscle testing, pain, and gait performance (Timed 25-Foot Walk, Timed Up & Go, 6 minute walk test, Mellen Center Gait Test). Subject satisfaction was evaluated by using a 9-item custom questionnaire. RESULTS: There was a statistically significant improvement of strength in the affected lower extremity at 8 and 12 weeks (effect size [ES]=0.63; ES=1.32, respectively), of pain at 12 weeks only (ES=-0.64), and of all gait tests at 8 and 12 weeks (ES range, 0.38-1.33). The overall mean satisfaction score at 12 weeks was 39 (maximum score, 45). No serious adverse events were recorded during the study. The most frequent side effect of the HFAO was low back pain (19%). No side effects led to discontinuation of the HFAO use during the study. CONCLUSIONS: The HFAO was safe and well tolerated. HFAO use was associated with significant improvement of gait performance as well as improvement of strength in the lower extremity fitted with the HFAO. Subjective reports suggest that there was an increase in daily life activity level. PMID- 18674996 TI - The steep ramp test: questions about sensitivity and reliability. PMID- 18674997 TI - Flawed analyses in driving pedal reaction time study. PMID- 18674998 TI - [Depression in the second half of life]. PMID- 18674999 TI - [Depressive moods during the second half of life. Introduction]. PMID- 18675001 TI - Vascular disease, depression and cognitive impairment. PMID- 18675002 TI - [Symptomatic kaleidoscope of the depression in the elderly persons]. PMID- 18675003 TI - [Getting rid of and rejecting depressive state, neurotic conflict seeking and old age crossing]. PMID- 18675004 TI - [Looking for functional imagery in the understanding of emotional regulation process]. PMID- 18675006 TI - New antidepressants for older people: a critical review of the evidence base. PMID- 18675008 TI - [The depressed elderly: between bereavement and the temptation to give up]. PMID- 18675007 TI - [Looking for oneself: theory and practice of auto-biographic memory in Alzheimer's disease]. PMID- 18675009 TI - [Dementia and resilience: narration or regression?]. PMID- 18675010 TI - Guest editorial: response to "preserving today with an eye on our future". PMID- 18675011 TI - Sweet home-medical or health? PMID- 18675012 TI - Comment on "ways of knowing hope: Carper's fundamental patterns as a guide for hope research with bereaved palliative caregiver.". PMID- 18675014 TI - Building social capital in healthcare organizations: thinking ecologically for safer care. AB - Research on patient safety and health human resources, 2 critical issues for 21st century healthcare, converges on similar findings. Specifically, it is apparent that along with the patients, families, and communities we serve, nurses and other healthcare professionals navigate a volatile health care system where persistent restructuring, market pressures, and workforce instability present ongoing threats to the delivery of safer care. Drawing from the fields of nursing, healthcare ethics, health systems management, and ecological restoration, we outline the role of social capital for organizational integrity, healthy workplace cultures, sustainable resource management, improved nurse retention, effective knowledge translation, and safer patient care. Nursing leaders can use ecological thinking to build the vital resource of social capital by taking concrete steps to commit the necessary human and material resources to: (1) forge relations to foster bonding, bridging and linking social capital; (2) build solidarity and trust; (3) foster collective action and cooperation; (4) strengthen communication and knowledge exchange; and (5) create capacity for social cohesion and inclusion. PMID- 18675015 TI - Telehealth: a policy analysis for quality, impact on patient outcomes, and political feasibility. AB - Telehealth is seen as a venue for improving healthcare access in vulnerable populations through use of electronic devices in the patient's home that monitor and assess for early complications. Major technological advances have improved telehealth services, prompting introduction of federal legislation for reimbursement of telehealth care. Yet, policymakers have stalled passage based on several concerns. This article conducted a policy analysis of HR1601, one of the most recent Congressional proposals for telehealth. The policy analysis was based on 3 criteria: quality of telehealth services, impact of telehealth on patient outcomes, and political feasibility. Overall, the concerns regarding quality, impact on patient outcomes, and feasibility are outweighed by the benefits of using telehealth to extend care. Federal funding of telehealth through passage of legislation, such as HR1601, is a good first step to establishing further evidence for its capabilities, and it could lead to a more comprehensive approach to quality care. PMID- 18675016 TI - Using clinical simulations in geriatric nursing continuing education. AB - Accompanying an increased number of older adults in the population is the need for knowledgeable and clinically competent nurses to provide care for this group, especially those experiencing sudden changes in health status. A continuing education program was designed to improve geriatric nursing competencies through the use of clinical simulations. Three-day nurse educator institutes and one-day workshops for registered and licensed practical nurses were offered to 312 nurses. The clinical simulations that were developed specifically focused on acute health events or conditions. Specific geriatric clinical competencies were also emphasized. Different types of clinical simulations included unfolding cases, use of a human patient simulator, and online case studies. Geriatric nursing knowledge significantly increased and clinical simulations were well received. Clinical simulations involving the human patient simulator were highly rated. Clinical simulations are an excellent teaching strategy to help nurses increase knowledge and skill in caring for older adults. PMID- 18675017 TI - Shared faculty governance: a decision-making framework for evaluating the DNP. AB - The concept of shared governance was used to frame the dialogue and decision making process around the Doctor of Nursing Practice (DNP) at a leading School of Nursing. This article reports the process used to support dialogue around the DNP so that an informed decision could be made. The shared governance process promoted reflective conversation within the entire faculty and, in the end, supported the decision-making related to the DNP. PMID- 18675018 TI - Innovative options for the doctoral dissertation in nursing. AB - The doctoral dissertation is the capstone event of doctoral study. While developing their dissertations, graduate students discover the cutting edge of the discipline, learn the intricacies of the research process, and engage in scientific dialogue with experts in the field. Traditional dissertation formats are confined to the Dissertation Abstracts International and are not easily accessed by clinicians, administrators, and researchers who are most in need of the information. Consequently, the number of universities offering options to the traditional dissertation format is growing. This article describes and compares 2 formats: the traditional dissertation and the publication option. The benefits and challenges of each format to doctoral students and faculty are discussed. The discussion reflects changing trends in the nursing profession as an academic discipline. PMID- 18675019 TI - "It's what we're here for:" nurses caring for military personnel during the Persian Gulf Wars. AB - Military nursing service during wartime represents significant contributions to a unique type of health care. The purposes of this study were to: (1) generate themes that elucidate combat nursing experiences, (2) honor nurses who served by sharing their stories, and (3) permanently archive accounts of nursing personnel who served during the Persian Gulf Wars during the late 20th and early 21st centuries. Eleven military nurses who provided health care to American troops in the Persian Gulf participated in a historical study as part of the ongoing Nurses at War oral history project documenting the experiences of nurses during times of armed conflict. The overriding theme, "It's what we're here for," demonstrates the commitment of nurses: a commitment to care and to sacrifice. Other themes drawn from the study included lessons learned from their wartime nursing experiences, sacrifices made, and chronicles of caring. During armed conflict in the Persian Gulf Wars, military nurses' personal stories demonstrated the importance of being engaged in making meaningful professional and historical contributions. These nurses displayed professional commitment and hardiness in the face of difficult life circumstances, saying, "We did what we had to do." PMID- 18675020 TI - Commentary: gerontological nursing graduate education: creating a future beyond "the tipping point". PMID- 18675021 TI - Pediatric Resuscitation. Preface. PMID- 18675022 TI - Background and epidemiology of pediatric cardiac arrest. AB - Pediatric cardiac arrest is not a single problem. Although most episodes of pediatric cardiac arrest occur as complications and progressions of respiratory failure and shock, sudden cardiac arrest may result from sudden arrhythmias. With better understanding of the epidemiology of pediatric cardiac arrest, clinicians can better tailor therapy to optimize outcome. PMID- 18675023 TI - CPR-why the new emphasis? AB - The importance of high quality, prompt cardiopulmonary resuscitation (CPR) for patients in cardiac arrest is receiving new attention and emphasis. This extends to CPR for children. In this article, the authors examine the differences in pediatric anatomy and the mechanisms of blood flow during CPR. Additionally, new evidence on the frequent poor performance of CPR and mechanisms to improve it are presented. PMID- 18675024 TI - Airway management. AB - The pediatric airway and respiratory function differ from those in adults. Optimum management requires consideration of these differences, but the application of adult principles is usually sufficient to buy time in an emergency until specialist pediatric help is available. Simple airway opening techniques such as head tilt and jaw thrust are usually sufficient to open the child's airway, but there is now a range of equipment available to bypass supraglottic airway obstruction-the strengths and weaknesses of such devices are explored in this article. The role of endotracheal intubation is also discussed, along with the pros and cons of the use of cuffed endotracheal tubes in children, and methods of confirming tracheal placement of the tube. PMID- 18675025 TI - Tachyarrhythmias and defibrillation. AB - Although cardiac arrhythmias less commonly cause hemodynamic compromise in children than in adults, prompt recognition and treatment of arrhythmias remain an important part of pediatric resuscitation because of the availability of specific, effective therapies. This article summarizes the 2005 American Heart Association Guidelines for Pediatric Advanced Life Support regarding tachyarrhythmias, including treatment with antiarrhythmics and direct current countershock therapy, and provides an update of recent literature since the guidelines were published. PMID- 18675026 TI - Vascular access and drug therapy in pediatric resuscitation. AB - Using the evidence brought together through the 2005 International Liaison Committee on Resuscitation evidence evaluation process and the subsequent 2005 American Heart Association Guidelines for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care, the role for specific drug therapy in pediatric cardiac arrest is outlined. The drugs discussed include epinephrine, vasopressin, calcium, sodium bicarbonate, atropine, magnesium, and glucose. The literature addressing how best to deliver these drugs to the critically ill child is also presented, specifically looking at the use of intraosseous and endotracheal drug therapy. PMID- 18675027 TI - Extracorporeal cardiopulmonary resuscitation in refractory pediatric cardiac arrest. AB - The purpose of this article is to discuss the indications for extracorporeal cardiopulmonary resuscitation (ECPR), physiologic and mechanical issues that arise in patients managed with ECPR, and optimal patient selection for ECPR. ECPR can provide very good outcomes for some children who, in all likelihood, would otherwise have died. Having the capability to routinely offer ECPR represents an enormous institutional commitment of people and resources. For ECPR to be successful, it must be rapidly deployed, patients must be selected with care, and consistently excellent conventional CPR must take place while awaiting ECPR. PMID- 18675028 TI - Postresuscitation care. AB - Cardiac arrest in infants and children is a rare but critical event that typically follows a period of respiratory or circulatory compromise and has a low survival rate. The only intervention demonstrated to increase survival rate is the provision of bystander CPR. This article examines the pathophysiology of the postarrest reperfusion state; postresuscitation care of the respiratory and cardiovascular systems; postresuscitation neurologic management; therapeutic hypothermia; blood glucose control; immunologic disturbances and infections; coagulation abnormalities; and gastrointestinal and hepatic dysfunction, among other topics. PMID- 18675029 TI - Outcome following cardiopulmonary arrest. AB - This article summarizes the current state of outcomes and outcome predictors following pediatric cardiopulmonary arrest with special emphasis on neurologic outcome. The authors briefly describe the factors associated with outcome and review clinical signs, electrophysiology, neuroimaging, and biomarkers used to predict outcome after cardiopulmonary arrest. Although clinical signs, imaging, and somatosensory evoked potentials are best associated with outcome, there are limited data to guide clinicians. Combinations of these predictors will most likely improve outcome prediction, but large-scale outcome studies are needed to better define these predictors. PMID- 18675030 TI - Medical emergency and rapid response teams. AB - Hospitals that care for children are establishing medical emergency or rapid response teams as system solutions for preventing unexpected but foreseeable respiratory and cardiac arrest on inpatient units. Typically, an experienced team of doctors and nurses responds quickly to a direct request by any level of staff or even a parent for assistance with a child whose physiologic parameters meet predetermined criteria or whose condition causes concern to them. Several pediatric studies comparing outcomes before and after introduction of these rapid response systems reported reductions in rates of respiratory or cardiac arrest and death but no prospective study has compared pediatric hospitals that have implemented rapid response teams to hospitals that have not. PMID- 18675031 TI - Teamwork during resuscitation. AB - Effective resuscitation requires the integration of several cognitive, technical, and behavioral skills. Because resuscitation is performed by teams of health care professionals, these individuals must be able to work together in a coordinated and efficient manner, making teamwork a critical skill for care of patients in distress. Despite the importance of teamwork in health care, little consensus exists as to what it is, how it can most effectively be learned, and how it should be assessed. This article reviews current knowledge on the measurement, training, and importance of teamwork in pediatric resuscitation. PMID- 18675032 TI - Resuscitation education: narrowing the gap between evidence-based resuscitation guidelines and performance using best educational practices. AB - Recent data from in- and out-of-hospital cardiopulmonary arrests reveal that health care teams frequently deviate from American Heart Association guidelines during resuscitation efforts. These discrepancies between the current state of evidence-based resuscitation guidelines and the quality of basic and advanced life support actually delivered represent a missed opportunity and provide a significant target for optimizing patient outcomes through improved educational effectiveness. This article presents discussion of the quality of resuscitation delivered to patients, a brief history of the development of cardiopulmonary resuscitation and attempts to translate the science of resuscitation to the bedside through effective educational strategies, a review of educational best practices that relate to resuscitation education, and discussion of the role of medical simulation in resuscitation training. PMID- 18675033 TI - Future directions in cardiocerebral resuscitation. AB - Outcomes from pediatric cardiac arrest and cardiopulmonary resuscitation (CPR) seem to be incrementally improving. The past 2 decades have brought advances in the understanding of the pathophysiology of cardiac arrest and ventricular fibrillation, better treatment strategies, and a more robust standard for CPR epidemiology and research reporting. The evolution of practice based on an improved understanding of the pathophysiology and timing, intensity, duration, and variability of the hypoxic-ischemic insult should lead to goal-directed therapy gated to the phase of cardiac arrest and the postarrest period encountered. By strategically focusing therapies to specific phases of cardiac arrest and resuscitation and to the evolving pathophysiology and by implementing evidence-based practice, there is great promise that critical care interventions can lead the way to more successful cardiopulmonary and cerebral resuscitation in children. PMID- 18675034 TI - [Cerebral imaging in healthy aging: contrast with Alzheimer disease]. AB - With age, the brain undergoes both structural and functional alterations. Overall, the literature consistently reports global brain atrophy in normal adults, generally more pronounced in frontal areas. As a result of different methodologies and inclusion criteria, other brain areas have been the matter of conflicting findings, notably the hippocampus. Regarding resting-state PET studies, they have consistently highlighted a metabolic deterioration of the frontal and anterior cingulated cortices. By contrast, relatively few investigations have sought to identify those areas that remain intact with aging, or undergo the least deterioration. We report a study designed to establish a comprehensive profile of both structural and functional changes in the aging brain, using up-to-date voxel-based methodology in 45 optimally healthy subjects aged 20-83 years. One of the main findings is that the lesser structural deterioration of the anterior hippocampal region, together with the lesser functional alteration of the posterior cingulate cortex, appear to mark the parting of the ways between normal aging and Alzheimer's disease, which is characterized by early and prominent deterioration of both structures. This paper also deals with studies set out to establish the relationship between changes in episodic memory retrieval in normal aging on the one hand and gray matter volume and 18FDG uptake on the other hand. Frontal areas dysfunction is involved in memory decline in older subjects, at least in some conditions, a finding which clearly contrasts with that found in Alzheimer's disease where the hippocampus plays a key role. Finally, compensatory mechanisms are reviewed through activation studies which often show supplementary activations in old subjects compared to young as well as in Alzheimer's disease patients compared to healthy elderly subjects. Paradoxically, those mechanisms seem to be underpinned, at least partially, by frontal areas in both populations, but researches are needed to better identify which subregions are involved. PMID- 18675035 TI - [The rapid development of child neuropsychology]. AB - The past few years have seen important progress and new developments in the field of child neuropsychology. Children can exhibit acquired or learning disorders, but in all cases such deficits take place in a developmental trajectory that requires taking into account cerebral maturation and plasticity processes. Child neuropsychology finds its specificity in the perspective of developmental neuropsychology and has close connections with health and education. This article focuses on two pathologies - childhood epilepsies and developmental dyslexia - that highlight the recent progress in this specialty and its perspectives. PMID- 18675036 TI - [Knowing without remembering: the contribution of developmental amnesia]. AB - The organization of episodic and semantic memory is currently debated, and especially the rule of the hippocampus in the functioning of these two systems. Since theories derived from the observation of the famous patient HM, that highlighted the involvement of this structure in these two systems, numerous studies questioned the implication of the hippocampus in learning a new semantic knowledge. Among these studies, we found Vargha-Kadem's cases of developmental amnesia. In spite of their clear hippocampal atrophy and a massive impairment of episodic memory, these children were able to acquire de novo new semantic knowledge. In the present paper, we describe a new case of developmental amnesia characteristic of this syndrome. In conclusion, the whole published data question the implication of the hippocampus in every semantic learning and suggest the existence of a neocortical network, slower and that needs more exposures to semantic stimuli than the hippocampal one, which can supply a massive hippocampal impairment. PMID- 18675037 TI - [Frontal lobe, executive functions and cognitive control]. AB - The approach of executive functions began with the early description of behavioural disorders induced by frontal damage. The development of neuropsychology has led to the description of a large variety of cognitive disorders. The interpretation in cognitive terms of these disorders has emphasized the critical impairment of executive functions which are involved in non-routine situations. The role of working memory, initially suggested by animal studies, has also largely influenced theoretical approaches. Numerous studies have been interpreted within the theoretical frameworks developed by Shallice and by Baddeley. A large amount of studies have allowed the specification of control processes (initiation, inhibition...). Recent studies are beginning to investigate cognitive disorders underlying behavioural changes such as deficits of emotional, social and metacognitive processes. In addition these studies seriously question the assumption of a unique central control system. These studies have deeply influenced the clinical approach, the assessment and the diagnosis of executive syndrome. For clinical practice, these data lead to favour specific assessment of some key behavioural and cognitive deficits based on a battery of tests and structured interview of an informant. PMID- 18675038 TI - [Neuropsychology and psychopathology: cognitive analysis during medically refractory epilepsy surgery]. AB - In psychopathology, few studies have been focused on the psychiatric complications of medically refractory mesial temporal lobe epilepsy (MTLE). The aim of the present study was to study NG's capacities, who presented emotional change after right temporal epilepsy surgery with phobias and empathy disorders. NG was examined in two emotional judgment tasks: one explicit and another implicit. For negative stimuli, NG had attraction in the explicit task and dependency in the implicit task. This study suggests that surgical intervention might be one of the causes of postoperative psychiatric disorders in patients with MTLE. MTLE patients have to be explored with neuropsychological paradigms. PMID- 18675039 TI - [Unilateral spatial neglect: 30 years of research, discoveries, hope, and (especially) questions]. AB - Unilateral spatial neglect (USN) was defined 30 years ago as "a failure to report, respond, or orient to stimuli that are presented contralateral to a brain lesion, provided that this failure is not due to elementary sensory or motor disorders" by Heilman and Valenstein (1979). Even though this definition still holds, the last 30 years have been characterized by a profusion of clinical descriptions, neuroanatomical investigations and theoretical models of neglect. This article summarizes the wealth of neuroanatomical, clinical and experimental data concerning USN, by focusing on attentional and spatial deficits. Finally, some perspectives on neglect research are outlined. PMID- 18675040 TI - [From perception to learning]. AB - In the field of perception, learning is formed by a distributed functional architecture of very specialized cortical areas. For example, capacities of learning in patients with visual deficits - hemianopia or visual agnosia - from cerebral lesions are limited by perceptual abilities. Moreover a visual deficit in link with abnormal perception may be associated with an alteration of representations in long term (semantic) memory. Furthermore, perception and memory traces rely on parallel processing. This has been recently demonstrated for human audition. Activation studies in normal subjects and psychophysical investigations in patients with focal hemispheric lesions have shown that auditory information relevant to sound recognition and that relevant to sound localisation are processed in parallel, anatomically distinct cortical networks, often referred to as the "What" and "Where" processing streams. Parallel processing may appear counterintuitive from the point of view of a unified perception of the auditory world, but there are advantages, such as rapidity of processing within a single stream, its adaptability in perceptual learning or facility of multisensory interactions. More generally, implicit learning mechanisms are responsible for the non-conscious acquisition of a great part of our knowledge about the world, using our sensitivity to the rules and regularities structuring our environment. Implicit learning is involved in cognitive development, in the generation of emotional processing and in the acquisition of natural language. Preserved implicit learning abilities have been shown in amnesic patients with paradigms like serial reaction time and artificial grammar learning tasks, confirming that implicit learning mechanisms are not sustained by the cognitive processes and the brain structures that are damaged in amnesia. In a clinical perspective, the assessment of implicit learning abilities in amnesic patients could be critical for building adapted neuropsychological rehabilitation programs. PMID- 18675041 TI - [Hemispheric specialisation versus inter-hemispheric communication]. AB - The first part of this article covers the main discoveries that led to the concept of hemispheric specialisation, from Egyptian antiquity to present times, through the pivotal XIXth century period that saw the attribution of specific cognitive functions to the left and right hemispheres. Next, this dichotomous conception of cerebral function, attributing a given process to a hemisphere and hypothesising callosal transmission, is discussed in the light of recent studies on language comprehension. Present day knowledge suggesting an alternative to the structuralist view of hemispheric specialisation in the form of dynamic, complementary sharing of labour, and of cooperation through transcortical neural networks, is then considered. Finally, the role of the corpus callosum in interhemispheric communication is briefly covered. An emphasis is placed on the diversity of this structure that is at the origin of highly different functions (fibre size, homotopic vs heterotopic connections). Ultimately, we contrast the view of a corpus callosum serving as an information transmitting channel with that of a fibre tract co-activating the non-engaged hemisphere and preparing it for potential stimulation. In this manner, the corpus callosum minimises disparities in the distribution of attention between the two hemispheres. PMID- 18675042 TI - [Cognitive rehabilitation]. AB - During these last 30 years, cognitive rehabilitation has accomplished dramatic improvement. In this paper, we review progress in four main domains: the development of pragmatic and ecological approaches in neuropsychological rehabilitation; the development of computerised rehabilitation; rehabilitation of executive functions; cognitive rehabilitation in degenerative dementia. Finally, we present a single-case study, recently published elsewhere, showing the effectiveness of rehabilitation of verbal working memory in a patient with left hemisphere stroke. In addition, future issues for rehabilitation research are presented. The development of bottom-up rehabilitation strategies as well as the use of inter-hemispheric interactions appears as future promising tracks for clinical rehabilitation. PMID- 18675043 TI - [Clinical neuropsychology: a method and discipline aimed at healing thought and thought processes]. PMID- 18675044 TI - [Revue neurologique. Preface]. PMID- 18675045 TI - [Neurology]. PMID- 18675046 TI - [Functional neuroimaging and the treatment of aphasia: speech therapy and repetitive transcranial magnetic stimulation]. AB - Functional imaging has provided new evidence of the neurobiological impact of the treatment of aphasia, including speech therapy, through the alteration of the activated language neural network. In such a way, speech therapy has proved its impact. The role of each hemisphere is still very unclear. Some of the authors link the left-lateralisation of activations to the therapeutic improvement of language and the right-activated network to a maladaptative strategy, whereas others consider the latter as a useful compensatory network for speech disorders. Repetitive trans-cranial magnetic stimulation (rTMS), first used to determine cortical activity, is now used to directly interfere with cerebral activity. In the years to come, rTMS should be developed as an adjuvant therapy for aphasia. PMID- 18675047 TI - [The evolution of adult clinical neuropsychology in Paris from 1957 to 2000]. AB - This sketchy account of the history of French-speaking clinical neuropsychology from 1957 to 2000 first recalls the Salpetriere school leaded by Alajouanine and his disciples, and second the Ste Anne psychiatric hospital, with on one hand the Chair of Mental and Brain Diseases, and on the other hand a number of research teams gathered around disciples of Dejerine such as Jean Lhermitte. Because of the large scope of this topic, the authors could not elude the need for a bibliographic selection that may seem questionable. PMID- 18675048 TI - [The French-Speaking Neuropsychological Society: 30 years young]. AB - On the occasion of a one-day scientific conference staged at the College de France on December 7th 2007 to mark the thirtieth birthday of the French-Speaking Neuropsychological Society (SNLF for Societe de Neuropsychologie de Langue Francaise), the authors retrace the landmark events in the history of this learned society, beginning with its founding by Henri Hecaen and a group of French-speaking clinicians and research scientists. Emphasis is placed on its increasingly influential status and on its noteworthy contribution to the development of clinical and cognitive neurosciences at both scientific and institutional levels. With a current membership of five hundred, the SNLF engages in a range of activities, including publishing, and holds several meetings each year. It played an active part in setting up the Federation of the European Societies of Neuropsychology (FESN), but has also been keen to reinforce its links with Canadian researchers and clinicians. Loyal to the stated aims of its founder members, the SNLF advocates as high a theoretical level of debate as possible, while working to promote constructive discussions and exchanges of ideas between clinicians, researchers and academics from varied professional backgrounds. PMID- 18675049 TI - [The twenty-first century as a neuropsychology era]. AB - This article reviews how neuropsychology, in the French-speaking world, has evolved as a discipline focused on research, teaching and clinical work. The article targets the last 30 years as this corresponds to the time at which the Societe de Neuropsychologie de Langue Francaise (French-Speaking Neuropsychological Society) was created. The review addresses how the cognitive neuropsychology approach and the advent of brain imaging have shaped the field of neuropsychology in recent years. It presents the status of the discipline in the main French-speaking countries (where neuropsychology is currently developed) including France, Belgium, Switzerland and Canada. It also analyzes a number of indicators that reflect the vitality of the discipline and its cohesion as a science and as a clinical domain. These indicators include the creation of specialized journals, organization of scientific meetings, accessibility to training programs in neuropsychology, development of discipline-oriented clinical programs, and the increase in scientific productivity. The Quebec academic environment is used as an illustration, whereby structured clinical doctoral training programs that meet national standards in neuropsychology were implemented to train clinical neuropsychologists. Finally, the authors emphasize the major role that the discipline is likely to play at different levels of society in the near future. PMID- 18675050 TI - [Alexia without agraphia: an exemplary deficit, cherished by neuropsychologists]. AB - Alexia without agraphia is the result of a left inferotemporal cortical lesion. This unilateral lesion provokes an almost irrecoverable word reading disability. This anatomical-clinical observation has identified as crucial an area on the left fusiform gyrus, which represents a sort of "letter cranial bump". This area, located near the left V4, has been labeled the visual word form area (VWFA). The VWFA ensures visual word processing whatever the visual field stimulated. In normal subjects, imaging methods show an activation of the VWFA during reading. In patients, the alexia that follows a VWFA lesion proves that this area is an essential relay for cerebral word reading processing. Therefore, pure alexia is no longer explained by a posterior interhemispheric disconnection. PMID- 18675051 TI - [The reader brain: natural and cultural story]. AB - The report of cases of pure alexia suggests that some regions of the neural system are dedicated to reading-specific visual processing abilities. Pure alexia results from the disruption or the disconnection of the visual word form area, a region reproducibly located within the left occipito-temporal sulcus and encoding the abstract identity of strings of visual letters. The functional specialization of this area suggests that it is initially plastic and becomes attuned to the orthographic regularities that constrain letter combination during the acquisition of literacy. The visual word form area further belongs to the network of areas that are consistently implicated in studies of people with developmental dyslexia. Developmental dyslexia is typically interpreted as resulting from a core phonological disorder and most neuroimaging studies showed reduced activity in the left perisylvian regions which have a role in phonological processes. Although low level visual and/or visual attentional disorders have been consistently reported suggesting a visual basis of developmental dyslexia, these disorders typically co-occurred with phonological problems so that the phonological deficit was viewed as the most plausible cause of the poor reading outcome of dyslexic children. In the last years however, dissociations have been reported in developmental dyslexia between phonological processing deficits and a particular kind of visual disorder, a visual attention span deficit characterised by a reduction in the number of distinct orthographic units which can be processed simultaneously in a single fixation. Large sample studies revealed that a non trivial number of dyslexic children exhibit a visual attention span disorder and that this disorder typically dissociates from phonological impairments in the dyslexic population. Neuroimaging studies suggest involvement of the parietal lobes - in particular the superior parietal lobules - in visual attention span and these brain regions are less active in people with developmental dyslexia. A visual attention span disorder thus appears as a second core disorder related to a parietal dysfunction in developmental dyslexia. Further studies are required to determine whether the phonological disorder or the visual attention span disorder independently contribute to the development of the visual word form area during literacy acquisition. These neurobiological dysfunctions are further modulated by environmental factors such as language characteristics, remedial interventions or socio-economic status. Future studies would help better understanding the interactions between neurobiological and environmental factors and the potential influence of the later on the development of the visual word form area. PMID- 18675052 TI - [Communication, language and the brain: from past anterior to near future]. AB - The goal of this article is to share some reflection on the astonishing evolution of the ideas in communication neuroscience and neurolinguistics over the last 30 years, since the founding of the societe de neuropsychologie de langue francaise. in particular, a number of conceptual and methodological advancements have characterized this period, many of which have been lead or heavily contributed by french-speaking research groups in that field. among the advances discussed, are (a) the widening to discourse and pragmatic components of the concept of language, as well as the theoretical and clinical implication of this conceptual extension, (b) the unique contribution of anatomical and functional neuroimaging, (c) the massive impact of the cognitive revolution on theoretical frameworks of language components, and its influence on therapy, (d) the disappearance and the re-appearance of the brain in the main stream research on language over this period, (e) the new perspectives offered though attention put on social aspects of language and the social participation of the individual with language disorders, (f) the emergence of a genuine science of aphasia rehabilitation, and (g) the rediscovery of inter-individual characteristics both genetically- and environmentally- determined. The authors then risk themselves in trying to envision what could characterize the evolution in the field for the next 30 years. Though perilous, this speculative exercise allowed to pinpoint to a number of anticipated advancements including (a) a probable reconciliation between cognitive and connectionist models to the benefit of both visions, (b) an increased contribution of social and emotional neurosciences, (c) some inescapable technological advancements, including light portable neuroimaging devices which will allow to study the neurobiological bases of communication in natural contexts, (d) the integration between communication and aging in cognitive neuroscience and (e) the development of new research fields at some unexpected cross-roads (e.g., physical activity and communication ?) as well as the description of new types of aphasia (e.g., towards a chat aphasia !). Taken together, the advancements made over the 30 last years, and the perspective of future development, confirms the premier role of cognitive neuroscience of language and neurolinguistics in setting the pace through innovative approaches that allow unveiling the mysteries of the mutual relationships between brain and cognition. PMID- 18675053 TI - [Episodic memory, frontal functioning, and aging]. AB - Episodic memory is commonly defined as the kind of memory that renders possible conscious recollection of personal happenings and events from one's personal past. Although it is classically assumed that episodic memory is subserved by a distinct neurocognitive system including mediotemporal cortex and hippocampus, recent evidence also supports the idea of a close relationship between episodic memory and frontal cortex. This view assumes that the frontal cortex plays a critical supervisory role in empowering encoding and retrieval episodic memory operations. In recent years, this view had significantly influenced research in the field of normal memory aging. Indeed, different data have highlighted that age-related cognitive differences, most particularly age-related memory differences, might be explained by the decline of executive-frontal functioning that accompanies aging. In this article, we provide studies on aging and episodic memory that, in support of the executive hypothesis of aging episodic memory, have provided evidence that age-related differences in strategies implemented at encoding and retrieval in this type of memory are mediated by the executive functioning difficulties of older adults. PMID- 18675054 TI - [Early episodic memory impairments in Alzheimer's disease]. AB - Patients with Alzheimer's disease (AD) show early episodic memory impairments. Such deficits reflect specific impairments affecting one or several stages of encoding, storage and retrieval processes. However, AD patients not only have great difficulty retrieving memories and information but also suffer from distortions of memory, as intrusions and false recognitions. Intrusions can be defined as the unintentional recall of inappropriate information in a laboratory learning tasks such as word-list recall and story recall. False recognition refers to the erroneous recognition of information that was not previously presented. The first objective of this review is to present studies from the literature that allowed a better understanding of the nature of episodic memory deficits in AD, and to examine recent research on false memories. The second part of this review is aimed at presenting recent research conducted on prospective memory (PM) in Alzheimer's disease. Prospective memory situations involve forming intentions and then realizing those intentions at some appropriate time in the future. Everyday examples of prospective memory include remembering to buy bread on the way home from work, remembering to give friends a message upon next encountering them, and remembering to take medication. Patients suffering from AD show difficulties in performing prospective tasks in daily life, according to the complaints of their care givers, and these difficulties are massively present at the first stages of the disease. Nevertheless, very few studies have been dedicated to this subject, although the evaluation of PM could be helpful for the early diagnosis of AD. PMID- 18675056 TI - Tissue procurement and transplantation: a Tuscany perspective. AB - Tissue procurement and transplantation are rarely taken into account as indicators of the efficiency of a regional donor procurement network. We present herein a retrospective review on Tuscany tissue procurement activities from 2004 until 2006. In 2003 the Tuscan Regional Government appointed a transplantation service authority to reorganize all regional donation and transplantation activities: the Organizzazione Toscana Trapianti (OTT). The regional tissue procurement network was based on either brain death (BD) and cardiac death (CD) donors under the responsibility of in-hospital transplantation coordinators (IHTCs). From 2004 to 2006, a total of 397 tissue donors were procured in Tuscany, and 4151 tissue transplantations were performed: 2909 skin grafts, 1209 bone grafts, and 33 heart valves. Over the same period, a total of 2116 cornea donors were procured; 4117 corneas were retrieved; 1779 were fit for transplantation, and 1418 were transplanted. Based on our experience, implementation of tissue procurement requires use of BD donors and paramount organizational efforts from IHTCs. PMID- 18675057 TI - Model for End-Stage Liver Disease (MELD) system to allocate and to share livers: experience of two Italian centers. AB - BACKGROUND: The use of the Model for End-stage Liver Disease (MELD) score to prioritize patients on liver waiting lists and to share organs among centers was effective according to US data, but few reports are available in Europe. MATERIALS AND METHODS: We evaluated the outcome of 887 patients listed between April 2004 and July 2006 in a common list by two transplant centers (University of Bologna [BO] and University of Modena [MO] ordered according to the MELD system. Patients with hepatocellular carcinoma had a score calculated according to their real MELD, tumor stage, and waiting time. RESULTS: Five hundred eighty six (67%) patients were listed from BO and 291 (33%) from MO. The clinical features of recipients (sex, age, blood group, and real MELD) were comparable between centers. The number of liver transplantations performed was 307, and 273 (89%) recipients had a calculated MELD >or=20. Liver transplantations were equally distributed according to the number of patients listed: 215 out of 586 (36.7%) for BO and 92 out of 291 (31.6%) for MO. The median real MELD of patients transplanted was 20, and 246 out of 307 (80.1%) grafts transplanted were functioning. The dropouts from the list were 124 (14%), and 87 (70%) of these patients had a calculated MELD >or=20. CONCLUSION: The MELD system was effective to share livers among the two Italian centers. According to this policy, livers were allocated to the recipients with the highest probability of dropout and who had a satisfactory survival after liver transplantation. PMID- 18675058 TI - The quality assurance program of organ donation in Tuscany. AB - Constant monitoring is paramount in order to detect the criticalities and improve the results of the deceased donation process. Concomitant with the institution of a regional transplantation service authority--Organizzazione Toscana Trapianti- in 2003, Tuscany adopted a program of quality assurance of the deceased donation process by compulsory reporting of all encephalic deaths from local intensive care units to the regional transplant office in Florence. The indicators we adopted were the efficiency of deceased donor (DD) identification, expressed as the ratio of encephalic deaths (ED) to total deaths with encephalic lesions (EL) (ie, ED/EL); the efficiency of DD reporting, expressed as the ratio of reported potential DD (RPDD) to total ED (ie, RPDD/ED); the efficacy of the DD process, as the ratio between actual DD (ADD) to total ED (ie, ADD/ED); the conversion rate; the percent of opposition to donation; and the incidence of DD maintenance failures. Data were collected prospectively, stratified by regional hospital consortia (Aziende Sanitarie Locali) and compared with international benchmarks. In the period 2003-2006 the mean efficiency of DD identification was 48.3%+/-4.4% (range 42.6%-53.2%); the mean efficiency of DD reporting was 95.2%+/-2.5% (range 92.5%-98.5%); the mean efficacy of the deceased donation process was 51.8%+/-2.4% (range 48.6%-54.4%); the mean conversion rate was 59.6%+/-2.2% (range 57.6% 62.7%); the mean opposition rate was 31.9%+/-1.1% (range 30.6%-33.2%); and the incidence of DD maintenance failure was 5%+/-2.9% (range 2.2%-8.7%). The breakdown analysis revealed wide interhospital variability in terms of efficiency of DD identification (from a low of 25% to a high of 80%); efficacy of the donation process (from a low of 22% to a high of 79%); and conversion rate (from a low of 29% to a high of 79%). Our results highlight that the donation process gets started in about 50% of eligible cases. Further strategies are favored to address this critical area. PMID- 18675059 TI - Legal aspects of organ transplantation in Italy. AB - Informed consent is of paramount importance in any field of surgery, both from the ethical and the legal points of view. Concerning organ transplantation, potential recipients are fully informed before entering the waiting list. However, according to Italian law, they have to sign another informed consent form before entering the operating room. In our opinion, not only should recipients be informed of the quality of the donor and of the particular organ(s) they are going to receive, but also before entering the waiting list they should accept or refuse the future possibility of receiving an organ from a so-called marginal or extended criteria donor (ECD) and/or a non-heart-beating donor (NHBD). PMID- 18675060 TI - Evaluation of research products released during a national project funded by the Italian Ministry of Health. AB - Evaluation of research products released during activities that have received public funds should be a mandatory practice. In this study, we evaluated the research products released during the research program "Innovative Strategies to Expand Cadaveric Donor Pool for Liver Transplantation" (SITF Project), funded by the Italian Ministry of Health. The SITF Project prefigured the participation of Italian transplantation centers with more advanced experience in the field of liver transplantation. The research products released during the SITF Project were evaluated according to Guidelines for Research Evaluation of the Committee for Evaluation of Research (CIVR) from the Italian Ministry of Education, University and Research. Thus, we considered as the research products released during the SITF Project the number of articles published in English language in peer-reviewed scientific journals by each operative unit (OU), as returned by PubMed. The articles were included on the basis of relationship with the research lines of the SITF Project and availability of the impact factor (IF) calculated by Thomson Scientific for each journal. Following these criteria, 62 research products were released during the executive phase of the SITF Project (January 1, 2004-December 31, 2005). All research products were original articles, with no reviews or letters. The articles were published in 10 different scientific journals (mean IF, 2.64+/-1.74). Spearman's rank test found a weak negative correlation between journal IF and number of published articles (r= -.2919; P= .413). Although the OU involved in the coordination of the SITF Project released a relatively high number of articles (n=8; IF, 0.923+/-0.352), several other OUs obtained better results for number of articles (9-11) and/or IF (3.071+/-2.248 or 2.959+/-1.779). In projects that benefit from public funds, the potential negative impact of coordination activities to scientific production should be adequately considered. PMID- 18675061 TI - Evidence of liver histological alterations in apparently healthy individuals evaluated for living donor liver transplantation. AB - BACKGROUND: Living donor liver transplantation (LDLT) represents an important therapeutic option for patients with end-stage liver disease (ESLD). It has been reported that steatosis may be a serious problem in patients who donate a part of their liver. Liver biopsy represents an accepted method to assess the rate of steatosis and the possible risk to the donor. Nonetheless, some histological abnormalities have been documented in the specimens from potential donors. The aim of this study was to evaluate the possible hepatic histological alterations among apparently healthy candidates for liver donation who did not show serological or ultrasound (US) evidence. MATERIALS AND METHODS: From January 1, 2005 until October 15, 2006, we performed virological, biochemical, and tumor marker evaluations and liver biopsies on 20 LDLT donor candidates. At histological evaluation we classified the evidence of steatosis (5%-10% or 10% 20%), fibrosis (absent or 1-3 portal space), inflammation, iron deposition, biliary neoductulation, and portal vein vascular alterations. RESULTS: Among the 20 subjects, serological markers did not show any pathological alterations. At liver biopsy we found: steatosis (5%-10%) in 6 individuals (about 30%) with 1 ranging from 10% to 20%; iron deposition in 4 (20%); biliary neoductulation in 3 (about 16%); fibrosis in 4 (20%); inflammation in 5 (25%); and portal vein dilatation in 10 (50%). CONCLUSIONS: Our data showed that apparently healthy individuals who did not display serological markers or US evidence of pathology had liver histological abnormalities. This result suggested that in absence of clinical or laboratory alterations, liver biopsy may represent a useful diagnostic tool for living donor candidates. Long-term follow-up results for the laboratory data among those patients should be performed even though they were not qualified for LDLT. PMID- 18675062 TI - Donor affected by hemosiderosis: is kidney transplantation possible? A case report. AB - Marginal donors (advanced age, comorbidities, and so on) provide an increasing contribution to the kidneys used to alleviate the relative organ shortage. We describe the evaluation process and clinical outcome of two kidneys with hemosiderosis used as a double graft. The donor was a 59-year-old hypertensive man, known to have a mechanical mitral valve, who died from a cerebral hemorrhage, with a normal serum creatinine (SCr) and kidneys with normal appearances at sonography. A protocol donor biopsy showed a Karpinsky score of 5 for both kidneys. A double graft was therefore scheduled. The recipient was a 59 year-old man, on dialysis because of chronic glomerulonephritis. HLA match was incompatibility 4/6; immunosuppression was based on steroids, cyclosporine, and mycophenolate mofetil with basiliximab as induction therapy. The grafts showed delayed function with dialysis treatments performed from postoperative day (POD) 1. On POD 2, a magnetic resonance imaging (MRI) study showed the typical appearance of siderosis. Pearl's staining performed on a protocol biopsy confirmed the presence of widespread iron deposits. On POD 5, a recipient renal biopsy showed a superimposed severe acute tubular necrosis. Renal function recovered slowly; SCr at discharge on POD 22 was still 4.2 mg/dL. Two months later, the SCr was 2.2 mg/dL. A second MRI performed at 3 years and 6 months after transplantation confirmed a progressive removal of iron overload while the patient had stable renal function (glomerular filtration rate) of 33 mL/min and SCr: 2.3 mg/dL. We concluded that donors with hemosiderosis should be treated as marginal donors and may be grafted based on a pretransplant biopsy. PMID- 18675063 TI - "Deviceless" hand-assisted laparoscopic donor nephrectomy. AB - Hand-assisted laparoscopic nephrectomy (HLN) in living donors is a minimally invasive surgical modality that uses classic laparoscopic techniques combined with the surgeon's hand as a support tool during renal dissection. We describe our experience with 14 donors undergoing HLN with a novel "deviceless" technique (DL-HLN). We used a midline or a paramedian incision. The first 10-mm trocar (camera) was inserted near the umbilicus and another 10-mm trocar placed under laparoscopic vision at the level of the anterior axillary line above the iliac crest. DL-HLN was performed in 14 patients (11 women and 3 men) of overall mean age of 40 years (range=33-60). Left nephrectomy was performed in all cases. Mean surgical time was 105 minutes (range=60-150). Estimated blood loss was 50 to 800 mL (mean=200 mL). Mean warm ischemia time was 3.5 minutes (range=2-11). Mean hospital stay was 4 days (range=3-6). In one case, uncontrollable hemorrhage developed due to a renal vein lesion at the level of the adrenal vein outlet, requiring conversion to open surgery. As to graft function, recipient serum creatinine on day 7 ranged from 0.9 to 2.6 mg/dL (mean=1.6). We used no device in our technique. The pneumoperitoneum was maintained by the sealing effect of the muscular fascia around the surgeon's wrist. Moreover, the kidney was removed through the hand port without an Endobag. Our modified HLN technique avoids the use of costly disposables and offers the advantages of a smaller incision. PMID- 18675064 TI - Hyperimmunized patients awaiting cadaveric kidney graft: is there a quick desensitization possible? AB - On all kidney waiting lists the 10% to 20% of patients who have antibodies against more than 80% of a panel of HLA antigens (panel reactive antibody [PRA] >80%) are difficult to transplant. The best solution for these patients is to find a compatible donor, ideally a full match, who yields a negative crossmatch test (CMX). If this is not possible, desensitization treatment (high-dose) intravenous immunoglobulin (IVIG) or plasmapheresis (PP) + low-dose IVIG is possible with good results in living donor kidney transplantation mainly if the antibody titer is low. It may also be offered to patients awaiting cadaveric donors too after a long waiting time; however, when applied for several months, it has the obvious disadvantage of giving the patient the risk for long-lasting immunologic weakness without the certitude of finding a kidney. In one of our recent cases of combined liver plus kidney transplantation, a positive CMX became negative 8 hours after the liver operation; the kidney was transplanted with a good result which lasted over 3 years. This observation suggested the possibility of a quick desensitization protocol in selected patients with a large (but not strong) immunization who probably are the majority. Patients sensitized to IVIG and with low titer PRA could be given a single PP + low-dose IVIG (what can be done within the time limit of cadaveric donor kidney transplantation) with good probability of turning an initial positive CMX to negative with the possibility of performing the operation and the advantage of giving the immunosuppression only when the kidney is present. PMID- 18675065 TI - Clinical significance of a positive flow crossmatch on the outcomes of cadaveric renal transplants. AB - Pretransplantation crossmatching is an integral part of kidney transplantation. Flow cytometric crossmatch (FCXM) is more sensitive than complement-dependent cytotoxic crossmatch (CDC-XM). However, the clinical significance of positive FCXM with negative CDC-XM is controversial. We evaluated FCXM in 455 consecutive deceased donor renal transplants. All had a negative CDC-XM. There were 341 T cell and B-cell FCXM negative and 38 T-cell and B-cell positive. There was a higher percentage of retransplantations and HLA mismatches (26.3% vs 8.2%, P= .002 and 2.45 vs 1.99, P= .02, respectively) in the FCXM-positive group compared with the FCXM-negative group; 65.8% of the FCXM-positive patients had rejection compared with 49.3% of the FCXM-negative patients (odds ratio [OR]=1.89, P= .06). FCXM-positive patients had a higher incidence of vascular rejection (28.9% vs 12.6%, OR=2.68, P= .008). One- and 5-year graft survivals were 84% and 66% in the FCXM-positive group vs 90% and 75% in the FCXM-negative group. Censoring for patient death, 1- and 5-year graft survivals were 84% and 73% in the FCXM positive group vs 94% and 82% in the FCXM-negative group. There was no difference in renal function between the 2 groups. In conclusion, a positive T-cell and B cell FCXM transplant with a negative CDC-XM is associated with a higher incidence of rejection, twice the risk of vascular rejection, and a trend toward poorer graft survival. PMID- 18675066 TI - Evaluation of an information system for kidney transplantation in adult and pediatric recipients using the RAND/UCLA Appropriateness Method. AB - In the mid-1980s, RAND Corporation and University of California Los Angeles (UCLA) developed the RAND/UCLA Appropriateness Method (RAM) to evaluate the correctness of medical and surgical procedures. In this study, the RAND/UCLA Appropriateness Method was used to evaluate the appropriateness of a dataset concerning kidney transplantation in adult and pediatric recipients for an information system funded by the Italian Ministry of Health. The original dataset was obtained using an interdisciplinary pool of regional experts (n=60). This dataset held 514 items about kidney transplantation in adult (n=268) and pediatric (n=246) recipients. The items were stratified as 3 main groups: pretransplantation items (adult, n=141; pediatric, n=122), transplantation items (adult, n=49; pediatric, n=45), and early posttransplantation and follow-up items (adult, n=78; pediatric, n=79). In the second round, the dataset was subjected to an extraregional panel of independent experts (n=9) to assess each item using a score ranging from 1 to 9 based on increasing appropriateness. The expert-opinion process returned for adult and pediatric kidney recipient items whole mean scores of 8.52+/-0.32 and 8.65+/-0.32, respectively. Overall agreement, uncertainty, and disagreement between experts about item appropriateness concerning adult kidney recipients were 94.6%, 5.4%, and 0%, respectively. For pediatric kidney recipients, overall agreement, uncertainty, and disagreement between experts about item appropriateness were 96.9%, 2.35%, and 0.07%, respectively. This study supported the use of a structured expert-opinion process as an effective strategy to evaluate the appropriateness of large datasets for kidney transplantation in both adult and pediatric recipients. PMID- 18675067 TI - Clinical operational tolerance after kidney transplantation: a short literature review. AB - The clinical era of solid organ transplantation started with a renal transplantation (RT) performed between identical twins in Boston in 1954. The patient did not receive any immunosuppression, thus representing the very first case of operational tolerance (Tol). However, more than half a century later, we must admit the inadequacy of our knowledge regarding such a fundamental aspect of transplant immunology, as demonstrated by the fact that Tol has never been achieved in an intention-to-treat protocol. Herein we aim to shortly review the worldwide experience on clinical operational Tol after RT. Thus far, reports on successful cases of Tol after RT have been anecdotal: the largest series included no more than 10 individuals. We will understand that Tol can develop even in the presence of either HLA mismatches or blood group incompatibility at baseline, in the presence of anti-HLA antibodies during follow-up, as well as in patients having experienced acute rejection. Despite the lack of robust evidence, the fact that Tol is often accidentally discovered by transplant physicians during follow up in noncompliant patients justifies the hypothesis that the real number of Tol cases might be much higher than currently reported. PMID- 18675068 TI - Pediatric kidney transplantation: a snapshot 10 years later. AB - Herein we report the outcomes of pediatric kidney recipients who underwent transplantation at least 10 years prior. A cohort of 36 patients (mean age, 26.4+/-6 years) with a mean follow-up time of 14.2+/-4 years was selected for the study. Immunosuppression consisted of cyclosporine and steroids. Actuarial patient and graft survivals 15 years after the transplantation were 97% and 86%, respectively. Only 1 patient died due to a complicated sclerosant peritonitis. Graft function was good with a mean serum creatinine of this selected cohort of 1.5+/-0.6 mg/dL. Eighteen percent were class 1, 33% class 2, and 49% chronic kidney disease. Hypertension was treated in almost 80% of the patients. The majority of patients were smaller than the average population with a final height (between 0 and -2) standard deviation score (HSDS) but only 27% had a severe growth impairment (HSDS>-2). Regarding nutritional status, fewer than 30% were overweight and only 1 patient was obese with a body mass index (BMI) >30. The majority of patients, except 2 mentally retarded individuals, are or have been attending normal school and achieved full-time employment. In conclusion, long term survivors of a kidney transplant received during childhood reached a high degree of rehabilitation despite a long period of immunosuppression. PMID- 18675069 TI - Primary nonfunction: timing retransplantation versus hemodynamic parameters and kidney function. AB - BACKGROUND: Patients with acute liver failure (ALF) show an aggravated hyperdynamic circulation. We evaluated potential changes in systemic hemodynamics and improved kidney function induced by the molecular adsorbent recirculating system (MARS) in a group of patients with primary nonfunction (PNF). PATIENTS AND METHODS: In the intensive care unit we treated 18 patients with PNF (6 females and 12 males) after orthotopic liver transplantation (OLT) of overall mean age 47.8 years (range, 28-60 years). Continuous MARS treatment was performed on all patients with a kit change every 8 hours during a mean of 10 days (range, 1-20 days). Double-lumen catheter type veno-venous access was used for the blood supply. The blood flow rate was 150 to 250 mL/min, depending on the hemodynamic situation of the patient. Blood passed through an albumin nonpermeable, high flux dialysis membrane. During MARS treatment we monitored the hemodynamic condition, using a series of parameters: heart rate (HR), mean arterial pressure (MAP), cardiac index (CI), systemic vascular resistance index (SVRI), and pulmonary vascular resistance index (PVRI) before (baseline value) as well as after 1 hour (T1), 3 hours (T2), and the end of treatment (T3). RESULTS: There was a progressive decrease in positive inotropic support (dobutamine, norepinephrine) and significant improvement in hemodynamic parameters, such as MAP (P< .01), PVRI/SVRI/V(mean) (P< .002), and KARI (P< .01). The improved kidney functions were shown by significant improvements in serum creatinine (P< .03), urea (P< .02), and urine volume (P< .005). Eleven patients were alive: 6 with OLT and 5 without OLT. Seven patients died: 4 after OLT and 3 before OLT due to multiorgan failure. CONCLUSIONS: The MARS device significantly improved the hemodynamic parameters and kidney function that also determine patient survival in ALF (61.1%) with PNF while awaiting retransplantation presumably by removal of certain vasoactive substances. PMID- 18675070 TI - Excellent long-term results in de novo renal transplant recipients treated with proliferation signal inhibitors and reduced calcineurin inhibitors exposure. AB - BACKGROUND: A new class of immunosuppressants, proliferation signal inhibitors (PSI)--sirolimus and everolimus--has the potential to prevent chronic allograft nephropathy (CAN). This retrospective analysis reports a 6-year practice using PSI at a single center, comparing a regimen based on reduced-dose calcineurin inhibitors (CNI) and PSI versus full-dose CNI and mycophenolic acid (MPA). METHODS: The study population included 70 patients (group A) who received de novo PSI therapy in combination with reduced dose of CNI, standard steroids, and basiliximab induction, and 216 patients (group B) with full-dose CNI, MPA, steroids, and basiliximab induction. RESULTS: No statistically significant differences were recorded in the baseline donor and recipient characteristics. A difference was observed in cold ischemia time, which could represent a bias for the analysis. No differences were recorded in actuarial patient survival, delayed graft function, biopsy-proven acute rejection rates, and renal function analysis. A significant difference was recorded in the actuarial graft survival rate at years 2, 3, and 4 (P< .01), as well as overall graft survival rates (P= .025). DISCUSSION: The reduction of cold preservation time seemed to be an important factor to improve both short- and long-term renal function. This regimen revealed a long-term trend toward better renal function and graft survival. The use of PSI with reduced doses of CNI seems to be indicated for suboptimal grafts, especially when a reduced quality of the kidney is associated with prolonged cold ischemia time. PMID- 18675071 TI - Patient mortality after graft failure reduces kidney transplant patient survival only in the long term: an "intention to treat" analysis. AB - The benefits of kidney transplantation over dialysis on patient survival have been demonstrated without considering the outcomes of patients with graft loss. To determine whether mortality after graft failure reduced the transplantation advantage in patient survival, we retrospectively reviewed the outcomes of 918 first-deceased renal transplant recipients from May 1979 to August 2005. Patient survivals were 88% and 72% at 10 and 20 years; cancer (26%) and cardiovascular disease (25%) were the major causes of death. Graft survivals were 72% and 50% at 10 and 20 years; chronic rejection was the major cause of graft loss (50%). Patient outcomes after return to dialysis were reviewed in 224 of 240 patients. The survivals were 97%, 83%, and 70% at 1, 5, and 10 years, respectively; cardio cerebrovascular disease (56%), infections (9%), cachexia (9%), and cancer (8%) were the major causes of death. Mortality correlated with patient age at transplantation (P< .001). Re-listed patients (96 of 224) were younger (32+/-10 vs 43+/-11 years; P< .001), had a shorter dialysis period pretransplant (3.2+/ 3.1 vs 4.3+/-3.9 years; P< .03), and a better survival at 10 years (98% vs 56%; P< .001). Ten-year mortality for patients who returned to dialysis was 20% higher than for patients with a functioning graft (P< .001). The reduction in overall patient survival was 2.2% at 10 years (P=NS), 5% at 15 years (P=NS), and 14% at 20 years (P< .05). The same results have been demonstrated for patients >50 years at transplantation. In conclusion, the mortality rate after return to dialysis did not influence the long-term benefits of kidney transplantation. PMID- 18675072 TI - Quality of life in renal transplant patients over 60 years of age. AB - Kidney transplantation not only drastically improves the life-expectancy of hemodialyzed patients, but it also affords psychological and social advantages with improvements in short- and long-term personal and working lives. Quality of life (QoL) is one of the parameters of psychological well-being. There is an improvement of QoL from pre- to posttransplant, but it is not to the level of healthy samples. The aim of this study was to examine QoL in older renal transplant recipients. All recipients older than age 60 were included, with a minimum follow-up of 12 months. To measure QoL, the nationally standardized ShortForm-36 (SF-36) questionnaire was administered. The SF-36 responses by our patients were compared with national age- and gender-appropriate norms, and also between genders. The enrolled population included 19 women (36.5%) and 33 men (63.5%), with a mean age of 66.8 years (range, 60-73 years). Enrolled women reported significant limitations compared to gender- and age-matched norms in social activities (42.11 vs 70.58), perception of pain (22.11 vs 59.17), and general health perception (39.58 vs 48.69). Enrolled men reported significant limitations compared to gender- and age-matched norms in social activities (46.59 vs 78.35), perception of pain (18.18 vs 73.62), psycho-physical energy (50.15 vs 67.88), and general health perception (37.33 vs 61.66). No significant differences were noted between the genders. This study clearly showed how the psychological state was not as good as the clinico-physical recovery following renal transplantation in older recipients. PMID- 18675073 TI - Kidney transplantation combined with other organs in Bologna: an update. AB - BACKGROUND: We retrospectively reviewed our experience in combined liver-kidney (L-KT) and heart-kidney (H-KT) transplantations. PATIENTS AND METHODS: Between January 1997 and April 2007, we performed 25 L-KT and 5 H-KT. Patient mean age was 51+/-8 years in L-KT and 43+/-11 years in H-KT. The main cause of liver failure was chronic viral hepatitis (14 cases). Etiology of heart failure was dilated cardiomyopathy and hypertrophic cardiomyopathy (4 and 1 patients, respectively). The main causes of renal failure in L-KT were chronic glomerulonephritis (n=8) and polycystic disease (n=7). Etiology of renal failure in H-KT was interstitial nephropathy (n=2), vascular nephropathy (n=2), and chronic glomerulonephritis (n=1). RESULTS: Mean follow-up was 32+/-26 months in L KT and 24+/-17 months in H-KT. Immunosuppression was cyclosporine-based (n=4) or tacrolimus-based (n=21) in L-KT and cyclosporine-based in H-KT. Acute rejection rate was 8% for both liver and kidney in L-KT; 80% (mild) for heart and 40% for kidney in H-KT. In the L-KT group, there was no primary graft nonfunction (PGNF). Two patients experienced liver delayed graft function (DGF); 1 patient required postoperative dialysis. One-year graft and patient survivals were both 84% and overall graft and patient survival was 76%. In the H-KT group, 3 patients needed postoperative dialysis and 1 required a cardiac assistance device for 48 hours; overall graft and patient survival was 100% with good cardiac and renal functions. CONCLUSION: Our experience confirmed that H-KT and L-KT are safe procedures, offering good long-term results. PMID- 18675074 TI - Multicenter study on double kidney transplantation. AB - BACKGROUND: Marginal organs not suitable for single kidney transplantation are considered for double kidney transplantation (DKT). Herein we have reviewed short and long-term outcomes of DKT over a 7-year experience. PATIENTS AND METHODS: Between 2001 and 2007, 80 DKT were performed in the transplant centers of Bologna, Parma, and Modena, Italy. Recipient mean age was 61+/-5 years. The main indications were glomerular nephropathy (n=33) and hypertensive nephroangiosclerosis (n=14). Mean HLA A, B, and DR mismatches were 3.1+/-1.2. Donor mean age was 69+/-8 years and mean creatinine clearance was 75+/-27 mL/min. Almost all kidneys were perfused with Celsior solution. Mean cold ischemia time was 17+/-4 hours and mean warm ischemia time was 41+/-17 minutes. Mean biopsy score was 4.4. Immunosuppression was based on tacrolimus (n=52) or cyclosporine (n=26). RESULTS: Fifty (62.5%) patients displayed good postoperative renal function. Thirty (37.5%) experienced acute tubular necrosis and required postoperative dialysis treatment; 8 acute rejections occurred. Urinary complications were 13.7% with 8/11 requiring surgical revision. There were 6 surgical reexplorations: intestinal perforation (n=2), bleeding (n=3), and lymphocele (n=1). Two patients lost both grafts due to vascular and infectious complications at 7 or 58 days after transplantation. Two patients underwent intraoperative transplantectomy due to massive vascular thrombosis. Four (5%) patients underwent transplantectomy of a single graft due to vascular complications (n=2), bleeding (n=1), or infectious complications (n=1). Graft and patient survivals were 95% and 100% versus 93% and 97% at 3 versus 36 months, respectively. CONCLUSIONS: DKT is a safe approach for organ shortage. The score used in this study is useful to determine whether a kidney should be refused or accepted. PMID- 18675075 TI - In situ elongation patch in right kidney transplantation. AB - BACKGROUND: A short right renal vein complicates transplantation causing traction and difficulties during anastomosis. When we perform a kidney transplantation from a cadaveric donor, this problem may be resolved by using the vena cava to create a venous duct. This elongation technique is proposed to be performed during bench surgery. We propose a small change in the technique: execution of an "elongation patch" during harvesting and under cold perfusion. MATERIALS AND METHODS: From January 2004 to June 2006, we performed 12 in situ (during the harvesting procedure) vena cava elongation patches. In these cases, the right renal vein was too short. The elongation patch was used in only 8 transplantations. In the other 4 cases we sacrificed the patch to perform a direct venous anastomosis due to favorable recipient anatomical characteristics. RESULTS: The 8 transplantations performed with the elongation patch did not show vascular complications and the venous anastomosis was easy to perform. CONCLUSIONS: The "extension patch" was not associated with a greater incidence of vascular complications. Using the elongation patch during the harvest showed some advantages: performed during cold perfusion with a reduction in bench ischemia; the anatomical relationships are preserved so we can perform a calibrated suture; the perfusion of the organ allows us to observe the integrity of the anastomosis. This technique did not significantly increase the harvesting time. PMID- 18675076 TI - Infective complications in renal allograft recipients: epidemiology and outcome. AB - INTRODUCTION: Successful renal transplantation strictly depends on good control of rejection and better prevention and treatment of infections, which remain serious threats. METHODS: This retrospective, observational study of 245 renal allograft recipients who underwent transplantation between January 2002 and December 2005 included a 21+/-10 months follow-up. RESULTS: A total of 110 (44.9%) patients developed an infective process during the posttransplantation period, namely, 232 infective processes. Eighty patients developed at least 1 episode of urinary tract infection (UTI) 11 patients (4%) had a wound infection, and 30 patients (12%) had pneumonia. We diagnosed 35 cases of bacteremia (35%), whereas cytomegalovirus (CMV) infection was demonstrated in 40 patients (16%). CONCLUSIONS: Immunosuppressive therapy, necessary to avoid acute and chronic rejection, exposes patients to a higher rate of infectious complications. The immunosuppressive protocols led to a relatively low incidence of infectious complications, mainly of little clinical significance. The highest incidence was evident by the sixth month after transplantation, when the immunosuppressive regimen exercised its most depressive effects on patient immune systems. PMID- 18675077 TI - Human papilloma virus infection and cervical intraepithelial neoplasia in transplanted patients. AB - Progress in diagnosis and treatment has led to an increased number of transplantation patients who consequently have immunological depression and emergence of tumors. The incidence of cervical neoplasia, according to previous studies, is 11%; this tumor is the only one that can be investigated by screening before and after a graft. Our purpose was to evaluate whether transplanted patients showed an increased incidence of genital human papilloma virus (HPV) infection and whether this infection produced greater progression of disease in cases of low-risk HPV infections. Our study involved 151 transplant patients who underwent Papanicolaou (Pap) and HPV tests. Patients listed for grafts underwent Pap and HPV tests 6 months before and 6 months after transplantation. All patients had negative Pap tests before their grafts. After their grafts 16 patients (10.59%) had negative Pap tests, but positive viral typing. Eleven patients (7.28%) showed positive Pap tests, 6 of whom had low-grade squamous intraepithelial lesion (SIL) and 5 patients high-grade SIL. The final HPV infection incidence (15.23%) was consistent with the literature. The incidence of lower female genital tract intraepithelial lesions (7.28%) was higher than the healthy population or analogous studies (4.5%-8.5%). We showed a constant association between high-risk HPV infection and gynecologic intraepithelial neoplasia, whereas there was no association between low-risk broods HPV infection and neoplasia. In conclusion, screening should start at almost 6 months before grafting to avoid an irreversible situation that is difficult to treat. PMID- 18675078 TI - Incidence of urinary tract infections caused by germs resistant to antibiotics commonly used after renal transplantation. AB - BACKGROUND: The inadequate utilization of antibiotics is responsible for the development of urinary tract infections (UTI) after renal transplantation (RT), through the induction of resistance to the antibiotics themselves. The purpose of this study was to evaluate the incidence of resistance to cefotaxime (CEF) and trimethoprim/sulfamethoxazole (TMP-SMX), routinely used for surgical perioperative prophylaxis and prevention of Pneumocystis carinii, respectively. MATERIALS AND METHODS: We enrolled all adult patients having received an RT from 2001 to 2006 and having a minimum follow-up of 6 months. Urine cultures (UC) were routinely performed at every outpatient clinic control and whenever required by the onset of significant clinical signs/symptoms. UTI was diagnosed by the presence of a positive UC. The endpoint of the study was the emergence of bacterial strains resistant to either CEF or TMP/SMX. RESULTS: We recorded 169 UTI in 76 patients (38 men/38 women, 33%) over a mean follow-up of 779.9+/-523.3 days. Thirty-nine patients (51%) developed more than 1 UTI episode. When gram negative bacteria were considered, 102/144 (70.8%) tests showed resistance to TMP/SMX, while data were available in about only 7 gram-positive infections (5/7, 71%). CEF was tested less frequently with 21/43 (49%) germs resistant to this molecule. CONCLUSIONS: The onset of bacterial resistance to either TMP/SMX or CEF is frequent after RT. A wiser stricter utilization of antibiotics is mandatory. Standard antibiotic protocols should be revised. PMID- 18675079 TI - New-onset diabetes mellitus after kidney transplantation: the role of immunosuppression. AB - BACKGROUND: Complications related to posttransplantation immunosuppressive therapy remain common. New-onset diabetes mellitus after transplantation (PTDM) is a well-recognized complication associated with reduced graft and patient survival. The type of immunosuppression may be responsible for more than two thirds of PTDM. We retrospectively reviewed our experience in a population of 284 kidney transplant recipients, evaluating the incidence of PTDM with regard to the type of immunosuppression. PATIENTS AND METHODS: From January 2001 to December 2005, 284 kidney transplantations were performed using tacrolimus-based (TAC) immunosuppression in 192 patients and a cyclosporine-based (CyA) regimen in 62 patients, whereas 30 patients received sirolimus-based immunosuppression. RESULTS: The overall incidence of PTDM was 4.9%. Among the immunosuppression protocols, 8 patients (4.1%) received TAC and 6 patients (9.6%) received CyA, whereas no patients treated with sirolimus developed PTDM. Graft and patient survival rates were 93% and 100%, respectively. CONCLUSIONS: The overall risk of PTDM with recent immunosuppressive protocols is low, but it is increased among calcineurin inhibitor (CNI)-treated kidney transplant recipients. Sirolimus did not increase the risk of PTDM, allowing potential clinical application in diabetic recipients and in patients affected by PTDM. PMID- 18675080 TI - Analysis of posttransplant diabetes mellitus prevalence in a population of kidney transplant recipients. AB - AIM: The onset of posttransplant diabetes mellitus (PTDM) among kidney recipients is associated with an increased risk of graft failure and death. Minimizing the risk of PTDM is a priority for long-term improvement in survival rates. We sought to evaluate the prevalence of PTDM and impaired fasting glucose (IFG) among a population of kidney transplant recipients to identify the risk factors and to evaluate graft and patient survivals. METHODS: We analyzed 250 consecutive Caucasian patients who received kidney allografts in our center between May 2000 and December 2005, with a median follow-up of 32 months (range, 1-78 months). RESULTS: We observed altered glucose metabolism in 17% of patients; specifically, the prevalences of PTDM and IFG were 12.2% and 4.8%, respectively. Patients who developed PTDM or IFG were overweight (BMI, 26.4+/-3.4 and 28.1+/-3.4 kg/m(2), respectively), whereas the normal glucose (NG) group's BMI was 23.8+/-3.5 kg/m(2) (P= .002 and P= .004, respectively). Prevalence of acute rejection was higher in the PTDM and IFG patients compared with the NG patients (60.7%, 63.6%, and 32.1%, respectively; P= .006; P< .04), while no difference was observed in terms of graft and patient overall survival. CONCLUSION: In our series of patients, we showed that being overweight represents a major risk factor for the development of PTDM, which results in an increased acute rejection rate. These results confirmed the importance of appropriate weight control among patients undergoing kidney transplantation, which should also be strictly monitored for all risk factors associated with the development of impaired glucose metabolism. PMID- 18675081 TI - Arterial changes in children undergoing renal transplantation. AB - INTRODUCTION: Pathological changes of large arterial walls and the heart have been described in patients with chronic renal failure. The aim of the study was to verify the incidence of arterial changes among our series of pediatric patients with end-stage renal disease undergoing transplantation. PATIENTS AND METHODS: From January 2004 to December 2006, 26 patients (15 boys and 11 girls) of overall mean age of 13.12 years (range=3-27 years) underwent renal transplantation in our department. The pretransplant dialysis treatment was peritoneal in eight and hemodialysis in 18 cases. All patients were divided in two groups according to primary renal disease: group A were 18 patients with congenital urinary malformations; and group B, eight patients with acquired glomerular diseases. In each case, a sample of artery from both donor (aortic patch of kidney) and recipient (iliac patch of graft allocation) was obtained during renal transplantation. The donors were considered to be the control group. RESULTS: Light microscopy showed pathological changes in 12/26 recipient arteries: nine showed light fragmentation of the internal elastic lamina; two, more severe fragmentations; and one, fragmentation of the internal elastic lamina associated with mucopolysaccharide deposits. Pathological changes were more evident in group A than B, but the difference was not significant (P> .05). Among the donor group, 11 patients showed light fragmentation of the internal elastic lamina, but there was no significant difference with the recipients (P> .05). CONCLUSIONS: Among our group of patients, we observed only slight modifications of the arterial wall. These changes were nonspecific, similar to those in a control group of donors matched for age. A possible explanation of these findings may depend on the short time of dialysis before transplantation. PMID- 18675082 TI - Gastrointestinal perforations following kidney transplantation. AB - This study reports major gastrointestinal (GI) complications among a group of 1611 patients following kidney transplantation. The immunosuppressive regimen changed somewhat during the course of the study but included azathioprine, prednisolone, antilymphocyte globulin, cyclosporine, tacrolimus, mycophenolate mofetil, and sirolimus. Perforations occurred in the colon (n=21), small bowel (n=15), duodenum (n=6), and stomach (n=4). Nearly 50% of the complications occurred while patients were being given high-dose immunosuppression to manage either the early postoperative period or acute rejection episodes. Of the 46 patients affected, 11 (24%) died as a direct result of the GI complication. This high mortality appeared to be related to the effects of the immunosuppression and the associated response to sepsis. Reduction of these complications may be achieved by improved surgical management, preventive measures, prompt diagnosis, and a reduced immunosuppressive protocol. PMID- 18675083 TI - Donor risk index and organ patient index as predictors of graft survival after liver transplantation. AB - In liver transplantation the identification of risk factors and the risk quantification for each single case represent a field of great interest. There are donor-related and recipient-related risk factors. Donor risk index (DRI) was retrospectively calculated in 223 liver transplant cases. We did not include patients with preoperative diagnosis of hepatocarcinoma and retransplants. The cases were stratified into two classes according to the DRI (low risk, DRI<1.7, and high risk, DRI >or= 1.7). A new index, namely the organ patient index (OPI) was calculated adding the Model for End-stage Liver Disease (MELD) score to the DRI. Patients were stratified into two classes according to the OPI (low risk, OPI 2.85). The cases with low DRI (n=144) showed better survival than the cases with high DRI (n=82; P< .02). The cases with low OPI (n=173) showed better survival than cases with high OPI (n=50; P< .01). The OPI predicted outcomes better than DRI, increasing the gap in the long-term graft survival between the low- and the high-risk class. The inclusion of the MELD in the new index allowed better prediction of graft survival. PMID- 18675084 TI - Model for End-Stage Liver Disease (MELD) score and organ allocation from cadaveric donors for 198 liver transplantation procedures performed in a single center. AB - Since February 2002, the United Network for Organ Sharing (UNOS) proposed to adopt a modified version of the Model for End-Stage Liver Disease (MELD) to assign priority on the waiting list for orthotopic liver transplantation (OLT). In this study, we evaluated the impact of MELD score on liver allocation in a single center series of 198 liver recipients (mean age of patients, 52.21+/-8.92 years), considering the relationship between clinical urgency derived from MELD score (overall MELD, 18.7+/-6.83; MELD <15 in 69 patients, MELD >or=15 in 129 patients) and geographical distribution of cadaveric donors (inside/outside Liguria Region, 125/73). The waiting time for OLT was 230+/-248 days, whereas the 3-month and 1-year patient survivals were 87.37% and 79.79%, respectively. No difference was observed for MELD score retrospectively calculated for patients who underwent OLT before February 2002 (n=71) compared with MELD score calculated for patients who received a liver thereafter (18.26+/-6.68 vs 18.94+/-6.92; P= .504). No significant difference was found in waiting time before and after adoption of MELD score (213+/-183 vs 238+/-278 days; P= .500), or by stratifying patients for MELD <15/>or=15 (225+/-234 vs 232+/-256 days; P= .851). Using the geographical distribution of donors as a grouping variable (outside vs inside Liguria Region), no significance occurred for MELD score (19.68+/-7.42 vs 18.17+/ 6.42; P= .135) or waiting time (211+/-226 vs 242+/-261 days; P= .394). In our series, more OLTs were performed among sicker patients and no differences were found in the management of livers procured from cadaveric donors outside or inside Liguria Region. However, further efforts are needed to reduce the waiting time among patients with higher MELD scores. PMID- 18675085 TI - Model for End-Stage Liver Disease (MELD) score system to evaluate patients with viral hepatitis on the waiting list: better than the Child-Turcotte-Pugh (CTP) system? AB - BACKGROUND: The Model for End-Stage Liver Disease (MELD), based on creatinine, bilirubin, and International normalized ratio (INR), has been shown to be superior to the Child-Turcotte-Pugh (CTP) score in predicting 3-month mortality among patients on the transplant waiting list due to end-stage liver disease (ESLD). An additional advantage of MELD is the possibility to add "adjustment points" for exceptional patients at risk for death because of liver disease not identified by changes in the used parameters, as occurs in the case of hepatocellular carcinoma (HCC). Although it is useful, MELD has some important limitations: There are no differences for patients with or without ascites, and for the absence of other laboratory parameters involved in the etiology of disease. In this study, we evaluated dropouts of patients on the waiting list for orthotopic liver transplantation (OLT) based upon the characteristics of these subjects before and after introduction of the MELD score. METHODS: All patients on the OLT waiting list from June 1, 2006 to June 30, 2007 were enrolled in the MELD group (A) and evaluated with CHILD and MELD score, while those listed from January 1, 2004 to May 31, 2005 were enrolled in pre-MELD group (B) to be evaluated with CHILD. In these subjects we assessed the drop out frequency and waiting time and we compared the results to assess possible differences (U Mann Whitney Test; P<.05). RESULTS: The total number of patients included in this study was 176: 116 patients in Group A and 60 in Group B. We had a drop-out frequency of 21% with a median of 9+/-6 S.E. months in Group A, while 9% with a median of 15+/-8 months S.E. in Group B. The dropout frequencies were as follows: Group A--16 deaths (1 HCC--15 disease complications) while in Group B we had 13 drop outs, 10 exitus (4 HCC and 6 disease complications) and three exclusions for nonmedical reasons. In Group A we had a higher number of deaths due to disease complications than in group B (P<.05). Further, we had 32 OLTx in Group A and 45 in Group B. Survival rate did not show any differences between the two groups while number needed to harm was 11. CONCLUSIONS: The use of MELD score in this group of patients produced an advantage for HCC, but seemed to cutoff patients with viral hepatitis complications during the waiting time. Particularly, about one in every 11 patients may receive an harm using this score system. Other parameters should be introduced as adjustment points to make the MELD score suitable also for patients with infectious liver diseases. PMID- 18675086 TI - What is the biopsychosocial role of human immunodeficiency virus positivity in patients with end-stage liver disease who undergo orthotopic liver transplantation? AB - INTRODUCTION: Since 2003 the National Research Program for Solid Organ Transplantation in patients with human immunodeficiency virus (HIV) is active at our liver transplantation center. Patients with HIV who enter this protocol are assessed by the Consultation Liaison Psychiatry Service. The aim of the present study was to evaluate their psychiatric comorbidity. METHODS: An observational prospective study was conducted comparing end-stage liver disease (ESLD) patients with and without HIV. After the assessment, the psychiatrist compiled the Transplant Evaluation Rating Scale (TERS) and the Montgomery Asberg Depression Rating Scale (MADRS). Baseline evaluation was made before inclusion on the OLT waiting list and the follow-up evaluation was made 12 months later. RESULTS: From January 2003 to December 2006 we assessed 553 patients: 39 (6%) with HIV and 361 (94%) without HIV. The 2 groups were homogeneous for gender (75% of male patients; P=not significant [NS]) but not for age (46+/-5 vs 56+/-9; P=NS). Psychiatric history was negative in 176 (49%) patients without HIV and in 6 (15%) patients with HIV (P< .001). At baseline psychiatric comorbidity was present in 33 HIV patients (85%) and in 148 non-HIV patients (41%; P< .001). At follow-up MADRS highlighted an improvement in all of the items for HIV patients. In the non HIV group, the variation was as follows: baseline, 7.10; follow-up, 8.15. In the HIV group, the variation was as follows: baseline, 10.20; follow-up, 4.09 (P< .001). The average score at TERS was higher among patients with HIV (43+/-9 vs 35+/-9; P=NS). CONCLUSIONS: At baseline HIV patients with ESLD showed a higher rate of psychopathology, but they improved at follow-up; the contrary happened in the non-HIV group. PMID- 18675087 TI - Endolymphatic immunotherapy in inoperable hepatocellular carcinoma. AB - INTRODUCTION: We report the preliminary results of endolymphatic immunotherapy in patients with inoperable hepatocellular carcinoma (HCC). METHODS: From 2003 to 2005 we enrolled 31 patients with inoperable HCC. The patients underwent monthly endolymphatic injections of 15-30 x 10(6) interleukin-2 (IL-2)-activated peripheral autologous lymphocytes (LAK) and 250 IU of IL-2. Follow-up included blood biochemistry every 3 months and imaging studies every 6 months. To assess therapy efficacy we considered 12 biochemical parameters, vascular invasion or thrombosis, Child-Pugh scoring system, histological grading, lymphadenopathy, viral state, and alpha-fetoprotein. RESULTS: Sixteen patients completed at least 3 cycles, and 10 patients completed more than 6. No clinically significant adverse reactions occurred. Imaging studies showed no significant decrease in tumor mass. However, the survival of patients who completed 12 therapy cycles was significantly higher than survival of patients with fewer than 12 cycles. Both are significantly higher than that of untreated patients. All patients with 12 completed cycles showed an improvement of 9 parameters or more. DISCUSSION: Endolymphatic immunotherapy is safe, easily performed, inexpensive, and effective in terms of survival. This study should encourage future large-scale investigations so as to reach a firmer conclusion and define uniform inclusion criteria. PMID- 18675088 TI - Microvascular autonomic dysfunction may justify false-positive stress myocardial perfusion imaging in patients with liver cirrhosis undergoing liver transplantation. AB - BACKGROUND: Up to 15% of liver transplant candidates have asymptomatic coronary artery diseases, which increase the risk of cardiac complications during and after transplantation. The aim of this study was to prospectively investigate the usefulness of an integrated cardiological approach in cirrhotic patients undergoing liver transplantation. METHODS: Twenty-four consecutive patients undergoing evaluation for liver transplantation were studied by assessing risk factors for coronary artery diseases, electrocardiogram with QTc interval determination, chest X-ray, echocardiography, 24-hour Holter monitor, myocardial perfusion scintigraphy (99mTc)MIBI-GSPECT at rest and after dipyridamole infusion. Cardiac (123)I-metaiodobenzylguanidine (MIBG) scan and coronarography were performed in patients with myocardial perfusion defects. Twenty three of 24 patients underwent successful liver transplantation; one patient died on the waiting list. RESULTS: Before liver transplantation, 29% of patients were diabetic and 41% were smokers. Eleven of 24 patients had a prolonged QTc interval, and 3/24 had positive myocardioscintigraphy after dipyridamole infusion: in two coronarography was negative, while the (123)I-MIBG washout was altered. No cardiac events were recorded during the short-and long-term follow-up after surgery. CONCLUSIONS: Predictive value of positive cardiac (99mTc)MIBI GSPECT in patients with liver cirrhosis is low, and this may be due to alterations of cardiac microvascular tone as showed by cardiac (123)I-MIBG scan. PMID- 18675089 TI - Treatment of chronic hepatitis C virus infection with pegylated interferon and ribavirin in cirrhotic patients awaiting liver transplantation. AB - Successful treatment of chronic hepatitis C virus (HCV) infection can prevent reinfection after orthotopic liver transplantation (OLT). Pegylated interferon (PEG-IFN) may ameliorate virological response (VR), making the risk-to-benefit ratio of therapy favorable in waiting list patients. From January 2001 to April 2006, we treated 15 HCV cirrhotics with PEG-IFN alpha-2b (1.5 microg/kg/week) and ribavirin (RIBA; >or=10.6 mg/kg/d). Their mean age was 51.5 years. There were 9 men. In 6 cases the genotype was 1b. With Child-Pugh scores >or=9 (range 9-12) and Model for End-Stage Liver Disease (MELD) scores >or=14 (range, 14-22). Adverse events occurred in all subjects: thrombocytopenia (<40,000/microL) in 8; neutropenia (<700/microL) in 10; anemia (Hb <8.5 g/dL) in 1; grade III hepatic encephalopathy in 2; pelvic infection in 1; variceal hemorrhage in 1; and hepatocellular carcinoma (HCC) recurrence in 1. Adverse events caused treatment withdrawal in 6 (40.0%) and RIBA and/or PEG-IFN dose reduction in 10 (66.6%). Early VR (EVR) was obtained in 9 subjects (60.0%), end-of-treatment (EOT) VR in 7 (46.6%), and sustained VR (SVR) in 3 (20.0%). Three subjects--2 nonresponder and 1 breakthrough--were transplanted at 25, 23, and 16 months after the EOT, respectively. Three subjects died at 6, 8, and 15 months after the EOT due to HCC, spontaneous bacterial peritonitis, and liver failure. Nine patients are awaiting OLT. The risk-to-benefit ratio is against PEG-INF and RIBA treatment of severely decompensated cirrhotics infected with genotype 1 awaiting OLT, but therapy is probably beneficial in genotype 2 subjects, due to an expected SVR rate of more than 40%. However, one must carefully consider the high risk for severe adverse events. PMID- 18675090 TI - Pediatric acute liver failure with molecular adsorbent recirculating system treatment. AB - BACKGROUND: The prognosis of pediatric acute liver failure (PALF) has been significantly improved by emergency orthotopic liver transplantation (OLT). Since 2004, the molecular adsorbent recirculating system (MARS) has been proposed as a bridging procedure. The aim of our study was to assess its efficacy in children with PALF. PATIENTS AND METHODS: Since 1999 we performed treatment of 39 fulminant hepatic failure (FHF) cases with MARS. Since September 2004 we treated 6 pediatric patients with FHF who were of mean age 10.6 years (range, 3-15 years) including 4 females and 2 males. In 3 cases the cause of FHF was unknown; in 2 cases, it was induced by paracetamol overdose; and in 1, by acute hepatitis B virus. Inclusion criteria were: bilirubin >15 mg/dL; creatinine >or=2 mg/dL; encephalopathy grade >II; and International normalized ratio (INR) >2.5. Other estimated parameters were: AST and ALT serum levels, lactate, and urine volume. Neurological status was monitored using the Glasgow Coma Scale (GCS). Continuous MARS treatment was performed in all patients with a kit change every 8 hours. Intensive care unit (ICU) treatment was applied to optimize regeneration and to prevent cardiovascular complications. RESULTS: We observed a significant improvement among levels of bilirubin (P< .009), ammonia (P< .005), creatinine (P< .02), GCS (P< .002), and predictive criteria and as Sequential Organ Failure Assessment (SOFA) and Pediatric End-Stage Liver Disease (PELD). Three children underwent OLT: 1 died after 5 days due to primary nonfunction and 2 children are alive after a median follow-up of 14 months. In 2 children the MARS treatment led to resolution of clinical status without liver transplantation. One child died before OLT due to sepsis and multiorgan failure. CONCLUSIONS: We concluded that application of the MARS liver support device in combination with experienced ICU management contributed to improve the clinical status in children with PALF awaiting liver transplantation. PMID- 18675091 TI - Hemodynamic improvement as an additional parameter to evaluate the safety and tolerability of the molecular adsorbent recirculating system in liver failure patients. AB - BACKGROUND: The molecular adsorbent recirculating system (MARS) is an extracorporeal acute liver failure (ALF) support system method using albumin enriched dialysate to remove albumin-bound toxins. PATIENTS AND METHODS: Since 1999 we performed 2027 MARS treatments in 191 patients: 39 fulminant hepatic failure (FHF), 16 primary nonfunction (PNF), 21 delayed function (DF), 94 acute on-chronic liver failure (AoCHF), 7 post-hepatic resection, and 14 intractable pruritus. RESULTS: We divided the complications by the AoCHF versus the ALF populations. Among 83 ALF patients, we observed worsening of hemodynamic parameters in 16 patients: 3 with PNF, 2 with DF without retransplantation, 9 with FHF, and 2 after hepatic resection. Among 94 AoCHF patients, 42 showed hemodynamic instability requiring intensive care unit support. Our study did not note significant adverse effects (1.8%), except for infections and hemorrhage from the central venous catheter not due to MARS treatment. The thrombocytopenia was controlled through administration of platelets before the start of treatment when a patient showed a level under 30,000 mm(3). CONCLUSION: Our results confirmed that nonbiological hepatic support by MARS was safe and tolerable. PMID- 18675092 TI - Acute decompensation and absence of brain and kidney dysfunction predict long term efficacy of plasma exchange in hyper-bilirubinemic cirrhotic patients awaiting liver transplantation. AB - Various artificial liver support systems are currently used in patients with decompensated chronic liver disease or acute liver failure as a bridge to recovery or to orthotopic liver transplantation (OLT). Between June 2004 and September 2006, 9 subjects were treated with plasma exchange (PE) for acute decompensation on chronic liver disease or chronic decompensation in end-stage liver disease. All of them were awaiting OLT or were listed at the moment of decompensation. Grade II to III hepatic encephalopathy (HE) was present in 4 patients, significant renal dysfunction in 3 patients, and ascites in 6 patients. Baseline serum total bilirubin was 35.1+/-11.2 mg/dL (mean value+/-SD). The patients underwent a mean of 12.1 2-hour exchanges over 1 to 8 weeks. The 3 who recovered were alive after a mean follow-up of 22.7+/-10.3 months. There were 3 patients who underwent transplantation and 3 who died due to liver failure during treatment. Only subjects with acute decompensation and without HE or significant renal dysfunction survived without OLT. PE did not significantly modify the grade of HE or the renal function. PE seemed to be a safe, long-term, effective therapeutic option for acute decompensation among subjects with chronic liver disease without brain or renal dysfunction. PMID- 18675093 TI - Twenty seven years of experience in pediatric liver transplantation in Strasbourg: focus on the ex situ split techniques. AB - INTRODUCTION: Despite the well-known controversies about split-liver procedures, since 1979 we have utilized an ex situ instead of an in situ technique because of its feasibility. However, we sought to prove the equality of the results of these two procedures. Herein, we have presented our experience after 27 years' follow up. MATERIALS AND METHODS: Between March 1979 and June 2006, we transplanted 84 livers in 67 pediatric recipients including 37 ex situ split livers implanted into 28 patients. RESULTS: We recorded demographic characteristics, transplantation, and retransplantation indications, age difference between donors and recipients, comorbidities, cold ischemia times, surgical times and complications, graft/recipient body weight ratios, organ recovery times, and overall survivals after 1, 5, and 15 years follow-up. We have herein reported 1, 5, and 15 years of patient versus organ survivals of 88.9.1%, 84.5%, 62.1% versus 78.6%, 74.2%, 57.4%, respectively. CONCLUSION: We have concluded that an ex situ split liver may be a valid alternative to in situ techniques to achieve good grafts for pediatric transplantation. PMID- 18675094 TI - Temporary porto-caval shunt utility during orthotopic liver transplantation. AB - INTRODUCTION: In liver transplantation (OLT) a porto-caval shunt is a well defined technique practiced by many surgeons in several centers. METHODS: We considered 186 cadaveric OLT patients who underwent a cavo-cavostomy-type reconstruction; they were divided into two groups: those in whom we performed a porto-caval shunt (group A) and those in whose we did not (group B). We evaluated several variables: warm and total ischemia time, intraoperative blood and fresh frozen plasma transfusions, crystalloid and colloid requirements, blood loss, operative duration, hemodynamic intraoperative changes and diuresis, length of hospital stay, and creatinine values at days 1 and 2, and at discharge day. RESULTS: Total and warm ischemic time differed significantly between the two groups. Infusion of blood, fresh frozen plasma, colloid, and crystalloid did not significantly differ. Blood loss was lower, and intraoperative diuresis was not significantly increased in group A subjects. Postoperative hospitalizations were 16.5 and 17.8 days and operative times, 504 and 611 minutes in the two groups. Both cardiac index and ejection fraction values during the anhepatic phase were significantly greater among group A than group B patients. PAD at the two phases was greater in group B. The PAS was significantly different only at reperfusion time. Creatinine values were significantly different at discharge. Better survival was shown for group A patients over group B subjects. CONCLUSION: The results presented herein confirmed that a porto-caval shunt during OLT was a safe, useful expedient contributing to an improved hemodynamic status and a better time distribution in the various phases of liver transplantation. PMID- 18675095 TI - Technical solutions for venous outflow reconstruction in damaged liver grafts during procurement: case reports. AB - The incidence and clinical consequences of hepatic injuries (parenchymal, vascular, and biliary) due to surgical handling during multiorgan procurement are still underestimated. Surgical damage to liver grafts may lead to an increased mortality and graft dysfunction rate; therefore, multiorgan procurements require a high level of expertise and training. We report our experience in two cases of accidental venous outflow damage during liver procurement focusing on our repair strategies. In one case, a short suprahepatic inferior vena cava (IVC) was extended by a venous cuff obtained from a long infrahepatic IVC from the same liver graft. In the second case, we observed a complete transection of the middle hepatic vein during in situ splitting procedure. The damage was reconstructed by cadaveric iliac vein interposition. In both cases, liver transplantation was successfully performed without venous complication. An adequate surgical technique in liver procurement and venous reconstruction during living donor and domino liver transplantation are formidable tools to achieve successful liver transplantation with a damaged graft. PMID- 18675096 TI - Adult living donor liver transplantation with right lobe graft: the venous outflow management in the Milan-Niguarda experience. AB - In right lobe living donor liver transplantation (ALDLT), reconstruction of middle hepatic vein (MHV) tributaries is often necessary to avoid severe graft congestion. From March 2001, we performed 36 right lobe ALDLT (segments 5, 6, 7, and 8) without MHV and one pediatric transplant (segments 2 and 3). In the presence of MHV tributaries larger than 5 mm, we intraoperatively evaluated the need for reconstruction. At a mean follow-up of 848 days (range=8-2412), 33/37 transplanted patients are alive with overall patient and graft survivals of 89.2% and 83.8%, respectively. Large MHV tributaries (>5 mm) were present in 10 cases, and inferior right hepatic veins (IRHV) draining segment 6 in 11 cases. In 10 cases, we performed an end-to-side anastomosis between the IRHV and the side of the recipient vena cava. In three cases, the MHV tributaries were end-to-end anastomosed to the stump of the recipient MHV. In all other cases, the vein tributaries were not reconstructed. A computed tomography scan performed from 1 to 3 months after surgery did not show any congested area in the liver parenchyma. In our experience, reconstruction of the MHV tributaries was not always necessary when graft-to-recipient weight ratio is >0.8. Pre- and intraoperative evaluation of the segmental branches of the hepatic vein is crucial to decide about reconstructing these collaterals. Anastomosis of V5 or V8 to the stump of the recipient MHV reduces the number of vascular anastomosis and maintains a physiological angle between these collaterals and the caval vein. PMID- 18675097 TI - Effects of everolimus monotherapy on hematological parameters and iron homeostasis in de novo liver transplant recipients: preliminary results. AB - INTRODUCTION: Anemia after orthotopic liver transplantation (OLT) is a common complication due to several reasons. Immunosuppressive drugs play an important role in anemia occurring at 1 month or more after OLT. Several studies describe myelosuppression immunosuppressants such as the mammalian target of rapamycin inhibitors. METHODS: We performed a single-center, prospective trial consisting of a short 30-day course of cyclosporine (CsA) associated with everolimus (EVL) from postoperative day 10 (Group EVL) versus a CsA immunosuppressive regimen (Group CsA) in de novo OLT patients. We explored the influence of immunosuppressive drugs on hematological parameters comparing EVL versus CsA. RESULTS: Twenty-eight patients were enrolled in the EVL and 12 in the CsA Groups. After OLT, hemoglobin (Hb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), white blood cell (WBC), platelets (PLT), transferrin saturation (TSAT), iron, ferritin, and transferrin did not differ significantly between the 2 groups at any time point. Among the patients who reached 6-months of follow-up, 5 (41.7%) EVL and 4 (80%) CsA subjects were anemic (P=not significant [NS]). Only anemia in patients enrolled in Group EVL showed a trend toward the features of microcytic, hypochromic anemia. DISCUSSION: Our results demonstrated that de novo anemia in OLT patients treated with EVL monotherapy showed the same incidence as in patients treated with CsA. Hb values remained similar during the entire follow-up. Moreover, overall myelosuppression in the EVL Group was not significantly different from patients in the CsA Group. PMID- 18675098 TI - Sirolimus therapy in liver transplant patients: an initial experience at a single center. AB - Sirolimus (SRL) is an mTOR inhibitor that has been shown, in contrast to calcineurin inhibitors (CNI), to inhibit cancers in experimental models. Since February 2005, we introduced SRL in liver transplant patients in group a, in whom the primary disease was hepatocellular carcinoma (HCC) associated with hepatitis B virus (HBV), hepatitis C virus (HCV), alcoholic or autoimmune liver cirrhosis, and group b, HCC-negative patients who developed posttransplantation cancers de novo. Of 18 patients in group a, 11 received SRL ab initio (subgroup a1), starting for 10 patients at 66.1+/-29.2 days after surgical healing and after 10 days in 1 case; the remaining 7 patients (subgroup a2) received SRL at 31.2+/ 24.2 months. Three patients in group b, included 1 with Kaposi's sarcoma, 1 with bladder cancer, and 1 with thyroid cancer. In this group, SRL was introduced at 80.8+/-40.4 months. In all patients but one, who received a single 5 mg loading dose, SRL was started at 2 mg/d and adjusted to 6 to 8 ng/mL blood levels. CNI drugs, present as primary therapy, were gradually tapered to low levels and eventually stopped. The following observations were drawn from this initial experience: (1) 4/21 (19.0%) patients had to discontinue SRL because of early and late side effects: thrombocytopenia (n=2) and headache with leukopenia and leg edema associated with knee joint arthralgia (n=2); (2) 14 patients (11 in group a and 3 in group b) are still on SRL monotherapy; (3) 1 HCC recurrence and 1 de novo pancreatic adenocarcinoma were observed at 14 and 16 months, respectively (at the time of transplantation, both patients were beyond the MIlan HCC criteria), and (4) 1 patient, from subgroup a1, died after 99 days due to pneumonitis and possible relation to SRL lung toxicity. In conclusion, SRL appeared to be an effective immunosuppressant that could be used as monotherapy in liver transplant patients. Any conclusion on SRL anticancer effects can only come from randomized large studies after long follow-up. PMID- 18675099 TI - The role of basiliximab induction therapy in adult-to-adult living-related transplantation and deceased donor liver transplantation: a comparative retrospective analysis of a single-center series. AB - AIM: The aim of this study was to report our single-center experience with the use of basiliximab, in combination with a steroid and tacrolimus-based regimen in adult to adult living-related liver transplantation (ALRLT) and in deceased donor liver transplantation (DDLT). MATERIALS AND METHODS: Seventy-seven consecutive ALRLT recipients (group 1) and 244 DDLT recipients (group 2) were analyzed. All patients received 2 20-mg doses of basiliximab (days 0 and 4 after transplantation) followed by tacrolimus (0.15 mg/kg/d; 10-15 ng/mL target trough levels) and a dose regimen of steroids. Follow-up ranged from 4-1972 days after transplantation in group 1 and from 1-2741 days in group. RESULTS: In group 1, 89.32% of the patients remained rejection-free during follow-up, with an actuarial rejection-free probability of 93.51% within 3 months. Actuarial patient survival rate at 3 years was 84.49%. In group 2, 86.07% of the patients remained rejection-free during follow-up, with an actuarial rejection-free probability of 93.04% within 3 months. Actuarial patient survival rate at 3 years was 87.69%. We observed 14 cases of hepatitis C virus (HCV) recurrence in group 1 (prevalence of 26.92%) and 80 cases in group 2 (prevalence of 54.05%). CONCLUSION: Basiliximab in association with tacrolimus and steroids is effective in reducing episodes of acute cellular rejection (ACR) and increasing ACR-free survival after ALRLT and DDLT. No difference in patient and graft survival was found between group 1 and 2, nor was there any difference in the incidence of ACR between the 2 groups. However, less risk of HCV recurrence was present in the LRLT group. PMID- 18675100 TI - A risk score and a flowchart for liver retransplantation. AB - Rates of overall graft survival after liver retransplantation (RETX) are still 20% lower than those after primary liver transplantation (TX). On the basis of previous mathematical approaches from other authors who tried to identify prognostic variables for survival and prognostic risk scores for liver RETX, we studied 12 categorical and 17 continuous variables from the donor, the recipient, and the surgical procedure, among patients who underwent liver retransplantation. Data were retrieved in a retrospective study over the last 12 years, in order to overcome the possible gap of other series that often included RETX performed many years ago. We considered 394 consecutive cadaveric liver TXs in adult patients, namely, 351 primary TXs and 43 RETXs. Using multivariate logistic regression, we calculated the following equation for 1-year risk of death for patients undergoing liver RETX: log(Odds)= -4.81+2.23 x Recipient Sex + 1.86 x Donor Age + 1.60 x MELD Score (where: Recipient Sex: F=0, M=1; Donor Age (years): <40=0, 40 59=1; 60+ =2; MELD Score: <26=0, 26+ =1). With this formula, we built a decision tree to predict the individual risk of death based on the subject's profile. Keeping in mind that mathematical models can only help our decisional process and are not conclusive, our data needs to be validated on a larger scale. PMID- 18675101 TI - Liver transplantation for hepatitis B virus patients: long-term results of three therapeutic approaches. AB - The indications for liver transplantation among patients with post-hepatitis B virus (HBV)-related cirrhosis have changed over the past 35 years. We reviewed the long-term results of 47 patients treated with liver transplantation for HBV related cirrhosis. Patients were classified into 3 groups according to the perioperative regimen. In the initial experience, no immunoprophylaxis was adopted (no-IP; n=5). From 1988-1996, an immunoprophylaxis scheme was adopted (HBIg; n=16). From 1997-2007, we adopted the combination of lamivudine and HBIg (LAM-HBIg; n=26). We calculated the prevalence of serological reinfection and patient survival at 1 to 20 years, using the 3 regimens. The recurrence rate was 75% in the group of untreated patients; 30% in the HBIg group; and 9% in the LAM HBIg group. The overall survival was 67% at 5 years, and 64% at 10 and 20 years. The long-term survival for each of the 3 therapeutic approaches, namely, for the patients who did not receive any treatment, for the HBIg group, and for the LAM HBIg group, were 20%, 50%, and 84%, respectively. We suggest to use the LAM-HBIg combination. PMID- 18675102 TI - Human immunodeficiency virus and liver transplantation: our point of view. AB - INTRODUCTION: Highly active antiretroviral therapy (HAART) has been able to improve the immune system function and survival of HIV patients with a consequent increase in the number of HIV patients affected by end-stage liver disease (ESLD). Between June 2003 and October 2006, 10 HIV-positive patients underwent liver transplantations in our center. METHODS: All patients were treated with HAART before transplantation; treatment was interrupted on transplantation day and was restarted once the patients' conditions stabilized. Five patients were hepatitis C virus (HCV)-positive, 3 were hepatitis B virus (HBV)-positive, and 2 were HBV-HCV coinfected. HIV viral load before transplantation was <50 copies/mL in all cases. CD4+ cell count before transplantation ranged between 144 and 530 c/microL. Immunosuppression was based on Cyclosporine (CyA) and steroid weaning for 8 patients, and on Tacrolimus and steroid weaning for 2 patients. RESULTS: Five patients were cytomegalovirus (CMV)-positive pp65 antigenemia posttransplantation, and 1 patient was EBV-positive; 2 patients had a coinfection with HHV6. Four patients suffered from a cholestatic HCV recurrent hepatitis treated with antiviral therapy (peginterferon and Ribavirin). Three patients died after transplantation. DISCUSSION: The outcome of liver transplantation in HIV patients was influenced by infections (HCV, CMV, and EBV) and Kaposi's Sarcoma. HCV recurrence was more aggressive, showing a faster progression in this patient population. Drug interaction between HAART and immunosuppressants occurs; longer follow-up and better experience may improve the management of these drug interactions. PMID- 18675103 TI - Preliminary results of liver transplantation for hepatocellular carcinoma among allocation organ policy strategies, neoadjuvant treatments, and intention-to treat analysis. AB - We retrospectively evaluated the impact of our strategy for patients with hepatocellular carcinoma (HCC) according to an intention-to-treat analysis and drop-out probability. We evaluated only patients within the Milan criteria. We analyzed the outcomes of neoadjuvant strategies for HCC, organ allocation policy, and systematic application of strategies to increase the deceased donor pool as the current tendency to expand transplantability criteria for those patients. Kaplan-Meier survival probability rates at 1, 3, and 5 years according to an intention-to-treat analysis were 87.02%, 74.53%, and 65.93% for transplanted patients (n=108), and 50%, 14.29%, and 14.29% for the excluded or waiting list group (n=13), respectively (P< .0001). Drop-out risk at 3, 6, and 12 months was 2.40%, 8.59%, and 16.54%, respectively. During the same period, the mortality probability rates at 3, 6, and 12 months among patients without HCC awaiting orthotopic liver transplantation (OLT) were 3.60%, 9.50%, and 18.34%, respectively. Drop-out rate was lower among patients treated before OLT (P< .0001). On the basis of the neoadjuvant treatment results to reduce drop-out risk, we suggest avoiding the high priority for the HCC cohort, particularly within the first 6 months from entrance on the waiting list, because this approach can reduce the chances of patients with end-stage liver disease (ESLD) alone. PMID- 18675104 TI - HCV histological recurrence and survival following liver transplantation in patients with and without hepatocellular carcinoma. AB - BACKGROUND AND AIM: Hepatitis C virus (HCV)-related cirrhosis is one of the leading indication for liver transplantation (LT) and a major risk factor for the development of hepatocellular carcinoma (HCC). HCV recurrence after LT is universal. This study evaluated HCV recurrence and survival in patients transplanted for HCV and HCC. METHODS: We evaluated all adults transplanted for HCV cirrhosis between January 1999 and December 2006, HCC was diagnosed on the explant and HCV recurrence confirmed on protocol liver biopsies performed at 6 months and yearly after LT. The sustained viral response (SVR) was defined as HCV RNA undetectable at 6 months after therapy discontinuation. The patient survival rates were assessed with Kaplan-Meier curves and the chi-square test was used when appropriate. RESULTS: Two hundred sixteen patients underwent LT for HCV including 153 men and 63 women of mean age 54 years with a mean follow-up of 35 months. There were 71 (33%) HCC(+) patients. At 1, 3, and 5 years from LT severe fibrosis (Scheuer 3-4) due to the HCV recurrence was reported in 18%, 14%, and 11% for HCC(+) and 14%, 16%, and 28% for HCC(-) patients respectively (P=NS). HCC recurred only in 3 (4%) patients at a mean follow-up of 3 years. Patients who received antiviral treatment after LT were 10% HCC(+) and 12% HCC(-) patients (P=NS). SVR was seen in 3/7 (43%) of HCC(+) and in 10/18 (55%) of HCC(-) patients (P=NS). At 1, 3, and 5 years the patient survivals was 91%, 86%, and 86% for HCC(+) and 94%, 86%, and 83% for HCC(-) patients, respectively (P=NS). CONCLUSIONS: Severe fibrosis due to HCV recurrence, which increases over time, involves one third of transplanted patients at 5 years after LT. The long-term survival was identical among HCC(+) compared to HCC(-) recipients. The recurrence of HCC was negligible and did not affect patient survival. PMID- 18675105 TI - Liver transplantation using donors older than 80 years: a single-center experience. AB - AIM: The shortage of organs for orthotopic liver transplantation (OLT) has forced transplantation centers to expand the donor pool by using donors traditionally labeled as "extended criteria donors." One such example is OLT using a donor with advanced age. MATERIALS AND METHODS: We retrospectively evaluated 10 patients who received a liver graft from cadaveric donors older than 80 years. We analyzed pretransplantation donor and recipient characteristics, as well as the evolution of the recipients. RESULTS: All 10 donors were older than 80 years (median age, 83.5; range, 80-93). No steatosis (>30%) was accepted in the older donor group. Medium follow-up was 19.5 months. The most frequent cause for OLT was hepatitis C virus (HCV) cirrhosis (8/10 patients). We had 1 case of primary nonfunction, 1 patient died immediately after surgery because of extrahepatic complications (cardiac arrest), and 2 other patients had a severe HCV recurrence and died after 1 and 2 years from OLT, respectively. Five patients had HCV recurrence and biliary complications were present in 60% of the patients. No cases of acute or chronic rejection were described. Overall survival rates after 1 and 3 years were 80% and 40%, respectively. CONCLUSIONS: Old donor age is not an absolute contraindication to OLT. Liver grafts from donors older than 80 years can be used knowing that there is a high risk of postoperative complications. Furthermore, the increased risk of developing severe HCV recurrence, related to older donor age, suggests that such livers should be used in HCV-negative recipients. PMID- 18675106 TI - Noninvasive ventilation in adult liver transplantation. AB - Noninvasive ventilation (NIV) has proven to be a safe and effective technique in the treatment of respiratory failure complicating various medical and surgical diseases. In recent years, a growing interest has emerged in its adoption for ventilatory assistance in immunocompromised patients, such as those undergoing bone marrow, liver, lung, cardiac, and kidney transplantation. Weaning from the ventilator after liver transplantation can take longer because of unsatisfactory gas exchange during various attempts of T-piece trials. Rapid extubation followed by an immediate NIV application should be considered in this setting to shorten and accelerate the weaning process in those recipients who do not completely fulfill the criteria for safe extubation. By adding the pressure support (PS) mode with a continuous positive end expiratory pressure (PEEP), NIV could prevent the loss of vital capacity and impede severe lung derecruitment following extubation. Clinical experience has shown that properly delivered NIV mostly benefits moderately dyspneic recipients in acute respiratory failure, while it appears less promising and efficient in patients ventilated for extended periods of time. It has proven safe and efficient mainly as (1) a tool to promote an early ventilatory discontinuation and extubation; (2) a prophylactic strategy for preventing postoperative pulmonary complications; and (3) a simple method to start with in cases of acute hypoxic and/or hypercapnic respiratory failure. The improvements in arterial hypoxemia, the decreased ventilatory demand provided with an inspiratory support, as well as the scarcity of hemodynamic repercussions are among the major benefits of this method. PMID- 18675107 TI - The "You Are Not Alone" care program for liver transplantation. AB - Current clinical practice is based on the principles of efficacy, appropriateness, efficiency, quality, and safety. Compliance with these tenets requires experienced medical and nursing staff, and active participation of patients and their families in the planned therapeutic program. To match patients' expectations on quality and safety of care and spur active participation in the transplant care process, we set up an integrated, multiphase, multidisciplinary care program devoted to liver transplantation (LT) candidates, engrafted patients, and their families: the "Non Sei Solo" care program (You Are Not Alone). The basic principle of the care program was that, to provide efficient and effective education to their patients, health care professionals need to learn how to teach and what to teach, acquire successful communication skills, and monitor the process of education. The methodology encompassed 5 distinct phases: phase 1, exploration of patients' needs, by means of a questionnaire devoted to waitlisted and engrafted patients and their care givers; and phase 2, creation of 16 patient-oriented educational brochures directed to patients and their families. Once created, the educational brochures were presented, discussed, and amended during a consensus meeting involving all transplantation nurses and physicians (phase 3). To acquire the necessary skills and ease communication with patients, the transplantation nurses, physicians, surgeons, and anesthesiologists attended a 6-month counseling course under the tutorial of an expert counselor (phase 4). Finally, in June 2007 the program started officially with monthly meetings with patients and their families, guided hospital tours on patient request, and activation of a toll-free phone number to provide support to patients and answer their questions. PMID- 18675108 TI - Incidence and clinical outcomes of ventilator-associated pneumonia in liver transplant and non-liver transplant surgical patients. AB - The aim of this study was to compare the incidence of ventilator-associated pneumonia (VAP) and clinical outcome among patients undergoing orthotopic liver transplantation (OLT) admitted to our surgical intensive care unit (ICU). Patients with an ICU stay longer than 4 days who had undergone surgery within 48 hours of admission were included in the study. Patients were subdivided into a liver transplant group (OLT) and no-liver transplant group (noLT). Diagnosis of VAP was based on microbiological data with a positive culture from a sample collected >or=48 hours after admission. VAP was defined as early if the positive culture occurred within the 4th day of admission, and late if after the 4th day. Three hundred seventy-three noLT and 71 OLT patients showed no differences in sex, mean severity score on admission (SAPS II), length of stay, and outcomes. The incidence of VAP was also similar in the 2 groups (27.3% in the noLT group vs 25.3% in the OLT group). Both in the OLT and noLT groups, the VAP patients showed higher (P< .05) SAPS II scores on admission, length of ICU stay, and mortality rates than the non-VAP patients, without any difference between the 2 groups. VAP is a frequent complication in ICU surgical patients, particularly those with high severity scores on admission. In an ICU surgical population, liver transplantation per se does not seem to increase the patients' risk either for VAP acquisition or for bad outcomes. PMID- 18675109 TI - Recombinant activated factor VII in critical bleeding after orthotopic liver transplantation. AB - Critical bleeding throughout the intraoperative phase of orthotopic liver transplantation (OLT) strongly increases patient mortality and intensive care unit (ICU) stay. The aim of this study was to report our experience on the use of recombinant activated factor VII (rFVIIa) in postoperative critical bleeding after OLT. In 7 patients with persistent severe bleeding after application of a standard transfusion protocol, we administered a 90 microg/kg bolus of rFVIIa and if necessary eventually repeated it after 3 hours. We recorded the blood loss and the need for transfusions before and after the rFVIIa therapy. Blood losses and need for platelets significantly decreased after rFVIIa administration; a nonsignificant decrease in red blood cells and fresh frozen plasma transfusions also occurred. In 6 patients treatment with rFVIIa was effective; only 1 patient died because of hemorrhagic shock and no thromboses were detected among the treated patients. Awaiting stronger evidence from randomized controlled trials, we suggest that in some challenging cases of massive bleeding rFVIIa should be considered a useful option to control bleeding. PMID- 18675110 TI - Emergency use of wearable left ventricular assist devices in patients not eligible for heart transplantation. AB - BACKGROUND: We reviewed our experience concerning the clinical outcomes of emergency implantation of left ventricular assist devices (LVAD) in patients not eligible for heart transplantation (HTX). PATIENTS AND METHODS: Between January 1998 and September 2006, 62 patients were referred for emergency HTX including 35 (55%) who received cardiac allografts; 9 (14%) died on the waiting list. The other 18 subjects (31%) did not meet listing criteria due to pulmonary hypertension with massive transpulmonary gradients (TPG), severe diabetes, or another factor; 7 patients had wearable LVAD implants and the remaining 11 who had contraindications both to HTX and to LVAD implantation remained on medical therapy. RESULTS: Twelve month actuarial survivals were 47% in LVAD G1 vs 1% in medical therapy G2 (P< .005). Four G1 patients died either due to cerebrovascular vascular episodes or to thromboembolic events. The beneficial use of LVAD in 3 patients led to consistent improvement in hemodynamic performance allowing listing for HTX. Two of them have been successfully transplanted; 1 is still waiting. Due to refractory heart failure, 90% of G2 died within the first months after evaluation. CONCLUSION: Our limited experience indicated that emergency use of LVAD ensued a satisfactory outcome allowing better actuarial survival. PMID- 18675111 TI - Outcome of patients listed for heart transplantation after a failed surgical ventricular restoration procedure. AB - Patients with end-stage ischemic cardiomyopathy (IHD) and left ventricular (LV) dilatation are increasingly treated by means of surgical ventricular restoration (SVR). In some patients, SVR can delay heart transplantation (HTX). We retrospectively analyzed our experience, trying to ascertain whether HTX after a failed SVR (fSVR) carried a greater mortality risk. Since 1985, we performed 742 HTX. Since June 1999, 133 IHD patients were listed for HTX. We assigned them to 3 groups: (A) not a redo (n=54); (B) redo after coronary artery bypass grafting (n=54); and (C) redo after fSVR (n=25). Respectively, 37, 33, and 12 patients underwent HTX with in-hospital mortality after HTX of 4/37 (10.8%), 12/33 (36.4%), and 2/12 (16.7%). Mortality on the list was 9/54 (16.7%), 11/54 (20.4%), and 7/25 (28.0%) respectively. Removal from the list occurred in 4, 5, and 2 patients, and 4, 5, and 4 patients are still awaiting HTX, respectively. In group C, the mean time from SVR to HTX list was 45.6+/-43.3 months, and list mortality occurred after 5.83+/-5.81 months. In-hospital mortality in both patients of group C was due to the occurrence of multisystem organ failure; 10/12 were extubated after 19.3+/-9.6 hours and discharged from the intensive care unit after 3.9+/-1.6 days. The recorded complications were: 3 acute renal failure, 1 pericardial effusion, and 2 episodes of acute rejection. Since only 5/25 patients with fSVR had undergone SVR at our institution, we cannot establish which patients were really eligible for HTX at the time of SVR. Our experience showed that patients listed for HTX displayed a high list mortality, but that HTX after a failed SVR did not seem to have a poorer outcome than HTX after previous conventional CABG. PMID- 18675112 TI - The tailored medical therapy in patients with advanced heart failure referred for cardiac transplantation. AB - INTRODUCTION: Optimal pharmacologic management of heart transplant (HT) candidates is required prior to evaluation so as to obtain a reliable prognostic stratification and to address the donor shortage. The aim of this study was to determine whether a tailored medical approach was effectively achieved before HT waiting list enrollment. MATERIALS AND METHODS: This study concerned 40 consecutive patients referred for HT evaluation who underwent a clinical assessment, including hemodynamic, echocardiographic, and brain natriuretic peptide determinations. Medical therapy was optimized according to the clinical assessment to improve neurohormonal and hemodynamic profiles. We analyzed the distribution of the different drugs between the first and the following evaluation to demonstrate whether a significant improvement of medical therapy could be achieved in advanced chronic heart failure (ACHF). RESULTS: The mean age was 53 years, including 93% males. The etiology of disease was ischemic in 40% and idiopathic in 45%. The mean left ventricular ejection fraction was 23%, mean values of hemodynamic data were cardiac index (CI) 2+/-0.6 L/min/m(2), mean pulmonary arterial pressure (mPAP) 30+/-10 mm Hg, wedge pressure (PWP) 23+/-8 mm Hg; mean BNP was 618 pg/mL. Median follow-up was 397 days; 82% of candidates underwent HT waiting-list enrollment. The medical treatment was modified as follows: beta-blockers were introduced or uptitrated in 32%, angiotensin receptor blockers (ARB) were introduced in 7.5%, spironolactone was started in 42%, nitrates were introduced in 20%, and diuretics were uptitrated in 35% of patients. CONCLUSION: In patients with ACHF referred for HT, a further effort in the assessment of the medical treatment is strongly recommended. PMID- 18675113 TI - Lung transplantation for cystic fibrosis: ten years of experience. AB - Lung transplantation represents the only therapeutic option for patients affected by end-stage cystic fibrosis (CF). We performed 76 lung transplantations in 73 patients from 1996-2007. The mean time on the waiting list was 10+/-6 months. The median follow-up after the transplantation was 69.3 months. Twenty-one transplants (27.6%) were performed under cardiopulmonary bypass. Perioperative mortality, excluding retransplants, was 16.4% (12 patients) and the causes of death were sepsis, primary graft failure, and myocardial infarction. The overall survival was 74.5%+/-5%, 62.9%+/-5%, 54.1%+/-6%, and 43.4%+/-6% at 1, 3, 5, and 10 years, respectively. The accurate selection of potential recipients and the correct timing of referral and transplantation are factors that play crucial roles to obtain satisfactory results in term of improvement of quality of life and long-term survival. PMID- 18675114 TI - Lung transplantation for cystic fibrosis in Italy. AB - Lung transplantation (LT) is the only effective form of therapy for cystic fibrosis (CF) associated with end-stage pulmonary failure. In Italy, the management of CF is regulated by national law, which has instituted regional centers for care and follow-up of all CF patients. LT has been performed since 1992 in only nine LT certified centers. The structured national organization has led to a unified database for LT for CF. As of December 2006, 197 bilateral LT (96 male and 94 female patients; 7 retransplants) have been performed. Of these, four had also liver or heart and liver transplantation, and three are long-term survivors. Overall median survival is 7 years. Mean age at transplantation is 26.5 years, and the mortality on the waiting list is 33.6%. Patients listed for transplant either received a suitable donor within a mean of 10 months or died within a mean of 5.5 months. The most frequent cause of death is bronchiolitis obliterans syndrome (BOS). Our nationwide database indicates the excellent results obtained by LT in FC. Still, mortality on the waiting list remains a challenge and long-term outcome is limited by BOS. PMID- 18675115 TI - Lung transplantation for emphysema. AB - Emphysema is a major cause of morbidity and mortality worldwide. Despite optimal medical therapy, smoking cessation, and pulmonary rehabilitation, a large number of patients remain symptomatic with a poor quality of life. A pool of patients with end-stage disease can benefit from surgical treatments like bullectomy, lung volume reduction, or lung transplantation. Emphysema represents the most common indication leading to lung transplantation. A functional improvement and better quality of life are clear benefits deriving from lung transplantation, while a survival advantage has not yet been proven. PMID- 18675116 TI - Single versus double lung transplantation in pulmonary fibrosis: a debated topic. AB - Idiopathic pulmonary fibrosis (IPF) represents the second most frequent indication for lung transplantation after chronic obstructive pulmonary disease. Survival rate after transplantation is poorer compared with other lung diseases for reasons that are not completely clear. Medical therapy with anti-inflammatory drugs may improve symptoms and quality of life, but it does not influence the survival rate. Lung transplantation is the best therapy for end-stage IPF. The debate regarding the superiority of double lung transplantation (DLT) compared with single lung transplantation (SLT) is still ongoing. Until some years ago, SLT was almost uniformly utilized for this indication. In the most recent years, a larger application of DLT has been observed worldwide, probably related to higher 1-year and 5-year survivals. The unanswered question is whether it is ethical to use two lungs for the same patient, considering the donor shortage, when a single lung would suffice. Many reports have demonstrated that SLT offers acceptable pulmonary function and satisfactory early and intermediate survival. Probably DLT should be reserved for younger recipients, for those with concomitant or possible chronic infection of the contralateral lung, or cases of marginal donors. Further studies will be needed to formulate recommendations regarding the preferred surgical approach in IPF. PMID- 18675117 TI - Combined cytomegalovirus prophylaxis in lung transplantation: effects on acute rejection, lymphocytic bronchitis/bronchiolitis, and herpesvirus infections. AB - Lung transplantation recipients are at high risk for herpesvirus infections. We evaluated the effect of combined cytomegalovirus (CMV) prophylaxis on CMV pneumonia, acute rejection episodes (ARE), lymphocytic bronchitis/bronchiolitis (LB), and obliterans bronchiolitis (OB) diagnosed in 180 transbronchial biopsies (TBB) of lung transplant recipients. At our center, 25 patients (control group; 1999-2002) received acyclovir for 12 months and 21 recipients (study group; 2003 2007) received combined CMV prophylaxis consisting of CMV-IG (Cytotect Biotest) for 12 months and ganciclovir or valganciclovir from postoperative day 21 for 3 weeks. Among the study group (since 2005), CMV shell vial viral culture and Epstein-Barr virus (EBV), human herpesvirus-6 (HHV-6), and HHV-7 DNA were determined on BAL specimens. In the study group, the number of LB was significantly lower than in the control group (2% vs 11%; P= .04). Similar results were obtained for ARE (6% vs 17%; P= .04). No difference was observed in OB (5% vs 5%; P= .53, NS). A reduction trend was found in CMV pneumonia (2% vs 7%; P= .23, NS). Logistic regression analysis showed a relationship between prophylaxis and a reduced prevalence of ARE (odds ratio [OR] 3.25, confidence interval [CI] 1.12-9.40; P= .03). Finally, in the study group, BAL EBV-DNA positivity and EBV-CMV coinfections were low (6% and 0%, respectively) compared with other herpesviruses and with the literature. Our data suggested the efficacy of combined CMV prophylaxis to prevent ARE and LB, 2 risk factors for chronic rejection, and a possible role to reduce the trend toward CMV pneumonia and EBV infections. PMID- 18675118 TI - Role of sildenafil in acute posttransplant right ventricular dysfunction: successful experience in 13 consecutive patients. AB - BACKGROUND: Superimposed acute right ventricular dysfunction in the setting of preexisting pulmonary hypertension is a nearly fatal complication after heart transplantation. The optimal treatment modality remains a matter of debate. Recently, sildenafil citrate, a nonselective pulmonary vasodilator, has gained popularity in the treatment of pulmonary hypertension in transplant candidates. METHODS: Herein we have presented a series of 13 patients in whom sildenafil was used to treat right ventricular dysfunction and pulmonary hypertension as detected by transesophageal echocardiography and Swan-Ganz right heart catheterization after heart transplant. Their characteristics were mean age 49+/ 11.4 years; 38.4% with previous cardiac procedures, 30.8% status I, basal pulmonary vascular resistance index 10.4+/-4.6 WoodU, mean transpulmonary gradient 18.7+/-5.4 mmHg. In addition to conventional inodilator support, we administered 1 to 3 mg per kilogram of sildenafil. Complete hemodynamic measurements were obtained before and after the institution of the therapy and at 1-month follow-up. RESULTS: Within the first 72 hours, acute right ventricular dysfunction resolved in all cases without untoward side effects or significant systemic impact. Sildenafil significantly decreased the transpulmonary gradient and pulmonary vascular resistance index relative to baseline values; 5.6+/-1.82 versus 10.4+/-4.6 WU, (P< .05), 13.5+/-3.4 mm Hg versus 18.7+/-5.4 mm Hg (P< .05), respectively. Improved indices of right ventricular function were observed on echocardiographic monitoring. After 1 month, sildenafil treatment was discontinued. CONCLUSION: Management of acute right ventricular dysfunction in heart transplant recipients with pulmonary hypertension using sildenafil proved safe and effective. PMID- 18675119 TI - Application of the RAND/UCLA Appropriateness Method to evaluate an information system for kidney/pancreas transplantation in adult recipients. AB - With the aim to evaluate the correctness of medical and surgical procedures, RAND Corporation and University of California Los Angeles (UCLA) developed the RAND/UCLA Appropriateness Method (RAM). In this study, the RAM was applied to evaluate the appropriateness of a dataset concerning kidney/pancreas transplantation in adult recipients for an information system funded by the Italian Ministry of Health. The original dataset was obtained using an interdisciplinary pool of experts (n=60) involved in kidney/pancreas transplantation activity in the Liguria Region. This dataset held 291 items, stratified as pretransplantation items (n=158), transplantation items (n=49), and early posttransplantation and follow-up items (n=84). In the second round, the dataset was subjected to an extraregional panel of independent experts (n=9) to assess each item using a score ranging from 1 to 9 based on increasing appropriateness. The expert-opinion process returned a whole mean score of 8.47+/ 0.43 (95% confidence interval [CI] 8.30-8.63). Overall agreement, uncertainty, and disagreement between experts about item appropriateness were 98.5%, 1.49%, and 0%, respectively. Agreement/uncertainty for pretransplantation, transplantation, and posttransplantation items were 99.87%/0.12%, 100%/0%, and 96.37%/3.62%, respectively. This study supported the utility of a structured expert-opinion process as an effective strategy to evaluate the appropriateness of large datasets for kidney/pancreas transplantation in adult recipients. PMID- 18675120 TI - Simultaneous pancreas-kidney transplantation: a single-center experience and prospective analysis. AB - In patients with end-stage chronic kidney disease (CKD) and type 1 diabetes mellitus (DM 1), simultaneous pancreas-kidney (SPK) transplantation is currently considered the gold standard therapy. The aim of this study was to analyze and report the long-term clinical outcomes of the 23 SPK transplantations performed at our institution over an 84-month period (January 1, 2000 to December 31, 2006). A prospective analysis of these patients included donor, recipient, and transplantation characteristics. The only requirements for transplantation were blood group compatibility and a negative cross-match. Bladder drainage via pancreaticoduodenocystostomy was performed in all of the patients. Due to a pulmonary embolus 1 patient (4.3%) died at 2 months. The actuarial patient survival rates at 3 months and 1, 3, and 5 years were 95.6%. Causes for the renal graft loss were chronic allograft nephropathy in 3 cases (13%) and death of the patient in 1 case (4.3%). The actuarial censored renal allograft survival rates at 3 months and at 1 year were 100%, and at 3 and 5 years were 91.3%. Causes for the renal graft loss were chronic rejection in 1 case (4.3%) and patient death in 1 case (4.3%). The actuarial censored pancreatic allograft survival rates at 3 months and at 1 and 3 years were 100%, and at 5 years was 95.6%. The results of this work add further evidence that SPK is the gold standard therapy for selected patients with end-stage CKD due to DM 1. PMID- 18675121 TI - Pancreas transplantation inside Emilia-Romagna, Italy: referral pattern, demand forecasting, and organ availability. AB - In Italy, referral of diabetic patients for pancreas transplantation (PT) is an unstructured process, resulting in a low rate of activity and late referrals, often when the patient has already undergone dialysis. In addition, the continuous improvement in pancreas transplant alone, offering the opportunity to reduce cardiovascular risk due to proteinuria and reduced glomerular filtration rate (GFR), is rarely appreciated. We therefore analyzed (1) referral activity to PT during the time frame 2001-2005 in Emilia-Romagna, Italy (four million inhabitants), by collecting ICD 9 CM codes (55.69 + 52.80; 52.86 and 52.80 alone) by residence of the patient; (2) demand for PT among a sample population of 1670 diabetes patients, whose charts were reviewed for the type of diabetes and presence of overt diabetic nephropathy (DN: proteinuria >300 mg/24 h and/or GFR <60 mL/min); (3) potential pancreas availability as the ratio between pancreas and hearts utilized (UP/HR) in different areas of our country. As a results, (1) referral activity reached 8.4 PT per million people in 5 years in the whole region, ranging from 2.6 in the province where a PT program is active, to a maximum value of 20.7 in the province where a devoted outpatient clinic is operated by nephrologists. (2) Prevalence of overt DN was 6% in our cohort, corresponding to 510 D1 patients worthy of evaluation for PT inside Emilia Romagna region. (3) During 2006, UP/HR was 0.58 in Associazione Inter-Regionale Trapianti agency, 1.16 in Tuscany, 0.30 in Piedmont, and 0.26 in our region. Taken together, our data showed that (1) the referral of D1 to PT has to be empowered, keeping in touch with all patients suffering from diabetic nephropathy; (2) the outpatient clinic devoted to evaluation and recruitment of D1 with nephropathy plays the key role in this program of timely and widespread referral; (3) the availability of pancreata can be increased by utilizing broader criteria for harvesting, increased consent rate to donation and increased the demand for PT (recipient pool). Pancreas grafts need to increase, since the current low demand produces underutilization of the pancreas resource, due to the frequent lack of a suitable recipient. PMID- 18675122 TI - Fluid management of patients undergoing intestinal and multivisceral transplantation. AB - Small bowel transplantation can be associated with large fluid shifts due to massive blood loss, dehydration, vascular clamping, long ischemia times, intraoperative visceral exposure, intestinal denervation, ischemic damage, and lymphatic interruption. Fluid management is the major intra- and postoperative problem after small bowel and multiple organ transplantation, because of the highly variable fluid and electrolyte needs of the transplant recipient. Third space fluid requirements can be massive; inadequate replacement leads to end organ dysfunction, particularly renal failure. Several liters of fluid may be required in the initial 24 to 48 hours postoperatively to simply maintain an adequate central pressure to provide a satisfactory urine output. During this time patients may develop extensive peripheral edema, which dissipates over the next few days as the fluids are mobilized and requirements stabilize. Based on our experience in 29 cases of intestinal transplantation and 4 cases of multivisceral transplantation, we have herein described the intraoperative fluid management and hemodynamic changes. Our study confirmed a large quantity of fluid administration during and after small bowel transplantation that required adequate volume monitoring. PMID- 18675123 TI - A new swine training model of hand-assisted donor nephrectomy. AB - INTRODUCTION: Despite the described advantages of hand-assisted laparoscopic donor nephrectomy (HALDN), the learning curve risks discourage many transplant centers to switch from the traditional technique to the laparoscopic approach. Considering that the learning curve risk may be softened with practice on a training model the aim of this study was examine a low-cost, high-fidelity model of HALDN in pigs. METHODS: Ten female white pigs underwent a left and then a right HALDN in the same session for a total of 20 procedures. For each nephrectomy, we assessed operative times and intraoperative complications. All nephrectomies were performed by a single senior transplantation surgeon. RESULTS: All animals that survived bilateral nephrectomy were sacrificed. Two right HALDNs were converted to open procedures due to bleeding. One spleen lesion and one lumbar vein injury were treated laparoscopically. Considering only the 18 HALDN completed, we registered a mean total operative time of 75.4 min (range=52 to 120). DISCUSSION: The in vivo training model described herein made it possible to reproduce the positions and operative difficulties similar to those encountered in clinical practice. Moreover, the costs can be considerably reduced by performing two procedures in each animal employing reusable instruments. Our model represented a valid high-fidelity training procedure that was useful and convenient to achieve skills for HALDN that may help transplantation centers adopt this technique to reduce the learning curve risk. PMID- 18675124 TI - Liver apoptosis following normothermic ischemia-reperfusion: in vivo evaluation of caspase activity by FLIVO assay in rats. AB - Normothermic liver ischemia-reperfusion (I-R) may induce hepatocellular apoptosis. Caspase activation is involved in the initiation and execution of apoptosis. The aim of this study was to determine in vivo caspase activity in normothermic liver I-R in rats. Segmental normothermic ischemia of the liver was induced for 120 minutes in rats. After intravenous injection of the green probe FLIVO, in vivo caspase-3- and -7-specific activity was determined using fluorescence microscopy, in either nonischemic or ischemic liver lobes at 3 and 6 hours after reperfusion. Liver apoptosis was assessed by the deoxynucleotide transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay. Fluorescence microscopy showed that in vivo caspase-3- and -7-specific activities were significantly increased (P< .005) in ischemic lobes at 3 and 6 hours of reperfusion, compared with nonischemic liver lobes. Quantitative analysis of apoptotic cells measured by the TUNEL method showed a significant increase among apoptotic cells in ischemic lobes at 3 and 6 hours after reperfusion (P< .005), compared with nonischemic liver lobes. In conclusion, 120-minute normothermic liver I-R resulted in increased caspase-3- and -7-specific activities and in liver cell apoptosis. PMID- 18675125 TI - Liver HIF-1 alpha induction precedes apoptosis following normothermic ischemia reperfusion in rats. AB - Apoptosis plays an important role in ischemia-reperfusion (I-R) injury during liver transplantation. The hypoxia-inducible factor alpha (HIF-1alpha) may trigger liver apoptosis following I-R through the induction of hypoxically regulated genes. The aim of this study was to evaluate the effect of normothermic liver I-R on HIF-1alpha expression and apoptosis in rats. Segmental normothermic ischemia of the liver was induced in rats for 120 minutes. Liver extracts from either ischemic or nonischemic lobes were prepared at 0, 1, 3, and 6 hours after reperfusion. Liver HIF-1alpha protein expression was examined by Western blot analysis. Liver apoptosis was quantified using terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick end labeling assay. Normothermic I-R resulted in a significant (P< .05) increase in liver HIF-1alpha protein levels 1 and 3 hours after reperfusion. Liver apoptosis was significantly (P< .005) increased at 3 and 6 hours after reperfusion. In conclusion, normothermic liver I-R leads to increased liver expression of HIF-1alpha and apoptosis. PMID- 18675126 TI - Extracorporeal portal vein oxygenation improves outcome of acute liver failure in swine. AB - BACKGROUND: Portal vein arterialization (PVA) has shown efficacy to treat acute liver failure (ALF) in preclinical studies. The next step is to perform large animal studies that propose a clinically acceptable method of PVA. In this study, we assessed the efficacy of PVA using an extracorporeal device to treat 2 ALF models in swine. MATERIALS AND METHODS: The 2 ALF swine models were carbon tetrachloride toxic ALF and subtotal hepatectomy using 8 animals per group. PVA was performed with an extracorporeal device that may be suitable for future clinical studies. Arterial blood was drawn from the iliac artery and delivered into the portal vein for a 6-hour treatment. We analyzed biochemical, blood gas, and histological parameters as well as 1-week survival rates. RESULTS: In both models, ALF was successfully achieved. Control group animals deteriorated biochemically, dropping their prothrombin times and increasing the liver enzymes. In contrast, treated animals improved with a survival rate of 75% at 7 days compared with 0% for the former group. CONCLUSIONS: PVA using an extracorporeal device was feasible and effective to treat both toxic and resective ALF in swine. PMID- 18675127 TI - Peritoneal implantation of cryopreserved encapsulated porcine hepatocytes in rats without immunosuppression: viability and function. AB - The bioartificial liver (BAL) represents a promising approach to cell transplantation without immunosuppression as a method to support patients with hepatic insufficiency. The aim of this study was to assess viability and function of cryopreserved encapsulated porcine hepatocytes implanted intraperitoneally in rats without immunosuppression. Isolated porcine hepatocytes were cryopreserved at -196 degrees C for 1 month. Four groups were created: group 1 (n=10), freshly encapsulated porcine hepatocytes cultured in albumin-free medium for 10 days; group 2 (n=10), freshly encapsulated porcine hepatocytes implanted in the rat peritoneum without immunosuppression for 1 month and cultured for 10 days after explantation; group 3 (n=10), cryopreserved encapsulated porcine hepatocytes cultured for 10 days; group 4 (n=10), cryopreserved encapsulated porcine hepatocytes implanted in the rat peritoneum without immunosuppression for 1 month and cultured for 10 days after explantation. We assessed urea and albumin production and hepatocyte viability. The hepatocytes of all groups retained the capacity to produce urea and albumin, although the albumin synthesis was significantly decreased among hepatocytes of group 4 (P< .01). Encapsulated cryopreserved porcine hepatocytes explanted from rat peritoneum after 1 month appeared morphologically viable; their ultrastructure was preserved. In conclusion, long-term cryopreservation of porcine hepatocytes resulted in retention of their biological activity and in significant viability when transplanted into the rat peritoneum without immunosuppression. PMID- 18675128 TI - BKV reactivation in renal transplant recipients: diagnostic and therapeutic strategy--case reports. AB - The Polyomaviridae family includes several viruses that are ubiquitous with specific host spectra. The human polyoma viruses BK and JC were discovered in 1971. Following primary infection, transmitted by the respiratory and probably the oral route, BK remains latent in uroepithelial cells, in B lymphocytes, or in other tissues (spleen, brain). Reactivation with asymptomatic viruria may occur in both immunocompetent subjects and immunocompromised patients. In renal transplant recipients, BKV replication may cause tubulointerstitial nephropathy (BKVAN) with increasing prevalence rates--1% in 1995, 8% in 2007--leading to the loss of the transplanted organ in 30% to 80% of cases. With the availability of diagnostic programs (decoy cells in urine, amplification of viral DNA by polymerase chain reaction (PCR) on serum and urine, real time (RT)-PCR test for mRNA VP1 urine (mRNA-VP1), and renal biopsy accompanied by reduction in immunosuppression, administration of leflunomide, cidofovir (after hydration), and N-acetylcysteine, as well as immunoglobulin by intravenous injection (IVIg), the incidence of renal loss caused by BKVAN infection has been reduced by 10% to 80%. In this study, we have described 12 patients: 6 treated with tacrolimus (FK), mycophenolate mofetil (MMF), and steroids, and 6 treated with cyclosporine or with mTOR inhibitors. Two patients from the first group showed BKVAN about 3 months posttransplantation. Early diagnosis and therapeutic intervention (cidofovir + IVIg) led to reduction in the viral load, with improvement and stabilization in renal function. Considering the high positive predictive value (98%) of mRNA VP1, it should be possible to avoid renal biopsy. The level of immunosuppression--rather than the immunosuppressive drug itself (FK and MMF)- seemed to be associated with BKV reactivation. PMID- 18675129 TI - Idiopathic severe elevation of serum alkaline phosphatase following adult renal transplantation: case reports. AB - Transient hyperphosphatasemia (TH) in infancy is a benign condition characterized by elevated alkaline phosphatase (ALP) levels severalfold the adult upper limits, occurring mainly in children under 5 years, without evidence of liver or bone disease, and a return to normal ALP levels by 4 months. Herein we have reported 3 cases of TH in adults following renal transplantation. The first case, a 47-year old woman, blood group AB positive, had hypertensive renal disease. Five months after successful renal transplantation from a deceased donor she had a 50-fold increase in ALP. The second case, a 34-year-old man, blood group A positive, had renal failure due to IgA nephropathy. Nine weeks after a second renal transplant from a deceased donor a 25-fold increase in ALP was noted. The third case, a 45 year-old woman, blood group A positive, experienced renal failure 15 years earlier of unknown etiology. Thirteen years after her second renal transplant a 12-fold increase in ALP was observed during a routine follow-up. In all cases, the isolated ALP serum levels returned to normal limits within 12 weeks. Bone scans and abdominal ultrasounds during these periods were normal with no evidence of bone or liver disease. ALP isoenzyme electrophoresis revealed a pattern characteristic of TH of infancy and childhood. The 3 cases reported highlight the occurrence of benign TH in adults, with renal transplantation. However, liver disease, bone disease, and infection should be excluded first in these susceptible individuals on immunosuppression before establishing the diagnosis of TH. PMID- 18675130 TI - Incidentally discovered yellowish lesions in a renal graft from a deceased donor. AB - The opening of Gerota's fascia, soon after harvesting the kidney, is a standard kidney donor procedure in Italy to exclude a renal cell carcinoma (RCC), a frequent finding in older donors. Herein we have reported our experience with the diagnosis and management of subcapsular yellow areas suggestive of RCC on the kidney surface during back-table procedures. From 2001 to 2006, 12/445 grafts showed a single yellowish subcapsular nodule during the back-table procedure which was excised for frozen section (FS) to rule out RCC. The affected donors were 7 males and 5 females of overall mean age of 60 years (range, 25-77 years). The mean nodule diameter was 0.75 cm (range, 0.3-1.2 cm), and all lesions were located in the upper renal pole. In 5 cases, a diagnosis of RCC could not be excluded by FS, and both kidneys were discarded. The final histology confirmed RCC in only 3 cases, and adrenal heterotopia (AH) in the other 2. In the remaining 7 cases, FS showed AH in 4, 1 angiomyolipoma, and 2 areas of infarction confirmed by histology. The adrenal foci consisted of clear cells and scattered cells with eosinophilic, granular cytoplasm and small round nuclei, some with small nucleoli. Immunostains for cytokeratins, CD10, and epithelial membrane antigen were negative, confirming the adrenal origin. AH is the most common pathological yellowish lesion in the upper kidney pole found incidentally during back-table preparation. A histological differential diagnosis with RCC at FS is difficult, relying on the distinction of normal corticoadrenal spongiocytes from Fuhrman grade 1 clear cancer cells. In Italy, for any renal mass suggestive of RCC, a graft discard is mandatory, even if several reports have described cases of renal transplantation performed after back-table excision of small unifocal tumors. PMID- 18675131 TI - Renal transplant compartment syndrome: a case report. AB - An unusual case of early double kidney transplant dysfunction due to abdominal compartment syndrome is herein reported. A 62-year-old woman on peritoneal dialysis underwent dual kidney transplantation. The grafts were positioned extraperitoneally in both iliac possae using standard techniques. Surgical procedures and immediate postoperative period were uneventful. The urine output was immediate and the creatinine decreased, but in a few days she developed severe ascites with reduced urine output, increased creatinine, and progressive changes on Doppler ultrasound. The patient underwent paracentesis: the kidney function recovered as well as the Doppler ultrasound. Kidney biopsy was negative for rejection or renal pathology. Graft dysfunction was related to the presence of ascites. A catheter inserted in the abdomen measured intra-abdominal pressure (IAP) of 14 mm Hg. IAP correlated with renal function showing that IAP probably explained renal flow modifications. PMID- 18675132 TI - Donor-recipient MELD-based match in a patient who required three liver grafts in the era of nonstandard donors: case report. AB - In recent studies, nonstandard donors and high Model for End-stage Liver Disease (MELD) values have been indicated as risk factors for both graft survival and patient survival. A recent debate concerns which donor and recipient match guarantees the best results in terms of early and late survival. To emphasize the role of the donor-recipient match, we have reported herein a complex case of a patient who changed his preoperative risk status, being transplanted three times using donors of different risk levels. At each transplant, the patient moved to a higher MELD class: first transplant MELD=22; second transplant MELD=37; third transplant MELD=38. Only at the third transplant did the patient recover. Besides the liver, almost all his organs (kidneys, heart, lungs) recovered in a few weeks, as well. Unfortunately, severe cortical and subcortical brain damage remained a crucial limiting impairment, leading to death 5 months later, due to pulmonary infection, yet with a perfectly working liver. We underlined the role of donor factors to predict the outcome after liver transplantation in the MELD era. PMID- 18675133 TI - Activated recombinant protein C in septic shock early after liver transplantation: a case report. AB - Severe infectious diseases after liver transplant are associated with high risk of multiorgan failure and mortality. Septic shock is difficult to manage in this setting since it is often unresponsive to conventional aggressive therapy. Adjuvant therapies have been proposed in association with full combination treatment to sustain the failing organs and improve outcomes in severe sepsis. Recombinant human activated protein C drotrecogin alfa, Xigris) has been occasionally administered to treat posttransplant sepsis to modulate and downregulate the complex network of inflammatory and coagulopathic processes. Herein we have reported on a patient who was given drotrecogin alfa 15 days following liver transplant for acute septic shock originating from a nosocomially acquired pneumonia. Recombinant activated drotrecogin alfa, associated with conventional aggressive treatment, was efficacious to revert the life-threatening "slippery slope" of vasoplegia and uncontrolled diffuse inflammation. PMID- 18675134 TI - De novo autoimmune hepatitis following liver transplantation: a case report. AB - De novo autoimmune hepatitis (AIH), a rare disorder first described in 1998, appears in patients with liver transplants due to autoimmune and nonautoimmune etiologies. De novo AIH occurs in 2.5% to 3.4% of allografts; children seem to have a predilection for this syndrome. We have present herein a case of a liver allograft recipient who developed chronic hepatitis associated with autoimmune features outlining the clinical course, liver histology, and response to treatment. PMID- 18675135 TI - Combined liver-kidney transplantation in polycystic disease: case reports. AB - Polycystic disease causes a progressive decrease in renal function and liver degeneration. The progression of the disease evolves separately between organs and transplantation options vary: simultaneous or sequential liver-kidney transplantation or single-organ transplantation. From September 2006 to June 2007 3 combined liver kidney transplantations (CLKT) were performed for polycystic disease with end-stage renal disease: 2 with polycystic liver disease, and 1 with hepatic failure due to congenital hepatic fibrosis. The widest dimensions of the polycystic liver of 50 and 60 cm diameter were due to extensive cystic degeneration. We performed 1 simultaneous CLKT and 2 sequential transplantations: 1 liver after kidney, and 1 kidney after liver. At present all patients are alive with 100% graft function. Median creatinine level at discharge was 0.9 mg/dL (ranges, +/-0.2). Good liver graft function was reported in all 3 cases. Transplant benefit in polycystic liver-kidney disease has been already demonstrated; conservative surgical options may result in a high incidence of complications in highly involved polycystic livers. Delaying transplantation results in a more difficult surgical technique, a higher rate of postoperative complications, and a disturbance of optimal graft retrieval because of the worse preoperative condition of the patients. PMID- 18675136 TI - Use of recombinant factor IX and thromboelastography in a patient with hemophilia B undergoing liver transplantation: a case report. AB - Hemophilia B is a congenital recessive disorder caused by deficiency of coagulation factor IX (FIX). Surgical procedures can be performed in patients with hemophilia using high-purity and/or recombinant FIX, which has been shown to be safe and effective in surgical hemostasis. Liver transplantation is the only potentially curative treatment available for these patients, providing a long term phenotypic cure for hemophilia. End-stage liver disease together with hemophilia exposes patients to greater risks of bleeding complications during the perioperative period with consequent difficulties in managing coagulopathy. The limited experiences reported by different investigators and the various strategies for clotting factor replacement make it difficult to define a single approach with respect to the optimal dose and method of administering FIX to achieve perioperative hemostasis. The limits of plasma-based coagulation tests- prothrombin time, activated partial thromboplastin time--have made thromboelastography a valid alternative in this kind of surgery. It has been demonstrated to be a useful tool for real-time analysis of clot formation using a whole-blood assay format. Further, it accurately illustrates the clinical effects of procoagulant or anticoagulant interventions. In this article, we have described the usefulness of thromboelastography to monitor the ability of high purity FIX supplementation to restore a normal coagulation state and to guide the perioperative administration of blood products in a successful orthotopic liver transplantation in a hemophilic patient with deficiencies of factors IX and X, presenting with hepatitis C virus-related cirrhosis and hepatocellular carcinoma. PMID- 18675137 TI - [What's new in dermatological research? Contributions from the 2006 literature]. AB - A lecture is given every year at the annual meeting of the French Society of Dermatology in Paris by a dermatologist who has reviewed the scientific literature for the previous year. This manuscript is based on the 2007 lecture and reviews topics felt to be of potential interest to understanding of cutaneous biology, pathophysiology of skin diseases and treatments related to dermatology. It includes the following areas of interest: Wnt signaling, genetic disorders, rosacea, acne, itch, autoimmune bullous diseases, skin-barrier function, psoriasis and new therapies. PMID- 18675138 TI - [What's new in medicine in 2007?]. AB - What's new in medicine in 2007? Find out what has changed in medicine in 2007 in the field of inflammatory and autoimmune diseases. The year has been productive with many fundamental advances. 2007 was the year of genetics in autoimmune diseases (rheumatoid arthritis, lupus, scleroderma, vascularities, etc.), coming notably from work by the WTCCC. The comprehension of inflammatory diseases is based on increasingly detailed knowledge of epigenetic regulation and posttranscriptional mechanisms. The small RNA play a fundamental role and potentially offer many therapeutic applications.Sex and stress play a role in the appearance of inflammatory diseases. The mechanisms are progressively being better understood.More and more illnesses have autoimmune mechanisms, such as emphysema in the ex-smoker or pustulosis palmaris et plantaris. The revolution in biological therapies is continuing to grow. Medications that modulate innate immunity are being evaluated in inflammatory, allergic, and neoplastic diseases. Other, sometimes surprising options such as arsenic are now possible. PMID- 18675139 TI - [What's new in clinical dermatology?]. AB - For this What's new in clinical dermatology?, I have selected some important publications mainly in the field of the infectious dermatosis and drugs allergy. The Chikungunya virus which is responsible of several epidemics since 2005 seems to be a virus with a cutaneous tropism. Sexually transmitted diseases are still a public health problem and we shall review the situation for syphilis, lymphogranuloma venereum and the new dynamic profile of HIV in France. Lyme disease has been the subject of consensus conference in France and in the United States. Original clinical presentations in relation with infectious agent will be also presented. We shall give a large place to what I call the toxidermatology. During the year 2007, the role of gadolinium in the nephrogenic systemic fibrosis has been confirmed, many publications have reported the side effects of biotherapies on the skin and we shall emphasize the paradoxical of some of them. Hypersensitivity syndrome and notably the controversial role of viral reactivations has been reported. The relationship between psoriasis and metabolic syndrome or cardiovascular risks has been discussed since 2006. New clinical features have been described and will be presented during this session. PMID- 18675140 TI - [What's new in pediatric dermatology?]. AB - The literature review in paediatric dermatology for 2007 is very rich and covers the principal themes of the speciality: eczema, angiomas, genetic diseases, and juvenile and teenage systemic diseases. The role of environmental factors in infant eczema was particularly studied, as well as the problem of therapeutic adherence in the case of topical treatments. The treatment of severe forms of angioma, such as hemangioma and vascular dysplasia were also dealt with. Several publications dealt with the paediatric specificities of auto-immune diseases: dermatomyositis, scleroderma, bullous pemphigoid. Genetic diseases with a cutaneous expression were the basis for many publications. These publications deal with the genomic aspect but also the psycho-social aspect of these chronic diseases which greatly affect the patient's quality of life. PMID- 18675141 TI - [Skin cancer: what's new in clinical research?]. AB - Every year skin cancer is the subject of many publications. In this review of the recent literature, we selected epidemiological, clinical, and therapeutic data published between October 2006 and September 2007. Some of the studies presented in conferences in 2007 are also mentioned. New epidemiological data have been provided regarding the incidence of different types of skin cancers, the effect of certain risk factors or drug or food protective factors, the overall improvement of survival in melanoma, the epidemiological, clinical, and/or therapeutic particularities of rapidely growing melanomas and thick melanomas, and the relations between sun exposure, skin cancers, and other solid tumors. New prognostic studies in melanoma have improved the evaluation of prognosis in specific situations such as thin melanoma, acrolentiginous melanoma, and melanoma with positive sentinel lymph nodes (depending on the type of positivity) or negative sentinel lymph nodes. Important results on adjuvant interferon in melanoma have been presented. Clinical trials evaluating new therapeutic approaches in stage III and IV melanoma are going on. Several studies confirmed the clinical benefit of adjuvant irradiation on the tumor site in Merkel cell carcinoma. New treatments are being studied or have been approved in refractory forms of cutaneous T-cell lymphoma and in inoperable forms of squamous cell carcinoma and dermatofibrosarcoma protuberans. PMID- 18675142 TI - [What's new in dermatological therapy?]. AB - A review of the medical literature and of the regulatory agencies website was performed to identify new information about dermatological therapy from October 2006 to October 2007. Care was taken to prioritize results from randomized controlled trials and epidemiological studies of acceptable methodology. In the last year, significant advances have been made in the field of psoriasis, pemphigus, prevention of infection with oncogenic papillomaviruses, leg ulcers, evaluation of lasers and photodynamic therapy. The availability of biological agents for the treatment of psoriasis, auto-immune disease and skin cancer will certainly induce major changes in our therapeutic strategies in the near future. The dermatologist needs to keep-up with new therapies in order to address the therapeutic needs of patients with skin diseases. PMID- 18675143 TI - Health care worker pertussis immunization requirements and patient safety. PMID- 18675144 TI - Implementation of a pertussis immunization program in a teaching hospital: an argument for federally mandated pertussis vaccination of health care workers. AB - BACKGROUND: As pertussis disease becomes more common, health care-associated outbreaks have been reported with increasing frequency. Often, these clusters are costly and labor intensive to investigate and contain. It is clear that health care workers are among the adults who transmit pertussis to susceptible infants. Recent focus on patient safety, together with a concern for protecting employees in the workplace and those they expose elsewhere, has spurred interest in optimizing measures to prevent infection and disease transmission. Shortly after a tetanus toxoid, reduced diphtheria toxoid and acellular pertussis vaccine adsorbed booster was licensed and became available, we designed, launched, and analyzed a campaign to immunize the employees of our institution against pertussis. METHODS: To optimize acceptance of a tetanus toxoid, reduced diphtheria toxoid and acellular pertussis vaccine adsorbed booster by employees, we adopted a program consisting of a 3-phase publicity and educational model and a 3-phase vaccine delivery approach. RESULTS: Despite extraordinary resources dedicated to this program, and our institution's better than average annual uptake of influenza vaccine, less than one third of our eligible employees were immunized. A significant number of employees declined to be vaccinated for inappropriate reasons. CONCLUSION: A campaign of this kind is quite labor intensive and expensive, yet limited overall vaccine uptake was achieved. A federal mandate to require pertussis immunization of all health care workers appears to be a more effective way to protect our patients, employees, families, and society. PMID- 18675145 TI - Why don't doctors wash their hands? A correlational study of thinking styles and hand hygiene. AB - BACKGROUND: The World Health Organization has identified cognitive determinants of hand hygiene as an outstanding research question. This study investigated whether doctors' preferences for a rational thinking style or an experiential thinking style are associated with hand hygiene compliance. METHODS: This was an observational study of hand hygiene practices of 32 doctors in 2 teaching hospitals in South Australia. Compliance rates were correlated with self-reported thinking styles. The doctors were observed by a trained observer during a ward round or outpatient clinic and were unaware that hand hygiene was under observation. The main outcome measures were hand hygiene compliance (hand hygiene compliance tool) and thinking style (Rational-Experiential Inventory). RESULTS: An overall mean compliance rate of 7.6% (standard deviation +/- 7.2%) was found. Compliance was significantly positively correlated with experiential/automatic thinking (r = .46; P = .004) and the observational setting of ward rounds (vs clinics) (r = -.47; P = .003). No significant relationship was found between compliance and a rational/deliberate thinking style (r = -.01; P = .472). CONCLUSIONS: Hand hygiene is more experiential than rational. Findings suggest that certain promotional strategies appealing to the experiential thinking mode may improve compliance, and that traditional approaches based on logic and reasoning alone probably will not work. PMID- 18675146 TI - Impact of 2 different levels of performance feedback on compliance with infection control process measures in 2 intensive care units. AB - BACKGROUND: Performance monitoring and feedback of infection control process measures is an important tool for improving guideline adherence. Different feedback strategies may lead to distinctive outcomes. OBJECTIVES: Our objective was to determine the relative impact of 2 different levels of feedback on compliance in an intensive care unit (ICU) setting. METHODS: Proportion of head of bed (HOB) elevation, hand hygiene (HH) compliance, and proportion of femoral catheter (FC) to all central venous catheter-days were observed in a medical ICU and a surgical ICU. After a 3-month baseline observation phase (phase 1; P1), we provided quarterly feedback on these process measures and major health care associated infections (HAIs) to unit leaders from July 2004 to June 2005 (P2). From July 2005 to June 2006 (P3), feedback parameters were also provided to unit leaders and to all staff via 48 x 72-inch color posters in ICU personnel-only areas. At the end of the study, a survey was performed to assess the influence of the posters and HH observations. RESULTS: The analysis of IC process measures included 6948 HOB elevation observations, 1576 HH opportunities, and 16,591 catheter-days. In P2, the overall compliance with HOB elevation and the proportion of FC use significantly improved from 51% to 88% (P < .001) and 13% to 7% (P < .001), respectively. No significant difference in HH compliance was observed during this phase (40% vs 47%, respectively; P = .28). Comparing P3 with P2, HH compliance significantly improved from 47% to 71% (P < .001), and there was a slight improvement in HOB elevation rate from 88% to 93% (P < .001). There was no significant change in FC use in P3. There were 53 survey respondents. Sixty percent reported that the poster information changed their practices. Nearly all respondents (92%) knew that their HH behavior was being observed; however, 61% claimed that HH compliance was not influenced by observation. CONCLUSION: Feedback of infection control process measures and major HAIs to unit leadership significantly improved compliance with HOB elevation rate and FC use but not HH. Multilevel feedback significantly improved HH compliance and delivered a satisfactory level of compliance with HOB and FC use in both ICUs during the study period. PMID- 18675147 TI - Characteristics of persons and jobs with needlestick injuries in a national data set. AB - BACKGROUND: Physicians, nurses, and others are at risk of needlesticks, yet little national information is available regarding incidence across demographic and occupational categories. METHODS: Analysis was conducted on national data on occupational injuries for 1992-2003 from the Bureau of Labor Statistics (BLS). Because BLS data were limited to cases with 1 or more days of work loss, and reasons related to reporting of incidents, the data only reflected a subset of all needlesticks. Nevertheless, the data were internally consistent across categories so that relative magnitudes were reliable. Statistical tests for differences in proportions were conducted that compared needlesticks with all other occupational injuries and employment. RESULTS: Cases with 1 or more days of work loss numbered 903 per year, on average, from 1992 through 2003. Women comprised 73.3% (95% CI: 72.5%-74.2%) of persons injured. For those reporting race, white, non-Hispanic comprised 69.3% of the total (95% CI: 68.1%-70.4%); black, non-Hispanic, 14.8% (95% CI: 13.9%-15.6%); and Hispanic, 13.8% (95% CI: 12.9%-14.6%). The age bracket 35 to 44 years had the highest percentage of injuries at 34.0% (95% CI: 33.1%-34.9%). Ages over 54 years reported smaller percentages of needlestick injuries than either all other injuries or employment. Occupations with greatest frequencies included registered nurses, nursing aides and orderlies, janitors and cleaners, licensed practical nurses, and maids and housemen. Occupations with greatest risks included biologic technicians, janitors and cleaners, and maids and housemen. Almost 20% (95% CI: 18.88%-20.49%) of needlesticks occurred outside the services industry. Seven percent (95% CI: 6.56% 7.53%) of needlesticks resulted in 31 or more days of work loss in contrast to 20.46% (95% CI: 20.44%-20.48%) of all other injuries. CONCLUSION: In this nationally representative sample, the most frequent demographic and occupational categories were women; white, non-Hispanic; ages 35 to 44 years; and registered nurses. PMID- 18675148 TI - Point-of-use water filtration reduces endemic Pseudomonas aeruginosa infections on a surgical intensive care unit. AB - BACKGROUND: Endemic infections because of Pseudomonas aeruginosa were observed on a surgical intensive care unit (ICU) for a period of >24 months. Tap water probing revealed persistent colonization of all ICU water taps with a single P aeruginosa clonotype. METHODS: Water outlets of the ICU were equipped with disposable point-of-use water filters, changed in weekly and, later, 2-week intervals. To delineate the effect of the filters, 4 study approaches were followed: (1) a descriptive analysis of the incidence of P aeruginosa colonizations and infections, (2) microbiologic examinations of tap water before and after installation of the filters, (3) a comparative cohort analysis of representative patient samples from the prefilter and postfilter time periods, and (4) an analysis of general ward variables for the 2 periods. RESULTS: (1) The mean monthly rate (+/-SD) of P aeruginosa infection/colonization episodes was 3.9 +/- 2.4 in the prefilter and 0.8 +/- 0.8 in the postfilter period. P aeruginosa colonizations were reduced by 85% (P < .0001) and invasive infections by 56% (P < .0003) in the postfilter period. (2) Microbiologic examinations of tap water revealed growth of P aeruginosa in 113 of 117 (97%) samples collected during the prefilter period, compared with 0 of 52 samples taken from filter-equipped taps. (3) In the comparative cohort analysis, a number of patient-related variables were significantly associated with P aeruginosa colonization/infection. Considering these variables in a multivariate analysis, belonging to the postfilter cohort was the factor most strongly associated with a reduced risk of P aeruginosa positivity (relative risk, 0.04; P = .0002). (4) General ward variables such as bed occupancy, personnel-to-patient ratio, or microbiologic culturing density did not differ significantly between the 2 periods. CONCLUSION: Taking into account various patient-related and general ward variables, point-of use water filtration was associated with a significant reduction of chronically endemic P aeruginosa colonizations/infections on a surgical ICU. PMID- 18675150 TI - Varicella zoster virus in health care workers in northern Israel: seroprevalence and predictive value of history of varicella infection. AB - Health care workers (HCWs) are susceptible to hospital acquired varicella zoster virus (VZV). We evaluated seroprevalence and predictive value of a history of varicella disease (VD) with VZV serology in HCWs in northern Israel. A total of 200 HCWs were enrolled. A high rate of seropositivity for VZV-IgG was found: 98.5% seropositive and only 1.5% seronegative. A positive history of VD was an excellent predictor for the presence of VZV-IgG; however, a negative history of VD does not rule out the presence of VZV-IgG. PMID- 18675149 TI - Impact of a standardized hand hygiene program on the incidence of nosocomial infection in very low birth weight infants. AB - BACKGROUND: This study examined the effects of a standardized hand hygiene program on the rate of nosocomial infection (NI) in very low birth weight (VLBW) infants (birth weight < 1500 g) admitted to our neonatal intensive care unit (NICU). METHODS: We compared the rate of NI in VLBW infants in 2 separate periods. In the first period, staff were encouraged to perform handwashing using a plain fluid detergent (0.5% triclosan). In the second period, a standardized hand hygiene program was implemented using antimicrobial soap (4% chlorhexidine gluconate) and alcohol-based hand rubs. RESULTS: NI after 72 hours of life was detected in 16 of the 85 VLBW infants in the first period and in 5 of the 80 VLBW infants in the second period. The rate of central venous catheter colonization was significantly lower in the second period (5.8%) than in the first period (16.6%). CONCLUSION: In our NICU, the incidence of NI in VLBW infants was significantly reduced after the introduction of a standardized handwashing protocol. In our experience, a proper hand hygiene program can save approximately 10 NI episodes/year, at a cost of $10,000 per episode. Therefore, improving hand hygiene practice is a cost-effective program in the NICU. PMID- 18675151 TI - Needlestick and sharps injuries in a tertiary hospital in the Republic of Korea. AB - BACKGROUND: The high incidence of hepatitis B virus (HBV) in the Republic of Korea has focused attention on monitoring the occurrence and characteristics of needlestick and sharps injuries (NSIs) as part of an effort to reduce the occupational exposure to bloodborne pathogens such as HBV. This study investigated NSIs reported in a tertiary referral hospital in Busan, Republic of Korea over a 6-year period (2001 to 2006). METHOD: Data on the number of NSIs, places where NSIs occurred, devices causing injury, purpose of using sharps, and circumstances surrounding NSIs were collected from the study hospital's NSI database. The incidence of NSIs per 100 full-time equivalent (FTE) employees was calculated by year and by profession. RESULTS: A total of 221 NSI cases were reported during the study period. Overall incidence was 2.6 cases per 100 FTE employees per year, with the highest rate occurring in interns (17.7 cases per 100 FTE interns per year). Some 34% of cases occurred in the ward, needles were the most common device causing injury (73%), and the most common circumstance surrounding an NSI was after sharps use and before disposal (24%). CONCLUSION: The pattern of NSI occurrence found in this study was comparable to that reported in previous studies. However, the overall incidence of NSIs was significantly lower than that in previous studies, apparently related to underreporting of NSIs. Further research to investigate reasons for this underreporting is recommended. Considering the high incidence of NSIs in interns, in-service training for this group should be enhanced. PMID- 18675152 TI - Molecular epidemiology of Acinetobacter baumannii spread in an adult intensive care unit under an endemic setting. AB - BACKGROUND: Acinetobacter baumannii (Ab) clones I, III, and IV were recovered in several Buenos Aires City hospitals. We investigated the prevalence of these clones with epidemic behavior (EB) in our intensive care unit (ICU) under an endemic setting and its spread. METHODS: A 10-week prospective cohort study including surveillance cultures of newly admitted patients was conducted. Air, environment, and staff hands were weekly screened. In the seventh week, a new environmental cleaning protocol and a staff hand hygiene reeducation program were implemented. RESULTS: Almost 15% of all screening samples (159/1042) were Ab positive. Up to the seventh week, carbapenem-resistant clone If was the main one recovered from patients, environmental frequently touched surfaces (EFTS), and staff hands screening samples. Few air samples were Ab positive. Clone I was also isolated from patients at admission. After the seventh week, a significant reduction of EFTS contamination and of clone If isolation was observed. During the last 3 weeks, clone I was no longer isolated from patients. Instead, the newly identified clone IVb was mainly cross transmitted. It was also recovered from staff hands and from EFTS. In the last week, clone If was again isolated from 1 bed rail. CONCLUSION: Patients with EB clones-positive culture at admission provide verification that interhospital patient transfers play a role in these clones spread. However, subtypes such as clone If seem to be endemic in our ICU. EFTS showed to have potential for EB clones transmission via transient staff hand carriage. Transmission did not involve airborne route. PMID- 18675153 TI - A Chryseobacterium meningosepticum outbreak observed in 3 clusters involving both neonatal and non-neonatal pediatric patients. AB - Three clusters of Chryseobacterium meningosepticum infections in a tertiary health center in July 2006 and January 2007 involving 8 newborns and 5 older children were investigated. The index patient was from the neonatal intensive care unit, and the older patients were from other pediatric wards. Cultures were obtained from the environment and from health care workers' hands as part of an outbreak investigation. C meningosepticum was isolated from hand cultures obtained from a senior resident and from environmental cultures obtained from powdered infant formula, an electrical button, a computer keyboard, phone, a doorknob, and an Ambu bag. Antibiogram typing and enterobacterial repetitive intergenic consensus sequence polymerase chain reaction indicated that all of the isolates were epidemiologically related. Nine patients improved on antimicrobial treatment, and 4 premature infants died after the infection. C meningosepticum is a well-known etiologic agent for nosocomial infections involving newborns and immunocompromised patients. Wet and dry environmental surfaces and equipment may act as a source or play a role in disseminating the microorganism. Outbreaks may be controlled with strong emphasis on infection control measures. PMID- 18675154 TI - A pilot study to isolate Staphylococcus aureus and methicillin-resistant S aureus from environmental surfaces in the home. AB - BACKGROUND: The major sources of Staphylococcus aureus, including methicillin resistant S aureus (MRSA), in the home are colonized or infected individuals and pets, such as cats and dogs; however, the occurrence of MRSA on surfaces in healthy homes is not well documented. METHODS: A convenience sample comprising 35 homes of health care and non-health care workers, each with a child in diapers and either a cat or dog in the home, was recruited from the Boston area between January and April 2006. In each home, a total of 32 surfaces were sampled in kitchens, bathrooms, and living areas. RESULTS: S aureus was found in 34 of the 35 homes (97%) and was isolated from all surfaces in 1 or more homes, with the exception of the kitchen chopping board and the child training potty. MRSA was isolated from 9 of 35 homes (26%) and was found on a variety of household surfaces, including the kitchen and bathroom sinks, countertops, kitchen faucet handle, kitchen drain, dish sponge/cloth, dish towel, tub, infant high chair tray, and pet food dish. A positive correlation was indicated for the presence of a cat and the isolation of MRSA from surfaces. CONCLUSIONS: This study has shown the presence of MRSA at hand-contact surfaces in healthy homes. This provides further evidence for the potential for infection transmission via inanimate surfaces and underscores the need for good hygiene practice in the home. PMID- 18675155 TI - Active surveillance cultures are not required to control MRSA infections in the critical care setting. AB - Although the effectiveness of active surveillance cultures to identify and isolate patients colonized with methicillin-resistant Staphylococcus aureus (MRSA) remains debated, hospitals are under increasing pressure to begin active surveillance programs. We analyzed our data on device-associated MRSA infections in the intensive care unit setting over a 4-year period during which multiple evidence-based interventions to reduce hospital-acquired infections were introduced without performing active surveillance cultures for MRSA. We observed reductions in all infections, including those caused by MRSA, and conclude that control of MRSA in the critical care setting does not require active surveillance cultures. PMID- 18675156 TI - [SIGAPS score: time to publish in the Annales de pathologie]. PMID- 18675157 TI - [Sessile serrated polyps of the colorectum: how to distinguish a sessile serrated adenoma from a hyperplastic polyp in a daily practice?]. AB - AIMS: Serrated polyps of the colorectum are a heterogenous group of mucosal lesions including hyperplastic polyps (HP) and sessile serrated adenomas (SSA), but their morphologic distinction is not always straightforward. However, it is important for the pathologist to identify SSA because recent data show that they might be the precursors of serrated adenocarcinomas which are probably involved in the serrated pathway. MATERIAL AND METHODS: We selected 102 serrated colorectal polyps resected by colonoscopy and evaluated the following parameters: location, size, number of biopsies per polyp, superficial or tangential biopsies, type of resection, location of the serrated feature, branching, horizontalisation, dilatation and herniation of crypts through the muscularis mucosae, cellular type, epithelial tufts, cytoplasmic eosinophilia and dysplasia. RESULTS: There were 81 HP (79%), seven SSA (7%) of which one showed foci of dysplasia, five traditional serrated adenomas (5%) and three mixed polyps (HP and tubulous adenoma: 3%). Only six serrated polyps could not be classified. The main architectural criterion for diagnosing SSA was a serrated pattern throughout the crypt axis and the rarity of undifferentiated cells at the base of the crypts. Moreover, clinical characteristics were also helpful, since sessile serrated adenomas were significantly more often located in the right colon and larger (median: 11mm versus 4mm) than HP. CONCLUSION: SSA can be distinguished morphologically from HP in a daily practice. The presence of foci of dysplasia in one case of SSA supports the hypothesis that these polyps have a carcinogenetic potential and should have the same clinical follow-up as traditional adenomas. PMID- 18675158 TI - [Placental mesenchymal dysplasia]. AB - Placental mesenchymal dysplasia (PMD) is a recently described entity. Because of the presence of an embryo and a multicystic placenta, it has been initially misinterpreted as a partial mole on ultrasound as well as on gross examination. However, careful microscopic examination, cytogenetic analyses, molecular studies and follow-up of children allowed to better understand this placental abnormality, allowing an antenatal diagnosis. Our purpose was to describe ultrasonographic, macroscopic and microscopic features in PMD on the basis of a literature review and our own experience. We also detailed clinical presentation, complications of pregnancy and fetopathologic associations. We discussed ethiopathogenic hypotheses as well as differential diagnoses. PMID- 18675159 TI - [Morphological and immunohistochemical pitfalls in pathology. Introduction]. PMID- 18675169 TI - [Vaginal adenoid cystic carcinoma: a case report]. AB - Adenoid cystic carcinoma generally arises from the salivary glands and is rarely found in the female genital tract. Infection with HPV is implicated in this cervical lesion. Differential diagnosis includes adenoid basal carcinoma, polymorphous low-grade adenocarcinoma and basaloid squamous cell carcinoma. Only one case of vaginal localisation was previously described. We report a case of adenoid cystic carcinoma in a 48-year-old woman with previous cervical HPV infection. Histological examination revealed nests of cells with peripheral palisading organisation and glandular lumina containing material produced by the tumor cells. PMID- 18675170 TI - [Pancreatic perivascular epithelioid cell tumor (PEComa)]. AB - Neoplasms with perivascular epithelioid-cell differentiation (PEComas) are rare tumors with a distinctive immunoreactivity for melanocytic markers. They have been described in various organs. We report an intrapancreatic PEComa discovered in a 46-year-old woman during a workup for diarrhea. CT scan showed a 1.7cm nodule in the body of the pancreas with slight-contrast enhancement at arterial time and isodense at portal time. The aspect was suggestive of an endocrine tumor despite negative somatostatin-receptor scintigraphy. Enucleation was performed. Pathologic evaluation showed a well-circumscribed intrapancreatic tumor consisting of a population of clear to eosinophilic spindle cells and a less abundant population of epithelioid cells arranged around blood vessels. Tumor cells expressed vimentin, HMB45 and actin and only focally S-100 protein, KL1, CD117 and CD34. These features were consistent with a PEComa. Pancreatic PEComas are rare, but should be included in the differential diagnostic of pancreatic clear cells tumors or pancreatic spindle- and epithelioid-cells tumors. PMID- 18675171 TI - [Uncommon retroperitoneal and bone lesions: Erdheim-Chester disease]. AB - We report a case of Erdheim-Chester disease, revealed by a polyuropolydipsic syndrome. During the patient's work-up, osteocondensing lesions were found; the biopsy of these lesions showed an infiltration by spumous cells of histiocytic lineage, CD68+, CD1a-, associated with a lymphoid infiltrate within an extensive fibrosis. Lung and retroperitoneal lesions were discovered. The surgical resection of the involved ureter was required. Histological examination of the resected specimen showed the same pattern of histiocytic infiltration. Our case report underlines the variety of lesions associated with Erdheim-Chester disease and the importance of a complete exploration. PMID- 18675172 TI - [Late-onset vaccination-induced subcutaneous pseudolymphoma]. AB - Persistent subcutaneous nodules arise on rare occasions at sites of injection of aluminium hydroxide-adsorbed vaccine. We report a case following a diphtheria, tetanus and pertussis vaccination. The late onset of the lesion, four years after the injection, led to an uncertain preoperative diagnosis. Histopathologic examination showed features of a subcutaneous pseudolymphoma. The demonstration of aluminium by Morin staining and atomic absorption spectrometry on a paraffin embedded tissue probe supported the diagnosis of a vaccination-induced pseudolymphoma. PMID- 18675173 TI - [A gastric tumor with unusual features]. PMID- 18675174 TI - [A renal tumor]. PMID- 18675175 TI - [A painful mass of the epidydimis]. PMID- 18675176 TI - [A perineal nodule in a woman]. PMID- 18675177 TI - [Misleading signet-ring cells...]. PMID- 18675178 TI - [A rare cervical tumor]. PMID- 18675179 TI - Investigator-initiated research in intensive care: achievement through collaboration. PMID- 18675181 TI - [Hepatitis C, cirrhosis and hepatocellular carcinoma]. AB - The screening for the detection of hepatocellular carcinoma is based on ultrasound sonography which should be realised in patients with post-hepatitis C cirrhosis with a delay between 3 and 6 months according to the most identified risk factors, in particular age and sex male. In the case of discovery of hypoechogen nodule < or = 1cm, a follow-up is mandatory because it is usually untypical by ultrasound sonography and to propose a liver biopsy in the case of an increasing in size is shown. The ultrasound guided cutting biopsy can precise the histological characteristics of the nodule, the grade, and indicate prognostic factors. The liver biopsy is also mandatory in the case of a nodule > 2 cm and when the ultrasound sonography is not contributive, especially when the nodule is between 1 and 2 cm in size. PMID- 18675180 TI - [Hepatitis C: which strategy in case of treatment failure ?]. AB - After a treatment by peginterferon alpha and ribavirin, the percentages of non response and relapse are approximatively 33 and 18 % respectively. These treatment failures may be due either to viral resistance or to an insufficient treatment. The prevention of treatment failure is based on a good knowledge of the predictive factors of failure before and during the treatment. Among the patients who did not respond to interferon alpha and ribavirin, a new treatment with peginterferon alpha-2b and ribavirin makes it possible to obtain 45 % of sustained virological response (SVR) among the relapsers and 17 % of SVR among the non responders. Among the patients who did not respond to peginterferon alpha and ribavirin, a new treatment with peginterferon alpha-2b and ribavirin makes it possible to obtain 36 % of SVR among the relapsers and only 4 % of SVR among the non responders. New therapeutic strategies are necessary for the non responders. Until now no new therapeutic strategy allowed a significant benefit in term of SVR. Protease inhibitors are currently tested in non responders but there are some concerns about the risk of selection of multi-resistant strains. PMID- 18675182 TI - [Management of chronic hepatitis C in 2008]. PMID- 18675183 TI - [Management of chronic hepatitis in 2008]. PMID- 18675184 TI - [Alcohol, steatohepatitis, insulin resistance and hepatitis C]. AB - Patients with chronic hepatitis C have frequently other morbidities, either because they are frequent in the general population (metabolic syndrome) and/or because the route of contamination (chronic alcohol consumption succeeding to drug abuse). These co-morbidities have a harmfull impact on fibrosis progression during the natural history of HCV infection and reduce the efficacy of antiviral treatments. Thus, it is crucial to diagnose early and treat these different diseases which may be combined. They are the metabolic syndrome and/or chronic alcohol consumption resulting in insuline resistance, infection by the human immune deficiency virus or by the hepatitis B virus as well as chronic tobacco use or excessive consumption of cannabis. An optimal is based on a multidisciplinary approach to reduce fibrosis progression and improve the efficiency of antiviral therapies. However, the hepatologist has to come back to a global care, which is mandatory at the individual level as well as for the public health. PMID- 18675185 TI - [HIV and hepatitis C virus coinfection]. AB - Managing chronic hepatitis C in patients coinfected with the human immunodeficiency virus is a significant challenge. Treatment is influenced by a number of viral and host characteristics, including hepatitis C virus genotype, baseline viremia, and adherence to medication. Accelerated progression of liver disease, immunodeficiency, and hepatotoxicity of antiretroviral drugs are additional concerns in coinfected patients. According to the results of 5 randomized clinical trials, 27 %-55 % of coinfected patients who receive therapy with peginterferon alfa plus ribavirin attain sustained virologic response. These studies also confirm the importance of early virologic response as a predictor of treatment outcome and reveal the considerable proportion of patients who experience hematologic tolerability issues. Effective management strategies that encompass patient and viral factors are necessary to improve the long-term outlook for coinfected patients. PMID- 18675186 TI - [Non-invasive evaluation of liver fibrosis in hepatitis C]. AB - In 2007, the << Haute Autorite de Sante >> recommended FibroScan, FibroTest or liver biopsy for the initial diagnosis of fibrosis in patients with hepatitis C without co morbidities. These methods have to be interpreted according to the clinical situation, keeping in mind negative and positive false results. For FibroTest, hemolysis, Gilbert syndrome or acute inflammation can modify the result. Pre-analytical and analytical conditions of FibroTest have to be respected according to manufactory recommendations. For FibroScan, the numbers of measurements, the rate of successful measurements, and the interquartile range have to be correct. In case of suspicious results, FibroTest or FibroScan have to be done again. The liver biopsy, FibroTest, and FibroScan are less relevant for the distinction of two adjacent stages of fibrosis. However, their performances are excellent for the diagnosis of severe fibrosis or cirrhosis compared to moderate fibrosis. PMID- 18675187 TI - [Adherence to pegylated combination therapy in patients with chronic hepatitis C. Importance of the hepatologist, general practitioner, and nurse]. AB - Adhesion to pegylated combination therapy is a key factor for therapeutic success in patients HCV infected. To optimize it, goals to reach are to limit dose reduction and premature discontinuation of treatment due to adverse events ; to improve the patient compliance to treatment, particularly during the first three months, particularly to ribavirin. Therapeutic education, management of psychiatric adverse events, epoetin alfa, have demonstrated their benefit in terms of sustained virologic response or quality of life. Preparing the treatment with the patient and a multi-disciplinary team, setting successive therapeutic goals with the predictive value of the early virologic response will promote adhesion to treatment. A hepatitis C training program for general practitioners (GP) allows an efficient follow-up of treated patients by a trio hepatologist - GP - nurse and a concrete implication of GP in the field of hepatitis C. Further developments are needed for : taking in account the patient quality of life during treatment to anticipate premature discontinuation, promotion of therapeutic education by specialized nurses, standardization of the diagnosis of depression during treatment, and regular updating of general practitioners on antiviral C treatment. PMID- 18675188 TI - American Journal of Orthodontics and Dentofacial Orthopedics. Editor's choice. PMID- 18675189 TI - New business model for better access to care. PMID- 18675190 TI - Fourth and forty? PMID- 18675191 TI - To be commercially available or not to be: that is the question. PMID- 18675192 TI - Moment-to-force ratio. PMID- 18675193 TI - Friction and anchorage loading. PMID- 18675196 TI - Effectiveness of lingual retainers bonded to the canines in preventing mandibular incisor relapse. AB - INTRODUCTION: A retainer bonded to the lingual surfaces of the mandibular canines (3-3 retainer) is a widely used type of retention. Our aim in this study was to assess the effectiveness of the 3-3 mandibular lingual stainless steel retainer to prevent relapse of the orthodontic treatment in the mandibular anterior region. METHODS: The sample consisted of the dental casts of 235 consecutively treated patients (96 boys, 139 girls) from the archives of the Department of Orthodontics and Oral Biology, Radboud University Nijmegen Medical Center, The Netherlands, who received a 3-3 mandibular lingual stainless steel retainer at the end of active orthodontic treatment. The casts were studied before treatment (Ts), immediately after treatment (T0), and 2 years (T2), and 5 years (T5) posttreatment. RESULTS: The main irregularity index decreased significantly from 7.2 mm (SD, 4.0) at Ts to 0.3 mm (SD, 0.5) at T0; it increased significantly during the posttreatment period to 0.7 mm (SD, 0.8) at T2 and 0.9 mm (SD, 0.9) at T5. The irregularity index was stable during the 5-year posttreatment period (T0 T5) in 141 patients (60%) and increased by 0.4 mm (SD, 0.7) in 94 patients (40%). The intercanine distance increased 1.3 mm between Ts and T0 and remained stable during the posttreatment period. CONCLUSIONS: The 3-3 mandibular lingual stainless steel retainer (bonded to the canines only) is effective in preventing relapse in the mandibular anterior region in most patients, but a relatively high percentage will experience a small to moderate increase in mandibular incisor irregularity. PMID- 18675197 TI - Effects of sandblasting and silica coating on the bond strength of rebonded mechanically retentive ceramic brackets. AB - INTRODUCTION: The aim of this study was to determine the bond strength of rebonded mechanically retentive ceramic brackets after treatment with 2 abrasive techniques. METHODS: In addition to a group of new brackets, 3 groups were treated according to the following conditions of debonded ceramic bracket bases: sandblasting, sandblasting + silane, and silica coating + silane (15 in each group). Treated ceramic brackets were rebonded on premolars. The samples were stored in distilled deionized water for 24 hours at 37 degrees C in an incubator and then thermocycled for 1000 times between 5 degrees C and 55 degrees C. Shear force was applied to the enamel-adhesive interface until debonding. RESULTS: The highest bond strength values were in the silica coating + silane and the new bracket groups (12.7 and 12.0 MPa, respectively), followed by the sandblasting + silane group (10.5 MPa). The sandblasting group had a significantly lower bond strength value (4.5 MPa). No enamel fracture was noted in any sample tested. In the new bracket and the sandblasting + silane groups, 20% of the samples had adhesive remnant index scores of 2, and 80% had scores of 3. In the sandblasting group, all specimens debonded at the bracket-adhesive interface. The silica coating + silane group showed mixed failures. CONCLUSIONS: Sandblasting + silane and silica coating + silane applications on debonded ceramic bracket base can produce bond strengths comparable with new brackets. PMID- 18675198 TI - Mandibular asymmetry diagnosis with panoramic imaging. AB - INTRODUCTION: Our objective was to determine the diagnostic capability of panoramic imaging in any of its forms to detect mandibular asymmetry. METHODS: Several electronic databases were searched. Abstracts that appeared to fulfill the initial selection criteria (mandibular or craniofacial asymmetry, panoramic imaging, and evaluation of validity or accuracy of diagnosis with an appropriate gold standard) were selected, and full articles from these abstracts were gathered. The references from these articles were also examined for possible articles that could have been missed. RESULTS: Only 11 articles met all criteria. Vertical measurements, although more accurate than horizontal or angular measurements, are still not true representations of the real objects they correspond to. Magnification values reported by the manufacturer might not correspond to the calculated magnification value and be uniform across the panoramic imaging entire area. CONCLUSIONS: Caution is advised when using conventional or digital panoramic images to assess mandibular asymmetry. Understanding the limitations of every form of panoramic imaging is required. Future studies with panoramic images generated from 3-dimensional data are required to determine whether they overcome the limitations of conventional and digital panoramic images. PMID- 18675199 TI - Laser etching of enamel for orthodontic bonding. AB - INTRODUCTION: The purposes of this study were to assess the shear bond strength, the surface characteristics, and the adhesive remnant index scores of teeth prepared for bonding with erbium, chromium doped: yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser etching, and to compare laser etching with phosphoric-acid etching and a self-etching primer. METHODS: Sixty-four premolars, extracted for orthodontic purposes, were randomly divided into 4 groups, and a different method was used to prepare the tooth enamel in each group for bonding: irradiation for 15 seconds with a 0.75-W Er,Cr:YSGG laser; irradiation for 15 seconds with a 1.5 W Er,Cr:YSGG laser; etching with 37% phosphoric acid; application of a self etching primer. After surface preparation, standard edgewise stainless steel premolar brackets were bonded; 1 tooth in each group was not bonded and was examined under a scanning electron microscopic. The brackets were debonded 24 hours later; shear bond strengths were measured, and adhesive remnant index scores were recorded. RESULTS: Irradiation with the 0.75-W laser produced lower shear bond strengths than the other methods. No statistically significant differences were found between 1.5-W laser irradiation, phosphoric-acid etching, and self-etching primer. Adhesive remnant scores were compared with the chi square test, and statistically significant differences were found between all groups; when the 0.75-W laser irradiation group was excluded, no statistically significant differences were observed. CONCLUSIONS: Irradiation with a 1.50-W laser produced sufficient etching for orthodontic bonding, but irradiation with the 0.75-W laser did not. PMID- 18675200 TI - Comparison of shear bond strengths of orthodontic brackets bonded to deciduous and permanent teeth. AB - INTRODUCTION: This study was conducted (1) to compare the bond strengths of identical orthodontic brackets bonded to deciduous and permanent teeth with either of 2 primer/adhesive systems and (2) to evaluate the modes of bracket/adhesive failure. METHODS: Thirty-four permanent teeth and 34 deciduous teeth were collected. To these freshly extracted teeth, metal brackets were bonded with 2 primer/adhesive systems. These specimens were divided into 4 groups. In group I, consisting of 17 permanent teeth, the brackets were bonded with an acid-etching adhesive system; in group II, the self-etching adhesive system was used for bonding the brackets to 17 permanent teeth; in group III, consisting of 17 deciduous teeth, the acid-etching adhesive system was used; and in group IV, the self-etching adhesive system was used for bonding the brackets to 17 deciduous teeth. Shear bond strength was measured with a universal testing machine, and the mode of bracket/adhesive failure was determined according to the adhesive remnant index. RESULTS: The shear bond strengths of all 4 groups exceeded what is believed to be clinically sufficient strength of 6 to 8 MPa. Whether deciduous or permanent teeth, there were no significant differences in shear bond strength between the acid-etching and self-etching adhesive systems. The shear bond strengths of the brackets bonded to the deciduous teeth with either adhesive system were lower than those to the permanent teeth. Bond failure occurred at the enamel-adhesive interface more frequently in the self-etching adhesive system than in the acid-etching adhesive system. CONCLUSIONS: Bond strengths of brackets were lower with deciduous teeth but were high enough for clinical purposes. PMID- 18675201 TI - Effect of self-etchant pH on the shear bond strength of orthodontic brackets. AB - INTRODUCTION: Self-etching primers (SEPs) have been used successfully during bonding to reduce technique sensitivity while minimizing the etching of enamel. Although serving the same purpose, SEPs differ in acidity and aggressiveness. Thus, the purpose of this study was to determine whether the pH of SEPs affects the shear bond strength of orthodontic brackets. METHODS: Forty-five molars were cleaned, mounted, and randomly divided into 3 groups with different SEPs: in group 1 (control), 15 orthodontic brackets were bonded to the teeth with Transbond Plus (3M Unitek, Monrovia, Calif) with a pH of about 1.0; in group 2, 15 brackets were bonded with Adper Prompt L-Pop (3M ESPE, St Paul, Minn) with a pH of 0.9 to 1.0; in group 3, 15 brackets were bonded with Clearfil S3 Bond (Kuraray America, New York, NY), with a pH of 2.7. All teeth were bonded with Transbond XT paste (3M Unitek). The teeth were debonded within half an hour after initial bonding by using a universal testing machine. The residual adhesive on each tooth was evaluated. ANOVA was used to compare the shear bond strength (SBS) of the 3 groups, and the chi-square test was used to compare the adhesive remnant index (ARI) scores for the 3 groups. RESULTS: ANOVA indicated significant differences between the groups. The Duncan multiple range test indicated that Clearfil S3 Bond produced a significantly stronger mean SBS (6.5 +/- 1.9 MPa) than the Transbond Plus system (mean, 4.2 +/- 1.9 MPa). No significant differences were seen between the SBS of the brackets bonded with Adper (mean, 5.9 +/- 3.4 MPa) and the other 2 groups. The comparisons of the ARI scores between the 3 groups indicated that bracket failure mode was not significantly different. CONCLUSIONS: The SEP with the highest pH (least acidic), Clearfil S3 Bond, bonded brackets successfully and with the strongest SBS; this suggests that the pH of the SEP is not the primary determinant of the SBS. The clinician should be aware that some SEPs can leave the enamel surface healthier after debonding. PMID- 18675202 TI - Assessing skeletal maturity by using blood spot insulin-like growth factor I (IGF I) testing. AB - INTRODUCTION: Accurate determination of skeletal maturity and remaining growth is crucial to many orthodontic, orthognathic, and dental-implant timing decisions. Cervical vertebral stages and hand-wrist radiographs are currently used to identify peak mandibular bone growth. These are highly subjective techniques that not only involve radiographic exposure but also lack the ability to determine the intensity of the growth spurt and the end of growth. Insulin-like growth factor I (IGF-I) is a circulating growth hormone-dependent factor whose level correlates with sexual maturity; it is used to diagnose growth hormone deficiency and excess. We hypothesized that IGF-I levels would also correlate with cervical skeletal maturity and would be highest at the cervical stages that correspond to the greatest amount of facial growth. METHODS: We measured mean blood spot IGF-I levels in a cross-sectional study of 83 patients (44 female, 39 male) on recall to begin orthodontic treatment, in active treatment, or in posttreatment follow up. RESULTS: Mean blood spot IGF-I levels were significantly higher in the late pubertal stages than in the prepubertal, early pubertal, and postpubertal stages. Linear correlation showed that IGF-I levels had a significant positive correlation with cervical skeletal maturity from the prepubertal to the late pubertal stages, and a significant negative correlation from the late pubertal to the postpubertal stages. In the postpubertal stage, IGF-I levels had a negative linear correlation with increasing time since the onset of puberty and with chronological age. CONCLUSIONS: Blood spot IGF-I could be used as a skeletal maturity indicator and might be useful in detecting residual mandibular growth in young adults. PMID- 18675203 TI - Skeletal maturation in Indonesian and white children assessed with hand-wrist and cervical vertebrae methods. AB - INTRODUCTION: The purpose of this study was to describe the stages of skeletal maturity of Deutero-Malay Indonesian children according to the hand-wrist and cervical vertebrae methods and to compare them with white children. METHODS: The study included 2167 patients with hand-wrist radiographs and lateral cephalometric radiographs. Of these, there were 648 Indonesian boys, 303 white boys (age range of boys, 10-17 years), 774 Indonesian girls, and 442 white girls (age range of girls, 8-15 years). The skeletal maturation index (SMI) was used to evaluate the stages of skeletal maturity from hand-wrist radiographs, and the cervical vertebrae maturation (CVM) index was used to evaluate the stages of skeletal maturity from lateral cephalometric radiographs. One observer made all observations, and a repeatability study was undertaken. RESULTS: Box-and-whisker plots were used to show the age distribution on attainment of each maturation stage based on the SMI and CVM. On average, both the SMI and the CVM showed that white children attained each maturation stage about 0.5 to 1 year earlier than their Indonesian peers, although the differences were less obvious in girls than in boys. Multiple regression analysis was used to predict the SMI from the chronologic age. Both the Indonesian and the white boys groups showed a good relationship between predicted SMI and chronologic age (R(2) = 0.728 and 0.739, respectively), as did the Indonesian and white girls groups (R(2) = 0.755 and 0.748, respectively). Further multiple regression analyses used to investigate the differences in the ages of attainment of skeletal development between Indonesian and white subjects indicated that, across the age ranges investigated, on average for a particular age, the white boys were 1 SMI stage ahead of the Indonesian boys, and the white girls were about 0.5 SMI stage ahead of their Indonesian peers. Because the CVM has only 5 categories, it was not considered appropriate to use this form of multiple regression analysis. CONCLUSIONS: The findings confirmed marked variations in the chronologic ages for each skeletal maturity stage and also showed differences between the timing of skeletal maturity with both the SMI and the CVM between the sexes and the ethnic groups. These differences should be considered during orthodontic diagnosis and treatment planning. PMID- 18675204 TI - Discriminatory ability of the skeletal maturation index and the cervical vertebrae maturation index in detecting peak pubertal growth in Indonesian and white subjects with receiver operating characteristics analysis. AB - INTRODUCTION: In this study, we aimed to determine the effectiveness of the skeletal maturation index (SMI) and the cervical vertebrae maturation (CVM) index in discriminating between patients who have yet to attain their peak pubertal growth, compared with those who have reached or passed it. An additional aim was to determine whether there was any significant difference in the ability of the 2 methods to predict peak pubertal growth. METHODS: The study included 2167 patients with hand-wrist and lateral cephalometric radiographs. There were 648 Indonesian boys and 303 white boys (age range, 10-17 years), and 774 Indonesian girls and 442 white girls (age range, 8-15 years). The SMI was used to evaluate the stages of skeletal maturity from hand-wrist radiographs, and the CVM index was used to evaluate skeletal maturity from lateral cephalograms. Several commonly used cephalometric parameters representing maxillary and mandibular dimensions were also measured to construct growth curves and calculate peak velocity. RESULTS: Receiver operating characteristic (ROC) analysis was performed for the craniofacial morphology parameters for both sex and ethnic groups. The percentages of correctly classified subjects into the appropriate maturational stages for the mandibular parameters, as well as the stages with high sensitivity values for the maxillary parameters, showed that both the CVM index and the SMI have good discriminatory ability. On average, the results of the area under curve (AUC) for the SMI (AUC >0.9) were greater than for the CVM (AUC >0.8), and the differences between them were also statistically significant (P <0.05 for all parameters investigated). However, the curves for both CVM and SMI approached the top left corner of the ROC graph, suggesting that both tests have good discriminatory ability, and the differences between the methods were only between 1% and 7%. CONCLUSIONS: Both the CVM index and the SMI are valid methods to discriminate between patients who have not yet attained peak pubertal growth and those who have reached or passed it. The differences in discriminatory ability between the SMI and the CVM index in detecting peak pubertal growth were small. These results question the necessity of taking hand-wrist radiographs and exposing a child to additional radiation when the discriminatory ability is similar with the CVM index, which is readily derived from most lateral cephalograms. PMID- 18675205 TI - Effects of posttreatment skeletal maturity measured with the cervical vertebral maturation method on incisor alignment relapse. AB - INTRODUCTION: Our aim was to test the hypothesis that relapse of incisor alignment is associated with skeletal maturity at the end of treatment, as assessed with the cervical vertebral maturation (CVM) method. METHODS: This was a case-control study with information from the postretention database at the University of Washington. Mandibular incisor irregularity (II) at least 10 years out of retention (T3) was used to define the subjects (II >6 mm, relapse group) and the controls (II <3.5 mm, stable group). The following model measurements were made: II at pretreatment (T1), II at posttreatment (T2), and intercanine width at T1 and T2. On cephalograms taken T2, the CVM status was determined. Logistic regression analyses were used to determine the association between relapse and CVM status after treatment. The models were adjusted for potentially confounding variables (II at pretreatment and posttreatment, intercanine width change during treatment, sex, age at T2, and treatment alternatives). RESULTS: No association between CVM stage at T2 and relapse was found (P = 0.89). Both groups had similar distributions of the CVM stages (P >0.05). Pretreatment II and postretention time were found to be correlated with long-term incisor stability (P = 0.007 and 0.034, respectively). Sex was not related to relapse (P = 0.33). CONCLUSIONS: Maturity of craniofacial structures at the end of treatment evaluated with the CVM method is not associated with long-term stability of incisor alignment. PMID- 18675206 TI - Is postadolescent mandibular anterior growth rotation a risk factor for relapse of incisor alignment in males? AB - INTRODUCTION: The purpose of this study was to test the hypotheses that (1) pronounced postadolescent anterior mandibular rotation is associated with relapse of mandibular incisor alignment and (2) mandibular anterior rotation is conducive to more severe relapse. METHODS: Based on the amount of mandibular anterior rotation posttreatment, 2 groups of orthodontically treated males were selected: 1 with pronounced anterior rotation (Max AR group; n = 27), defined as maximal decrease of sella-nasion/gonion-menton angle (SN/GoMe), and 1 with minimal anterior rotation (Min AR group; n = 29). Cephalograms and study models made before treatment, after treatment (T2), and at a minimum of 10 years postretention (T3) were evaluated, and SN/GoMe, the irregularity index (II), and intercanine width were measured. RESULTS: The incisor II increased in both groups from T2 to T3, but independent t tests failed to detect an intergroup difference (P = 0.969). Moderate crowding (II >or=3.5 and 120 ms. CONCLUSION: HRT is a potent risk predictor for both heart failure and arrhythmic death in patients with class II and III CHF. PMID- 18675218 TI - A turbulent era for congestive heart failure patients? PMID- 18675219 TI - Mechanical dyssynchrony in advanced decompensated heart failure: relation to hemodynamic responses to intensive medical therapy. AB - BACKGROUND: In patients with heart failure, the degree of intraventricular mechanical dyssynchrony (intra-VMD) at baseline may predict reversal of cardiac remodeling with cardiac resynchronization therapy (CRT). OBJECTIVE: The purpose of this study was to determine the prevalence and clinical significance of intra VMD in patients admitted for advanced decompensated heart failure (ADHF). METHODS: We prospectively enrolled 50 patients with ADHF without previous CRT implantation who had been admitted to a specialized heart failure intensive care unit because of hemodynamic derangements. All patients underwent comprehensive echocardiographic evaluation within 12 hours of admission and after 48 hours from baseline evaluation after intensive medical therapy. Intra-VMD was assessed by the opposing wall time-to-peak myocardial velocity intervals in a four-segment model using color-tissue Doppler imaging. RESULTS: In our study cohort (mean age 57 +/- 11 years, left ventricular ejection fraction 26 +/- 10%, QRS width 127 +/- 31 ms, cardiac index 2.0 +/- 0.6 L/min/m(2)), significant intra-VMD (>or=65 ms) was present in 44% of subjects at baseline, and 56% of patients presented with QRS >120 ms. There was no correlation between QRS width and extent of intra-VMD. Intensive medical therapy was associated with a significant reduction in intra VMD (85 +/- 23 ms vs. 39 +/- 19 ms; P <.001). Significant hemodynamic improvement in cardiac index, filling pressures, and systemic and pulmonic vascular resistance was seen only in patients with intra-VMD. CONCLUSION: A substantial subset of patients admitted with ADHF and hemodynamic derangements demonstrate evidence of intra-VMD, which is reduced at follow-up and independent of underlying QRS width. Nevertheless, the presence of significant intra-VMD is associated with a more reversible hemodynamic profile. PMID- 18675220 TI - Persistent left ventricular dilatation in tachycardia-induced cardiomyopathy patients after appropriate treatment and normalization of ejection fraction. AB - BACKGROUND: Tachycardia-induced cardiomyopathy (TIC) seems to be a form of reversible cardiomyopathy. With recurrence, TIC can be more severe and may increase the risk for sudden cardiac death. OBJECTIVE: We postulate that negative remodeling persists even though ejection fraction (EF) normalizes after appropriate treatment in these patients. METHODS: We analyzed 2-dimensional echocardiographic parameters of 24 patients with TIC (male: 21; age: 64.1 +/- 15.2 years; atrial arrhythmias: 92%) that improved significantly with treatment (mean time between pretreatment and posttreatment echocardiography: 14 +/- 6 months) and compared them with that of age-, gender-, and ejection fraction matched control subjects without a history of TIC. RESULTS: The majority of posttreatment echocardiographic parameters showed a significant improvement (P <.05) with treatment in patients with TIC, including left ventricular (LV) ejection fraction (31.2% +/- 8.2% to 55.0% +/- 5.7%) and LV end systolic volume index (55 +/- 21 ml/m(2) to 33 +/- 13 ml/m(2)). There was no significant difference in LV end diastolic volume index (78 +/- 22 ml/m(2) to 72 +/- 22 ml/m(2), P = .15). However, when compared with age-, gender-, and ejection fraction-matched control subjects, posttreatment echocardiographic parameters in TIC patients showed significant differences (TIC vs control group) in LV end systolic volume index (33 +/- 13 ml/m(2) vs 22 +/- 5 ml/m(2)), LV end diastolic volume index (72 +/- 22 ml/m(2) vs 51 +/- 12 ml/m(2)), and cardiac index (2.6 +/- 0.8 l/min/m(2) vs 1.8 +/- 0.6 l/min/m(2)). CONCLUSION: Although the majority of echocardiographic parameters, including EF, improved significantly with treatment in TIC patients, LV dimensions and volumes remained significantly elevated when compared with control subjects, indicating persistence of negative LV remodeling, even after appropriate treatment and normalization of EF at a mean follow-up of 14 months. PMID- 18675222 TI - Three-dimensional reconstruction of the anatomic course of the right phrenic nerve in humans by pace mapping. AB - BACKGROUND: Endocardial catheter ablation at the right atrial to superior vena cava junction or right pulmonary vein isolation, especially in balloon-guided procedures, is associated with risk for phrenic nerve palsy. OBJECTIVE: The purpose of this study was to obtain a three-dimensional reconstruction of the individual phrenic nerve course. METHODS: Electroanatomic reconstruction (CARTO, Biosense Webster) of the superior vena cava, right atrium, left atrium, and right superior and inferior PVs was performed. The phrenic nerve pace map was obtained using maximal stimulation output (10 V, 2.9 ms) and annotated on the three dimensional map. RESULTS: Eighteen patients with paroxysmal atrial fibrillation (12 men and 6 women, mean age 61 +/- 10 years) were included in the study. The phrenic nerve course was reconstructed over a mean craniocaudal length of 40 +/- 17 mm (mediolateral 11 +/- 4 mm). Median phrenic nerve distance to the right superior PV ostium was 12 mm (range 2-39 mm). The phrenic nerve could be captured in 16 of 18 patients but only in 7 of 18 patients from the right superior PV. The complete electroanatomic map was performed in 48 +/- 23 minutes, including the phrenic nerve pace map (14 +/- 6 min). CONCLUSION: Pace mapping of the phrenic nerve using electroanatomic mapping can be performed within a reasonable time frame and provides important information on the true anatomic course of the phrenic nerve. The three-dimensional display of its position in relation to potential ablation targets may be of value in preventing phrenic nerve palsy, particularly when innovative ablation energies are used. However, phrenic nerve pacing exclusively from the right superior PV appears insufficient to exclude a close anatomic relationship. PMID- 18675221 TI - Feasibility of endoscopic guidance for nonsurgical transthoracic atrial and ventricular epicardial ablation. AB - BACKGROUND: Epicardial catheter ablation via subxiphoid percutaneous access currently is used upon failure of endocardial catheter ablation. The safety, efficacy, and applicability of epicardial catheter ablation likely will improve with direct visualization of the pericardial space. OBJECTIVE: The purpose of this study was to assess the feasibility of percutaneous endoscopic guidance for identification of epicardial anatomic landmarks and epicardial catheter ablation. METHODS: Dual subxiphoid epicardial access and femoral venous and arterial access were obtained in six healthy swine. The endoscope and electrophysiology catheter were advanced to the pericardial space. Anatomic landmarks were identified via endoscopy and confirmed by multiview fluoroscopic assessment of proximity to endocardial catheters in the area of interest. Radiofrequency ablation of selected anatomic targets was performed under endoscopic guidance. Targeting of lesions was assessed by pathologic examination of the target and surrounding structures. RESULTS: Dual large-bore subxiphoid epicardial access was obtained without complications in all animals. The coronary sinus, left anterior descending coronary artery, left atrial appendage, and pulmonary veins were easily visualized in all animals. Catheter ablation of anatomic targets, including the right ventricular outflow tract, left atrial appendage, and pulmonary veins, was performed successfully under direct endoscopic observation. Endoscopic guidance of point and linear lesions near coronary vessels also was assessed. Pathology revealed successful targeting of lesions. CONCLUSION: Endoscopic guidance of percutaneous epicardial electrophysiologic procedures is feasible. Direct visualization of epicardial structures, catheters, and lesions may improve the safety and efficacy of epicardial catheter ablation and reduce staff and patient radiation exposure. PMID- 18675223 TI - Spatial and temporal stability of complex fractionated electrograms in patients with persistent atrial fibrillation over longer time periods: relationship to local electrogram cycle length. AB - BACKGROUND: Atrial fibrillation (AF) ablation guided by complex fractionated electrograms (CFE) has been described, but the spatial and temporal stability of the electrograms (EGMs) has been questioned. OBJECTIVE: The purpose of this study was to prospectively assess the spatial and temporal stability of CFE in patients with persistent AF. METHODS: Twenty-four patients were studied. For 12 patients, two high-density CFE maps were performed during AF at baseline (0 minute) and 20 minutes later using the EnSite NavX system. Six-second bipolar EGMs were collected throughout the left atrium (LA) using a circular mapping catheter. Automated software measured the time between discrete local EGM deflections yielding a mean local cycle length (CL). EGMs with mean CL <120 ms were considered CFE. The LA was divided into six regions. Spatial distribution of EGMs at 0 and 20 minutes was compared in each region across three different CL ranges (A = 50-120 ms, B = 121-200 ms, C = 200-500 ms). The 0- and 20-minute CFE maps were directly superimposed offline in MATLAB, and the mean CL value for each point that was sampled in both maps was compared in each CL range (A-C). For the other 12 patients, repetitive measurements (1-minute intervals for 5 minutes) of mean CL were obtained at a sample point for each CL range (A-C) in each patient and compared for consistency. RESULTS: A total of 324 +/- 65 points were collected per map. Globally in the LA, the distribution of CLs did not change from 0 to 20 minutes (A: 47.1% vs 45.0%; B: 35.3% vs 36.5%; C: 16.0% vs 16.9%; P = .6). The CL distribution in each of the six regions of the LA also did not change significantly from 0 to 20 minutes. There was no significant change in repetitive CL measures for range A (mean DeltaCL 12 +/- 4 ms, P = .4). However, there was significant variation over 5 minutes for ranges B and C (mean DeltaCL 39 +/- 19 ms and 48 +/- 22 ms, respectively, P <.05 for both). Superimposing maps showed 74.7% point-to-point match for range A, 39.3% for range B, and 14.2% for range C (within 30 ms), with a significant correlation seen only for range A (r = 0.82, P <.001). CONCLUSION: CFE show a high degree of spatial and temporal stability. Greater temporal variation is seen in progressively longer CL regions that are outside of the CFE region of interest. PMID- 18675224 TI - Standardization and validation of an automated algorithm to identify fractionation as a guide for atrial fibrillation ablation. AB - BACKGROUND: Atrial fibrillation catheter ablation is frequently guided by identification of fractionated electrograms, which are thought to be critical for maintenance of the arrhythmia. Objective automated means for identifying fractionation independent of physician interpretation have not been standardized or validated. OBJECTIVE: The purpose of this study was to standardize and validate an automated algorithm to rapidly identify fractionated electrograms for high-density atrial fibrillation fractionation mapping. METHODS: Left and right atrial fractionation maps were generated by EnSite NavX 6.0 software, using standardized ablation catheters in eight patients with atrial fibrillation. Two blinded electrophysiologists interpreted all electrograms as either fractionated or not fractionated. A stepwise approach was used to optimize automated settings to accurately identify fractionation. High-density fractionation maps were generated with a 20-pole mapping catheter in eight other patients. Two blinded electrophysiologists interpreted all electrograms as near field or far field. The algorithm was refined to optimize settings to exclude far-field signals and retain near-field signals. The sampling segment length was adjusted to optimize recording time to ensure reproducibility. RESULTS: Using 1,514 points, the automated software achieved sensitivity of 0.75 and specificity of 0.80 for identification of fractionated electrograms. Using 725 points collected via multipole catheters with optimal automated settings, 94% of near-field fractionated electrograms were accurately identified. A 6-second sampling length was needed for reproducible fractionation measurements. CONCLUSION: Standardized settings of EnSite NavX 6.0 software with 6-second data collection per point can rapidly and accurately generate high-density fractionation maps independent of physician electrogram interpretation. This may allow for an automated, standardized approach to atrial fibrillation fractionated ablation. PMID- 18675225 TI - Standardization of criteria used in automated algorithms for detection of fractionated electrograms to guide substrate-based ablation of atrial fibrillation: no rest for the wicked. PMID- 18675226 TI - Right atrial substrate properties associated with age in patients with typical atrial flutter. AB - BACKGROUND: Data detailing the age-related difference in the atrial substrate for formation of typical atrial flutter (AFL) are sparse. OBJECTIVE: The purpose of this study was to characterize the difference in the right atrial substrate related to aging using noncontact mapping of the right atrium. METHODS: A total of 54 patients (23 young [<60 years; 45 +/- 12 years] and 31 old [>or=60 years; 74 +/- 6 years]) with typical AFL who underwent three-dimensional noncontact mapping of typical AFL were enrolled in the study. The atrial substrate was characterized according to (1) regional wavefront activation mapping, (2) regional conduction velocity, and (3) regional voltage distribution by dynamic substrate mapping. RESULTS: During activation mapping of the crista terminalis, two activation patterns were observed: (1) around the upper end of the crista terminalis (67%) and (2) through a gap in the crista terminalis. The presence of a crista terminalis gap was associated with a high incidence of induced atypical AFL/atrial fibrillation (P <.001). The conduction velocities of the medial cavotricuspid isthmus were slower in the old group than in the young group. In regional activation mapping of the AFL, the location of the slowest conduction shifted from the lateral cavotricuspid isthmus (71%) in the young group to the medial cavotricuspid isthmus (40%) in the old group. More cases with a low voltage zone (10% or more prevalence). A search for a better diagnostic test that can be applied in field situations in China is essential and should be given high priority. PMID- 18675245 TI - Electrochemical immunosensor of platelet-derived growth factor with aptamer primed polymerase amplification. AB - A new method for the determination of platelet-derived growth factor BB (PDGF-BB) was developed using an electrochemical immunosensor with an aptamer-primed, long strand circular detection probe. Rabbit anti-human PDGF-B polyclonal antibody was immobilized on the electrode to serve as the capture antibody. The detection probe was synthesized via polymerase extension along a single-stranded circular plasmid DNA template with a primer headed by the anti-PDGF-B aptamer. In the presence of the analyte, the aptamer-primed circular probe was captured on the electrode via the formation of an antibody/PDGF-BB/aptamer sandwiched complex. The electroactivity indicator methylene blue was adsorbed on the electrode surface via the analyte-sandwiched complex with long-strand circular DNA, thus yielding a strong electrochemical signal for the quantification of PDGF-BB. This strategy allowed electrochemical detection with enormous signal amplification arising from the long-strand localized circular probe. The oxidation peak current of methylene blue in square wave voltammetric measurements showed a linear dependence on the concentration of PDGF-BB in the range from 50 to 500 ng mL(-1), with a detection limit as low as 18 pg mL(-1). PMID- 18675246 TI - PASK (proline-alanine-rich Ste20-related kinase) binds to tubulin and microtubules and is involved in microtubule stabilization. AB - Proline-alanine-rich Ste20-related kinase (PASK, also referred to as SPAK) has been linked to ion transport regulation. Here, we report two novel activities of PASK: binding to tubulin and microtubules and the promotion of microtubule assembly. Tubulin binding assay showed that full-length PASK and its kinase domain bound to purified tubulin whereas the N-terminal or C-terminal non catalytic domains of PASK did not. The full-length PASK and its kinase domain were sedimented with paclitaxel-stabilized microtubules by ultracentrifugation. These results indicate that the kinase domain of PASK can interact directly with both microtubules and soluble tubulin in vitro. Truncated PASK lacking the N terminal non-catalytic domain promoted microtubule assembly at a subcritical concentration of purified tubulin. FLAG-PASK expressed in COS-7 cells translocated to the cytoskeleton when the cells were stimulated with hypertonic sodium chloride, and stabilized microtubules against depolymerization by nocodazole. Our findings suggest that PASK may regulate the cytoskeleton by modulating microtubule stability. PMID- 18675247 TI - Membrane interactions and lipid binding of casein oligomers and early aggregates. AB - Caseins constitute the main protein components in mammalian milk and have critical functions in calcium transport and prevention of protein aggregation. Fibrillation and aggregation of kappa-casein, a phenomenon which has only recently been detected, might be associated with malfunctions of milk secretion and amyloidosis phenomena in the mammary glands. This study employs a newly designed chromatic biomimetic vesicle assay to investigate the occurrence and the parameters affecting membrane interactions of casein aggregates and the contribution of individual casein members to membrane binding. We show that physiological casein colloids exhibit membrane activity, as well as early globular aggregates of kappa-casein, a prominent casein isoform. Furthermore, inhibition of kappa-casein fibrillation through complexation with alphaS-casein and beta-casein, respectively, was found to go hand in hand with induction of enhanced membrane binding; these data are important in the context of casein biology since in secreted milk kappa-casein is found only in assemblies containing also alphaS-casein and beta-casein. The chromatic experiments, complemented by transmission electron microscopy analysis and fluorescence quenching assays, also revealed significantly higher affinity early spherical aggregates of k-casein to anionic phosphatidylglycerol-lipids, as compared to zwitterionic phospholipids. Overall, this study suggests that lipid interactions play important roles in maintaining the essential physiological functions of caseins in mammalian milk. PMID- 18675248 TI - Ubiquitin ligase Kf-1 is involved in the endoplasmic reticulum-associated degradation pathway. AB - Kf-1 was first identified as a gene showing enhanced expression in the cerebral cortex of a sporadic Alzheimer's disease patient. To date, however, the functional properties of Kf-1 protein remain unknown. In this study, immunohistochemical analysis showed that Kf-1 immunoreactivity was detected in rat hippocampus and cerebral cortex neurons. Interestingly, it was colocalized with endoplasmic reticulum (ER) marker. To investigate the specific function of Kf-1 protein, we generated Myc tagged wild type Kf-1 (Myc-Kf-1WT) and RING finger domain deletion mutant of Kf-1 (Myc-Kf-1DeltaR), and then transfected in HEK293 cells. Myc-Kf-1WT displayed a reticular pattern typical of ER localization, with large perinuclear aggregates and colocalized with ER marker, calnexin. Myc-Kf-1WT facilitated ubiquitination of endogenous proteins, whereas Myc-Kf-1DeltaR did not show ubiquitin ligase activity. In addition, we found that Kf-1 interacted with components of the ER-associated degradation (ERAD) pathway, including Derlin-1 and VCP. Taken together, these properties suggest that Kf-1 is an ER ubiquitin ligase involved in the ERAD pathway. PMID- 18675249 TI - Nuclear deformation and expression change of cartilaginous genes during in vitro expansion of chondrocytes. AB - Cartilaginous gene expression decreased when chondrocytes were expanded on cell culture plates. Understanding the dedifferentiation mechanism may provide valuable insight into cartilage tissue engineering. Here, we demonstrated the relationship between the nuclear shape and gene expression during in vitro expansion culture of chondrocytes. Specifically, the projected nuclear area increased and cartilaginous gene expressions decreased during in vitro expansion culture. When the nuclear deformation was recovered by cytochalasin D treatment, aggrecan expression was up-regulated and type I collagen (Col1a2) expression was down-regulated. These results suggest that nuclear deformation may be one of the mechanisms for chondrocyte dedifferentiation during in vitro expansion culture. PMID- 18675250 TI - Identification and characterisation of a nuclear localisation signal in the SMN associated protein, Gemin4. AB - Gemin4 is a ubiquitously expressed multifunctional protein that is involved in U snRNP assembly, apoptosis, nuclear/cytoplasmic transportation, transcription, and RNAi pathways. Gemin4 is one of the core components of the Gemin-complex, which also contains survival motor neuron (SMN), the seven Gemin proteins (Gemin2-8), and Unrip. Mutations in the SMN1 gene cause the autosomal recessive disorder spinal muscular atrophy (SMA). Although the functions assigned to Gemin4 predominantly occur in the nucleus, the mechanisms that mediate the nuclear import of Gemin4 remain unclear. Here, using a novel panel of Gemin4 constructs we identify a canonical nuclear import sequence (NLS) in the N-terminus of Gemin4. The Gemin4 NLS is necessary and independently sufficient to mediate nuclear import of Gemin4. This is the first functional NLS identified within the SMN-Gemin complex. PMID- 18675251 TI - Plectasin has antibacterial activity and no affect on cell viability or IL-8 production. AB - Animals and plants express endogenous peptide antibiotics called defensins. Defensins show broad-spectrum antimicrobial activity, even against bacteria that have resistance to conventional antibiotics, which has made them viable candidates for new antibiotics. However, human defensins have failed to reach the market because of their cytotoxic effects and non-antimicrobial bioactivities. Plectasin is a defensin that has shown promise but has not had its potentially negative effects clarified. To address this issue, we examined plectasin's cytotoxicity in human cells using an AlamarBlue reduction assay, its interleukin (IL)-8-inducing capacity using real-time PCR and ELISA, and measured its MIC against bacteria. We confirmed that plectasin has specific antibacterial activity against Streptococcus pneumoniae. Plectasin showed no cytotoxicity to A549 cells, normal human bronchial epithelial cells, or lung fibroblasts, and it did not induce IL-8 transcription or production in A549 cells. Our results suggest that plectasin could be an inoffensive alternative antibiotic for clinical application. PMID- 18675252 TI - PCSK9 binds to multiple receptors and can be functionally inhibited by an EGF-A peptide. AB - Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to low density lipoprotein receptor (LDLR) and induces its internalization and degradation. PCSK9 binding to LDLR is mediated through the LDLR epidermal growth factor-like repeat A (EGF-A) domain. We show for the first time that an EGF-A peptide inhibits PCSK9-mediated degradation of LDLR in HepG2 cells. In addition to LDLR, we show that PCSK9 also binds directly to ApoER2 and mouse VLDLR. Importantly, binding of PCSK9 to either LDLR or mouse VLDLR was effectively inhibited by EGF-A while binding to ApoER2 was less affected. In contrast, LDL receptor-associated protein (RAP), which interacts with LDL receptor repeat type A (LA) domains, inhibited PCSK9 binding to ApoER2 with greater efficacy than either LDLR or mVLDLR. These data demonstrate that while PCSK9 binds several receptors via its EGF-A binding domain, additional contacts with other receptor domains are also involved. PMID- 18675253 TI - Clathrin and Cx43 gap junction plaque endoexocytosis. AB - In earlier transmission electron microscopic studies, we have described pentilaminar gap junctional membrane invaginations and annular gap junction vesicles coated with short, electron-dense bristles. The similarity between these electron-dense bristles and the material surrounding clathrin-coated pits led us to suggest that the dense bristles associated with gap junction structures might be clathrin. To confirm that clathrin is indeed associated with annular gap junction vesicles and gap junction plaques, quantum dot immuno-electron microscopic techniques were used. We report here that clathrin associates with both connexin 43 (Cx43) gap junction plaques and pentilaminar gap junction vesicles. An important finding was the preferential localization of clathrin to the cytoplasmic surface of the annular or of the gap junction plaque membrane of one of the two contacting cells. This is consistent with the possibility that the direction of gap junction plaque internalization into one of two contacting cells is regulated by clathrin. PMID- 18675254 TI - Sumoylation of amyloid precursor protein negatively regulates Abeta aggregate levels. AB - The proteolytic processing of amyloid precursor protein (APP) to produce Abeta peptides is thought to play an important role in the mechanism of Alzheimer's disease. Here, we show that lysines 587 and 595 of APP, which are immediately adjacent to the site of beta-secretase cleavage, are covalently modified by SUMO proteins in vivo. Sumoylation of these lysine residues is associated with decreased levels of Abeta aggregates. Further, overexpression of the SUMO E2 enzyme ubc9 along with SUMO-1 results in decreased levels of Abeta aggregates in cells transfected with the familial Alzheimer's disease-associated V642F mutant APP, indicating the potential of up-regulating activity of the cellular sumoylation machinery as an approach against Alzheimer's disease. The results also provide the first demonstration that the SUMO E2 enzyme (ubc9) is present within the endoplasmic reticulum, indicating how APP, and perhaps other proteins that enter this compartment, can be sumoylated. PMID- 18675255 TI - Inhibitory interaction between P2X4 and GABA(C) rho1 receptors. AB - Reciprocal functional inhibition between P2X and GABA(A/C) receptors represents a novel mechanism fine-tuning neuronal excitability. However, the participating receptors and underlying mechanisms are not fully understood. P2X(4) receptor is widely found in neurons that express GABA(C) rho1 receptor. Thus, we co-expressed P2X(4) and rho1 receptors in HEK293 cells and, using patch-clamp recording, examined whether they have mutual functional inhibition. Currents evoked by simultaneous application of ATP and GABA (I(ATP+GABA)) were significantly smaller compared to the addition of I(ATP) and I(GABA). Furthermore, I(ATP) were strongly suppressed during rho1 receptor activation. Similarly, I(GABA) were greatly attenuated during P2X(4) receptor activation. Such mutual inhibition was absent in cells only expressing P2X(4) or rho1 receptor. Taken together, these functional data support negative cross-talk between P2X(4) and rho1 receptors. PMID- 18675256 TI - Novel roles for ceramides, calpains and caspases in kidney proximal tubule cell apoptosis: lessons from in vitro cadmium toxicity studies. AB - Apoptosis is a tightly regulated physiological process, which can be initiated by toxic stimuli, such as cadmium (Cd2+). Cd2+ (10-50 microM) induces a rapid increase in reactive oxygen species (ROS) (> or = 30 min) in a cell line derived from the S1 segment of rat kidney proximal tubule, without any apparent mitochondrial dysfunction. The sphingolipid ceramide is an important second messenger in apoptosis. Short exposure to Cd2+ (3h) causes an increase in ceramides, which occurs downstream of ROS formation, and may interact with cellular components, such as endoplasmic reticulum and mitochondria. Following apoptosis initiation, execution must take place. The classical executioners of apoptosis are caspases, a family of cysteine proteases. However, increasing studies report caspase-independent apoptosis, which questions the essentiality of caspases for apoptosis implementation. With low micromolar Cd2+ concentrations (< 10 microM), caspases are only activated after 24h and not at earlier time points, which supports the notion of caspase-independent apoptosis. Due to increased cytosolic Ca(2+) under pathological conditions, a role for the Ca2+-dependent proteases, calpains, has emerged. Calpain activation by Cd2+ (3-6h) seems to be regulated by ceramide levels, in order to induce apoptosis. Calpain and caspase substrates overlap but yield different fragments, which may explain their diverse downstream targets. Furthermore, calpains and caspases may interact with one another to enhance, as seen by Cd2+, or diminish apoptosis. In this review, we discuss novel roles for ceramides, calpains and caspases as part of Cd2+-induced apoptotic signalling pathways in the kidney proximal tubule and their in vivo relevance to Cd2+-induced nephrotoxicity. PMID- 18675257 TI - Summing it up: semantic activation processes in the two hemispheres as revealed by event-related potentials. AB - The coarse coding hypothesis suggests that semantic activation is broader in the right hemisphere, affording it an advantage over the left hemisphere for the activation of distantly related concepts or multiple meanings of lexically ambiguous words. Behavioral studies investigating coarse coding have yielded mixed results, perhaps in part because such measures sum across multiple processing stages. To more directly tap into the semantic activation processes that are the focus of the coarse coding hypothesis, the current study combined a visual half-field summation-priming paradigm with the measurement of event related potentials (ERPs). Two primes converged onto a lateralized, unambiguous target (e.g., lion-stripes-tiger) or diverged onto different meanings of a lateralized, ambiguous target (e.g., kidney-piano-organ); in both cases, the primes were related to one another only through the target. In two experiments, participants either made lexical decisions to the targets or made a semantic relatedness judgment between primes and target. Priming was measured as reductions in the amplitude of the N400, an ERP component that has been specifically linked to meaning activation and that showed semantic-level priming patterns in both of the tasks used in the present study. Counter to the predictions of the coarse coding hypothesis, equivalent N400 summation priming was observed for targets in the two visual fields, in both types of triplets and in both experiments. Thus, the current results fail to support the hypothesis that semantic activation patterns differ in the two hemispheres and point, instead, to other sources for observed asymmetries in verbal processing. PMID- 18675258 TI - Mesenchymal stem cells transplantation could be beneficial for treatment of experimental ischemic stroke in rats. AB - Cell therapy is prospective, modern attempt to ischemic stroke treatment. It has been being widely worked out recently. We suggest mesenchymal stem cells (MSC) as a cell therapy agent in the therapy of this disease. Experiments were carried out in inbred male Wistar-Kyoto rats. Animals were subjected to middle cerebral artery occlusion (MCAO). MSCs were isolated from rat bone marrow, expanded in culture and labelled with vital fluorescent dye PKH-26. Then 5 x 10(6) cells were injected into the tail vein on the day of MCAO and three days later. Control group animals received PBS injection (negative control). Cognitive function restoration was estimated by Morris Water Maze testing during 6 weeks after MCAO. Animals were sacrificed 1, 2, 3, 5 days and 1, 2, 4 and 6 weeks after operation. Intravenous MSC transplantation decreased post-operation mortality and benefited behavioural and neurological recovery. Experimental groups animals revealed changes in aseptic inflammation processes which were completed faster comparing to control group. That effect correlated with accelerated glial scar formation. Reduction of the infarct volumes and such post-stroke after-effects as border zone gliosis and liquor cysts formation accompanied by increased angiogenesis and subventricular zone cells proliferation were shown after cell therapy. The obtained results referred to both cell therapy groups. Thus, MSC injection benefited post-stroke rehabilitation irrespective of transplantation time. However, further investigation should be carried out in order to find out the mechanism of their action. PMID- 18675259 TI - The quantitative assessment of functional impairment and its correlation to infarct volume in rats with transient middle cerebral artery occlusion. AB - The purpose of this study was to quantitatively assess motor activity and sensory functions and to determine their relationships to infarct severity by measuring infarct volume in rats with transient ischemic stroke. Male Sprague-Dawley rats (11 weeks old, n = 42) were randomly divided into 4 separate groups; a sham operation group, and 1-h, 2-h and 3-h middle cerebral artery occlusion (MCAO) groups. Percent weights borne on paretic hind limbs were measured consecutively for 7 days starting from the day before the induction of ischemia. Fifty percent withdrawal threshold values of forepaws and hindpaws were measured using von-Frey hairs. Infarct volumes in the three ischemic groups, which were significantly different (p < 0.01) from each other, were found to increase in size with ischemic time. The percent weight borne on paretic hind limb in the three MCAO groups were significantly lower than that in the sham group, and this functional deficit remained significant throughout the observational period (p < 0.01). A significant correlation was found between infarct volumes and percent weight borne on paretic hind limbs after ischemia (-0.7 < rho < -0.4, p < 0.05). In contrast the 50% withdrawal threshold values of paretic forepaws and hindpaws in the three ischemic groups were not significantly different from those of the sham controls. Motor test findings, which were used to assess reductions in paretic hind limb weightbearing, were shown to be correlated with infarct volume. The present study shows that this test could be used as a tool to objectively and quantitatively assess functional outcome in MCAO rats. PMID- 18675260 TI - In utero PCP exposure alters oligodendrocyte differentiation and myelination in developing rat frontal cortex. AB - Several neurodevelopmental disorders, including schizophrenia, autism, ADD/ADHD and dyslexia are believed to originate during gestation and involve white matter abnormalities. Modulation of glutamate environments and glutamate receptors has also been implicated in alteration of oligodendrocytes, the myelin forming cells of the CNS. To begin to understand how modulation of the glutamate system affects the maturation of oligodendrocytes, developing rats were subjected to prenatal blockade of the NMDA receptor with phencyclidine (PCP). Oligodendrocyte development and differentiation were then examined postnatally by measuring markers for early, middle and late stage cells. The results indicate that, while the level of marker proteins for neurons and astrocytes remains the same, early oligodendrocyte progenitor cell markers are decreased in rat brains prenatally exposed to PCP. Labeling of cells of intermediate, immature cell stages is elevated. Late stage markers for myelinating oligodendrocytes are subsequently decreased. These data suggest that prenatal NMDA receptor blockade reduces the level of progenitors and that the surviving cells are arrested at an immature stage. This premature arrest appears to result in fewer fully differentiated, mature oligodendrocytes that are capable of producing myelin. These results have interesting implications for the role of glutamate and glutamate receptors in white matter abnormalities in neurodevelopmental disorders. PMID- 18675262 TI - Long-term assessment of enriched housing and subventricular zone derived cell transplantation after focal ischemia in rats. AB - The potential for using stem cells to treat stroke has garnered much interest, but stem cell therapies must be rigorously tested in animal models before transplantation studies progress to clinical trials. An enriched environment enhances transplanted subventricular zone (SVZ) cell migration and functional benefit following stroke in rats. However, the ability of SVZ cells to survive, migrate, differentiate and promote functional recovery at protracted survival times (e.g., 3 months) has not been investigated. The vasoconstrictive peptide endothelin-1 was injected adjacent to the middle cerebral artery to produce focal ischemia. Seven days later, cells derived from the SVZ of adult mice (800,000 cells/rat or vehicle injection) were transplanted into the sensory-motor cortex and striatum, and rats were then housed in enriched or standard conditions. Rats in enriched housing had access to running wheels once per week. Recovery was assessed in the forelimb-use asymmetry task (cylinder) at 1, 2, or 3 months after transplantation immediately prior to euthanasia. Transplanted cell survival and migration were quantified using stereology. Cell phenotype was determined with immunohistochemistry and confocal microscopy. Enriched housing did not enhance survival or migration of transplanted SVZ cells at protracted survival times, and the majority (~99%) of cells died within 2 months of transplantation. Cell survival was significantly, and negatively, correlated with microglial activation. Many surviving cells expressed an astrocytic phenotype. Functional recovery was not improved at any time. Therapies involving transplantation of SVZ cells following stroke must be further optimized in order to enhance long-term cell survival and thereby maximize functional benefit. PMID- 18675261 TI - Regulation of interleukin-1beta by the interleukin-1 receptor antagonist in the glutamate-injured spinal cord: endogenous neuroprotection. AB - Elevation of extracellular glutamate contributes to cell death and functional impairments generated by spinal cord injury (SCI), in part through the activation of the neurotoxic cytokine interleukin-1beta (IL-1beta). This study examines the participation of IL-1beta and its regulation by the endogenous interleukin-1 receptor antagonist (IL-1ra) in glutamate toxicity following SCI. Glutamate, glutamatergic agonists and SCI had similar effects on levels of IL-1beta and IL 1ra. Following spinal cord contusion or exposure to elevated glutamate, concentrations of IL-1beta first increased as IL-1ra decreased, and both then changed in the opposite directions. Applying the glutamate agonists NMDA and S AMPA to the spinal cord caused changes in IL-1beta and IL-1ra levels very similar to those produced by contusion and glutamate. The glutamate antagonists MK801 and NBQX blocked the glutamate-induced changes in IL-1beta and IL-1ra levels. Administering IL-1beta elevated IL-1ra, and administering IL-1ra depressed IL 1beta levels. Infusing IL-beta into the spinal cord impaired locomotion, and infusing IL-1ra improved recovery from glutamate-induced motor impairments. We hypothesize that elevating IL-1ra opposes the damage caused by IL-1beta in SCI by reducing IL-1beta levels as well as by blocking binding of IL-1beta to its receptor. Our results demonstrate that IL-1beta contributes to glutamate damage following SCI; blocking IL-1beta may usefully counteract glutamate toxicity. PMID- 18675263 TI - NMR-based molecular ruler for determining the depth of intercalants within the lipid bilayer Part I. Discovering the guidelines. AB - The development of "molecular rulers" would allow one to quantitatively locate intercalants within the liposomal bilayer. To this end, we have attempted to correlate the 13C NMR chemical shift of a polarizable "reporter" carbon (e.g., carbonyl) of the intercalant-with the ET30 polarity it experiences, and with its Angstrom distance from the interface. This requires families of molecules with the same two "reporter carbons" separated by a fixed distance, residing at various depths/polarities within the bilayer. The families studied included 4,4 dialkylcyclohexa-2,5-dienones 1, benzenediacetic esters 15, benzenedipropionic esters 17, 4-alkoxybenzaldehydes 19 and methyl 4-alkoxybenzoates 22. These compounds possessed the following characteristics: (1) a planar backbone; (2) polar/hydrophilic "head" groups; (3) modular hydrophobic tails; (4) large changes in the 13C NMR chemical shift (Deltadelta) of the reporter atoms with solvent polarity. These studies revealed a fifth requirement, namely: (5) the reporter carbons must not be strongly conjugated, lest it reflect the charge build-up at another site within the conjugated system. PMID- 18675264 TI - A p38 MAPK-CREB pathway functions to pattern mesoderm in Xenopus. AB - Dorsal-ventral patterning is specified by signaling centers secreting antagonizing morphogens that form a signaling gradient. Yet, how morphogen gradient is translated intracellularly into fate decisions remains largely unknown. Here, we report that p38 MAPK and CREB function along the dorsal-ventral axis in mesoderm patterning. We find that the phosphorylated form of CREB (S133) is distributed in a gradient along the dorsal-ventral mesoderm axis and that the p38 MAPK pathway mediates the phosphorylation of CREB. Knockdown of CREB prevents chordin expression and mesoderm dorsalization by the Spemann organizer, whereas ectopic expression of activated CREB-VP16 chimera induces chordin expression and dorsalizes mesoderm. Expression of high levels of p38 activator, MKK6E or CREB VP16 in embryos converts ventral mesoderm into a dorsal organizing center. p38 MAPK and CREB function downstream of maternal Wnt/beta-catenin and the organizer specific genes siamois and goosecoid. At low expression levels, MKK6E induces expression of lateral genes without inducing the expression of dorsal genes. Loss of CREB or p38 MAPK activity enables the expansion of the ventral homeobox gene vent1 into the dorsal marginal region, preventing the lateral expression of Xmyf5. Overall, these data indicate that dorsal-ventral mesoderm patterning is regulated by differential p38/CREB activities along the axis. PMID- 18675265 TI - Targeted disruption of the Pak5 and Pak6 genes in mice leads to deficits in learning and locomotion. AB - PAK6 is a member of the group B family of PAK serine/threonine kinases, and is highly expressed in the brain. The group B PAKs, including PAK4, PAK5, and PAK6, were first identified as effector proteins for the Rho GTPase Cdc42. They have important roles in filopodia formation, the extension of neurons, and cell survival. Pak4 knockout mice die in utero, and the embryos have several abnormalities, including a defect in the development of motor neurons. In contrast, Pak5 knockout mice do not have any noticeable abnormalities. So far nothing is known about the biological function of Pak6. To address this, we have deleted the Pak6 gene in mice. Since Pak6 and Pak5 are both expressed in the brain, we also generated Pak5/Pak6 double knockout mice. These mice were viable and fertile, but had several locomotor and behavioral deficits. Our results indicate that Pak5 and Pak6 together are not required for viability, but are required for a normal level of locomotion and activity as well as for learning and memory. This is consistent with a role for the group B PAKs in the nervous system. PMID- 18675267 TI - Different inflammatory reactions to vitamin D3 among the lateral, third and fourth ventricular choroid plexuses of the rat. AB - The four choroid plexuses in the brain ventricles are not identical, but differences among them have rarely been studied. The present work concerns the inflammatory and hemorrhagic choroid plexitis produced in Lewis rats by a single gavage of cholecalciferol (vitamin D(3)) or related steroids with vitamin D activity. Plexitis was very severe in the fourth ventricular plexus, somewhat less severe in the lateral ventricular plexuses, and almost absent in the third ventricular plexus. These findings were compared to the scanty data from the literature on differences among the plexuses. PMID- 18675266 TI - Antidepressant-like behavioral effects of IGF-I produced by enhanced serotonin transmission. AB - Previous research has suggested that mobilization of neurotrophic factors, such as insulin-like growth factor I (IGF-I), can be involved in the effects of antidepressant treatments. The current experiments showed that IGF-I leads to antidepressant-like effects in the modified rat forced swim test when tested 3 days, but not 1 day, after i.c.v. administration. These effects were sustained longer than the antidepressants paroxetine and desipramine. In addition, blockade of the IGF-I receptor with the IGF-I antagonist JB1 30 min before IGF-I administration prevented the antidepressant-like effects of IGF-I. However, when JB1 was administered 3 days after IGF-I administration and 30 min prior to testing, the antidepressant-like effects of IGF-I were still present suggesting that IGF-1 produces a long-term activation of neural systems involved in the antidepressant response. Because the pattern of antidepressant-like effects of IGF-I resembled those of selective serotonin reuptake inhibitors, the role of serotonin in the behavioral effects of IGF-I was studied. Depletion of serotonin, by the tryptophan hydroxylase inhibitor para-chlorophenylalanine, blocked the antidepressant-like effects of IGF-I. Administration of IGF-I increased basal serotonin levels in the ventral hippocampus and altered the effects of acute citalopram. IGF-I administration did not change hippocampal cell proliferation at the 3-day timepoint when behavioral effects were seen. In addition, IGF-I did not alter the expression of mRNA levels of tryptophan hydroxylase or SERT in the brain stem, or [3H] citalopram binding in the hippocampus or cortex. Thus, IGF-I administration initiates a long-lasting cascade of neurochemical effects involving increased serotonin levels that results in antidepressant-like behavioral effects. PMID- 18675268 TI - Presbyopia. Emerging from a blur towards an understanding of the molecular basis for this most common eye condition. AB - All people will be presbyopic by age 50, and we now understand something of the basis for this condition. It turns out to be a direct consequence of two features; first the design of the transparent lens and the way it must change shape to enable focussing by the human eye, and second the instability of proteins over a very long time period. The incremental changes that take place in the lens to render the central region inflexible by middle age and, as a consequence the person presbyopic, may also promote the subsequent development of cataract. Based on the most recent data, heat-induced denaturation of proteins in the lens appears to be a worthy topic for future investigation. Understanding such processes may allow us to glimpse the origin both of presbyopia and age related nuclear cataract. PMID- 18675269 TI - The presence of a woman increases testosterone in aggressive dominant men. AB - In line with the challenge hypothesis, this study investigated the effects of the presence of a woman on the testosterone (T) levels of young men. An informal contact with a woman of approximately 5 min resulted in an increase in salivary T among men. These effects occurred particularly in men with an aggressive dominant personality. In addition, higher salivary T levels were related to a more aggressively dominant personality, being sexual inactive for a month or more, and not being involved in a committed, romantic relationship. The most important findings of this study are that the short presence of a woman induces specific hormonal reactions in men, and that these effects are stronger for aggressively dominant men. PMID- 18675270 TI - Discrimination between exosomes and HIV-1: purification of both vesicles from cell-free supernatants. AB - Although enveloped retroviruses bud from the cell surface of T lymphocytes, they use the endocytic pathway and the internal membrane of multivesicular bodies for their assembly and release from macrophages and dendritic cells (DCs). Exosomes, physiological nanoparticles produced by hematopoietic cells, egress from this same pathway and are similar to retroviruses in terms of size, density, the molecules they incorporate and their ability to activate immune cells. Retroviruses are therefore likely to contaminate in vitro preparations of exosomes and vice versa and sucrose gradients are inefficient at separating them. However, we have found that their sedimentation velocities in an iodixanol (Optiprep) velocity gradient are sufficiently different to allow separation and purification of both vesicles. Using acetylcholinesterase as an exosome marker, we demonstrate that Optiprep velocity gradients are very efficient in separating exosomes from HIV-1 particles produced on 293T cells, primary CD4(+) T cells, macrophages or DCs, with exosomes collecting at 8.4-12% iodixanol and HIV-1 at 15.6%. We also show that immunodepletion with an anti-acetylcholinesterase antibody rapidly produces highly purified preparations of HIV-1 or exosomes. These findings have applications in fundamental research on exosomes and/or AIDS, as well as in clinical applications where exosomes are involved, more specifically in tumour therapy or in gene therapy using exosomes generated from DCs genetically modified by transfection with virus. PMID- 18675271 TI - Detection of low avidity CD8(+) T cell populations with coreceptor-enhanced peptide-major histocompatibility complex class I tetramers. AB - The development of soluble recombinant peptide-major histocompatibility complex class I (pMHCI) molecules conjugated in multimeric form to fluorescent labels has enabled the physical quantification and characterization of antigen-specific CD8(+) T cell populations by flow cytometry. Several factors determine the binding threshold that enables visualization of cognate CD8(+) T cells with these reagents; these include the affinity of the T cell receptor (TCR) for pMHCI antigen. Here, we show that multimers constructed from peptide-human leukocyte antigen (pHLA) A0201 monomers engineered in the heavy chain alpha2 domain to enhance CD8 binding (K(D) approximately 85 microM) without impacting the TCR binding platform can detect cognate CD8(+) T cells bearing low affinity TCRs that are not visible with the corresponding wildtype pHLA A0201 multimeric complexes. Mechanistically, this effect is mediated by a disproportionate enhancement of the TCR/pMHCI association rate. In direct ex vivo applications, these coreceptor enhanced multimers exhibit faithful cognate binding properties; concomitant increases in background staining within the non-cognate CD8(+) T cell population can be resolved phenotypically using polychromatic flow cytometry as a mixture of naive and memory cells. These findings provide the first validation of a novel approach to the physical detection of low avidity antigen-specific CD8(+) T cell populations; such coreceptor-enhanced multimeric reagents are likely to be useful in a multitude of settings for the detection of auto-immune, tumor-specific and cross-reactive CD8(+) T cells. PMID- 18675272 TI - Virulence of Hypocreales fungi to pecan aphids (Hemiptera: Aphididae) in the laboratory. AB - There is need for efficacious biocontrol agents for aphids in commercial orchards. As a preliminary step to this end we determined the virulence of several Hypocreales fungi to pecan aphids. In the first experiment we tested the virulence of Isaria fumosorosea (ARSEF 3581) blastospores to three pecan aphids Monellia caryella, Melanocallis caryaefoliae, and Monelliopsis pecanis under laboratory conditions. Rates of 1x10(7) or 1x10(8) spores per ml were applied in 2 ml via a spray tower to 90 mm Petri dishes containing 10 aphids each. Mortality and mycosis were determined after 24, 48 and 72 h. Treatment effects were observed by 48 h post-application, and by 72 h the higher application rate caused >90% mortality and mycosis in M. caryella and M. caryaefoliae, whereas <70% was observed in M. pecanis. We conducted two subsequent experiments (Experiments 2 and 3), using the same methodology, to compare the virulence of several Hypocreales species and strains against the aphid of primary economic concern to most pecan growers, M. caryaefoliae. In Experiment 2, we compared blastospores and conidia of two I. fumosorosea strains (ARSEF 3581 and ATCC 20874 [= strain 97]). The blastospores of ARSEF 3581 and conidia of ATCC 20874 showed higher virulence than other treatments and thus were included in Experiment 3, which also compared the virulence of conidia of Beauveria bassiana (GHA strain) and Metarhizium anisopliae (F52 strain). Results in Experiment 3 indicated the highest virulence in I. fumosorosea 3581 blastospores and M. anisopliae (F52) followed by I. fumosorosea (20874) conidia. The detection of pathogenicity to pecan aphids establishes the potential for commercial usage and additional study. Results reported here will narrow treatments to test in future greenhouse and field trials. PMID- 18675273 TI - G-CSF exerts dual effects on endothelial cells--opposing actions of direct eNOS induction versus indirect CRP elevation. AB - Granulocyte-colony stimulating factor (G-CSF) has been shown to have protective effects in the heart and brain. However, it may also be involved in the acute inflammatory response which may be harmful. The effects of G-CSF on endothelial cells (ECs) and the vasculature are mostly unknown. To study the possible dual effects of G-CSF on ECs, we investigated whether G-CSF induces release of C reactive protein (CRP) by hepatocytes and whether the direct beneficial effects of G-CSF could protect ECs from the detrimental effects of CRP. G-CSF treatment significantly induced monocytes to produce IL-6, and culture supernatants of G CSF-stimulated monocytes induced CRP production in hepatocytes. On the other hand, G-CSF directly promoted EC proliferation and migration and reversed the deleterious effects of CRP. In mechanistic analyses, G-CSF increased not only the protein expression of endothelial nitric oxide synthase (eNOS), but also its transcription. Furthermore, it enhanced eNOS phosphorylation and activation, leading to increased production of NO. Thus, G-CSF reversed the attenuated production of NO by CRP. These effects of G-CSF on eNOS transcription, translation, and activation were blunted by the PI3K inhibitor, suggesting that EC protective effects of G-CSF were associated with the activation of the Akt/eNOS pathway. In conclusion, although G-CSF induces an inflammatory reaction leading to CRP production, it has direct beneficial effects protecting ECs from the deleterious effects of CRP through activation of Akt/eNOS pathway, leading to an increase in NO production. Our data suggests that G-CSF may exert dual opposing effects on endothelial cells. PMID- 18675274 TI - Crystal structure of glucose-6-phosphate isomerase from Thermus thermophilus HB8 showing a snapshot of active dimeric state. AB - Glucose-6-phosphate isomerase (GPI) is a glycolytic enzyme with ill-defined oligomeric state. In order to obtain insight into the correlation between oligomerization and the catalytic function of this enzyme, the crystal structure of GPI from the extreme thermophile Thermus thermophilus HB8 (TtGPI) has been determined at 1.95 A resolution. The crystallographic asymmetric unit contains an apparent dimer. The core fold of protomer and the interprotomer spatial arrangement of the dimer are similar to those of already reported crystal structures of other GPIs. The active site is located on the dimer interface, and putative catalytic residues are well conserved among the GPIs. These results suggest that the observed dimeric state of TtGPI in the crystal is biologically relevant and that this enzyme uses a common catalytic mechanism for the isomerase reaction. Gel-filtration chromatography, chemical cross-linking, sedimentation equilibrium by analytical ultracentrifugation, and dynamic light-scattering experiments indicate that TtGPI exists in a dynamic equilibrium between monomeric and dimeric states in solution. Several factors potentially contributing to the thermal stability of TtGPI protomer were identified: (i) a decrease in denaturation entropy by the shorter polypeptide length and by amino acid composition, including the increased number of proline residues and a higher arginine-to-lysine ratio; (ii) a larger number of ion pairs; and (iii) a reduction in cavity volume. From these results, it is suggested that transient dimer formation is sufficient for the catalytic function and that the TtGPI protomer itself has intrinsically higher thermal stability. PMID- 18675275 TI - The novel protein complex with SMARCAD1/KIAA1122 binds to the vicinity of TSS. AB - The SMARCAD1/KIAA1122 protein is structurally classified into the SWI2/SNF2 superfamily of DNA-dependent ATPases that are catalytic subunits of chromatin remodeling complexes. Although the importance of other members of the SWR1-like subfamily in chromatin remodeling (EP400, INOC1, and SRCAP) has already been elucidated, the biological function of SMARCAD1/KIAA1122 in transcriptional regulation remains to be clarified. To gain insight into the role of this protein, we generated a specific antibody against SMARCAD1/KIAA1122 and used it for chromatin and protein immunoprecipitation assays. We employed high-resolution genome tiling microarrays in chromatin immunoprecipitation and found the binding sites of SMARCAD1/KIAA1122 in the vicinity of the transcriptional start site of 69 candidate target genes. In the protein immunoprecipitation assay, we found that endogenous SMARCAD1/KIAA1122 binds with TRIM28, a recently highlighted transcriptional regulator in the cancer field. From these findings, we propose a novel model for gene regulation via the SMARCAD1/KIAA1122 protein complex. PMID- 18675276 TI - Statistical coupling analysis of aspartic proteinases based on crystal structures of the Trichoderma reesei enzyme and its complex with pepstatin A. AB - The crystal structures of an aspartic proteinase from Trichoderma reesei (TrAsP) and of its complex with a competitive inhibitor, pepstatin A, were solved and refined to crystallographic R-factors of 17.9% (R(free)=21.2%) at 1.70 A resolution and 15.8% (R(free)=19.2%) at 1.85 A resolution, respectively. The three-dimensional structure of TrAsP is similar to structures of other members of the pepsin-like family of aspartic proteinases. Each molecule is folded in a predominantly beta-sheet bilobal structure with the N-terminal and C-terminal domains of about the same size. Structural comparison of the native structure and the TrAsP-pepstatin complex reveals that the enzyme undergoes an induced-fit, rigid-body movement upon inhibitor binding, with the N-terminal and C-terminal lobes tightly enclosing the inhibitor. Upon recognition and binding of pepstatin A, amino acid residues of the enzyme active site form a number of short hydrogen bonds to the inhibitor that may play an important role in the mechanism of catalysis and inhibition. The structures of TrAsP were used as a template for performing statistical coupling analysis of the aspartic protease family. This approach permitted, for the first time, the identification of a network of structurally linked residues putatively mediating conformational changes relevant to the function of this family of enzymes. Statistical coupling analysis reveals coevolved continuous clusters of amino acid residues that extend from the active site into the hydrophobic cores of each of the two domains and include amino acid residues from the flap regions, highlighting the importance of these parts of the protein for its enzymatic activity. PMID- 18675277 TI - Transposition of Mboumar-9: identification of a new naturally active mariner family transposon. AB - Although mariner transposons are widespread in animal genomes, the vast majority harbor multiple inactivating mutations and only two naturally occurring elements are known to be active. Previously, we discovered a mariner-family transposon, Mboumar, in the satellite DNA of the ant Messor bouvieri. Several copies of the transposon contain a full-length open reading frame, including Mboumar-9, which has 64% nucleotide identity to Mos1 of Drosophila mauritiana. To determine whether Mboumar is currently active, we expressed and purified the Mboumar-9 transposase and demonstrate that it is able to catalyze the movement of a transposon from one plasmid to another in a genetic in vitro hop assay. The efficiency is comparable to that of the well-characterized mariner transposon Mos1. Transposon insertions were precise and were flanked by TA duplications, a hallmark of mariner transposition. Mboumar has been proposed to have a role in the evolution and maintenance of satellite DNA in M. bouvieri and its activity provides a means to examine the involvement of the transposon in the genome dynamics of this organism. PMID- 18675278 TI - Crystal structure of the intact archaeal translation initiation factor 2 demonstrates very high conformational flexibility in the alpha- and beta subunits. AB - In Eukarya and Archaea, translation initiation factor 2 (eIF2/aIF2), which contains three subunits (alpha, beta, and gamma), is pivotal for binding of charged initiator tRNA to the small ribosomal subunit. The crystal structure of the full-sized heterotrimeric aIF2 from Sulfolobus solfataricus in the nucleotide free form has been determined at 2.8-A resolution. Superposition of four molecules in the asymmetric unit of the crystal and the comparison of the obtained structures with the known structures of the aIF2alphagamma and aIF2betagamma heterodimers revealed high conformational flexibility in the alpha- and beta-subunits. In fact, the full-sized aIF2 consists of a rigid central part, formed by the gamma-subunit, domain 3 of the alpha-subunit, and the N-terminal alpha-helix of the beta-subunit, and two mobile "wings," formed by domains 1 and 2 of the alpha-subunit, the central part, and the zinc-binding domain of the beta subunit. High structural flexibility of the wings is probably required for interaction of aIF2 with the small ribosomal subunit. Comparative analysis of all known structures of the gamma-subunit alone and within the heterodimers and heterotrimers in nucleotide-bound and nucleotide-free states shows that the conformations of switch 1 and switch 2 do not correlate with the assembly or nucleotide states of the protein. PMID- 18675279 TI - Feasibility of communication in binary signaling games. AB - In signaling games the replicator dynamics does not almost always converge to states of perfect communication. A significant portion of the state space converges to components of Nash equilibria that characterize states of partial communication. Since these components consist of non-hyperbolic rest points, the significance of this result will depend on the dynamic behavior of specific perturbations of the replicator equations. In this paper we study selection mutation dynamics of signaling games, which may be considered as one plausible perturbation of the replicator dynamics. We find that the long term behavior of the dynamics depends on the mutation rates of senders and receivers and on the relevance of communication. PMID- 18675280 TI - Expression of the cytochrome P450 epoxygenase CYP2J2 in human monocytic leukocytes. AB - CYP2J2 is one of the cytochrome P450 epoxygenases involved in the metabolism of arachidonic acid. CYP2J2 has been identified in several tissues, especially cardiovascular tissues. CYP2J2 has cardiovascular effects, as epoxyeicosatrienoic acid, one of its metabolites, has anti-inflammatory and vasodilative activities. We investigated the expression of CYP2J2 in human leukocytes using reverse transcription-polymerase chain reaction, immunoblotting and immunostaining. Human monocytic cells, but not human neutrophils, exhibited constitutive expression of CYP2J2. Furthermore, the expression of CYP2J2 mRNA increased when the human monocytic cell line THP-1 cells and human monocytes were stimulated with phorbol 12-myristate 13-acetate and macrophage-colony stimulating factor in combination with granulocyte/macrophage-colony stimulating factor, respectively. These results suggest that expression of CYP2J2 was up-regulated when human monocytes differentiated into macrophages and that human monocytic cells and macrophages have a pathway to metabolize arachidonic acid using CYP epoxygenases. PMID- 18675281 TI - Biological substrates of reward and aversion: a nucleus accumbens activity hypothesis. AB - The nucleus accumbens (NAc) is a critical element of the mesocorticolimbic system, a brain circuit implicated in reward and motivation. This basal forebrain structure receives dopamine (DA) input from the ventral tegmental area (VTA) and glutamate (GLU) input from regions including the prefrontal cortex (PFC), amygdala (AMG), and hippocampus (HIP). As such, it integrates inputs from limbic and cortical regions, linking motivation with action. The NAc has a well established role in mediating the rewarding effects of drugs of abuse and natural rewards such as food and sexual behavior. However, accumulating pharmacological, molecular, and electrophysiological evidence has raised the possibility that it also plays an important (and sometimes underappreciated) role in mediating aversive states. Here we review evidence that rewarding and aversive states are encoded in the activity of NAc medium spiny GABAergic neurons, which account for the vast majority of the neurons in this region. While admittedly simple, this working hypothesis is testable using combinations of available and emerging technologies, including electrophysiology, genetic engineering, and functional brain imaging. A deeper understanding of the basic neurobiology of mood states will facilitate the development of well-tolerated medications that treat and prevent addiction and other conditions (e.g., mood disorders) associated with dysregulation of brain motivation systems. PMID- 18675282 TI - Excitatory afferents to CA3 pyramidal cells display differential sensitivity to CB1 dependent inhibition of synaptic transmission. AB - Recent advances in immunohistochemical techniques have, contrary to earlier reports, positively identified CB1 receptors on glutamatergic terminals in the hippocampus. Further work has implicated these receptors in modulation of susceptibility to kainic acid induced seizures. Based on these results, the current study was designed to test the hypothesis that both exogenous and endogenous cannabinoids can selectively modulate glutamatergic afferents to CA3 pyramidal cells, and that such modulation is mediated by cannabinoid type 1 (CB1) receptors. Towards that end we employed either conventional or two-photon guided minimal stimulation techniques to isolate mossy fiber and/or associational/commissural (A/C) inputs to CA3 pyramidal cells. We report that bath application of WIN55,212-2 selectively inhibits minimally evoked A/C inputs to CA3 pyramidal cells, without significantly altering simultaneously recorded mossy fiber inputs. Further, we find that WIN55,212-2 mediated inhibition of A/C inputs is completely blocked by the CB1 selective antagonist AM-251 and absent in CB1(-/-) animals, suggesting a dependence on CB1 receptors. Finally, we demonstrate that depolarization of CA3 pyramidal cells leads to calcium dependent release of endogenous cannabinoids that transiently inhibit A/C mediated responses, and that this effect is also sensitive to both AM-251 and the muscarinic acetylcholine receptor antagonist atropine. To our knowledge this represents the first demonstration of depolarization induced suppression of excitation in area CA3 of the hippocampus. Collectively, these results provide new information relevant to developing a thorough understanding of how ECs modulate excitatory transmission in an area that is both essential for the acquisition of new memories and intimately involved in epileptogenesis. PMID- 18675284 TI - Effect of frontal lobe lesions on the recollection and familiarity components of recognition memory. AB - Single-process theories assume that familiarity is the sole influence on recognition memory with decisions being made as a continuous process. Dual process theories claim that recognition involves both recollection and familiarity processes with recollection as a threshold process. Although, the frontal lobes of the brain play an important role in recognition memory, few studies have examined the effect of frontal lobe lesions on recollection and familiarity. In the current study, the nonverbal recognition memory of 24 patients with focal frontal lesions due to tumour or stroke was examined. Recollection and familiarity were estimated using the receiver operating characteristic (ROC) method. A secondary analysis was also conducted using standard signal detection theory methodology. Both analyses led to similar conclusions where only the familiarity component of recognition memory was impaired in frontal patients compared to healthy controls whilst the recollection type (or variance ratio) processes remained intact. PMID- 18675285 TI - Episodic, but not semantic, autobiographical memory is reduced in amnestic mild cognitive impairment. AB - Amnestic mild cognitive impairment (aMCI) is characterized by decline in anterograde memory as measured by the ability to learn and remember new information. We investigated whether retrograde memory for autobiographical information was affected by aMCI. Eighteen control (age 66-84 years) and 17 aMCI (age 66-84 years) participants described a personal event from each of the five periods across the lifespan. These events were transcribed and scored according to procedures that separate episodic (specific happenings) from semantic (general knowledge) elements of autobiographical memory. Although both groups generated protocols of similar length, the composition of autobiographical recall differentiated the groups. The aMCI group protocols were characterized by reduced episodic and increased semantic information relative to the control group. Both groups showed a similar pattern of recall across time periods, with no evidence that the aMCI group had more difficulty recalling recent, rather than remote, life events. These results indicate that episodic and semantic autobiographical memories are differentially affected by the early brain changes associated with aMCI. Reduced autobiographical episodic memories in aMCI may be the result of medial temporal lobe dysfunction, consistent with multiple trace theory, or alternatively, could be related to dysfunction of a wider related network of neocortical structures. In contrast, the preservation of autobiographical semantic memories in aMCI suggests neural systems, such as lateral temporal cortex, that support these memories, may remain relatively intact. PMID- 18675286 TI - Confabulations in alcoholic Korsakoff patients. AB - Besides forgetting, memory is also prone to distortions, errors and illusions. Confabulation is one type of memory distortion that may occur in cases of brain damage. Although confabulations are described anecdotally in patients with alcoholic Korsakoff syndrome (KS), there are few systematic investigations of the presence and nature of these types of false memories in KS. Moreover, it is unclear whether KS patients' confabulations evenly affect all types of memories, or whether certain memory domains are more susceptible. Our study attempted to clarify two questions: first, whether confabulations are a critical feature of the cognitive impairment associated with long-term KS in a large sample of patients (N=42). Second, we investigated which memory domain is most likely affected by confabulations in KS. To elicit confabulations, we used a Confabulation Interview containing questions from different memory domains. We found that KS patients overall confabulated more compared to a group of healthy subjects. Furthermore, we found that patients confabulated most within the episodic/autobiographical memory domain. Our results imply that besides pronounced memory deficits typically associated with KS, confabulation can also be regarded as a clinical feature of the disease. The preponderance of episodic confabulation obtained here by using a standardized test, confirms anecdotic reports that KS patients confabulate in everyday life mainly with respect to their personal past and present. Thus, for a detailed description of the memory profile of KS patients, the screening of confabulation tendencies may be a useful supplementary clinical tool. PMID- 18675287 TI - Resting metabolic rate in young women classified as restrained or unrestrained eaters. AB - Restrained eating involves a mainly cognitive control of eating behaviour and has been supposed to be a predisposing factor for the development of eating disorders. Biological correlates of the behaviour are expected, too. The present study investigated resting metabolic rate in 33 female restrained and unrestrained eaters. Measured by indirect calorimetry RMR showed a significant negative relationship to the degree of dietary restraint (controlled for BMI, partial r=-.46, p<.01). The lower energy requirements of restrained eaters might be genetically predisposed, but also might have severe consequences in nonclinical and clinical groups. PMID- 18675288 TI - Attenuation of maternal behavior in virgin CD-1 mice by methylphenidate hydrochloride. AB - The administration of methylphenidate hydrochloride (MPH) to girls and women has increased in the last decade and the potential for mothers to receive this medication has also increased. Because substances that alter the dopaminergic systems can also disrupt maternal behavior, and MPH acts on dopaminergic neurons, we evaluated the influence of MPH on maternal behavior. The maternal induction paradigm allowed us to assess changes in spontaneous maternal behavior as a result of repeated exposure to MPH without exposing pups to the drug. Virgin female CD-1 mice received MPH (5 mg/kg) or saline daily, starting 3 days before pup exposure, and for the duration of the 10-day test period. Naive groups of three pups were placed with the female each day and maternal behavior was assessed during 10-minute observation periods 1 h post-injection. MPH-treated females showed significantly less maternal behavior, including reduced pup licking and crouching over pups, compared to saline treated females. MPH-treated females also exhibited higher activity levels than saline treated females. Given the disruption in spontaneous maternal behavior of MPH-treated mice, further research examining the relationship between maternal behavior and MPH exposure is warranted. PMID- 18675289 TI - Action of (R)-sila-venlafaxine and reboxetine to antagonize cisplatin-induced acute and delayed emesis in the ferret. AB - The chemotherapeutic drug cisplatin is associated with severe gastrointestinal toxicity that can last for several days. A recent strategy to treat the nausea and emesis includes the combination of a 5-HT3 receptor antagonist, a glucocorticoid, and an NK1 receptor antagonist. The present studies explore the use of the selective noradrenaline reuptake inhibitors, (R)-sila-venlafaxine, (R,R)-reboxetine and (S,S)-reboxetine to prevent cisplatin (5 mg/kg, i.p.) induced acute (0-24 h) and delayed (24-72 h) emesis in ferrets. The positive control regimen of ondansetron and dexamethasone, both at 1 mg/kg/8 h, reduced acute and delayed emesis by 100 (P<0.001) and 61% (P<0.05). (R)-sila-venlafaxine at 5 and 15 mg/kg/4 h reduced acute emesis by 86 (P<0.01) and 66% (P<0.05), respectively and both enantiomers of reboxetine at 1 mg/kg/12 h also reduced the response by approximately 70-90% (P<0.05). Out of the reuptake inhibitors, only (R)-sila-venlafaxine at 15 mg/kg/4 h was active to reduce delayed emesis (a 57% reduction was observed (P<0.05)); its terminal plasma levels were positively correlated with an inhibition of emesis during the delayed phase (P<0.05). (R) sila-venlafaxine was also examined against a higher dose of cisplatin 10 mg/kg, i.p. (3 h test) and it dose-dependently antagonized the response (maximum reduction was 94% at 10 mg/kg, p.o.; P<0.01) but it was ineffective against apomorphine (0.125 mg/kg, s.c.) and ipecacuanha (2 mg/kg, p.o.)-induced emesis (P>0.05). In conclusion, the studies provide the first evidence for an anti emetic potential of noradrenaline reuptake inhibitors to reduce chemotherapy induced acute and delayed emesis. PMID- 18675290 TI - Distinct position assignment mechanisms revealed by cross-order motion. AB - Motion perception influences perceived position. It has been shown that first order (luminance defined) motion shifts perceived position across a wide range of spatial and temporal frequencies. On the other hand, second-order (contrast defined) motion shifts perceived position over a narrow range of temporal frequencies, regardless of spatial frequency [Bressler, D. W., & Whitney, D. (2006). Second-order motion shifts perceived position. Vision Research, 46(6-7), 1120-1128]. These results suggest the presence of distinct position assignment mechanisms for first- and second-order motion. We investigated whether the first- and second-order systems independently encode and assign the position of a moving stimulus. To measure motion induced position shift we presented two horizontally offset Gabors placed above and below a central fixation point, with sine wave carriers drifting in opposite directions. Subjects judged the position of the top Gabor relative to the bottom one. We used both first-order Gabors (sinusoidal luminance modulation of a dynamic noise carrier enveloped by a static Gaussian) and second-order Gabors (sinusoidal contrast modulation of a dynamic noise carrier enveloped by a static Gaussian). Results showed a strong position shift in the direction of the carrier motion when both Gabors were first-order, a weak position shift when both Gabors were second-order, and no appreciable position shift when one Gabor was first-order and the other was second-order (cross-order motion). The absence of a position shift using cross-order motion supports the hypothesis that the two motion systems independently encode and assign the position of a moving object. These results are consistent with those of experiments investigating global spatial interactions between static first-order and second-order Gabor patches, indicating a commonality in the underlying spatial integration processes. PMID- 18675291 TI - The influence of acute varenicline administration on smoking and eating behavior in humans. AB - Varenicline (Chantix) is a novel smoking-cessation agent that acts at a number of nicotinic acetylcholine receptors. The aim of this study was to determine the behavioral effects of acute varenicline administration in human subjects. The effects of doses of varenicline (0.5, 1 and 2 mg), methylphenidate (40 mg; positive control) and placebo were assessed in 8 (7 males, 1 female) cigarette smokers. Staggered, double blind dosing was used to examine eating and smoking behavior during the peak effects of varenicline and methylphenidate. Starting at the published time to peak plasma levels of these drugs, subjects were allowed to smoke and eat ad libitum for 4 h. Acute varenicline was devoid of behavioral effects. Methylphenidate produced prototypical stimulant-like effects (e.g., increased smoking behavior; decreased caloric intake). The present results indicate that acute varenicline administration does not alter smoking behavior although the low number of subjects limits the ability to detect small effects. Future research should examine the effects of chronic varenicline on smoking and eating behavior in humans, particularly using operant techniques to determine whether varenicline alters the reinforcing effects of cigarettes and food in humans. PMID- 18675292 TI - Persistent depressive state after chronic stress in rats is accompanied by HPA axis dysregulation and reduced prefrontal dopaminergic neurotransmission. AB - Exposure to stress is thought to play an important role in the etiology of depression. Dysregulation of the hypothalamo-pituitary-adrenal (HPA) axis characterized by glucocorticoid negative feedback resistance is frequently observed in human depressives. Additionally, dysfunctions of the dopaminergic and serotonergic systems in the prefrontal cortex (PFC) are thought to be involved in the development of a depressive state. In rats, chronic stress induces a behaviorally depressive state, concomitant with dysregulation of the HPA axis and reductions in dopaminergic and serotonergic transmissions in the PFC. Considering that dysregulation of the HPA axis is associated with relapse and persistency of depression, it is possible that the chronic stress-induced depressive state persists during long-term rest after its exposure. In the present study, we examined this possibility in rats and found that the behaviorally depressive state in the rotarod test, negative feedback resistance in the dexamethasone suppression test, and a decrease in the extracellular concentration of dopamine but not serotonin in the PFC persisted for 3 months following a 4-week stress session. These results suggest that dysregulation of the HPA system and reduced dopaminergic transmission in the PFC underlies persistent behavioral depression following chronic stress. PMID- 18675293 TI - Social and physical environments of sports and exercise reported among adults in the American Time Use Survey. AB - OBJECTIVE: Demographic and temporal patterns in the social and physical environments of sports and exercise in the American Time Use Survey (years 2003 2006) are described. METHOD: The sample consisted of adult respondents (ages 21+) reporting at least one bout of sports or exercise (N=8844). During the interview, participants reported where (e.g., outdoors, home, work) and with whom (e.g., alone, family, coworkers) each bout occurred. Sample-weighted multinomial logistic regression analyses estimated the adjusted proportion of bouts occurring in each environment by gender, age, education, race/ethnicity, season, weekend/weekday, and time of day. RESULTS: Among members of the oldest age group (ages 60+), more exercise bouts occurred alone and outdoors compared to younger age groups. Among college graduates, more exercise bouts occurred at a gym/health club compared to groups with lower levels of education. Exercise bouts occurring alone were most likely to happen in the winter, on weekdays, and in the morning. Exercise bouts occurring outdoors were most likely to happen in the summer, on weekend days, and in the morning. CONCLUSION: Future research and intervention efforts exploring where, when and with whom exercise bouts occur may prove beneficial to addressing public health concerns about physical inactivity. PMID- 18675294 TI - One-year follow-up evaluation of Project Towards No Drug Abuse (TND-4). AB - OBJECTIVES: This paper describes the one-year outcomes of the fourth experimental trial of Project Towards No Drug Abuse. Two theoretical content components of the program were examined to increase our understanding of the relative contribution of each to the effectiveness of the program. METHODS: High schools in Southern California (n=18) were randomly assigned to one of three conditions: cognitive perception information curriculum, cognitive perception information+behavioral skills curriculum, or standard care (control). The curricula were delivered to high school students (n=2734) by project health educators and regular classroom teachers. Program effectiveness was assessed with both dichotomous and continuous measures of 30-day substance use at baseline and one-year follow-up. RESULTS: Across all program schools, the two different curricula failed to significantly reduce dichotomous measures of substance use (cigarette, alcohol, marijuana, and hard drugs) at one-year follow-up. Both curricula exerted an effect only on the continuous measure of hard drug use, indicating a 42% (p=0.02) reduction in the number of times hard drugs were used in the last 30 days in the program groups relative to the control. CONCLUSIONS: The lack of main effects of the program on dichotomous outcomes was contrary to previous studies. An effect on an ordinal count measure of hard drug use among both intervention conditions replicates previous work and suggests that this program effect may have been due to changes in cognitive misperception of drug use rather than behavioral skill. PMID- 18675295 TI - The Groningen Active Living Model, an example of successful recruitment of sedentary and underactive older adults. AB - OBJECTIVE: Many physical activity interventions do not reach those people who would benefit the most from them. The Groningen Active Living Model (GALM) was successful in recruiting sedentary and underactive older adults. METHOD: In the fall of 2000 older adults in three municipalities in the Netherlands received written information, were visited at home and, if eligible according to the GALM recruitment criteria, filled in the Stages of Change questionnaire and the Voorrips physical activity questionnaire. RESULTS: By using the strategy we succeeded in including 12.3% of the older adults (315 of the 2551 qualifying participants), 79.4% of whom could be indeed regarded as sedentary or underactive. These results can be considered in line with results described in the literature. The cost of successfully recruiting an older adult was estimated at $84. CONCLUSIONS: The GALM recruitment strategy is a potentially useful and effective method for reaching community-dwelling sedentary and underactive older adults. PMID- 18675296 TI - Inequalities in breast and cervical cancer screening among urban Mexican women. AB - OBJECTIVE: To examine the equity of screening for breast and cervical cancer among women aged 50 and over living in Mexico City according to the Andersen behavioural model. METHODS: Logistic regressions were used to analyze the associations of predisposing factors, enabling resources and health needs with mammography and Pap testing among 1323 women aged 50 and over living in Mexico City. The data were taken from SABE, a survey conducted in Latin American and Caribbean cities in 1999-2000. RESULTS: Referring to the previous two years, use of mammography was less frequent (14.8%) than Pap testing (53.7%). Higher education and having private health insurance were associated with receiving both procedures. Being married was associated with receipt of Pap testing, while being unmarried was associated with mammography. Younger women were more likely to receive a Pap test. Other cancer prevention behaviours were strong predictors of mammography and Pap test use. CONCLUSIONS: There are inequities in access to cancer screening services among women aged 50 and over in Mexico. Cancer prevention programs need to implement strategies that guarantee free and accessible services independently of social conditions or type of health insurance. The observed clustering of screening services suggests that packaging screening programs for gynaecological cancer could be used to increase coverage. PMID- 18675297 TI - Attitudes and perceptions towards increasing cigarette price: a population-based survey in Italy. PMID- 18675298 TI - The influence of the support nature on the kinetics parameters, inhibition constants and reactivation of immobilized acetylcholinesterase. AB - Acetylcholinesterase (AChE) was immobilized on two different composite membranes constituted by a chemically modified poly-acrylonitrile (PAN) membrane plus a layer of tethered chitosan of different molecular weight, 10 kDa or 400 kDa. AChE was also directly immobilized on a chemically modified PAN membrane with NaOH and ethylenediamine (EDA) without chitosan. To know how the different supports affected the enzyme activity and the kinetic parameters, the AChE activity was studied in the soluble form and in the insoluble form with all the three types of modified PAN membranes. The best performance was obtained by the modified PAN membrane having the chitosan with the lower molecular weight. The results concerning the AChE inhibition by methyl-paraoxon and the subsequent reactivation by pyridine-2-aldoxime methochloride (2-PAM) are presented and discussed. The composite membrane having chitosan with the lower molecular weight appeared to be potentially useful for applications in the field of biosensors. PMID- 18675299 TI - Gustatory hedonic value: potential function for forebrain control of brainstem taste processing. AB - Among well-nourished populations, eating beyond homeostatic needs when presented with caloric-dense palatable food evidences the assertion that an increasing proportion of consumption is driven by pleasure, not just by the need for calories. This presents a major health crisis because the affective component of foods constitutes a behavioral risk factor that promotes over consumption [Sorensen, L.B., Moller, P., Flint, A., Martens, M., Raben, A., 2003. Effect of sensory perception of foods on appetite and food intake: a review of studies on humans. Int. J. Obes. Relat. Metab. Disord. 27, 1152-1166; Yeomans, M.R., Blundell, J.E., Leshem, M., 2004. Palatability: response to nutritional need or need-free stimulation of appetite? Br. J. Nutr. 92 (Suppl. 1), S3-S14]. Overweight or obese individuals have an increased risk of developing hypertension, stroke, heart disease, chronic musculoskeletal problems, type-2 diabetes, and certain types of cancers [Hill, J.O., Catenacci, V., Wyatt, H.R., 2005. Obesity: overview of an epidemic. Psychiatr. Clin. N. Am. 28, 1-23, vii]. The etiology of obesity is complex involving genetic, metabolic, and behavioral factors, but ultimately results from long-term energy imbalance. Evidence indicates that learned and some forms of unlearned control of ingestive behavior driven by palatability (i.e. hedonic value) are critically dependent on reciprocal interactions between brainstem gustatory nuclei and the ventral forebrain. This review discusses the current understanding of centrifugal control of taste processing in subcortical gustatory nuclei and the potential role of such modulation in hedonic responding. PMID- 18675301 TI - Localization of nerve fiber bundles by polarization-sensitive optical coherence tomography. AB - Diagnostic modalities that can distinguish brain tumors from eloquent cortices or nerve fiber bundles are important for neurosurgery. For identifying nerve fiber bundles, various techniques such as diffusion tensor imaging and subcortical stimulation mapping have been used. In this article, we propose localization of nerve fiber bundles by polarization-sensitive optical coherence tomography (PS OCT) for the first time. PS-OCT performs tomographic imaging by measuring the travel distance and the change in polarization of the backscattered light from the tissue at different traverse positions, enabling selective visualization of birefringent tissues such as nerve fiber bundles. We examined the imaging of nerve fiber bundles in blocks of fixed rat brains. Nerve fiber bundles in internal and external capsules of the rat brains appearing on the surfaces of the coronal, horizontal, and sagittal planes were identified as to their locations and orientations. The nerve fiber bundles were clearly visualized by PS-OCT. The image penetration depth of the PS-OCT images was about 1.0mm in gray matter and about 0.5mm in white matter, so the refractive indices of gray and white matters were assumed to be 1.4. PMID- 18675302 TI - LPS induces inflammatory responses in human aortic vascular smooth muscle cells via Toll-like receptor 4 expression and nitric oxide production. AB - Inflammation is an important event in the development of vascular diseases such as hypertension, atherosclerosis, and restenosis. In addition, the stimulation of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) induces the release of critical proinflammatory cytokines that activate potent immune responses. In this study, LPS was found to induce TLR4 expression and increased nitric oxide (NO) production by increasing the expression of inducible nitric oxide synthase (iNOS). Furthermore, LPS was found to induce interleukin (IL)-8 and vascular endothelial growth factor (VEGF) production, as well as intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression. Taken together, these results indicate that LPS induces inflammatory responses in HASMC. Moreover, NOS inhibitor (L-NAME) and anti-TLR 4mAb reduced the LPS-induced NO, IL-8 and VEGF production and ICAM-1 expression. Additionally, TLR4 expression was reduced by NOS inhibitor. Taken together, these results indicate that LPS induced inflammatory responses are regulated by TLR4 expression and NO production. PMID- 18675303 TI - A reliable IC One-step RT-PCR method for the detection of BBrMV to ensure safe exchange of Musa germplasm. AB - An immunocapture (IC) One-step RT-PCR assay was developed to improve the detection of Banana bract mosaic virus (BBrMV) in single and bulked samples of banana plants. In this paper, an atypical strain of BBrMV was described, the BBrMV "Ref" strain, and we showed that detection with available BBrMV tools using ELISA and RT-PCR approaches was not reliable. Primer sets Bract N1/NR and N2/NR specific to BBrMV were designed and used in RT-PCR and IC-RT-PCR assays with two commercial kits that allow the RT and the PCR reactions to take place simultaneously in the same tube. The new assay enabled detection of BBrMV in leaf extract diluted up to 1 x 10(-10) and in bulked samples of 10 plants, and was proposed as a new international standard to index BBrMV. PMID- 18675304 TI - Side-specific operant conditioning of antennal movements in the honey bee. AB - Operant conditioning of antennal movements in honey bees was used to investigate whether learned changes on one antenna influence antennal movements of the contralateral antenna. Conditioning of the right antenna did not alter antennal movements of the left antenna and subsequent conditioning of the left antenna did not abolish the previously learned change in the right antenna. Thus, the antennal systems on each side are largely independent from each other. PMID- 18675300 TI - Dysregulated sarcoplasmic reticulum calcium release: potential pharmacological target in cardiac disease. AB - In the heart, Ca(2+) released from the intracellular Ca(2+) storage site, the sarcoplasmic reticulum (SR), is the principal determinant of cardiac contractility. SR Ca(2+) release is controlled by dedicated molecular machinery, composed of the cardiac ryanodine receptor (RyR2) and a number of accessory proteins, including FKBP12.6, calsequestrin (CASQ2), triadin (TRD) and junctin (JN). Acquired and genetic defects in the components of the release channel complex result in a spectrum of abnormal Ca(2+) release phenotypes ranging from arrhythmogenic spontaneous Ca(2+) releases and Ca(2+) alternans to the uniformly diminished systolic Ca(2+) release characteristic of heart failure. In this article, we will present an overview of the structure and molecular components of the SR and Ca(2+) release machinery and its modulation by different intracellular factors, such as Ca(2+) levels inside the SR as well as phosphorylation and redox modification of RyR2s. We will also discuss the relationships between abnormal SR Ca(2+) release and various cardiac disease phenotypes, including, arrhythmias and heart failure, and consider SR Ca(2+) release as a potential therapeutic target. PMID- 18675305 TI - Microbial adhesion of Cryptosporidium parvum: identification of a colostrum derived inhibitory lipid. AB - We previously described an unidentified lipid purified from calf small intestine that inhibits the in vitro adhesion of Cryptosporidium parvum sporozoites to host cells [Johnson JK, Schmidt J, Gelberg HB, Kuhlenschmidt MS. Microbial adhesion of Cryptosporidium parvum sporozoites: purification of an inhibitory lipid from bovine mucosa. J Parasitol 2004;90:980-90]. Intestinal mucosa from some calves, however, failed to yield this bioactive lipid. Accordingly, we examined other potential sources, especially dietary sources, of the inhibitory lipid and discovered it was principally derived from bovine colostrum. Interestingly, fresh colostrum yielded little or no inhibitory lipid, however, the lipid was found in relatively large quantities following incubation of colostrum with the aqueous fraction of calf intestinal contents. Using FAB-MS and NMR analysis, the sporozoite inhibitory lipid (SIL) was identified as oleic acid, a monounsaturated fatty acid likely released from colostrum triglycerides and phospholipids by digestion in the lumen of the calf small intestine. Oleic acid dose-dependently inhibited in vitro sporozoite-host cell adhesion with an inhibitory constant (IC(50)) of approximately 5 microM. Comparison of oleic acid with other C-18 fatty acids revealed linolenic, but not stearic acid, also displayed potent inhibitory activity. Neither linolenic nor oleic acid, however, affect either sporozoite or host cell viability at concentrations that inhibit sporozoite adhesion. These results suggest certain colostrum-derived long-chain fatty acids may serve as natural inhibitors of the early steps in C. parvum sporozoite-host cell interactions. PMID- 18675306 TI - Multiplex PCRs for assignment of Staphylocoagulase types and subtypes of type VI Staphylocoagulase. AB - Staphylocoagulases (SCs) have been classified by the differences in antigenicity using a serological method. We have developed a system to classify them based on the nucleotide differences in SC genes (coa). The system was composed of three multiplex PCRs (M-PCRs): M-PCR:A, identifying types III, IV, VII, and VIII; M PCR:B, identifying types I, II, V, and VI; M-PCR:C, identifying three subtypes of type VI. In this study, we found that coa genes of the serotype VI were not identical, but classified into three subtypes based on the nucleotide differences, especially in D2 and the central region: VIa, the coa gene carried by stp12 from human; and VIb and VIc, the coa genes carried by strains IFH556 and IFH514 isolated from bovine raw milk. The primer pair used in M-PCR:B was designed to identify all three subtypes of type VI coa. The results showed that coa types of 154 out of 155 Staphylococcus aureus strains from various origins assigned by M-PCR:A and B were identical to those obtained by serological methods, leaving a serotype IV strain unclassifiable. All 73 type VI strains were classified into one of three subtypes by M-PCR:C. Furthermore, we found that type VIa and VIb strains carried characteristic pyrogenic toxin superantigen genes, while no toxin genes were identified in type VIc strains, suggesting the correlation between the subtype of type VI coa gene and the carriage of genomic islands. Our results showed that these M-PCRs are convenient methods for SC typing that might be useful for epidemiological studies. PMID- 18675307 TI - Unacknowledged contributions of Pavlov and Barcroft to Cannon's theory of homeostasis. AB - Cannon's theory of homeostasis is the first, major, American contribution to physiological thought. Although it is clear that Cannon's account of homeostasis is personal and based primarily on the work of his laboratory, Cannon made it easy for readers to mistake his 1929 paper and 1932 book for a comprehensive review of the literature relevant to homeostasis. This is unfortunate because Cannon never acknowledged the important contributions of two of his contemporaries, Ivan Pavlov and Joseph Barcroft. Since he did not mention them, their contributions are rarely discussed. This paper attempts to correct this historical problem in two ways. First, I describe the unacknowledged contributions of Pavlov and Barcroft. Then I consider the possible reasons why Cannon ignored them. PMID- 18675308 TI - Mannheimia haemolytica chimeric protein vaccine composed of the major surface exposed epitope of outer membrane lipoprotein PlpE and the neutralizing epitope of leukotoxin. AB - Mannheimia haemolytica is commonly identified in cattle with shipping fever pneumonia. Vaccines currently available do not provide complete protection against the disease. In an effort to develop a vaccine that delivers the immunogenic regions of leukotoxin (LKT) A and the outer membrane protein (OMP) PlpE, a total of four chimeric proteins were constructed. Mice were subcutaneously immunized with 25, 50 and 75 microg quantities of each chimeric protein. The specificity of the immune response was confirmed by Western blot analysis and enzyme-linked immunosorbent assays (ELISA). Moreover, the hyperimmune sera were bactericidal to M. haemolytica in the presence of complement and neutralized LKT. While all of the chimeric proteins induced some level of immune response two, SAC87 and SAC89, were most promising. These results demonstrate that a functional immune response against M. haemolytica can be induced by vaccination with recombinant chimeric proteins created from specific immunogenic regions of the LKT and PlpE proteins. PMID- 18675309 TI - Evolution of the dietary exposure to polycyclic aromatic hydrocarbons in Catalonia, Spain. AB - The concentrations of 16 polycyclic aromatic hydrocarbons (PAHs) were determined in samples of foodstuffs widely consumed by the population of Catalonia, Spain. All samples were randomly acquired in 12 cities of Catalonia between March and June of 2006, and analyzed by gas chromatography-mass spectrometry. The dietary intake of total and carcinogenic PAHs was estimated for various age/gender groups of population. In order to determine the temporal trend on the dietary exposure to PAHs, the current results were compared with those of a previous survey (2000). The highest individual PAH levels corresponded to phenanthrene (29.66 microg/kg), naphthalene (25.87 microg/kg) and fluoroanthene (13.66 microg/kg), while the lowest levels were benzo[a]pyrene (1.28 microg/kg), benzo[k]fluoranthene (1.31 microg/kg) and indeno[1,2,3-c,d]pyrene (1.44 microg/kg). According to food groups, the highest levels of total PAHs were detected in meat and meat products (25.56 microg/kg), oils and fats (23.48 microg/kg), and cereals (20.44 microg/kg). For an average male adult (70-kg body weight), the current dietary intake of the sum of PAHs was higher (12.0 microg/day) than that found in our 2000 survey (8.4 microg/day). PMID- 18675310 TI - Exploring the link between gender, sensation seeking, and family history of alcoholism in cortisol stress-response dampening. AB - Many studies have demonstrated an inverse association between cortisol and risk taking behaviors, with high-sensation seekers (HSS) showing lower cortisol levels. We investigated the potential link between sensation seeking (SS) and stress-induced stress responses, as well as alcohol-induced stress-response dampening (SRD) effects in cortisol. First, we hypothesized that HSS would show inverse SRD effects in cortisol. Second, we hypothesized that females would display similar SRD effects to males. Third, we hypothesized an independent relationship between SS and family history (FH) with regard to alcohol-induced SRD effects in cortisol. 86 healthy men and women participated in two laboratory sessions, receiving alcohol in one of the two. Experimental stress paradigms were administered and serum cortisol was measured. SRD effects in cortisol developed for both genders in low-sensation seekers (LSS), but not in HSS. This study contributes to current literature by (1) supporting the association between SS and cortisol, (2) demonstrating that SRD effects in cortisol of females is inversely related to SS, and (3) demonstrating an independent relationship between SS and FH with regard to alcohol-induced SRD effects in cortisol. PMID- 18675311 TI - In silico study of kinetochore control, amplification, and inhibition effects in MCC assembly. AB - Eukaryotic cells rely on a surveillance mechanism, the "Spindle Assembly Checkpoint"SACM in order to ensure accurate chromosome segregation by preventing anaphase initiation until all chromosomes are correctly attached to the mitotic spindle. In different organisms, a mitotic checkpoint complex (MCC) composed of Mad2, Bub3, BubR1/Mad3, and Cdc20 inhibits the anaphase promoting complex (APC/C) to initiate promotion into anaphase. The mechanism of MCC formation and its regulation by the kinetochore are unclear. Here, we constructed dynamical models of MCC formation involving different kinetochore control mechanisms including amplification as well as inhibition effects, and analysed their quantitative properties. In particular, in this system, fast and stable metaphase to anaphase transition can only be triggered when the kinetochore controls the Bub3:BubR1 related reactions; signal amplification and inhibition play a subordinate role. Furthermore, when introducing experimentally determined parameter values into the models analysed here, we found that effective MCC formation is not combined with complete Cdc20 sequestering. Instead, the MCC might bind and completely block the APC/C. The SACM might function by an MCC:APC/C complex rearrangement. PMID- 18675312 TI - Down-regulation of hepatic lipase expression by elevation of cAMP in human hepatoma but not adrenocortical cells. AB - Expression of hepatic lipase (HL) in the liver is reduced during prolonged fasting. This effect is mainly mediated via catecholamines, which signal through elevation of Ca(i)(2+) as well as cAMP. We have studied the effect of cAMP on HL expression in cell culture. Overnight incubation of HepG2 cells with 10-300microM 8-bromo-cyclic AMP resulted in a dose-dependent, up to 50% reduction in secretion of HL, but had no effect on secretion of alpha(1)-antitrypsin or overall protein synthesis. HL mRNA levels were decreased 1.5 fold, as determined by semi quantitative and real-time RT-PCR. In HepG2 cells transiently transfected with human HL (-685/+13) or rat HL (-446/+9) promoter-reporter constructs, cAMP induced a similar dose-dependent suppression of HL promoter activity. cAMP responsiveness in HepG2 cells was mediated by a conserved 10-bp response element at -45/-36, that represents a potential binding site for CCAAT/enhancer-binding protein beta (C/EBPbeta). cAMP reduced expression of the 45kDa C/EBPbeta protein and binding of C/EBPbeta to the proximal promoter region of the human HL gene by 50%, as determined by immunoblotting and chromatin immunoprecipitation assay, respectively. In human H295R adrenocortical cells, cAMP failed to suppress HL promoter activity, and only slightly reduced C/EBPbeta expression. We conclude that the fall in HL expression during prolonged fasting may be mediated through elevation of cAMP and lowering of C/EBPbeta expression. PMID- 18675313 TI - Purkinje-cell-preferential transduction by lentiviral vectors with the murine stem cell virus promoter. AB - Viral-vector-mediated gene delivery into Purkinje cells is a promising method for exploring the pathophysiology of the cerebellum; however, it is generally difficult to achieve sufficiently high levels of gene expression in Purkinje cells using viral vectors with a cell-type-specific promoter because of the weakness of transcriptional activity. In this study, we prepared lentiviral vectors that express GFP under the control of various ubiquitous promoters derived from murine stem cell virus (MSCV), cytomegalovirus (CMV), CMV early enhancer/chicken beta actin (CAG), and Rous sarcoma virus (RSV) and compared their potential to transduce Purkinje cells. Mice were sacrificed 7 days after lentiviral injection and the ratios of GFP(+) Purkinje cells to all transduced cells were determined. The highest transduction ratio was observed when we used lentivectors containing the MSCV promoter: approximately 70% of GFP(+) cells were Purkinje cells, the next highest ratio was for the CMV promoter (approximately 40%), then the CAG promoter (approximately 35%), and the lowest ratio was for the RSV promoter (approximately 10%). Moreover, the highest levels of GFP expression were also caused by the MSCV promoter. Thus, among the ubiquitous promoters examined, the MSCV promoter was the best for the expression of a foreign gene in Purkinje cells in vivo. PMID- 18675314 TI - Painful facet joint injury induces neuronal stress activation in the DRG: implications for cellular mechanisms of pain. AB - The cervical facet joint is implicated as one of the most common sources of chronic neck pain, owing to its rich nociceptive innervation and susceptibility to injurious mechanical loading. Injuries to the facet joint and its ligament can induce inflammation in the joint and spinal cord. Inflammatory molecules which are known to have a role in pain can also stimulate the integrated stress response (ISR). Therefore, we hypothesize that ISR is activated by facet joint injury in a rodent model of pain. To address this hypothesis, we assessed the expression of binding protein (BiP) (also known as growth-related protein 78 (GRP78)), a marker of endoplasmic reticulum stress response, in the dorsal root ganglion (DRG) after painful facet joint injury. In a rodent model of facet joint injury, dynamic distraction of the C6/C7 joint (injury, n=12) was imposed; sham procedures were performed separately (sham, n=8). Forepaw mechanical allodynia was assessed postoperatively for 7 days as a quantitative measure of pain symptoms. The C6 DRG was harvested and assessed for BiP expression using triple label immunofluorescent confocal microscopy and immunoblot analyses. BiP was significantly higher (p<0.001) in the DRG after injury than sham and was expressed predominantly in neurons. Similarly, quantification of BiP by immunoblot demonstrated a significant 2.1-fold increase (p=0.03) in injury compared to sham at day 7. Findings suggest neuronal stress activation is associated with painful facet joint injury, and that joint loading may directly mediate the behavior of DRG neurons in this class of injury. PMID- 18675315 TI - Electrophysiological correlates of adjustment process in anchoring effects. AB - Anchoring is a judgmental bias that final judgments are assimilated toward the starting point of the judge's deliberations. The anchoring-and-adjustment heuristic holds that anchoring bias is caused by insufficient adjustment. With the manipulation of some subjective factors, previous research found that anchoring is an effortful process. However, there is little evidence supporting that the effortful process is an adjustment process. In the present work, number accuracy was introduced as an objective factor which involves in an adjustment process. An event-related brain potential (ERP) experiment on young normal subjects examined the impact of number accuracy on anchoring processes responding to anchors which were generated by subjects themselves. A dot-image paradigm was firstly employed to explore anchoring effects. Behavioral results found less accurate anchors which determined a coarser mental scale diminished the anchoring biases responding to self-generated anchors. A positive deflection at 250-800 ms after target onset can be taken as a direct electrophysiological evidence of the adjustment process, whose amplitude was more positive on more accurate anchors condition. The present results further support that people adjust upwards or downwards on a mental scale from the self-generated anchor, which is consistent with the adjustment heuristics. PMID- 18675316 TI - Induction of cell proliferation and neuroblasts in the subgranular zone of the dentate gyrus by aqueous extract from Platycodon grandiflorum in middle-aged mice. AB - In the present study, we observed the neurogenic effects of an aqueous extract from the root of Platycodon grandiflorum (EPG) in middle-aged (12-month-old) mice. For this, 100mg/kg EPG was administered orally to mice for 30 days before sacrifice and 5-bromodeoxyguanosine (BrdU) was injected intraperitoneally every 8h for 24h on the day prior to sacrifice. The increase of neurogenesis was estimated by immunohistochemical staining for cellular proliferation markers (BrdU and Ki67) and a marker for neuroblasts (Doublecortin, DCX). These markers were detected in the subgranular zone of the dentate gyrus in vehicle- and EPG treated groups. The number of BrdU-, Ki67- and DCX-positive cells in the EPG treated group was significantly increased compared to that in the vehicle-treated group. In addition, DCX-positive cells in the EPG-treated group showed well developed processes. These results suggest that the number of neuroblasts is increased by the repeated treatment of EPG in middle-aged mice. PMID- 18675317 TI - Iron availability and infection. AB - BACKGROUND: To successfully sustain an infection, nearly all bacteria, fungi and protozoa require a continuous supply of host iron. METHODS: Literature review. RESULTS: Mechanisms of microbial iron acquisition are determinants for the kinds of cells, tissues and hosts in which pathogens can flourish. As a corollary, hosts possess an array of iron withholding devices whereby they can suppress or abort microbial invasions. GENERAL SIGNIFICANCE: Awareness of environmental and behavioral methods that can prevent iron loading plus development of pharmaceutical agents that can block microbial access to iron may help to reduce our dependence on antibiotics. PMID- 18675318 TI - Ectopic expression of mouse Sry interferes with Wnt/beta-catenin signaling in mouse embryonal carcinoma cell lines. AB - In mammals, Sry is the master regulator of male sex determination, although how it functions is still unclear. By contrast, female sex determination depends on the action of Rspo1 and Wnt4, the regulators of Wnt/beta-catenin signaling. To seek a possible interaction between male and female sex determination mechanisms, we examined whether Sry affects Wnt/beta-catenin signaling. Using the TOPFLASH reporter system to measure Lef/Tcf-dependent transcriptional activity, we showed that ectopic expression of mouse Sry strongly suppressed Wnt/beta-catenin signaling in mouse embryonal carcinoma and human embryonic kidney cell lines. This inhibition occurred downstream of beta-catenin but upstream of Lef/Tcf, and depended on both the HMG-box and the C-terminal transcriptional activation domain. By contrast, TOPFLASH was not inhibited by human SRY, which apparently lacks a transcriptional activation domain. However, a fusion construct consisting of human SRY attached to the C-terminal domain of mouse Sry was able to inhibit TOPFLASH effectively. Furthermore, Sry constructs carrying point mutations equivalent to those in human sex reversal mutations were less effective in inhibiting Wnt/beta-catenin signaling. Also, we showed that the action of Sry as a transcriptional activator was both necessary and sufficient to inhibit Wnt/beta catenin signaling, suggesting that the transcriptional targets of Sry are responsible for the inhibition of signaling. Sox9 is a potential transcriptional target of Sry, although quantitative RT-PCR analysis indicates that the expression of Sox9 was not up-regulated by the ectopic expression of mouse Sry in mouse embryonal carcinoma cells. While the present study demonstrates an impact of mouse Sry on Wnt/beta-catenin signaling at an in vitro level, it requires further investigations to assess whether such action also takes place in vivo to regulate male sex determination. PMID- 18675319 TI - Caenorhabditis elegans galectins LEC-1-LEC-11: structural features and sugar binding properties. AB - Galectins form a large family of beta-galactoside-binding proteins in metazoa and fungi. This report presents a comparative study of the functions of potential galectin genes found in the genome database of Caenorhabditis elegans. We isolated full-length cDNAs of eight potential galectin genes (lec-2-5 and 8-11) from a lambdaZAP cDNA library. Among them, lec-2-5 were found to encode 31-35-kDa polypeptides containing two carbohydrate-recognition domains similar to the previously characterized lec-1, whereas lec-8-11 were found to encode 16-27-kDa polypeptides containing a single carbohydrate-recognition domain and a C-terminal tail of unknown function. Recombinant proteins corresponding to lec-1-4, -6, and 8-10 were expressed in Escherichia coli, and their sugar-binding properties were assessed. Analysis using affinity adsorbents with various beta-galactosides, i.e., N-acetyllactosamine (Galbeta1-4GlcNAc), lacto-N-neotetraose (Galbeta1 4GlcNAcbeta1-3Galbeta1-4Glc), and asialofetuin, demonstrated that LEC-1-4, -6, and -10 have a significant affinity for beta-galactosides, while the others have a relatively lower affinity. These results indicate that the integrity of key amino acid residues responsible for recognition of lactose (Galbeta1-4Glc) or N acetyllactosamine in vertebrate galectins is also required in C. elegans galectins. However, analysis of their fine oligosaccharide-binding properties by frontal affinity chromatography suggests their divergence towards more specialized functions. PMID- 18675320 TI - Tactile allodynia can occur in the spared nerve injury model in the rat without selective loss of GABA or GABA(A) receptors from synapses in laminae I-II of the ipsilateral spinal dorsal horn. AB - Although there is evidence that reduced inhibition in the spinal dorsal horn contributes to neuropathic pain, the mechanisms that underlie this are poorly understood. We have previously demonstrated that there is no loss of neurons from laminae I-III in the spared nerve injury (SNI) model [Polgar E, Hughes DI, Arham AZ, Todd AJ (2005) Loss of neurons from laminas I-III of the spinal dorsal horn is not required for development of tactile allodynia in the SNI model of neuropathic pain. J Neurosci 25:6658-6666]. In this study we investigated whether there was a difference between ipsilateral and contralateral sides in the levels of GABA, the vesicular GABA transporter (VGAT), or the beta3 subunit of the GABA(A) receptor at synapses in the medial part of the superficial dorsal horn in this model. Tissue from rats that had undergone SNI 4 weeks previously was examined with an electron microscopic immunogold method to reveal GABA, following pre-embedding detection of GABA(A) beta3 to allow identification of GABAergic terminals. Assessment of labeling for the GABA(A) beta3 subunit and VGAT was performed by using immunofluorescence and confocal microscopy. We found no difference in the intensity of immunolabeling for any of these markers on the two sides of the superficial dorsal horn. These results suggest that there is no significant loss of GABAergic boutons from the denervated area after SNI (which is consistent with the finding that neuronal death does not occur in this model) and that there is no depletion of GABA or GABA(A) receptors at GABAergic synapses within this region. An alternative explanation for disinhibition after nerve injury is that it results from reduced excitatory drive to GABAergic dorsal horn neurons following loss of primary afferent input to these cells. PMID- 18675321 TI - NF-kappaB signaling in cerebral ischemia. AB - The transcription factor NF-kappaB is a key regulator of hundreds of genes involved in cell survival and inflammation. There is ample evidence that NF kappaB is activated in cerebral ischemia, mainly in neurons. Despite its well known role as an antiapoptotic factor, in cerebral ischemia NF-kappaB contributes to neuronal cell death, at least if the ischemia is severe enough to lead to irreversible brain damage. In contrast, NF-kappaB also seems to be responsible for the preconditioning effect of a transient and sublethal ischemia, perhaps by dampening its own subsequent full activation. Among the five NF-kappaB subunits, RelA and p50 are responsible for the detrimental effect in cerebral ischemia. Activation of NF-kappaB signaling is mediated by the upstream kinase inhibitor of kappaB kinase and is triggered by hypoxia, reactive oxygen species, and several inflammatory mediators. Interestingly, the complex NF-kappaB signaling pathway provides drug targets at several levels. Modulation of NF-kappaB signaling has the potential to interrupt multiple inflammatory and apoptotic mechanisms through one specific molecular target. PMID- 18675322 TI - A protein kinase C activity localized to neuropeptide Y-like neurons mediates ethanol intoxication in Drosophila melanogaster. AB - Neuropeptide Y (NPY) regulates acute ethanol sensitivity and voluntary alcohol consumption in rodents. In Drosophila melanogaster, NPY-like neuropeptide F (NPF) and its receptor NPFR1 display a parallel function, suggesting that an evolutionarily conserved mechanism may underlie similar behavioral effects of ethanol in diverse organisms. We have used the fly model to uncover novel genes and molecular pathways important for acute ethanol response. Here we report a critical role of the conserved protein kinase C (PKC) pathway in mediating the intoxicating effect of ethanol. Flies expressing a pseudo-substrate inhibitor of PKC, directed by npf-gal4, displayed decreased ethanol sensitivity. Furthermore, the RNA interference analysis suggests that a calcium-independent PKC isoform (PKC98E, related to mammalian novel PKCs) is largely responsible for the behavioral phenotype. Finally, we provide evidence that the NPF/PKC-dependent mechanism selectively affects acute sensitivity but not rapid tolerance to ethanol intoxication. These findings reveal an uncharacterized role of PKC in NPY/NPF-mediated acute ethanol sensitivity in flies and possibly mammals. PMID- 18675325 TI - Identification and characterization of Rhox13, a novel X-linked mouse homeobox gene. AB - Homeobox genes encode transcription factors whose expression organizes programs of development. A number of homeobox genes expressed in reproductive tissues have been identified recently, including a colinear cluster on the X chromosome in mice. This has led to an increased interest in understanding the role(s) of homeobox genes in regulating development of reproductive tissues including the testis, ovary, and placenta. Here we report the identification and characterization of a novel homeobox gene of the paired-like class on the X chromosome distal to the reproductive homeobox (Rhox) cluster in mice. Transcripts are found in the testis and ovary as early as 13.5 days post coitum (dpc). Transcription ceases in the ovary by 3 days post partum (dpp), but continues in the testis through adulthood. The Rhox13 gene encodes a 25.3 kDa protein expressed in the adult testis in germ cells at the basal aspect of the seminiferous epithelium. PMID- 18675323 TI - Distinct mechanisms of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyrimidine resistance revealed by transcriptome mapping in mouse striatum. AB - The etiology of idiopathic Parkinson's disease is thought to involve interplay between environmental factors and predisposing genetic traits, although the identification of genetic risk factors remain elusive. The neurotoxicant, 1 methyl-4-phenyl-1,2,3,6-tetrahydropyrimidine (MPTP) produces parkinsonian-like symptoms and pathology in mice and humans. As sensitivity to MPTP is genetically determined in mice this provides an opportunity to identify genes and biological mechanisms that modify the response to an exogenous agent that produces a Parkinson's disease-like condition. MPTP primarily targets dopaminergic nerve terminals in the striatum and elicits changes in striatal gene expression. Therefore, we used Affymetrix and qRT-PCR technology to characterize temporal mRNA changes in striatum in response to MPTP in genetically MPTP-sensitive, C57BL/6J, and MPTP-resistant Swiss Webster and BCL2-associated X protein (Bax)-/- mice. We identified three phases of mRNA expression changes composed of largely distinct gene sets. An early response (5 h) occurred in all strains of mice and multiple brain regions. In contrast, intermediate (24 h) and late (72 h) phases were striatum specific and much reduced in Swiss Webster, indicating these genes contribute and/or are responsive to MPTP-induced pathology. However, Bax-/- mice have robust intermediate responses. We propose a model in which the acute entry of MPP+ into dopaminergic nerve terminals damages them but is insufficient per se to kill the neurons. Rather, we suggest that the compromised nerve terminals elicit longer lasting transcriptional responses in surrounding cells involving production of molecules that feedback on the terminals to cause additional damage that results in cell death. In Swiss Webster, resistance lies upstream in the cascade of events triggered by MPTP and uncouples the acute events elicited by MPTP from the damaging secondary responses. In contrast, in Bax-/- mice resistance lies downstream in the cascade and suggests enhanced tolerance to the secondary insult rather than its attenuation. PMID- 18675324 TI - Activity of protein kinase C is important for 3alpha,5alpha-THP's actions at dopamine type 1-like and/or GABAA receptors in the ventral tegmental area for lordosis of rats. AB - In the ventral tegmental area, progestogens facilitate sexual receptivity of rodents via actions at dopamine type 1-like and/or gamma-aminobutyric acid type A receptors and activation of downstream signal transduction molecules. In the present study, we investigated whether effects of progesterone's metabolite, 3alpha,5alpha-THP, to enhance lordosis via actions at these receptors in the ventral tegmental area requires phospholipase C-dependent protein kinase C. The objective of this study was to test the hypothesis that: if progestogens' actions through dopamine type 1-like and/or gamma-aminobutyric acid type A receptors in the ventral tegmental area for lordosis require protein kinase C, then inhibiting protein kinase C in the ventral tegmental area should reduce 3alpha,5alpha-THP facilitated lordosis and its enhancement by dopamine type 1-like or gamma aminobutyric acid type A receptor agonists. Ovariectomized, estradiol (E(2); 10 microg s.c. at h 0)-primed rats were tested for their baseline lordosis responses and then received a series of three infusions to the ventral tegmental area: first, bisindolylmaleimide (75 nM/side) or vehicle; second, SKF38393 (100 ng/side), muscimol (100 ng/side), or vehicle; third, 3alpha,5alpha-THP (100, 200 ng/side) or vehicle. Rats were pre-tested for lordosis and motor behavior and then tested for lordosis after each infusion and 10 and 60 min after the last infusion. Rats were tested for motor behavior following their last lordosis test. As has been previously demonstrated, 3alpha,5alpha-THP infusions to the ventral tegmental area increased lordosis and effects were further enhanced by infusions of SKF38393 and muscimol. Infusions of bisindolylmaleimide to the ventral tegmental area attenuated 3alpha,5alpha-THP-, SKF38393-, and/or muscimol facilitated lordosis. Effects on lordosis were not solely due to changes in general motor behavior. Thus, 3alpha,5alpha-THP's actions in the ventral tegmental area through membrane receptors may require activity of protein kinase C. PMID- 18675326 TI - Two new members of the Tetrahymena multi-stress-inducible metallothionein family: characterization and expression analysis of T. rostrata Cd/Cu metallothionein genes. AB - We report the cloning and characterization of two new metallothionein (MT) genes (TrosMTT1 and TrosMTT2), isolated as cDNAs, from the ciliated protozoa Tetrahymena rostrata. The TrosMTT1 inferred protein has been identified as a CdMT and included into the 7a subfamily of Tetrahymena MTs, while TrosMTT2 has been identified as a CuMT (including it into 7b subfamily), due to its similarity to TpigMT-2 and its significant induction by copper. TrosMTT1 protein sequence reveals a remarkably regular and hierarchical modular organization, as it is known for other Tetrahymena CdMTs, showing a bi-modular structure. TrosMTT2 presents a structural organization based on CKCX(2-5)CKC repeats, like it occurs in other Tetrahymena CuMTs, indicating that an evolutionary history based on intra-gene duplications might be also possible. Both are also multi-stress inducible genes because they are induced by other heavy metals and stressors, as it has been shown by quantitative real-time RT-PCR. It is the first time that the gene expression of a putative Tetrahymena CuMT is analyzed by quantitative PCR, confirming it as a CuMT. These two new Tetrahymena MTs complete, at present, the actual view of this protein superfamily, and corroborate the unique features of ciliate MTs. Furthermore, both, a comparative analysis of relative gene expression values obtained by quantitative RT-PCR on other Tetrahymena MT genes and an analysis of the different Tetrahymena MTs based on the different Cys clusters of these proteins are carried out, which show an update view of Tetrahymena MT gene family. PMID- 18675327 TI - Molecular cloning and characterization of the sheep malic enzyme cDNA. AB - Malic enzyme catalyzes decarboxylation of malate to pyruvate and CO(2), providing de novo biosynthesis of fatty acids with NADPH. Since lipogenesis in ruminants, contrarily to some monogastric species like human and rodents, occurs predominantly in adipose tissue, the activity of many lipogenic enzymes is higher in adipose tissue compared to liver. Expression of malic enzyme is regulated by nutrition; refeeding after a period of starvation results to an induction of the enzyme. Here we present the nucleotide sequence of two transcripts of the ovine cytosolic malic enzyme gene that differ at the length of the 3' UTR. These are the first published cDNA sequences for ruminant species and share high similarity with the corresponding sequences of other species. Malic enzyme mRNA was present in every ovine tissue that was examined. In agreement with the fact that adipose tissue is the major lipogenic site for ruminants, mRNA levels in adipose tissue were higher than in liver. Refeeding after two weeks of caloric restriction resulted in a two-fold increase of the mRNA level of malic enzyme in adipose tissue. PMID- 18675328 TI - Molecular cloning and sequencing of metallothionein in squamates: new insights into the evolution of the metallothionein genes in vertebrates. AB - Metallothioneins are cysteine-rich, metal-binding proteins ubiquitously expressed in living organisms. In the last past years, a plethora of vertebrate metallothionein sequences have become available, but so far there has been an almost absolute lack of data about sequences of metallothionein of non-avian diapsida. In the framework of the investigations on structural and functional properties of non-mammalian metallothioneins, we have cloned and sequenced the cDNAs encoding for metallothioneins of 10 squamate reptiles, belonging to 5 different infraorders. These sequences have been used to gain insight into the evolutionary history of metallothioneins in reptiles. Phylogenetic analysis shows that reptilian metallothionein phylogeny is inconsistent with the species phylogeny. Such findings allow us to hypothesize that the identified metallothionein in each squamate species used for this study might be considered a paralogous gene derived from more events of gene duplication and losses occurred during the diversification of the squamate species. Finally, through vertebrate metallothionein comparisons and phylogenetic analysis, we also add a novel contribution to the understanding of the evolution of metallothionein genes along the major vertebrate lineages. PMID- 18675329 TI - Bindin genes of the Pacific oyster Crassostrea gigas. AB - When Crassostrea gigas oyster sperm acrosome react a ring of bindin protein is exposed that bonds the sperm to the egg vitelline envelope. The putative functional unit of bindin is a fucose lectin (F-lectin) domain that is structurally conserved among phyla. There is only one bindin gene in C. gigas, which can possess 1-5 tandem F-lectin repeats. Alternative splicing can alter the number of repeats per bindin mRNA. Recombination occurs in a highly variable intron in the middle of each F-lectin repeat to create many different lectin domain sequences [Moy, G.W., Springer, S.A., Adams, S.L., Swanson, W.J., Vacquier, V.D., 2008. Extraordinary intraspecific diversity in oyster sperm bindin. Proc. Natl. Acad. Sci. U.S.A. 105, 1993-1998]. Two bindin genes were sequenced to learn more about bindin introns. The first gene (6914 bp) contained one F-lectin repeat. The second gene (25,932 bp) contained three tandem F-lectin repeats. Four of the introns in this larger gene are conserved in size among individuals. However, the one intron in each F-lectin repeat is highly variable in size and sequence, indicating that it has been a hot spot for recombination. A retroposon with high reverse transcriptase homology is present in the three repeat gene immediately upstream of the first F-lectin repeat, suggesting that retroposition is one mechanism by which F-lectin repeats are duplicated. The retroposon is not present in the one F-lectin repeat bindin gene. Three GA microsatellites, one in each intron immediately upstream of the start of each F lectin repeat exon, and one downstream CT microsatellite, suggest that loopout strand hybridization can occur, and lectin repeats replicate and transpose within the gene. The CT microsatellite is not found in the one F-lectin repeat containing gene. Oysters appear to use every possible mechanism to create variation in the F-lectin domains of sperm bindin. This is presumably in response to sexual conflict that operates in the prevention of polyspermy. PMID- 18675330 TI - Activation of MAPK/AP-1 signaling pathway in lung injury induced by 2-chloroethyl ethyl sulfide, a mustard gas analog. AB - We reported earlier that the activation of free-radical-mediated tumor necrosis factor-alpha (TNF-alpha) cascade is the major pathway in the inflammatory lung disease induced by 2-chloroethyl ethyl sulfide (CEES), a mustard gas analog. TNF alpha induces activating protein 1 (AP-1) activation via phosphorylation of mitogen activated protein kinases (MAPKs). The present study examines the relationship between CEES induced lung injury and MAPKs signaling pathway. Adult guinea pigs received single intratracheal injection of different doses of CEES and were sacrificed at different time points. CEES exposure caused lung injury with evidence of fibrosis. The optimum activation of all members of the MAPKs family (ERK1/2, p38 and JNK1/2) was achieved at 0.5 mg/kg dose and at 1h. No significant change was observed beyond that time point. This led to an activation of AP-1 transcription factors associated with an increase in the protein levels of Fos, activating transcription factor (ATF) and Jun family members. To explore the involvement of AP-1 in cell proliferation, we determined the protein levels of cell cycle protein cyclin D1 and cell differentiation marker proliferating cell nuclear antigen (PCNA). An up regulation of these proteins was observed. Hence it is suggested that CEES exposure causes accumulation of TNF-alpha, which is associated with an activation of MAPK/AP-1 signaling pathway and cell proliferation. Further studies are needed to clarify whether the observed effects are the adaptive responses of the lung or they contribute to the lung injury. PMID- 18675331 TI - Effects of 3-benzidino-6-phenylpyridazine, as an acetylcholinesterase inhibitor, on outward potassium current in acutely isolated rat hippocampal pyramidal neurons. AB - As an acetylcholinesterase (AChE) inhibitor, the effects of 3-benzidino-6 phenylpyridazine (BPP) on outward potassium current including delayed rectifier potassium current (I(K(DR))) and transient outward potassium current (I(K(A))) in acutely isolated rat hippocampal pyramidal neurons were studied, using the whole cell patch-clamp technique. BPP reversibly inhibited electric eel AChE as an inhibitor, with IC(50) of 1.43 microM. BPP (0.10-100 microM) decreased I(K(DR)) and I(K(A)) in a concentration-dependent, voltage-independent and partial reversible manner, with IC(50) of 0.47 and 0.31 microM, respectively. 10 microM BPP did not affect steady-state activation of I(K(DR)) and I(K(A)). In addition, 10 microM BPP shifted the voltage dependence of steady-state inactivation of I(K(A)) towards negative potential. In conclusion, BPP potently inhibits I(K(DR)) and I(K(A)) in rat hippocampal pyramidal neurons, which may contribute to BPP's restoring the damaged central nervous system. PMID- 18675332 TI - Proteomic analysis of human breast cell lines using SELDI-TOF MS shows that mixtures of estrogenic compounds exhibit simple similar action (concentration additivity). AB - Endocrine modulating chemicals in the environment are possible causative agents of a number of human diseases. Many of these compounds act on the same molecular target, and hence risk assessment requires consideration of their possible combined effects. Here, we studied the combined effects of 17beta-estradiol, genistein, bisphenol A and endosulfan on MCF-7 cells. Full concentration-effect curves for cell proliferation were obtained for each compound and used to identify appropriate concentrations for investigating the effects of binary mixtures of the compounds. Protein profiling by surface-enhanced laser desorption/ionization time of flight mass spectrometry was performed to identify responsive proteins. Treatment with each of the compounds produced similar protein profile changes. Prominent and consistent changes were seen in 12 protein ions. Cell proliferation and protein levels responded monotonically to the estrogens, with identifiable no observable effect concentrations in all cases. Binary mixtures of the compounds produced effects on cell proliferation and on each of the responsive protein ions that were fully consistent with concentration additivity. Thus, no reason to deviate from the application of the principles of dose-response and dose additivity in the risk assessment of combinations of estrogenic compounds was found in this study. PMID- 18675333 TI - Role of complexes formation between drugs and penetration enhancers in transdermal delivery. AB - The use of chemical penetration enhancers (CPE) is growing due to their ability to improve drug delivery through the skin. A possible mechanism of penetration enhancement could involve the complex formation between drug and components in the pharmaceutical formulation, thus altering the physicochemical properties of the active substance. Here, modelling studies indicate that hydrocarbon and oxygen-containing terpenes (penetration enhancers) could form complexes with drugs. Satisfactory correlations have been obtained between the predicted molecular properties of enhancers and their enhancement effects. PMID- 18675334 TI - Long-living growth hormone receptor knockout mice: potential mechanisms of altered stress resistance. AB - Endocrine mutant mice have proven invaluable toward the quest to uncover mechanisms underlying longevity. Growth hormone (GH) and insulin-like growth factor (IGF) have been shown to be key players in physiological systems that contribute to aging processes including glucose metabolism, body composition and cellular protection. Examination of these mutant mice across several laboratories has revealed that differences exist in both the direction and magnitude of change, differences that may result in variation in life span. Growth hormone receptor knockout mice lack a functional GH receptor, therefore GH signaling is absent. These mice have been shown to lack the heightened oxidative defense mechanisms observed in other GH mutants yet live significantly longer than wild type mice. In this study, glutathione (GSH) and methionine (MET) metabolism was examined to determine the extent of variation in this mutant in comparison to the Ames dwarf, a mouse that exhibits delayed aging and life span extension of nearly 70%. Components of GSH and MET were altered in GHR KO compared to wild type controls. The results of these experiments suggest that these pathways may be partially responsible for differences observed in stress resistance and the capacity to respond to stressors, that in the long term, affect health and life span. PMID- 18675336 TI - Toxic effects of DDT and methyl mercury on the hepatocytes from Hoplias malabaricus. AB - Here, we examined the impact of dichlorodiphenyltrichloroethane (DDT) and monomethyl mercury (MeHg) on the redox milieu and survival of hepatocytes from Hoplias malabaricus (traira). After isolation and attachment of cells, we established one control and four treatments: DDT (50nM of DDT), MeHg I (0.25microM of MeHg), MeHg II (2.5microM of MeHg) and DDT * MeHg I (combination of 50nM of DDT and 0.25microM of MeHg). After four days the exposed hepatocytes presented significantly increased damage in lipids (all treatments), proteins (DDT * MeHg I and MeHg II) and reduced cell viability (all treatments). Also the antioxidant enzymes catalase, glucose-6-phosphate dehydrogenase (G6PDH), glutathione reductase and superoxide dismutase were affected. The current data showed that despite of some protective responses, the increased disturbs on membrane lipids and proteins, increased hydrogen peroxide levels, and decreased glutathione concentration and cell viability strongly indicate oxidative stress as the reason of hepatotoxicity due to DDT and MeHg exposure. In addition, DDT and MeHg together had greater effect than alone when G6PDH and glutathione-S transferase activities and lipids damage were considered. These findings are indicative of hepatotoxicity occurring at realistic concentrations of DDT and MeHg found in Amazonian fish tissues. PMID- 18675335 TI - A novel prfA mutation that promotes Listeria monocytogenes cytosol entry but reduces bacterial spread and cytotoxicity. AB - Listeria monocytogenes is an environmental bacterium that becomes a pathogen following ingestion by a mammalian host. The transition from environmental organism to pathogen requires significant changes in gene expression, including the increased expression of gene products that contribute to bacterial growth within host cells. PrfA is an L. monocytogenes transcriptional regulator that becomes activated upon bacterial entry into mammalian cells and induces the expression of gene products required for virulence. How PrfA activation occurs is not known, however several mutations have been identified that increase PrfA activity in strains grown in vitro (prfA mutations). Here we describe a novel prfA mutation that enhances extracellular PrfA-dependent gene expression but in contrast to prfA mutants inhibits the cytosol-mediated induction of virulence genes. prfA Y154C strains entered cells and escaped from phagosomes with an efficiency similar to wild type bacteria, however the mutation prevented efficient L. monocytogenes actin polymerization and reduced spread of bacteria to adjacent cells. The prfA Y154C mutation severely attenuated bacterial virulence in mice but the mutant strains did generate target antigen specific CD8(+) effector cells. Interestingly, the prfA Y154C mutant was significantly less cytotoxic for host cells than wild type L. monocytogenes. The prfA Y154C mutant strain may therefore represent a novel attenuated strain of L. monocytogenes for antigen delivery with reduced host cell toxicity. PMID- 18675337 TI - Use of the rat postimplantation embryo culture to assess the embryotoxic potency within a chemical category and to identify toxic metabolites. AB - The implementation of in vitro alternatives in the safety evaluation of chemicals in the animal intensive area of reproductive toxicity testing is highly desirable, but has been limited by issues around predictivity and applicability domains. The validation of alternatives may gain from a category approach, in which, rather than validating a test for the universe of chemicals, its predictive value is assessed for each class of chemicals for which the test represents relevant end point(s). We studied the embryotoxicity in rodent postimplantation whole embryo culture (WEC) of a series of phthalates and their metabolites. Phthalate diesters are widely applied industrial chemicals, their monoester derivatives being considered as their embryotoxic metabolites. The relative in vitro potency of three out of four monophthalates was found to mimick that of corresponding diphthalates tested in vivo. The phthalate that deviated from this ranking, monoethylhexylphthalate (MEHP), showed a relatively high in vitro toxicity as compared to in vivo data. This deviation could be explained through kinetic differences among phthalates, as shown between MEHP and monobutylphthalate. In addition, in vitro testing of specific secondary MEHP metabolites showed that they were all less potent than MEHP. This finding confirmed that MEHP in vitro embryotoxicity is most likely the best correlate to DEHP in vivo embryotoxicity. This study shows that a category approach in the assessment of the validation of in vitro alternatives is feasible, and can be improved when kinetic considerations are taken into account. PMID- 18675338 TI - Exploring the interaction of microcystin-LR with proteins and DNA. AB - The physiological role of microcystin-LR is still under discussion, and since binding of microcystin-LR to proteins different from their main cellular targets was described, we have performed experiments in order to explore this interaction. A non-specific interaction of microcystin-LR with a variety of soluble proteins in vitro is disrupted when using organic solvents such as methanol. The isoelectric point of proteins is not affected by their interaction with microcystin-LR, even though the presence of microcystin-LR alters the pool of peptides obtained by tryptic digestions. Under the conditions tested, microcystin-LR does not exhibit affinity for DNA. Although it is unlikely that the non-specific binding of microcystin-LR to proteins has a physiological meaning, one must be aware of the fact that determinations of the toxin extracted from any biological sample may be affected by the presence of proteins in the extracts. Consequently, we strongly recommend use organic solvents and to lyophilise the tissue samples to guarantee the accessibility of these organic solvents to microcystin-LR when performing experiments with tissue or cell extracts. PMID- 18675339 TI - Ascorbate sustains neutrophil NOS expression, catalysis, and oxidative burst. AB - Previous studies from this lab have demonstrated that in vitro ascorbate augments neutrophil nitric oxide (NO) generation and oxidative burst. The present study was therefore undertaken in guinea pigs to further assess the implication of ascorbate deficiency in vivo on neutrophil ascorbate and tetrahydrobiopterin content, NOS expression/activity, phagocytosis, and respiratory burst. Ascorbate deficiency significantly reduced ascorbate and tetrahydrobiopterin amounts, NOS expression/activity, and NO as well as free radical generation in neutrophils from scorbutics. Ascorbate and tetrahydrobiopterin supplementation in vitro, though, significantly enhanced NOS catalysis in neutrophil lysates and NO generation in live cells, but could not restore them to control levels. Although phagocytic activity remained unaffected, scorbutic neutrophils were compromised in free radical generation. Ascorbate-induced free radical generation was NO dependent and prevented by NOS and NADPH oxidase inhibitors. Augmentation of oxidative burst with dehydroascorbate (DHA) was counteracted in the presence of glucose (DHA uptake inhibitor) and iodoacetamide (glutaredoxin inhibitor), suggesting the importance of ascorbate recycling in neutrophils. Ascorbate uptake was, however, unaffected among scorbutic neutrophils. These observations thus convincingly demonstrate a novel role for ascorbate in augmenting both NOS expression and activity in vivo, thereby reinforcing oxidative microbicidal actions of neutrophils. PMID- 18675340 TI - Role of NADPH oxidase in the apoptotic death of cultured cerebellar granule neurons. AB - Cerebellar granule neurons (CGN) cultured in a medium containing 25 mM KCl and treated with staurosporine (ST) or transferred to a medium with 5 mM KCl (K5) die apoptotically. CGN death is mediated by an increase in reactive oxygen species (ROS) production. When CGN are treated with antioxidants all apoptotic parameters and cell death are markedly diminished, showing a central role for ROS in this process. Recently, it has been suggested that a possible ROS source involved in cell death is a NADPH oxidase. In that regard, we found expression in CGN of the components of NADPH proteins, p40phox, p47phox and p67phox, and p22phox, as well as three homologues of the catalytic subunit of this complex, NOX1, 2, and 4. The inhibition of NADPH oxidase with diphenylene iodonium or 4-(2 aminoethyl)benzenesulfonyl fluoride significantly reduced ROS production, NADPH oxidase activity, all the apoptotic events, and cell death induced by both K5 and ST. We conclude that ROS could be an early signal of apoptotic neuronal death and that NADPH oxidase, including NOX1, 2, and/or 4, could have a central role in apoptotic death induced by different conditions in these neurons. PMID- 18675342 TI - WITHDRAWN: SOCS3 mediates IL-1beta-dependent inhibition of IL-6-induced gene expression. AB - This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologises for any inconvenience this may cause. PMID- 18675341 TI - Dynamic interaction between Arf GAP and PH domains of ASAP1 in the regulation of GAP activity. AB - ASAP family Arf GAPs induce the hydrolysis of GTP bound to the Ras superfamily protein Arf1, regulate cell adhesion and migration and have been implicated in carcinogenesis. The ASAP proteins have a core catalytic domain of PH, Arf GAP and Ank repeat domains. The PH domain is necessary for both biological and catalytic functions of ASAP1 and has been proposed to be integrally folded with the Arf GAP domain. Protection studies and analytical ultracentrifugation studies previously reported indicated that the domains are, at least partly, folded together. Here, using NMR spectroscopy and biochemical analysis, we have further tested this hypothesis and characterized the interdomain interaction. A comparison of NMR spectra of three recombinant proteins comprised of either the isolated PH domain of ASAP1, the Arf GAP and ankyrin repeat domain or all three domains indicated that the PH domain did interact with the Arf GAP and Ank repeat domains; however, we found a significant amount of dynamic independence between the PH and Arf GAP domains, consistent with the interactions being transient. In contrast, the Arf GAP and Ank repeat domains form a relatively rigid structure. The PH-Arf GAP domain interaction partially occluded the phosphoinositide binding site in the soluble protein, but binding studies indicated the PIP2 binding site was accessible in ASAP1 bound to a lipid bilayer surface. Phosphoinositide binding altered the conformation of the PH domain, but had little effect on the structure of the Arf GAP domain. Mutations in a loop of the PH domain that contacts the Arf GAP domain affected PIP2 binding and the K(m) and k(cat) for converting Arf1 GTP to Arf1 GDP. Based on these results, we generated a homology model of a composite PH/Arf GAP/Ank repeat domain structure. We propose that the PH domain contributes to Arf GAP activity by either binding to or positioning Arf1 GTP that is simultaneously bound to the Arf GAP domain. PMID- 18675343 TI - Gene therapy of Hunter syndrome: evaluation of the efficiency of muscle electro gene transfer for the production and release of recombinant iduronate-2-sulfatase (IDS). AB - Mucopolysaccharidosis type II (MPSII) is an inherited disorder due to a deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS). The disease is characterized by a considerable deposition of heparan- and dermatan-sulfate, causing a general impairment of physiological functions. Most of the therapeutic protocols proposed so far are mainly based upon enzyme replacement therapy which is very expensive. There is a requirement for an alternative approach, and to this aim, we evaluated the feasibility of muscle electro gene transfer (EGT) performed in the IDS-knockout (IDS-ko) mouse model. EGT is a highly efficient method of delivering exogenous molecules into different tissues. More recently, pre-treatment with bovine hyaluronidase has shown to further improve transfection efficiency of muscle EGT. We here show that, by applying such procedure, IDS was very efficiently produced inside the muscle. However, no induced IDS activity was measured in the IDS-ko mice plasma, in contrast to matched healthy controls. In the same samples, an anticipated and rapidly increasing immune response against the recombinant protein was observed in the IDS-ko vs control mice, although reaching the same levels at 5 weeks post-injection. Additional experiments performed on healthy mice showed a significant contribution of hyaluronidase pre treatment in increasing the immune response. PMID- 18675344 TI - Relative bioavailability of griseofulvin lyophilized dry emulsion tablet vs. immediate release tablet: a single-dose, randomized, open-label, six-period, crossover study in healthy adult volunteers in the fasted and fed states. AB - The oral bioavailability of griseofulvin (GF) formulated as a fast disintegrating lyophilized dry emulsion (LDE) tablet was studied and compared to the commercially available immediate release (IR) tablet, as a reference, in both the fasted and fed states in nine healthy volunteers after a single oral dose (125 mg) in a crossover design. Furthermore the LDE tablets were ingested with and without water under both the fasted and fed states. In the fasted state, the rate of absorption was found to be significantly faster from LDE tablets, in the presence and absence of water, as shown by a higher C(max) (more than two times higher, p=0.0001) and a shorter t(max) (by more than 3h, p=0.0001) compared to IR tablets. The extent of absorption, expressed as AUC, from LDE tablets in the presence and absence of water was 65% and 77% larger and statistically significantly different relative to the mean AUC from IR tablets (p=0.006). In the fed state, C(max) from LDE tablets ingested with and without water was found to be about 30% and 50% higher, respectively, than the immediate release tablets. A shorter t(max) was also shown whether LDE tablets were ingested with or without water in the fed state as compared to immediate release tablets. The mean AUC from LDE tablets under fed conditions in the presence of water was about 21% larger and was not statistically significantly different from AUC from immediate release tablets (p=0.517). When ingested without water, AUC from LDE tablets was about 43% larger and statistically significantly different relative to AUC from IR tablets (p=0.033). The mean AUC from the LDE tablet ingested with water under fed conditions relative to AUC from LDE tablet ingested without water was not statistically significantly different (p=0.454). Results show that the food effect of the high fat meal is very pronounced in case of the immediate release tablets, Fulvin, than in case of LDE tablets whether given with or without water. PMID- 18675345 TI - Role of KIRs and KIR ligands in hematopoietic transplantation. AB - This review focuses on recent research demonstrating the role alloreactive natural killer (NK) cells play in adoptive immunotherapy of leukemia in allogeneic hematopoietic transplantation. For patients with hematologic malignancies and an indication to allogeneic hematopoietic transplantation who do not have a matched sibling donor, unrelated donor, or cord blood transplants are almost always available (as long as the patient's ethnicity is represented in the donor registries). However, up to one half of patients relapse and do not make it to transplant during the time required for the donor search, completion of donor HLA typing, bone marrow harvest, and shipment. Donor-versus-recipient NK cell alloreactivity is effected by a functional repertoire of NK cells which express inhibitory Killer-Cell Immunoglobulin-like Receptor(s) (KIR) for self class I ligand(s), sense missing expression of donor KIR ligand(s) in the recipient and mediate alloreactions. It improves outcomes of HLA haplotype-mismatched ('haploidentical') transplants by controlling acute myeloid leukemia relapse without causing graft-versus-host disease. It is hoped the dramatic improvements afforded by the discovery of the role of NK cell alloreactivity will extend the use of haploidentical transplants, as the donors are, unlike the unrelated, immediately available family members. PMID- 18675346 TI - Relevance of MICA and other non-HLA antibodies in clinical transplantation. AB - The clinical importance of HLA-specific antibodies for organ allograft outcome is well established. In the past few years, there has been an increasing interest in non-HLA antigens as targets of injury in organ transplant recipients. This increased interest has been spurred by the fact that HLA-identical kidney transplants also undergo immunological rejections. Polymorphisms within non-HLA genes associated with evoking an immune response to alloantigens are currently being studied for their association with transplant outcome. Non-HLA antigens, such as the polymorphic MHC class I-related chain A (MICA), expressed on endothelial cells have been implicated in the pathogenesis of hyperacute, acute and chronic organ allograft rejections. Use of endothelial cells as targets may clarify the specificities of other clinically relevant non-HLA antibodies in graft rejections. This review summarizes past and current knowledge of the clinical importance and specificities of non-HLA antibodies, and mechanisms by which these antibodies may contribute to graft destruction in clinical transplantation. The aims of current research into the role of non-HLA antigens and their genetics in predicting outcome are to develop an improved insight into the basic science of transplantation and to develop a risk or prognostic index for use in the clinical setting. Non-HLA antibody responses are receiving increasing interest in acute and chronic rejection and specificity, affinity, and pathogenicity need to be investigated to estimate their contribution. Undoubtedly, this will continue to be an area of interest in terms of fully understanding the role of non-HLA antigens as targets of immune-mediated injury and the potential for clinical intervention. PMID- 18675347 TI - Joergensenia, a new genus to accommodate Psoroma cephalodinum (lichenized Ascomycota). AB - The taxonomy of the genus Psoroma (lichenized Ascomycota) is currently under revision, as it has long been recognized as heterogeneous. The aim of the present study was to reconstruct the phylogeny of Psoroma and related genera. The ITS region of nu-rDNA and the mtSSU rDNA from 22 collections of Psoroma and Pannaria were amplified. Sequences from GenBank were also used. For the phylogenetic analysis, direct optimization was implemented, using the program POY, and standard MP using PAUP. Both analyses resulted in similar trees. Two main clades were obtained, one including Santesoniella and most Psoroma species and the other including Pannaria and two Psoroma species, indicating that Psoroma is polyphyletic. Based on its phylogenetic position and ascus type, P. implexum is transferred to the genus Pannaria, and the new combination P. implexa is proposed. Psoroma cephalodinum clusters with Parmeliella and Degelia as a basal group within the Pannariaceae. The ascus apex of P. cephalodinum has a strong amyloid reaction revealing a compact cap-shaped plug. This differs from most of the ascus apical structures observed in the family. Both ascus internal structure and phylogenetic position suggest that P. cephalodinum is distantly related to Psoroma or Pannaria, and thus a new genus Joergensenia is proposed. PMID- 18675348 TI - Do different species of Smittium (Harpellales, Legeriomycetaceae) influence each other in the host gut? AB - Smittium (Harpellales, Legeriomycetaceae) belongs to a cosmopolitan group of filamentous fungi (Trichomycetes, Zygomycota) that live as obligate commensals in the digestive tract of various marine, freshwater, and terrestrial arthropods. The outcome of the paired introductions of three species of Smittium was investigated within the individual hosts of the mosquito Aedes aegypti (Culicidae: Diptera). In the first set of experiments, the host was inoculated with a single species of Smittium to determine whether hyphae location within the host was species specific. In the second experiment the host was exposed to two species of Smittium to determine whether hyphae showed positional displacement when two species of fungi co-inhabited the same host gut. Single species introductions of Smittium resulted in 80-85% of hosts with hyphae present only in the rectum. In all three paired combinations of Smittium species examined, only 40-65% of host larvae had hyphae restricted to the rectum. This is first study to experimentally demonstrate that the microdistribution of Harpellaceae hyphae can be influenced by the presence of a second species of Harpellaceae, suggesting a competition of the symbionts within the host. PMID- 18675349 TI - Fatty acid profiles of polar and neutral lipids of ten species of higher basidiomycetes indigenous to eastern Canada. AB - Neutral and polar lipid contents of ten species of edible mushrooms indigenous to Eastern Canada belonging to the families Agaricaceae, Amanitaceae, Boletaceae, Coprinaceae, Ganodermataceae, and Lycoperdaceae were analysed. The total lipid content of the species analysed ranged from 3.1% (Ganoderma applanatum) to 16% (w/w) d.w. (Amanita vaginata) and averaged 8.6% (w/w) d.w. Polar lipids accounted for more than 50% of the total lipids in most species and differences were observed between neutral and polar lipid contents according to the species analysed. In both lipid fractions, high proportions of unsaturated fatty acids (FAs) ranging from 62.7 to 82.3% (polar lipids) and 59.8 to 82.5% (neutral lipids) of the total FAs were observed. Analysis of FA profiles showed that both neutral and polar lipids were mainly composed of linoleic (18:2 Delta9c,12c), oleic (18:1 Delta9c), and palmitic (16:0) acids. Significant differences (P<0.05) in the contents of specific FAs were observed between mushroom species. Among the 44 FAs detected in the species analysed, the occurrence of cis-11-heptadecenoic (17:1 Delta11c) acid is reported for the first time in basidiomycetes, while elaidic acid (18:1 Delta9t) is reported for the first time in fungi. PMID- 18675350 TI - Coevolution with higher taxonomic host groups within the Puccinia/Uromyces rust lineage obscured by host jumps. AB - Partial beta-tubulin 1 sequence data were obtained for 80 taxa of Pucciniaceae, with hosts from 33 angiosperm families, covering all major ordinal groups in the Angiosperm Phylogeny Group classification. As in previous studies, most species of Puccinia and Uromyces fell into two main clades (I and II), with P. glechomatis and P. psidii excluded from Pucciniaceae. Results suggest two processes; a coevolution of and hosts in each clade, as well as associated frequent jumps to ecologically close, but taxonomically distant, hosts. Clade I contained all rusts on Cyperaceae and Juncaceae, and most rusts on host orders from rosids to euasterids II. Clade II contained all rusts on Poaceae, and most on host orders from monocots to core eudicots. In both main clades, several well supported subclades were identified. The grouping in clade I, subclade E of rusts of Cyperaceae and Asteraceae and, in particular, of an Australian isolate of P. dioicae with rusts on Australian families of Asterales, suggested a local radiation, and supported the coevolutionary relationship between rusts on these two families seen with a different range of asteraceous rusts in the Northern Hemisphere. In clade I, two clades contained only rusts of Asteraceae and Fabaceae, respectively, and in clade II, subclade F contained only rusts of pooid hosts. Rusts on non-pooid hosts were separated from them in subclade G. Other subclades contained a range of rusts on distantly related angiosperm families. Urediniospore morphology was often, but not always, correlated with the molecular phylogeny. Most rusts with urediniospores having few (1-5) equatorial germ pores were in clade I, whereas most with spores having several (5-14) scattered pores were in clade II. The distribution of telial host families on the beta-tubulin rust phylogeny was not random. Aecial hosts of heteroecious rusts played an important role in the evolutionary process. Possible examples of host jumps were seen in rusts on Geraniaceae, Polygonaceae, and Apiaceae. Despite such jumps obscuring past host associations, possible ancestral hosts were identified by the pattern of host distribution at higher taxonomic levels along the ss-tubulin phylogeny. Results suggest that clade I diverged with Cyperaceae, Juncaceae, and the more advanced core eudicot orders (rosids and asterids), whereas clade II diversified with earlier angiosperm groups, such as monocots, Poaceae, and Ranunculales. Qualified support was given to the hypothesis that rusts can reveal taxonomic relationships between their hosts, at genus, family, and ordinal levels. PMID- 18675351 TI - An N-acetyl-beta-D-glucosaminidase gene, cr-nag1, from the biocontrol agent Clonostachys rosea is up-regulated in antagonistic interactions with Fusarium culmorum. AB - Clonostachys rosea is a widely distributed fungus that often acts as a parasite on other soil fungi. This fungus has also been reported as a potential parasite against nematodes and insects. The antagonistic activity is thought to be correlated with the secretion of cell wall-degrading enzymes, including chitinases. In this work, we identified and characterized an N-acetyl-beta-D glucosaminidase-encoding gene, cr-nag1, belonging to glycosyl hydrolase family 20, from the C. rosea strain IK726 using a degenerated primer strategy designed from conserved motifs. The complete gene, including its promoter region, was obtained by genomic walking. Southern analysis showed that cr-nag1 is present as a single copy gene in C. rosea. Phylogenetically, cr-nag1 showed the highest similarity to N-acetyl-beta-d-glucosaminidase genes from other mycoparasitic fungi. Enzymatic assays and RT-PCR showed that the NAGase activity of C. rosea is specifically repressed in medium containing a high glucose content and is expressed in media containing chitin or Fusarium culmorum cell walls as sole carbon sources. Macroscopic and microscopic observations indicated that the mycelial growth of F. culmorum and Pythium ultimum were inhibited during interactions with C. rosea. High expression of cr-nag1 was found in interactions between C. rosea and F. culmorum, whereas the expression of cr-nag1 in interactions between C. rosea and P. ultimum was similar to the control. This indicates that although C. rosea secretes chitin-hydrolysing agents in order to target the cell wall of F. culmorum, it seems to use another strategy for controlling the development of the oomycete P. ultimum. PMID- 18675352 TI - Identification of nitrogen mineralization enzymes, L-amino acid oxidases, from the ectomycorrhizal fungi Hebeloma spp. and Laccaria bicolor. AB - Amino acids are major nitrogen sources in soils and they harbour a central position in the nitrogen metabolism of cells. We determined whether Hebeloma spp. and Laccaria bicolor expressed the enzyme L-amino acid oxidase (LAO), which catalyses the oxidative deamination of the alpha-amino group of L-amino acids. We measured LAO activities from the mycelial extracts of seven laboratory-grown fungal strains with three methods, and we measured how LAO activities were expressed in one Hebeloma sp. strain grown on four nitrogen sources. Hebeloma spp. and L. bicolor converted L-phenylalanine, but not D-phenylalanine, to hydrogen peroxide, 2-oxoacid, and ammonia, suggesting that they expressed LAO enzymes. The enzymes utilized five out of seven tested L-amino acids as substrates. LAO activities were maximal at pH 8, where Michaelis constant (Km) values were 2-5mm. The LAO of Hebeloma sp. was expressed on every nitrogen source analysed, and the activities were the highest in mycelia grown in nitrogen-rich conditions. We suggest that LAO is a mechanism for cellular amino acid catabolism in Hebeloma spp. and L. bicolor. Many soil bacteria and fungi also express LAO enzymes that have broad substrate specificities. Therefore, LAO is a potential candidate for a mechanism that catalyses nitrogen mineralization from amino acids at the ecosystem level. PMID- 18675354 TI - Immunohistochemical detection and biological activities of CYP17 (P450c17) in the indifferent gonad of the frog Rana rugosa. AB - Sex steroids play a crucial role in the gonad differentiation in various species of vertebrates. However, little is known regarding the localization and biological activity of steroid-metabolizing enzymes during gonadal sex differentiation in amphibians. In the present study, we showed by real-time RT PCR analysis that the expression of CYP17, one of the key steroidogenic enzymes, was higher in the indifferent gonad during sex differentiation in male than in female tadpoles of Rana rugosa but that there was no difference detected in the 3betaHSD mRNA level between the male and female gonads. We next examined the localization of CYP17, 3betaHSD and 17betaHSD in the indifferent and differentiating gonads by using three kinds of antibodies specific for CYP17, 3betaHSD and 17betaHSD, respectively. Positive signals for CYP17, 3betaHSD and 17betaHSD were observed in somatic cells of the indifferent gonad of males and in the interstitial cell of the testis. The enzymatic activity of CYP17 was also examined in the gonad during sex differentiation in this species. [(3)H]Progesterone (Prog) was converted to [(3)H]androstenedione (AE) in the indifferent gonad in males and females, but the rate of its conversion was higher in males than in females. Moreover, fluorescence in situ hybridization (FISH) analysis revealed that the CYP17 gene was located on the q arm of chromosome 9, indicating that CYP17 was autosomal in R. rugosa. Taken together, the results demonstrate that the CYP17 protein is synthesized in somatic cells of the indifferent gonad during gonadal sex differentiation in R. rugosa and that it is more active in converting Prog to AE in males than in females. The data suggest that CYP17 may be involved in testicular formation during sex differentiation in this species. PMID- 18675353 TI - Fluorescence characterization of denatured proteins. AB - Characterization of unfolded states, while critical to a complete understanding of protein folding, is inherently difficult due to structural heterogeneity and dynamic interchange between states. The growing body of work focusing on single molecule fluorescence techniques for the study of protein folding, also highlights their potential for studies of unfolded proteins. These methods can obtain conformational information about individual subpopulations of molecules in an ensemble, and measure dynamics without the need for synchronization. The studies highlighted here demonstrate the promise of these techniques for obtaining novel information about unfolded states in vitro and in more physiologically relevant milieu. PMID- 18675355 TI - Differential promotion of hematopoietic chimerism and inhibition of alloreactive T cell proliferation by combinations of anti-CD40Ligand, anti-LFA-1, everolimus, and deoxyspergualin. AB - Allogeneic bone marrow (BM) engraftment for chimerism and transplantation tolerance may be promoted by combinations of costimulation blocking biologics and small molecular weight inhibitors. We showed previously in a mouse model that anti-CD40Ligand (anti-CD40L, CD154) combined with anti-LFA-1 or everolimus (40-O (2-hydroxyethyl)-rapamycin) resulted in stable chimerism in almost all BM recipients, whereas anti-LFA-1 plus everolimus conferred approximately 50% chimerism stability. Here, we investigated whether this lower incidence could be increased with deoxyspergualin (DSG) in place of or in addition to everolimus. However, DSG and everolimus were similarly synergistic with costimulation blockade for stable hematopoietic chimerism. This correlated with allospecific T cell depletion and inhibition of acute but not chronic skin allograft rejection. Different treatments were also compared for their inhibition of alloreactive T cell proliferation in vivo. While anti-CD40L did not impair T cell proliferation, anti-LFA-1 reduced both CD4 and CD8 T cell proliferation, and combining anti-LFA 1 with everolimus or DSG had an additive inhibitory effect on CD4 T cell proliferation. Thus, despite their strong inhibition of alloreactive T cell proliferation, combinations of anti-LFA-1 with everolimus or DSG did not reach the unique potency of anti-CD40L-based combinations to support stable hematopoietic chimerism in this system. PMID- 18675356 TI - Low-frequency stimulation of the tuberomammillary nucleus facilitates electrical amygdaloid-kindling acquisition in Sprague-Dawley rats. AB - Histamine plays a suppressive role in seizure. The tuberomammillary nucleus (TM) is the only locus of histaminergic neurons in the brain. To determine whether deep brain stimulation (DBS) of the TM provides protection against seizures, we tested the effects of low-frequency stimulation (LFS, 1 Hz), high frequency stimulation (HFS, 100 Hz), and electrolytic lesions of the TM on seizures generated by amygdaloid kindling, pentylenetetrazol (PTZ) and maximal electroshock (MES) in rats. LFS of TM accelerated the progression of behavioral seizure stage and increased the mean afterdischarge duration (ADD) during acquisition of amygdaloid-kindling seizures, but had no considerable anticonvulsive effect in fully kindled animals. It augmented the MES-induced seizures as well, but had no appreciable effects on PTZ-kindled seizures. In addition, both HFS and bilateral lesions of the TM exacerbated the progression of amygdaloid-kindling seizures. These results suggest that specific negative sites for DBS exist in the brain, such as the TM. This study indicates that it is crucial to choose a suitable target for DBS in the clinical treatment of epilepsy. PMID- 18675358 TI - Effect of cilostazol on delayed cerebral vasospasm after subarachnoid hemorrhage in rats: evaluation using black blood magnetic resonance imaging. AB - The purpose of this study was to use black blood magnetic resonance imaging (BB MRI) to assess delayed cerebral vasospasm (DCV) after subarachnoid hemorrhage (SAH) in rats, and evaluate whether delayed treatment with the anti-platelet agent cilostazol was effective on DCV. BA vasospasm was sequentially assessed at 1, 2, and 3 h, and 1-6 days after SAH by BB-MRI. BB-MRI clearly visualized biphasic vasospasm; early vasospasm at 1 h later and the maximal DCV at day 2. Cilostazol was perorally administered twice at day 1 after having confirmed significant DCV using BB-MRI. The effect of cilostazol on DCV was evaluated at day 2. Cilostazol significantly attenuated DCV and suppressed the levels of malondialdehide and 8-isoprostane in CSF after SAH. This study shows that BB-MRI is a useful and less invasive method for the evaluation of DCV, and cilostazol may be effective on DCV. PMID- 18675359 TI - Neuroinflammation in the generation of post-transplantation dyskinesia in Parkinson's disease. AB - The observation that neural grafts can induce dyskinesias has severely hindered the development of a transplantation therapy for Parkinson's disease (PD). We addressed the hypothesis that inflammatory responses within and around an intrastriatal graft containing dopamine neurons can trigger dyskinetic behaviors. We subjected rats to unilateral nigrostriatal lesions with 6-hydroxydopamine (6 OHDA) and treated them with L-DOPA for 21 days in order to induce abnormal involuntary movements (AIMs). Subsequently, we grafted the rats with allogeneic embryonic ventral mesencephalic tissue in the dopamine-denervated striatum. In agreement with earlier studies, the grafted rats developed dyskinesia-like AIMs in response to amphetamine. We then used two experimental approaches to induce an inflammatory response and examined if the amphetamine-induced AIMs worsened or if spontaneous AIMs developed. In one experiment, we challenged the neural graft hosts immunologically with an orthotopic skin allograft of the same genetic origin as the intracerebral neural allograft. In another experiment, we infused the pro-inflammatory cytokine interleukin 2 (IL-2) adjacent to the intrastriatal grafts using osmotic minipumps. The skin allograft induced rapid rejection of the mesencephalic allografts, leading to disappearance of the amphetamine-induced AIMs. Contrary to our hypothesis, the rejection process itself did not elicit AIMs. Likewise, the IL-2 infusion did not induce spontaneous AIMs, nor did it alter L-DOPA-induced AIMs. The IL-2 infusions did, however, elicit the predicted marked striatal inflammation, as evidenced by the presence of activated microglia and IL2Ralpha-positive cells. These results indicate that an inflammatory response in and around grafted dopaminergic neurons is not sufficient to evoke dyskinetic behaviors in experimental models of PD. PMID- 18675357 TI - Genetics of iron regulation and the possible role of iron in Parkinson's disease. AB - Parkinson's disease (PD) is acknowledged as the second most common neurodegenerative disorder after Alzheimer's Disease. Older age may be the only unequivocal risk factor for PD although the male to female ratio is consistently greater than 1 in populations of European ancestry. Characteristic features of PD include dopaminergic neuron death in the substantia nigra (SN) pars compacta, accumulation of alpha-synuclein inclusions known as Lewy bodies in the SN, and brain iron accumulation beyond that observed in non-PD brains of a similar age. In this review article, we will provide an overview of human and animal studies investigating the contributions of iron in PD, a summary of human studies of iron related genes in PD, a review of the literature on the genetics of iron metabolism, and some hypotheses on possible roles for iron in the pathogenic processes of PD including potential interactions between iron and other factors associated with Parkinson's disease. PMID- 18675360 TI - SSM-based electrophysiology. AB - An assay technique for the electrical characterization of electrogenic transport proteins on solid supported membranes is presented. Membrane vesicles, proteoliposomes or membrane fragments containing the transporter are adsorbed to the solid supported membrane and are activated by providing a substrate or a ligand via a rapid solution exchange. This technique opens up new possibilities where conventional electrophysiology fails like transporters or ion channels from bacteria and from intracellular compartments. Its rugged design and potential for automation make it suitable for drug screening. PMID- 18675361 TI - High performance computing in structural determination by electron cryomicroscopy. AB - Computational advances have significantly contributed to the current role of electron cryomicroscopy (cryoEM) in structural biology. The needs for computational power are constantly growing with the increasing complexity of algorithms and the amount of data needed to push the resolution limits. High performance computing (HPC) is becoming paramount in cryoEM to cope with those computational needs. Since the nineties, different HPC strategies have been proposed for some specific problems in cryoEM and, in fact, some of them are already available in common software packages. Nevertheless, the literature is scattered in the areas of computer science and structural biology. In this communication, the HPC approaches devised for the computation-intensive tasks in cryoEM (single particles and tomography) are retrospectively reviewed and the future trends are discussed. Moreover, the HPC capabilities available in the most common cryoEM packages are surveyed, as an evidence of the importance of HPC in addressing the future challenges. PMID- 18675362 TI - Neural correlates of successful encoding of semantically and phonologically mediated inter-item associations. AB - This experiment investigated whether the neural correlates of inter-item associative encoding vary according to study task. At study, pairs of unrelated words were subjected to either semantic or phonological relational judgments. Test items comprised studied word pairs (intact), pairs comprised of words belonging to different study pairs (rearranged), and novel pairs. The test requirement was to discriminate between these different classes of test item. fMRI was employed to contrast the neural activity elicited by studied pairs that were correctly endorsed as intact on the later associative recognition test, as opposed to pairs for which associative information was unavailable. In contrast to prior findings for the encoding of single items, there was no evidence that the loci of subsequent associative memory effects varied according to study task. Instead, in both tasks, pairs that were later correctly endorsed as intact elicited enhanced activity in mid- and ventral regions of the left ventrolateral prefrontal cortex (VLPFC). These findings were accompanied by extensive task invariant reversed subsequent memory effects in medial and lateral parietal and frontal cortices. The findings suggest that the left VLPFC may play a domain general role in the encoding of item-item associations, and in addition highlight the importance of elucidating the functional significance of reversed subsequent memory effects. PMID- 18675363 TI - Reliable assessment of lower limb motor representations with fMRI: use of a novel MR compatible device for real-time monitoring of ankle, knee and hip torques. AB - This study describes the use of a novel magnetic resonance imaging (MRI) compatible system capable of measuring isometric ankle, knee and hip joint torques in real-time during functional MRI (fMRI) testing in healthy volunteers. The motor representations of three isometric torques--ankle dorsiflexion, ankle plantarflexion and knee extension--were studied at two time points. The reliability of motor performance and fMRI-derived measures of brain activity across sessions was examined. Reproducible motor performance was observed for each of the tasks; torques of the requested amplitude, assisted by visual feedback, were generated at the relevant joint with good accuracy, both within and across the two sessions. Significant blood oxygen level dependent (BOLD) signal increases were observed in the left primary sensorimotor cortex (SM1) in the paracentral lobule and in secondary motor areas for all tasks. Within these areas there was substantial overlap of the motor representations though differential activation was observed in SM1, with greater activation of inferior paracentral lobule during knee extension than for either ankle task. Also, BOLD signal decreases were observed bilaterally within SM1 in the hand knob region for all tasks. No major session-related effects were identified at the group level. High intraclass correlation coefficients were observed for t-values of voxels in cortical motor areas for each contraction type for individuals, suggesting that fMRI-derived activity across time points was reliable. These findings support the use of this apparatus in serial studies of lower limb function. PMID- 18675364 TI - Testosterone biases automatic memory processes in women towards potential mates. AB - Female mate choice involves the comparative evaluation of potential mates. Females use a pooled comparison of sampled males to maximize the perceived reproductive fitness of their partner, implying the memorization of sampled males. However, hormonal and reproductive states influence female choosiness, and women's preference and memory for masculinity. Here, we investigated whether testosterone biases memory processes in women towards male faces using functional MRI. A single nasal testosterone dose was administered to healthy women in their early follicular phase, in a double-blind, placebo-controlled, crossover design. Testosterone increased the difference in reaction times to categorize male and female faces during encoding, without influencing subsequent recognition accuracy or response bias. The imaging results showed that testosterone shifted memory formation in the hippocampus and inferior temporal gyri from the encoding of female faces towards the encoding of male faces. In contrast, testosterone shifted memory formation in the left inferior frontal gyrus from the encoding of male faces towards the encoding of female faces. Furthermore, the hippocampal contribution to memory retrieval also shifted from female towards male faces. These results indicate that testosterone biases memory processes towards the relatively automatic encoding and retrieval of males in temporal brain regions and elaborate encoding of females in frontal brain regions, suggesting that testosterone may support female mate sampling and comparison by biasing automatic memory processes towards the encoding and retrieval of potential mates. PMID- 18675365 TI - Phylogeny and biogeography of bees of the tribe Osmiini (Hymenoptera: Megachilidae). AB - The Osmiini (Megachilidae) constitute a taxonomically and biologically diverse tribe of bees. To resolve their generic and suprageneric relationships, we inferred a phylogeny based on three nuclear genes (Elongation factor 1-alpha, LW rhodopsin and CAD) applying both parsimony and Bayesian methods. Our phylogeny, which includes 95 osmiine species representing 18 of the 19 currently recognized genera, is well resolved with high support for most basal nodes. The core osmiine genera were found to form a well-supported monophyletic group, but four small genera, Noteriades, Afroheriades,Pseudoheriades and possibly Ochreriades, formerly included in the Osmiini, do not appear to belong within this tribe. Our phylogeny results in the following taxonomic changes: Stenosmia and Hoplosmia are reduced to subgeneric rank in Hoplitis and Osmia, respectively, Micreriades is recognized as a subgenus in Hoplitis and the subgenus Nasutosmia is transferred from Hoplitis to Osmia. We inferred a biogeographic scenario for the Osmiini applying maximum likelihood inference and models of character evolution. We provide evidence that the Osmiini originated in the Palearctic, and that extensive exchanges occurred between the Palearctic and the Nearctic. The latter finding may relate to the fact that many osmiine species nest in wood or in stems, facilitating dispersal by overseas transport of the nests. PMID- 18675366 TI - The need for specifying species concepts: How many species of silvered langurs (Trachypithecus cristatus group) should be recognized? PMID- 18675367 TI - A differential phenotypic expression of a divergent spindle mutation in interspecific Brachiaria hybrids. AB - Several mutations are known to alter the normal progression of meiosis and can be correlated with defects in microtubule distribution. The dv mutation affects the spindle organization and chromosomes do not converge into focused poles. Two Brachiaria hybrids presented the phenotypic expressions of dv mutation but exhibited many more details in the second division. Bivalents were distantly positioned and spread over a large metaphase plate and failed to converge into focused poles. Depending on the distance of chromosomes at the poles, telophase I nuclei were elongated or the chromosomes were grouped into various micronuclei of different sizes in each cell. The first cytokinesis occurred. However, when there were micronuclei, a second cytokinesis immediately took place dividing the prophase II meiocytes into three or four cells. In each meiocyte, meiosis progressed to the second division. Slightly elongated nuclei or micronuclei were recorded in telophase II. After a third cytokinesis, hexads or octads were formed. Pollen grains of different sizes were generated. One of these hybrids presented a higher frequency of abnormal cells than when previously analyzed. The fate of these hybrids as genitors or as candidates for cultivars in the Brachiaria breeding program is discussed. PMID- 18675368 TI - NF-kappaB-dependency and consequent regulation of IL-8 in echinomycin-induced apoptosis of HT-29 colon cancer cells. AB - The present study was to see whether echinomycin-induced apoptosis would be NF kappaB-dependent and if so, whether echinomycin would activate or inhibit NF kappaB as well as resultant chemokine IL-8 expression. In HT-29 cells echinomycin activated NF-kappaB in time-dependent manner. EMSA in the presence of antibodies specific for p50 and p65 subunits indicated that echinomycin-induces the translocation of p50-p65 heterodimeric subunits of NF-kappaB. Levels of IkappaB were detected at initial echinomycin treatment and thereafter decreased, faintly seen after a 6h treatment. In contrast p-IkappaB levels were clearly detected throughout 6-24h of echinomycin treatment, albeit initially fainted. To clarify the role of NF-kappaB on IL-8 expression in echinomycin-mediated apoptosis of HT 29 cells, ELISA plus RT-PCR clearly showed that IL-8 production is inducible by echinomycin treatment. Using a specific inhibitor, IL-8 regulation at echinomycin treatment in HT-29 cells occurred via both caspase-3 and NF-kappaB-dependent signal pathway. To confirm whether two different pathways (NF-kappaB and caspase) would be coupled, only NF-kappaB inhibitor (PDTC) and caspase-3 specific inhibitor (Z-DEVD-FMK) together significantly attenuated echinomycin-initiated apoptosis of HT-29 cells, pretreatment of HT-29 cells with PDTC rarely affected echinomycin-induced caspase-3 activation. So echinomycin-induced apoptosis in HT 29 cells occurs via NF-kappaB activation independent of caspase-3 activation modulating the resultant-linked key chemokine IL-8 expression and echinomycin induced apoptosis is NF-kappaB-dependant and directly related to NF-kappaB activation, consequently regulating IL-8 expression. PMID- 18675369 TI - Bcl-2 and caspase-8 related anoikis resistance in human osteosarcoma MG-63 cells. AB - Detachment of adherent cells from extracellular matrix results in apoptosis, a process termed "anoikis". Resistance to anoikis is implicated in the progression of many malignancies by facilitating the migration and eventual colonization of distant sites. Human kidney epithelial cells 293T, human osteoblast cells hFOB 1.19 and human osteosarcoma cells Saos-2 significantly underwent anoikis when adherence was prevented. But human osteosarcoma MG-63 cells were distinctly anoikis resistant when detached. They formed large aggregates and showed little apoptosis compared to the other cells. When MG-63 cells were in suspension, caspase-8, physically associated with death receptor was activated by cell-matrix detachment, whereas. Caspase-3 and caspase-9 were not activated. Translational level of Bcl-2 significantly increased in a time-dependent manner, but the level of beta-catenin and PI3K did not. Caspase-8 participates in an anoikis-inducing process in MG-63 cells at an early time, and overexpression of Bcl-2 blocks activation of caspase-8 making MG-63 cells anoikis resistant. PMID- 18675370 TI - Posterior parietal cortex: an interface between attention and learning? AB - The posterior parietal cortex (PPC) of rats has most recently been defined based on patterns of thalamic and cortical connectivity. The anatomical characteristics of this area suggest that it may be homologous to the PPC of primates and contribute to similar functions. This review summarizes evidence for and against a role for the rat PPC in attention and working memory and evaluates how the function of the rat PPC compares to that of primates on these dimensions. Theories of how the rat PPC contributes to behavior are presented, including the notion that PPC may serve as an interface between attention and learning. Finally, several avenues for future research are considered. PMID- 18675371 TI - Cytoplasmic localization of sterol transcription factors Upc2p and Ecm22p in S. cerevisiae. AB - Ergosterol homeostasis is a critical process for fungal cells. Paralogous zinc cluster transcription factors Upc2p and Ecm22p are major regulators of ergosterol biosynthesis in Saccharomyces cerevisiae. Upc2p and Ecm22p sense and respond to sterol depletion but their mechanism of activation has not been defined. Subcellular localization and functional expression of Upc2p-GFP and Ecm22p-GFP was monitored by fluorescence microscopy and flow cytometry in live yeast cells. Both fusion proteins localized to intracellular membranes and to perinuclear foci. Perinuclear localization of Upc2p-GFP and Ecm22p-GFP was increased when ergosterol biosynthesis was blocked by azole drug treatment. Nuclear localization in response to sterol depletion is consistent with the hypothesis that Upc2p and Ecm22p are trafficked from a membrane to the nucleus as a post-translational mechanism of sterol sensing. PMID- 18675372 TI - Comparison of the effect of heat shock factor inhibitor, KNK437, on heat shock- and chemical stress-induced hsp30 gene expression in Xenopus laevis A6 cells. AB - In this study, we compared the effect of KNK437 (N-formyl-3, 4-methylenedioxy benzylidene-gamma-butyrolactam), a benzylidene lactam compound, on heat shock and chemical stressor-induced hsp30 gene expression in Xenopus laevis A6 kidney epithelial cells. Previously, KNK437 was shown to inhibit HSE-HSF1 binding activity and heat-induced hsp gene expression. In the present study, Northern and Western blot analysis revealed that pretreatment of A6 cells with KNK437 inhibited hsp30 mRNA and HSP30 and HSP70 protein accumulation induced by chemical stressors including sodium arsenite, cadmium chloride and herbimycin A. In A6 cells subjected to sodium arsenite, cadmium chloride, herbimycin A or a 33 degrees C heat shock treatment, immunocytochemistry and confocal microscopy revealed that HSP30 accumulated primarily in the cytoplasm. However, incubation of A6 cells at 35 degrees C resulted in enhanced HSP30 accumulation in the nucleus. Pre-treatment with 100 microM KNK437 completely inhibited HSP30 accumulation in A6 cells heat shocked at 33 or 35 degrees C as well as cells treated with 10 microM sodium arsenite, 100 microM cadmium chloride or 1 microg/mL herbimycin A. These results show that KNK437 is effective at inhibiting both heat shock- and chemical stress-induced hsp gene expression in amphibian cells. PMID- 18675373 TI - Physiological correlates of growth and condition in the yellow perch (Perca flavescens). AB - This study on yellow perch (Perca flavescens) examines a series of enzymatic markers and the relative weights of pyloric caeca and visceral lipids, their response to changes in feeding regime and their potential use to infer recent changes in growth rate and fish condition. Fish were exposed to four different feeding regimes for 12 weeks resulting in specific growth rates ranging from 0.3% to 3.5% (%/day). Growth and condition responded rapidly to changes in ration and the weight of pyloric caeca and visceral lipids reflected increased feed intake. Growth rate was correlated with muscle citrate synthase and caecal nucleoside diphosphate kinase activities, whereas condition was correlated with muscle citrate synthase and lactate dehydrogenase activities and with caecal glucose-6 phosphate dehydrogenase activity. Results showed that enzyme activities and biometric parameters responded rapidly to increased feed intake, but the response was slower when food intake decreased. Plateaus were attained for both condition and visceral lipid index, but the relative weight of pyloric caeca continued to increase throughout the experimental period. Results from this study could, in principle, be used to infer recent growth and energy status in wild yellow perch and thus provide an indicator of food availability in their environment. PMID- 18675374 TI - Mutual self-defence: the trypanolytic factor story. AB - Around 1900 Laveran and Mesnil discovered that African trypanosomes (prototype: Trypanosoma brucei brucei) do not survive in the blood of some primates and humans. The nature of the trypanolytic factor present in these sera has been the focus of a long-standing debate between different groups, but recent developments have allowed the proposal of a coherent model incorporating most seemingly divergent views and providing an interesting example of the complex interplay that continuously occurs between hosts and parasites. Possibly as an adaptation to their natural environment, great African apes and humans have acquired a new member of the apolipoprotein-L family, termed apoL1. This protein is the only one of the family to be secreted in the blood, where it binds to a subset of HDL particles that also contain another human-specific protein, haptoglobin-related protein or Hpr. T. b. brucei possesses a specific surface receptor for the haptoglobin-hemoglobin (Hp-Hb) complex, as a way to capture heme into hemoproteins that contribute to cell growth and resistance to the oxidative stress of the host. As this receptor does not discriminate between Hp and Hpr, Hpr-containing HDL particles of human serum are efficiently taken up by the parasite, leading to the simultaneous internalization of apoL1, Hpr and Hb derived heme. Once in the lysosome, apoL1 is targeted to the lysosomal membrane, where its colicin-like anionic pore-forming activity triggers an influx of chloride ions from the cytoplasm. Osmotic effect linked to this ionic flux leads to uncontrolled swelling of the lysosome, ultimately causing the death of the parasite. Two T. brucei clones, termed Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense, have managed to resist this lysis mechanism and, therefore, cause sleeping sickness in humans. While the mechanism of this resistance is still not known in the case of T. b. gambiense, the dominant factor responsible for resistance of T. b. rhodesiense has been identified. This protein, named SRA for Serum Resistance-Associated, is a truncated version of the major and variable surface antigen of the parasite, the Variant Surface Glycoprotein or VSG. Presumably due to its defective nature, SRA is not targeted to the plasma membrane as do regular VSGs, but ends up in the late endosomal compartment. In this location SRA is thought to neutralize apoL1 through coiled coil interactions between alpha-helices. We discuss the potential of these discoveries in terms of fight against the disease. PMID- 18675375 TI - Tryptophan synthase, an allosteric molecular factory. AB - Tryptophan synthase (TrpS) is a pyridoxal phosphate-containing bifunctional enzyme that catalyzes the last two steps in the biosynthesis of L-tryptophan. Indole, an intermediate generated at the active site of the alpha-subunit is channeled via a 25 A long tunnel to the beta-active site where it reacts with an aminoacrylate intermediate derived from L-serine. The two reactions are kept in phase by allosteric interactions between the two subunits. The recent development of novel alpha-site ligands and alpha-reaction transition state analogs combined with kinetic and crystal structure analysis of Salmonella typhimurium tryptophan synthase has provided new insights into the allosteric regulation of substrate channeling, the reaction mechanisms of the alpha and beta active sites, and the influence of structural dynamics. PMID- 18675376 TI - The fluorinase, the chlorinase and the duf-62 enzymes. AB - The fluorinase from Streptomyces cattleya and chlorinase from Salinispora tropica have a commonality in that they mediate nucleophilic reactions of their respective halide ions to the C-5' carbon of S-adenosyl-L-methionine (SAM). These enzyme reactions fall into the relatively small group of S(N)2 substitution reactions found in enzymology. These enzymes have some homology to a larger class of proteins expressed by the duf-62 gene, of which around 200 representatives have been sequenced and deposited in databases. The duf-62 genes express a protein which mediates a hydrolytic cleavage of SAM to generate adenosine and L methionine. Superficially this enzyme operates very similarly to the halogenases in that water/hydroxide replaces the halide ion. However structural examination of the duf-62 gene product reveals a very different organisation of the active site suggesting a novel mechanism for water activation. PMID- 18675377 TI - Microelectrodes for studying neurobiology. AB - Microelectrodes have emerged as an important tool used by scientists to study biological changes in the brain and in single cells. This review briefly summarizes the ways in which microelectrodes as chemical sensors have furthered the field of neurobiology by reporting on changes that occur on the subsecond time scale. Microelectrodes have been used in a variety of fields including their use by electrophysiologists to characterize neuronal action potentials and develop neural prosthetics. Here we restrict our review to microelectrodes that have been used as chemical sensors. They have played a major role in many important neurobiological findings. PMID- 18675378 TI - Mutagenicity of 2-[2-(acetylamino)-4-[bis(2-hydroxyethyl)amino]-5-methoxyphenyl] 5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-6) and benzo[a]pyrene (BaP) in the gill and hepatopancreas of rpsL transgenic zebrafish. AB - We examined the in vivo mutagenicity of 2-[2-(acetylamino)-4-[bis(2 hydroxyethyl)amino]-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-6) and benzo[a]pyrene (BaP) by using transgenic (Tg) zebrafish carrying the mutational target gene rpsL. PBTA-6 is one of the PBTA-type compounds that were recently identified in highly mutagenic river water in Japan. BaP is a well-known contaminant that is frequently found in polluted water. Both compounds are potent mutagens, as determined by using the Ames test employing S9 mix and Salmonella. Adult rpsL Tg zebrafish were exposed to 0, 7, or 10 mg/L PBTA-6 or 0, 1.5, or 3 mg/L BaP for 96 h in a water bath and the mutations in their gills and hepatopancreata were measured 2-4 weeks later. At 3 weeks after exposure, 3 mg/L BaP significantly increased the rpsL mutant frequency (MF) in the gill and hepatopancreas by 5- and 2.3-fold, respectively, as compared to control fish. Sequence analysis showed that BaP mainly induced G:C to T:A and G:C to C:G transversions, which is consistent with the known mutagenic effects of BaP. In contrast, despite its extremely high mutagenic potency in Salmonella strains, PBTA-6 did not significantly increase the MF in the zebrafish gill or hepatopancreas. Although PBTA-6 is 300 times more mutagenic than BaP in the Ames test [T. Watanabe, H. Nukaya, Y. Terao, Y. Takahashi, A. Tada, T. Takamura, H. Sawanishi, T. Ohe, T. Hirayama, T. Sugimura, K. Wakabayashi, Synthesis of 2 phenylbenzotriazole-type mutagens, PBTA-5 and PBTA-6, and their detection in river water from Japan, Mutat. Res. 498 (2001) 107-115], calculation of the mutagenicity per mole of compound indicated that PBTA-6 was 33- and <3.7-fold less mutagenic in the zebrafish gill and hepatopancreas, respectively, than BaP. PMID- 18675379 TI - Sphingosine-1-phosphate regulation of mammalian development. AB - Sphingosine-1-phosphate (S1P) was first identified as a lysophospholipid metabolite whose formation is required for the irreversible degradation of sphingolipids. Years later, it was discovered that S1P is a bioactive lipid that provokes varied cell responses by acting through cell-surface receptors to drive cell signaling. More recent findings in model organisms have now established that S1P metabolism and signaling are integrated into many physiological systems. We describe here the surprising breadth of function of S1P in mammalian development and the underlying biologic processes that S1P regulates. PMID- 18675380 TI - Classification models for the prediction of clinicians' information needs. AB - OBJECTIVE: Clinicians face numerous information needs during patient care activities and most of these needs are not met. Infobuttons are information retrieval tools that help clinicians to fulfill their information needs by providing links to on-line health information resources from within an electronic medical record (EMR) system. The aim of this study was to produce classification models based on medication infobutton usage data to predict the medication related content topics (e.g., dose, adverse effects, drug interactions, patient education) that a clinician is most likely to choose while entering medication orders in a particular clinical context. DESIGN: We prepared a dataset with 3078 infobutton sessions and 26 attributes describing characteristics of the user, the medication, and the patient. In these sessions, users selected one out of eight content topics. Automatic attribute selection methods were then applied to the dataset to eliminate redundant and useless attributes. The reduced dataset was used to produce nine classification models from a set of state-of-the-art machine learning algorithms. Finally, the performance of the models was measured and compared. MEASUREMENTS: Area under the ROC curve (AUC) and agreement (kappa) between the content topics predicted by the models and those chosen by clinicians in each infobutton session. RESULTS: The performance of the models ranged from 0.49 to 0.56 (kappa). The AUC of the best model ranged from 0.73 to 0.99. The best performance was achieved when predicting choice of the adult dose, pediatric dose, patient education, and pregnancy category content topics. CONCLUSION: The results suggest that classification models based on infobutton usage data are a promising method for the prediction of content topics that a clinician would choose to answer patient care questions while using an EMR system. PMID- 18675381 TI - Sustained cardioprotection: exploring unconventional modalities. AB - Since Murry et al. [Murry, C.E., Jennings, R.B., Reimer, K.A., 1986. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium. Circulation. 74, 1124-36.] initially reported on the powerful protective effects of ischemic preconditioning (PC), a plethora of experimental investigations have identified varied preconditioning protocols or mimetics to achieve cardioprotection. These stimuli predominantly act via archetypal mediators identified in associated signalling studies (including PI3-K, Akt, PKC, mitochondrial K(ATP) channels). Despite an intense research effort over the last 20 years, there remains a paucity of evidence that this protective paradigm is clinically exploitable. This may arise due to a number of drawbacks to conventional protection, including effects of age, disease, and interactions with other pharmacological agents. This encourages investigation of alternate strategies that trigger protection via unconventional signalling (distinct from conventional PC) and/or mediate sustained shifts in ischemic tolerance in hearts of varying age and disease status. This review considers briefly drawbacks to conventional PC, and focuses on alternate strategies for generating prolonged states of cardiac protection. PMID- 18675382 TI - Endothelin-1 (1-31): from chymase-dependent synthesis to cardiovascular pathologies. AB - The mast cell-derived serine protease chymase is importantly involved not only in degradation, but in synthesis of bioactive peptides as well. Several studies suggest that chymase is the predominant enzyme in the production of angiotensin II (Ang II) from angiotensin-I in interstitial tissues. Interestingly, chymase has also been suggested to mature endothelin-1 (ET-1) from its precursor, big-ET 1 in vitro. The lack of availability of specific chymase inhibitors, beyond the chymotrypsin-like inhibitor chymostatin, currently hampers the investigation of the chymase/ET-1/Ang II paradigm in physiology and cardiovascular diseases. Nonetheless, the recent advent of highly selective chymase inhibitors is shedding new light on the role of this enzymatic pathway in the several inflammatory prone vascular diseases as summarized in the present review. Considering increasing evidence towards significant interactions between Ang II and ET-1 in cardiovascular diseases, the present review will address the role of chymase in the production of those two peptides. Whether chymase-dependent production of ET 1 plays an important role in cardiovascular pathologies will also be discussed. PMID- 18675383 TI - Kinetoplastid genomics: the thin end of the wedge. AB - The completion of the genome sequencing projects for major pathogens Trypanosoma brucei, Trypanosoma cruzi and Leishmania major has enabled numerous studies that would have been difficult or impossible to perform otherwise. New technologies in sequencing and protein analyses promise further rapid expansion in our capabilities. The keys to successful use of these new tools are recognizing the power and limitations of studies performed thus far, grasping the unrealized potential of new and developing technologies, and creating access to a multidisciplinary set of skills that will facilitate research, particularly in the bioinformatic analysis of the reams of data that will be forthcoming. In this Discussion, we will provide an overview of kinetoplastid genomics studies with emphasis on studies advanced through genomic data, and a preview of what may come in the near future. PMID- 18675384 TI - The role of histone H2A and H2B post-translational modifications in transcription: a genomic perspective. AB - In eukaryotic cells, the genome is packaged with histones H2A, H2B, H3, and H4 to form nucleosomes. Each of the histone proteins is extensively post translationally modified, particularly in the flexible N-terminal histone tail domains. Curiously, while post-translational modifications in histone H3 and H4 have been extensively studied, relatively little is known about post translational modifications in the N-terminal domains of histone H2A and H2B. In this review, we will summarize current knowledge of post-translational modifications in the N-terminal domains of histone H2A and H2B, and the histone variant H2AZ. We will examine the distribution of these modifications in genomic chromatin, and the function of these modifications in transcription. PMID- 18675385 TI - Dual delivery of an angiogenic and an osteogenic growth factor for bone regeneration in a critical size defect model. AB - This study investigated the effects of dual delivery of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) for bone regeneration in a rat cranial critical size defect. Four groups of scaffolds were generated with VEGF (12 microg), BMP-2 (2 mug), both VEGF (12 microg) and BMP-2 (2 microg), or no growth factor released from gelatin microparticles incorporated within the scaffold pores. These scaffolds were implanted within an 8 mm rat cranial critical size defect (n=8-9 for each group). At 4 and 12 weeks, implants were retrieved and evaluated by microcomputed tomography (microCT) and histological scoring analysis. Additionally, 4 week animals were perfused with a radiopaque material to visualize and quantify blood vessel formation. Histological analysis revealed that for all groups at 4 weeks, a majority of the porous scaffold volume was filled with vascularized fibrous tissue; however, bone formation appeared most abundant in the dual release group at this time. At 12 weeks, both dual release and BMP-2 groups showed large amounts of bone formation within the scaffold pores and along the outer surfaces of the scaffold; osteoid secretion and mineralization were apparent, and new bone was often in close or direct contact with the scaffold interface. MicroCT results showed no significant difference among groups for blood vessel formation at 4 weeks (<4% blood vessel volume); however, the dual release group showed significantly higher bone formation (16.1+/-9.2% bone volume) than other groups at this time. At 12 weeks, dual release and BMP-2 groups exhibited significantly higher bone formation (39.7+/-14.1% and 37.4+/-18.8% bone volume, respectively) than either the VEGF group or blank scaffolds (6.3+/-4.8% and 7.8+/-7.1% bone volume, respectively). This work indicates a synergistic effect of the dual delivery of VEGF and BMP-2 on bone formation at 4 weeks and suggests an interplay between these growth factors for early bone regeneration. For the doses investigated, the results show that the addition of VEGF does not affect the amount of bone formation achieved by BMP-2 at 12 weeks; however, they also indicate that delivery of both growth factors may enhance bone bridging and union of the critical size defect compared to delivery of BMP-2 alone. PMID- 18675386 TI - Dextran sodium sulfate-induced colitis causes rapid bone loss in mice. AB - INTRODUCTION: Osteopenia is a common complication of human inflammatory bowel disease (IBD). We evaluated the contribution of colonic inflammation to osteopenia and its mechanism in a murine colitis model. METHODS: Colitis was induced by adding dextran sodium sulfate (DSS) to the drinking water for 2 weeks to nine-week-old Balb/C male mice. 5% DSS was added on the first week and was reduced to 2.5% on the second week. Age- and sex-matched Balb/C mice served as the control group. Indices of femoral bone mass and architecture were determined by micro computed tomography (muCT). Bone formation parameters and osteoclast number were determined by dynamic histomorphometry. The degree of colonic inflammation was assessed by a clinical disease activity index, and colonic mucosal myeloperoxidase activity. RESULTS: DSS-treated mice exhibited a significantly lower bone mass compared to controls as indicated by decreased trabecular bone volume (BV/TV) of 32%. This reduction was accompanied by decreased trabecular number (23%) and connectivity density (37%) compared to the controls. No changes were observed in cortical bone indices. Osteopenia resulted from suppressed bone formation, as indicated by decreased trabecular double labeled surface (dL%) of 90%, mineralizing surface (MS) of 62%, and bone formation rate (BFR) of 67%, and increased bone resorption as indicated by a 34% increase in osteoclast number in DSS-treated mice compared to the controls. Myeloperoxidase activity inversely correlated with trabecular BV/TV (r=-0.67, p=0.02), trabecular number (r=-0.86, p=0.0008) and connectivity density (r=-0.63, p=0.03). Myeloperoxidase activity inversely correlated with the bone formation indices: dL%, MS, and BFR (r=-0.79, p=0.007, r=-0.84, p=0.002, r=-0.83, p=0.003, respectively). CONCLUSIONS: DSS-induced colitis is associated with reduced femoral bone mass and altered micro architecture, which results from suppressed bone formation and increased bone resorption. The decrease in indices of bone mass, structure and formation are directly linked to the degree of colonic mucosal inflammation. DSS-induced colitis can be used to study pharmacological interventions for bone loss in colitis. PMID- 18675387 TI - Does advanced driver training improve situational awareness? AB - Over 70 years of experiential evidence suggests that a specific form of advanced driver training, one based on an explicit system of car control, improves driver situation awareness (SA). Five experimental hypotheses are developed. They propose that advanced driving should increase the number of information elements in the driver's working memory, increase the interconnection between those elements, increase the amount of 'new' information in memory as well as the prominence of existing information, and that finally, it should stimulate behaviours that help drivers evolve better situations to be aware of. An approach to SA based on Neisser's perceptual cycle theory is anchored to a network based methodology. This is applied within the context of a longitudinal on-road study involving three groups of 25 drivers, all of whom were measured pre- and post intervention. One experimental group was subject to advanced driver training and two further groups provided control for time and for being accompanied whilst driving. Empirical support is found for all five hypotheses. Advanced driving does improve driver SA but not necessarily in the way that existing situation focused, closed loop models of the concept might predict. PMID- 18675388 TI - A field study of sleep and fatigue in a regular rotating 12-h shift system. AB - The aim of this study was to examine a regular rotating 12-h shift system (2D2N4Off) at an Australian Smelter. Sleep behavior, subjective fatigue and neurobehavioral performance were investigated over a 14-day period for 20 employees. Activity monitors, sleep/wake diaries, and 5-min psychomotor vigilance tasks were used. Sleep data showed differences between day and night shifts. While sleep prior to night1 was increased relative to day shifts, a reduced sleep length carried into the period leading to night2. Total wakefulness at the end of shift, and subjective fatigue were increased for night shifts, particularly night1. Decrements in performance data supported these findings. Both prior wakefulness and prior sleep are important in a 12-h shift system. Employees may "sleep in" after day shifts, rather than taking extra sleep prior to night work. Thus, sleep between day and night shifts is based on recovery rather than preparation. PMID- 18675389 TI - Radiodensity and hardness of enamel and dentin of human and bovine teeth, varying bovine teeth age. AB - Studies have evaluated dental hard tissues characteristics from animal species in order to be used as a substitute for human teeth. The aim of this study was to evaluate the radiodensity and hardness of human and bovine enamel and dentin, varying bovine teeth age. Five specimens (1mm thick) were obtained from animals aged 20 (B20), 30 (B30), 38 (B38) and 48 (B48)months and from 20 to 30-years-old human third molars (H). The radiographic images were taken with a phosphor plaque digital system (Digora Optime). The radiodensity was obtained and Knoop hardness (KHN) was recorded (100g for 15s--5 indentations per specimen). Data were analyzed by one-way ANOVA following Tukey's HSD test and Dunnet's two-sided t test. Radiodensity was similar within enamel groups, but bovine dentin presented higher radiodensity than human one regardless of age groups. Enamel-KHN showed differences between B20-B30 and B38-B48-H, and dentin-KHN was similar within all groups. Enamel was always more radiodense than dentin and also presented higher KHN (p=0.001). The use of bovine enamel or dentin should take into consideration the teeth age, but as a general rule it should be recommended to select older bovine teeth due to better chances to find greater similarity with human teeth. PMID- 18675390 TI - Tooth size patterns in patients with hypodontia and supernumerary teeth. AB - AIMS: Anomalies of tooth number may not be isolated conditions but may have wider associations in the development of the dentition including tooth size. This study aimed to examine links between hypodontia, supernumerary teeth and crown size, considering the effect on the development of the whole dentition and so increase understanding of the aetiology of these conditions. METHODS AND RESULTS: The patients, who were all of European ancestry, were 60 young adults (30 males and 30 females) with hypodontia and 60 age and sex matched controls together with 60 young adults (39 males and 21 females) with supernumerary teeth and 60 age and sex matched controls. Hand measurements of mesiodistal and buccolingual dimensions were made of the teeth on dental study models using Mitutoyo electric callipers. The mean value of two measurements was used and intra-operator and inter-operator reliability determined. Patients with hypodontia had smaller teeth than the control group and this difference was statistically significant (p<0.05) for all teeth except the MD dimensions of 13, 23, 24 and 44. The difference in size was greatest for the BL dimensions in hypodontia patients. Further, the greater the number of missing teeth the smaller the tooth size. The hypodontia patients also showed higher variability in tooth dimensions than the control group. Patients with supernumerary teeth had larger teeth than the controls, with the greatest differences in the MD dimensions. In both hypodontia and supernumerary patients the differences in tooth size were generalised throughout the dentition. CONCLUSIONS: In anomalies of tooth number the size of teeth is also involved. In patients with hypodontia and supernumerary teeth the crown size of the whole dentition is affected. These findings are compatible with a multifactorial aetiology of these conditions. PMID- 18675391 TI - Forces and motor control mechanisms during biting in a realistically balanced experimental occlusion. AB - Temporomandibular joint and masticatory muscle forces generated during bilateral biting on an experimental device simulating a symmetrically balanced maximum intercuspation, are unknown. The basic motor control strategies during such tasks, executed either strictly controlled or developed rather habitually, are also quite unclear. The main goal of this study was to compare muscle and joint forces at various magnitudes of force under two experimental conditions: (1) generation of a bite force vector perpendicular to the maxillary occlusal plane, (2) development of a directionally unrestricted (quasi-habitual) bite force, both with identical magnitude. Additionally, the experimental results were evaluated on the basis of optimisation strategies displaying physiologically reasonable neuromuscular objectives for coordinated muscle contraction. In 10 normal subjects, the electric activities of all jaw muscles were recorded bilaterally. Intraoral force transfer and force measurement were achieved by a measuring device with one anterior and two posterior force transmission points. Prior to the experiments, the force transmission was balanced at a directionally unrestricted resultant bite force of 100N. Under visual feedback-control, the subjects generated resultant forces of 50, 100, 150, 200, 300, and 400N, respectively. Joint and muscle forces were calculated based on the electromyograms of all jaw muscles, lines of action, geometrical data of the skull, and physiological cross-sectional areas acquired from all subjects. To identify possible motor control strategies, various physiologically reasonable objective functions were applied. The results revealed significant differences in force patterns generated under the two experimental conditions. Directionally unrestricted biting created higher forces in nearly all muscles and in the jaw joints. Muscle forces normalised with the magnitude of the inherent resultant force, and the findings from the optimisation calculations indicate variable central control mechanisms under the two experimental conditions, both minimizing energy consumption. PMID- 18675392 TI - Effects of concurrent strength and endurance training on physical fitness and symptoms in postmenopausal women with fibromyalgia: a randomized controlled trial. AB - OBJECTIVE: To examine the effectiveness of concurrent strength and endurance training on muscle strength, aerobic and functional performance, and symptoms in postmenopausal women with fibromyalgia (FM). DESIGN: Randomized controlled trial. SETTING: Local gym and university research laboratory. PARTICIPANTS: Twenty-six women with FM. INTERVENTION: Progressive and supervised 21-week concurrent strength and endurance training. MAIN OUTCOME MEASURES: Muscle strength of leg extensors, upper extremities, and trunk; peak oxygen uptake (Vo(2)peak), maximal workload (Wmax), and work time; 10-m walking and 10-step stair-climbing time and self-reported functional capacity (Health Assessment Questionnaire); and symptoms of FM. RESULTS: After concurrent strength and endurance training, the groups differed significantly in Wmax (P=.001), work time (P=.001), concentric leg extension force (P=.043), walking (P=.001) and stair-climbing (P<.001) time, and fatigue (P=.038). The training led to an increase of 10% (P=.004) in Wmax and 13% (P=.004) in work time on the bicycle but no change in Vo(2)peak. CONCLUSIONS: Concurrent strength and endurance training in low to moderate volume improves the muscle strength of leg extensors, Wmax, work time, and functional performance as well as perceived symptoms, fatigue in particular. Concurrent strength and endurance training is beneficial to postmenopausal women with FM without adversities, but more extensive studies are needed to confirm the results. PMID- 18675393 TI - The role of order of practice in learning to handle an upper-limb prosthesis. AB - OBJECTIVE: To determine which order of presentation of practice tasks had the highest effect on using an upper-limb prosthetic simulator. DESIGN: A cohort analytic study. SETTING: University laboratory. PARTICIPANTS: Healthy, able bodied participants (N=72) randomly assigned to 1 of 8 groups, each composed of 9 men and 9 women. INTERVENTIONS: Participants (n=36) used a myoelectric simulator, and participants (n=36) used a body-powered simulator. On day 1, participants performed 3 tasks in the acquisition phase. On day 2, participants performed a retention test and a transfer test. For each simulator, there were 4 groups of participants: group 1 practiced random and was tested random, group 2 practiced random and was tested blocked, group 3 practiced blocked and was tested random, and group 4 practiced blocked and was tested blocked. MAIN OUTCOME MEASURES: Initiation time, the time from the starting signal until the beginning of the movement, and movement time, the time from the beginning until the end of the movement. RESULTS: Movement times got faster during acquisition (P<.001). The blocked group had faster movement times (P=.009), and learning in this group extended over the complete acquisition phase (P<.001). However, this advantage disappeared in the retention and transfer tests. Compared with a myoelectric simulator, movements with the body-powered simulator were faster in acquisition (P=.004) and transfer test (P=.034). CONCLUSIONS: Performance in daily life with a prosthesis is indifferent to the structure in which the training is set up. However, practicing in a blocked fashion leads to faster performance; in novice trainees, it might be suggested to practice part of the training tasks in blocks. PMID- 18675394 TI - Rehabilitation of older Chinese patients with different cognitive functions: how do they differ in outcome? AB - OBJECTIVE: To examine the effect of cognition on functional and motor gain in older Chinese patients undergoing geriatric inpatient rehabilitation. DESIGN: A retrospective cohort study. SETTING: Geriatric rehabilitation units of 2 convalescence hospitals in Hong Kong. PARTICIPANTS: Older Chinese patients (N=778). INTERVENTIONS: Comprehensive geriatric assessment and inpatient rehabilitation by a multidisciplinary team. MAIN OUTCOME MEASURES: Cognitive status was assessed with the Cantonese version of the Mini-Mental State Examination (C-MMSE). We measured the absolute functional gain and motor gain by using the Barthel Index and Elderly Mobility Scale (EMS) and expressed it as Barthel Index efficacy and EMS efficacy. Relative efficacy was assessed by the Montebello Rehabilitation Factor Score (MRFS). Relative efficiency was deduced by relative efficacy divided by the hospital length of stay (LOS). RESULTS: There were significant differences in the total Barthel Index and EMS on admission and at discharge, with lower discharge scores in low C-MMSE groups. The change of Barthel Index (Barthel Index efficacy) and EMS (EMS efficacy) during rehabilitation did not differ among different C-MMSE groups. Significant differences were observed in Barthel Index MRFS efficacy and efficiency as well as EMS MRFS efficacy and efficiency in different cognitive groups; those with better cognitive function had better results. Multivariate analysis showed that LOS (odds ratio [OR]=1.02, P=.002) and diagnosis of musculoskeletal problems (OR=2.24, P=.007) were positive predictors for a Barthel Index MRFS efficacy of .25 or higher. C-MMSE was not an independent predictor for a Barthel Index MRFS efficacy of .25 or higher. LOS (OR=1.02, P=.003), admission C-MMSE (OR=1.04, P=.001), body mass index (OR=1.05, P=.006), and diagnosis of musculoskeletal disorders (OR=1.75, P=.04) were significant positive predictors for an EMS MRFS efficacy of .25 or higher, whereas urinary incontinence was a negative predictor for EMS MRFS efficacy (OR=0.69, P=.04). CONCLUSIONS: There was a strong association between admission cognition and relative change in functional and motor efficacy and efficiency. Cognitive function was not associated with absolute functional and motor gain. Cognitive function was an independent predictor for satisfactory mobility outcome but not for functional outcome. Older patients with impaired cognitive function could still benefit from rehabilitation. PMID- 18675395 TI - Changes and determinants of life satisfaction after spinal cord injury: a cohort study in the Netherlands. AB - OBJECTIVE: To determine the impact of spinal cord injury (SCI) on life satisfaction of persons with SCI 1 year after discharge of inpatient rehabilitation. DESIGN: A cohort study. Life satisfaction before SCI was retrospectively measured at the start of active rehabilitation. One year after discharge from inpatient rehabilitation, current life satisfaction was measured. SETTING: Eight rehabilitation centers in The Netherlands. PARTICIPANTS: Persons (N=147) aged 18 to 65 and wheelchair-dependent at least for long distances. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The Life Satisfaction Questionnaire. RESULTS: Mean satisfaction with life +/- SD as a whole was 5.3+/-0 before SCI and 4.3+/-1.3 one year after inpatient rehabilitation. Sexual life, self-care, and vocational situation showed the largest impact of SCI (P<.05), whereas the social relationships domains appeared to be the least affected. Decrease of life satisfaction after SCI was larger when using the retrospective ratings than when using general population scores. Significant determinants of life satisfaction after SCI were high lesion level (beta=.31, P<.05), pain (beta=.19, P<.05), and secondary impairments (beta=.22, P<.05). CONCLUSIONS: Life satisfaction decreased in persons with SCI. Level of lesion and suffering secondary impairments or pain were associated with low life satisfaction 1 year after discharge from inpatient rehabilitation. PMID- 18675396 TI - Isolated lumbar extensor strengthening versus regular physical therapy in an army working population with nonacute low back pain: a randomized controlled trial. AB - OBJECTIVE: To evaluate the effectiveness of specific lumbar extensor training compared with regular physical therapy (PT) in workers with nonspecific nonacute low back pain (LBP). DESIGN: A multicenter randomized controlled trial with 1 year follow-up. SETTING: PT department in (military primary care) health centers. PARTICIPANTS: Predominantly male soldiers (N=129) with 4 weeks or more of low back complaints who were referred by the health center's general practitioner for PT (mean age, 35.9+/-10.8 y; range, 20-56 y), of whom 127 randomized participants were included in the analyses. One patient withdrew because of adverse effects during treatment. INTERVENTIONS: Participants were assigned to 1 of 2 treatment programs: (1) a 10-week device-supported isolated lumbar extension training, twice a week, or (2) regular PT, mainly consisting of exercise therapy and aerobic activities. MAIN OUTCOME MEASURES: Functional status (Roland-Morris Disability Questionnaire, Patient-Specific Functional Scale) and global perceived effect were assessed in the short term (5 wk, 10 wk) and long term (6 mo, 12 mo). RESULTS: Both groups showed a favorable development in main outcomes over time: short-term improvements (after 10 weeks of treatment) remained stable or even slightly increased throughout the 12-month follow-up. No significant differences between the 2 groups were shown for any of the outcome measures, at any time. CONCLUSIONS: Consistent with prior evidence, specific back strengthening does not seem to offer incremental benefits in LBP management compared with regular PT care that mainly consists of general exercise therapy. (ISRCTN identifier ISRCTN19334317.) PMID- 18675398 TI - All we need is a cue to remember: the effect of an extinction cue on renewal. AB - In the present study, it was investigated by employing a human fear conditioning paradigm whether an extinction retrieval cue can attenuate renewal of conditioned responding after an extinction treatment procedure, and if so, what the precise role of such an extinction cue comprises. It was hypothesized that such a cue can attenuate renewal and would function as a safety signal capable of directly inhibiting the expectancy of an aversive outcome and conditioned skin conductance responding to a conditioned stimulus. The results demonstrated that the extinguished expectancy of an aversive outcome was renewed when the CS was presented outside the extinction context and that an extinction cue attenuated this effect. This extinction cue, however, only transferred its inhibitory properties to other, non-extinguished stimuli when there was no contextual switch. This safety signal function was not observed after a switch in context. Possible functions of the extinction cue and its application in extinction-based exposure treatments are discussed. PMID- 18675397 TI - The utility of the A1 and A2 criteria in the diagnosis of PTSD. AB - In the field of posttraumatic stress disorder (PTSD), the revisions to the DSM-IV definition of a potentially traumatic event are contentious. Proponents praise the subjective emphasis, while others contend that the changes to the criterion broadened the conceptualization of PTSD. This study examined the predictive utility of Criterion A events, examining the stressor (A1) and subjective emotional response (A2) components of the definition of a traumatic event. Rates of Criterion A events and PTSD were calculated for three diverse samples, and predictive power, sensitivity, specificity, and ROC curves were computed to determine the predictive utility of Criterion A requirements for PTSD symptom, duration, and functional impairment diagnostic criteria. Across all samples, the current Criterion A requirements did not predict much better than chance. Specifically, A2 reports added little to the predictive ability of an A1 stressor, though the absence of A2 predicted the absence of PTSD-related symptoms, their duration, and impairment. Notably, the combination of three A1 and A2 criteria showed the best prediction. Confronted events also showed less predictive ability than experienced events, with more variable performance across samples. These results raise fundamental questions about the threshold or "gate" that Criterion A ought to play in our current nosology. PMID- 18675399 TI - The effect of stress on craving for methadone depends on the timing of last methadone dose. AB - BACKGROUND: This study examined the extent to which the timing of last methadone dose moderates the influence of a laboratory stressor on craving for methadone in a sample of methadone maintenance patients. METHODS: Methadone maintenance patients (N=41) completed a computerized stress manipulation with two levels (low and high stress) on two separate days: (1) immediately following the administration of their standard daily methadone dose and (2) immediately prior to receiving their standard daily dose. The impact of stress, timing of methadone dose and their interaction were examined on self-report measures of stress and craving for methadone. RESULTS: Craving ratings varied according to the timing of last methadone dose. Immediately following methadone dose (i.e., day 1), there was no effect for stress on craving (0.12; p=0.30). However, prior to receiving methadone dose (i.e., day 2), the high stress effect on craving was significant (1.51; p<0.001). DISCUSSION: The present findings indicate that patients currently receiving methadone maintenance treatment may be particularly vulnerable to stress-related relapse immediately prior to daily methadone dosing. PMID- 18675400 TI - A multi-session interpretation modification program: changes in interpretation and social anxiety symptoms. AB - Previous research suggests that socially anxious individuals interpret ambiguous social information in a more threatening manner compared to non-anxious individuals. Recently, studies have experimentally modified interpretation and shown that this subsequently affected anxiety in non-anxious individuals. If similar procedures can modify interpretation biases in socially anxious individuals, they may lead to a reduction in social anxiety symptoms. In the current study, we examined the effect of a computerized Interpretation Modification Program (IMP) on interpretation bias and social anxiety symptoms. Twenty-seven socially anxious individuals were randomly assigned to the IMP or a control condition. Participants completed eight computer sessions over four weeks. The IMP modified interpretation by providing positive feedback when participants made benign interpretations and negative feedback in response to threat interpretations. The IMP successfully decreased threat interpretations, increased benign interpretations, and decreased social anxiety symptoms compared to the control condition. Moreover, changes in benign interpretation mediated IMP's effect on social anxiety. This initial trial suggests that interpretation modification may have clinical utility when applied as a multi-session intervention. PMID- 18675401 TI - Styles of pulling in youths with trichotillomania: exploring differences in symptom severity, phenomenology, and comorbid psychiatric symptoms. AB - To date, no studies have examined possible phenomenological differences between the automatic and focused styles of pulling in youths with TTM. The aims of the current study were to examine differences in TTM severity, phenomenology, comorbid psychiatric symptoms, and functional impact across youths with varying degrees of these pulling styles. Youths between the ages of 10 and 17 years (and their parent) were recruited via an Internet-based survey. A total of 186 youths with chronic hair pulling was classified as "high-focused" or "low-focused" and either "high-automatic" or "low-automatic" based upon scores obtained on the Milwaukee Inventory for Styles of Trichotillomania-Child Version (MIST-C) using a median-split procedure. Results demonstrated significant differences between pulling styles. More specifically, "high-focused" pullers reported more severe TTM and greater symptoms of anxiety and depression than "low-focused" pullers, and "high-automatic" pullers reported greater symptoms of depression than "low automatic" pullers. Subsequent analyses suggest that, in comparison to youths with low levels of both automatic and focused pulling, those experiencing high levels of focused pulling but low levels of automatic pulling reported phenomenological differences and were more likely to engage in additional repetitive behaviors (i.e., skin picking, lip/cheek biting). Clinical and research implications, study limitations, and future areas of research are discussed. PMID- 18675402 TI - A randomized comparison of two motivationally enhanced Internet behavioral weight loss programs. AB - PURPOSE: Sixty-six women participated in this study, which compared two 16-week motivationally enhanced weight loss interventions. Both intervention groups, which are entitled "Minimal" and "Enhanced" throughout the description that follows, received a motivationally enhanced initial face-to-face weight loss session. The initial session was followed by a 16-week Internet behavioral weight loss program, which included a study website with weekly lessons, an online self monitoring diary form, web links to related sites, and separate message boards. The Enhanced group also received weekly moderated online chat group sessions led using motivational techniques. RESULTS: Both groups lost weight over time (p<0.001), however the group by time interaction was non-significant (p=0.19). The Minimal group lost 5.22+/-4.72 kg and the Enhanced group lost 3.71+/-4.46 kg. In the Enhanced group, online chat session attendance averaged 8.0 of the 16 chats offered. Greater program utilization was associated with greater weight loss in both groups. CONCLUSIONS: The addition of weekly motivationally enhanced online chats did not increase weight loss in this study. Further research is needed on the type and frequency of group support that is most desirable and beneficial in Internet weight loss interventions and could also explore building motivation for program utilization. PMID- 18675403 TI - Parenting-related childhood learning history and panic vulnerability: a test using a laboratory-based biological challenge procedure. AB - A small but growing literature highlights specific parenting behaviors in increasing panic vulnerability among offspring. The current study examined the association between parenting-related instrumental and observational learning of sick-role behavior during childhood and reactivity to a panic-relevant biological challenge procedure that has evidenced predictive validity in terms of panic onset. Participants were 93 physically and psychologically healthy young adults (39 females; M(age)=23.41 years). As expected, results indicated that instrumental learning experiences involving (panic-relevant) arousal-reactive symptoms predicted increased post-challenge anxiety, arousal, and negative affective valence, even after accounting for variability associated with other theoretically relevant variables (e.g., anxiety sensitivity). Consistent with prior work, this learning history effect was specific to arousal-reactive, as opposed to arousal non-reactive, symptoms. Unexpectedly, observational learning was not related to challenge responding. Findings are discussed in terms of the potential role of parenting in etiologic models of panic development. PMID- 18675404 TI - Crystal structure of cyclomaltoheptaose (beta-cyclodextrin) complexes with p aminobenzoic acid and o-aminobenzoic acid. AB - Crystal structures of cyclomaltoheptaose (beta-cyclodextrin) complexes with p aminobenzoic acid and o-aminobenzoic acid have been determined by single-crystal X-ray diffraction. The space group of the beta-cyclodextrin-p-aminobenzoic acid complex is P2(1) with a host:guest stoichiometry of 1:1, and that of the beta cyclodextrin-o-aminobenzoic acid complex is P1 with a stoichiometry of 2:3. The different structures of the guest molecules lead to the different molecular packing structures of the two complexes. Intermolecular hydrogen-bond interactions are the main force that stabilize the supramolecular systems. In both crystals, there are water molecules located near the cavity rims and in interstices between molecules of beta-cyclodextrin participating in formation of intermolecular hydrogen bonds. PMID- 18675405 TI - Novel carbonyl-group-containing dextran synthesis by pyranose-2-oxidase and dextransucrase. AB - The carbonyl polysaccharide, keto-dextran, was synthesized by the regioselective oxidation of sucrose and by the subsequent transfer reaction of the oxidized sucrose. The regioselective oxidation of sucrose was performed by bioconversion with pyranose-2-oxidase (EC 1.1.3.10). After 24h, the conversion percentage of sucrose into keto-sucrose was 100% as determined by a colorimetric method with dinitrophenylhydrazine. Converted keto-sucrose was polymerized to keto-dextran by dextransucrase (EC 2.4.1.5). Polymerization of keto-dextran was confirmed by the increase in molecular weight and amount of keto-dextran produced. The amount of keto-dextran produced decreased to 80% of the amount of dextran produced owing to the substrate recognition of DSase. From a Lineweaver-Burk reciprocal plot, the Michaelis constants for sucrose and keto-sucrose were 4.6 mmol L(-1) and 14.0 mmol L(-1), respectively. The keto-dextran had a carbonyl group in all glucose units. PMID- 18675406 TI - Catalytic properties and mode of action of endo-(1-->3)-beta-D-glucanase and beta D-glucosidase from the marine mollusk Littorina kurila. AB - A complex of the enzymes from the liver of the marine mollusk Littorina kurila that hydrolyzes laminaran was investigated. Two (1-->3)-beta-d-glucanases (G-I and G-II) were isolated. The molecular mass of G-I as estimated by gel-permeation chromatography and SDS-PAGE analysis was 32 and 40kDa, respectively. The G-II molecular mass according to SDS-PAGE analysis was about 200kDa. The pH optimum for both G-I and G-II was pH 5.4. The G-I had narrow substrate specificity and hydrolyzed only the (1-->3)-beta-d-glucosidic bonds in the mixed (1-->3),(1-->6)- and (1-->3),(1-->4)-beta-d-glucans down to glucose and glucooligosaccharides. This enzyme acted with retention of the anomeric configuration and catalyzed a transglycosylation reaction. G-I was classified as the glucan endo-(1-->3)-beta-d glucosidase (EC 3.2.1.39). G-II exhibited both exo-glucanase and beta-d-glucoside activities. This enzyme released from the laminaran glucose as a single product, but retained the anomeric center configuration and possessed transglycosylation activity. The hydrolysis rate of glucooligosaccharides by G-I decreased with an increase of the substrate's degree of polymerization. In addition to (1-->3)-beta d-glucanase activity, the enzyme had the ability to hydrolyze p-nitrophenyl beta d-glucoside and beta-d-glucobioses: laminaribiose, gentiobiose, and cellobiose, with the rate ratio of 50:12:1. G-II may correspond to beta-d-glucoside glucohydrolase (EC 3.2.1.21). PMID- 18675407 TI - TGF-beta expression by allogeneic bone marrow stromal cells ameliorates diabetes in NOD mice through modulating the distribution of CD4+ T cell subsets. AB - BMSCs could promote the regeneration of islet beta-cell, but the status of BMSCs under diabetes is still unknown. Our study verified the effect of allogeneic BMSCs (ICR) transferred into NOD mice on blood glucose and CD4+ T cells subsets function. In vivo experiment, BMSCs could decrease blood glucose, weaken lymphocytes proliferation. In vitro experiment, the distribution of CD4+ T cell subsets was changed after co-culture with BMSCs, resulting in a greater frequency of Treg cells and reduced representation of Th17 cells. After TGF-beta blockade, CD4+ T cells differentiated along a route favoring development of Th17, but not Treg cells. Thus, NOD can be treated by BMSCs which changes the distribution of CD4+ T cells, increases the number of Treg cells, and inhibits the differentiation of Th17 cells. And the positive effects of allogeneic BMSCs in the treatment of NOD mice depend on the regulation of TGF-beta secreted by BMSCs. PMID- 18675408 TI - Immediate effects of form-class constraints on spoken word recognition. AB - In many domains of cognitive processing there is strong support for bottom-up priority and delayed top-down (contextual) integration. We ask whether this applies to supra-lexical context that could potentially constrain lexical access. Previous findings of early context integration in word recognition have typically used constraints that can be linked to pair-wise conceptual relations between words. Using an artificial lexicon, we found immediate integration of syntactic expectations based on pragmatic constraints linked to syntactic categories rather than words: phonologically similar "nouns" and "adjectives" did not compete when a combination of syntactic and visual information strongly predicted form class. These results suggest that predictive context is integrated continuously, and that previous findings supporting delayed context integration stem from weak contexts rather than delayed integration. PMID- 18675409 TI - Visual working memory capacity for objects from different categories: a face specific maintenance effect. AB - The capacity of visual working memory was examined when complex objects from different categories were remembered. Previous studies have not examined how visual similarity affects object memory, though it has long been known that similar-sounding phonological information interferes with rehearsal in auditory working memory. Here, experiments required memory for two or four objects. Memory capacity was compared between remembering four objects from a single object category to remembering four objects from two different categories. Two-category sets led to increased memory capacity only when upright faces were included. Capacity for face-only sets never exceeded their nonface counterparts, and the advantage for two-category sets when faces were one of the categories disappeared when inverted faces were used. These results suggest that two-category sets which include faces are advantaged in working memory but that faces alone do not lead to a memory capacity advantage. PMID- 18675410 TI - Cognitive function and blood methylmercury in adults living near a deserted chloralkali factory. AB - We assessed the association between blood methylmercury (MeHg) and cognitive function in 240 adult residents living near a deserted chloralkali plant. Total mercury (T-Hg) in the blood, MeHg, and health and dietary related questionnaire were examined for all participants. The Cognitive Abilities Screening Instrument (CASI C-2.0) and Mini-Mental State Examination (MMSE) were used to assess the participants' cognitive functions. We found a significantly high correlation (r=0.979; p<0.001) between blood T-Hg (17.3+/-10.9 microg/L) and MeHg (15.3+/-9.2 microg/L). We also found significantly higher blood MeHg levels in participants with high local fish and seafood consumption, which revealed that dietary intake was the major exposure route of MeHg. All the participants were assigned to the high-MeHg (H-MeHg, 27.0+/-10.4 microg/L) or low-MeHg (L-MeHg, 11.6+/-4.7 microg/L) groups based on the 75th percentile of their blood MeHg (19.2 microg/L), and then matched for cognitive function confounders: age, gender, and education levels. Higher abnormality rates for remote memory (p=0.036), mental manipulation (p=0.013), and orientation (p=0.005) were found in the H-MeHg group than in the L-MeHg group. Long-term consumption of MeHg-contaminated fish and seafood by residents living near this contaminated area may have persistent effects on their cognitive function. We suggest a follow-up study to monitor the long-term health effects on the residents living near this deserted plant. PMID- 18675411 TI - Hyaluronic acid coated poly-epsilon-caprolactone nanospheres deliver high concentrations of cyclosporine A into the cornea. AB - The objective of this study was to determine cyclosporine A (Cy A) levels in ocular tissues and fluids after topical administration of poly-epsilon caprolactone (PCL)/benzalkonium chloride (BKC) nanospheres and hyaluronic acid (HA) coated PCL/BKC nanospheres onto healthy rabbit corneas. Nanospheres were prepared by nanoprecipitation and purified by gradient-rate centrifugation. Cy A (0.1%) in either castor oil solution (group 1), PCL/BKC nanosphere formulation (group 2) or HA coated PCL/BKC nanosphere formulation (group 3) was instilled onto rabbit corneas. Tear samples were adsorbed onto Schirmer tear strips. Cy A concentrations of fluid (blood, aqueous humor, tear) and specimen extracts (cornea, conjunctiva, iris/ciliary body) were determined by high performance liquid chromatography-mass spectrometry (LC-MS). The mean corneal Cy A concentration obtained at 0.5, 1, 2, 4, 8 and 24h following instillation of the formulations ranged between 0.12 and 1.2 ng/mg tissue for group 1, 5.9-15.5 ng/mg tissue for group 2 and 11.4-23.0 ng/mg for group 3 (one-way analysis of variance (ANOVA) and pairwise tests (SNK (Student-Newman-Keuls) and Tukey); p<0.05). Conjunctival Cy A levels of group 2 and 3 were not significantly different at any of the time points tested. However, there was a significant difference between Cy A concentration of castor oil formulation and that of PCL/BKC nanosphere formulation at 1 and 8h (p<0.05). The mean iris/ciliary body concentrations obtained with the three formulations were not significantly different at any time point with the exception of group 2 levels being higher than those of groups 1 and 3 at 1h (p<0.05). The lowest ocular tear Cy A concentrations (16-114 ng/ml) were found following the instillation of HA coated PCL/BKC nanoparticles (group 3) during the time period tested. Cy A loaded PCL/BKC and HA coated PCL/BKC nanospheres are able to achieve high levels of Cy A in the cornea that is 10-15 fold higher than that is achieved with Cy A solution in castor oil. Nanosphere formulation and HA may play an important role in delivering high levels of cyclosporine A into the cornea. PMID- 18675412 TI - Clinical pregnancy and delivery of a healthy baby by a 44-year-old woman after a successful in vitro fertilization frozen cycle using her own oocytes. AB - OBJECTIVE: To present a successful IVF frozen-embryo replacement cycle in a 44 year-old woman, using her own oocytes. DESIGN: Case report. SETTING: Assisted reproduction unit in a university hospital. PATIENT(S): A 44-year-old woman underwent IVF frozen-embryo replacement cycle after three unsuccessful IVF fresh cycles with her own oocytes. The couple were unable to achieve pregnancy for 4 years and were advised to have IVF because of the long duration of subfertility and woman's age. INTERVENTION(S): In vitro fertilization. MAIN OUTCOME MEASURE(S): Pregnancy, live birth, baseline FSH on day 3, and IVF outcome. RESULT(S): The woman achieved pregnancy through a frozen-embryo replacement cycle and gave birth to a healthy girl who weighed 3,330 g. CONCLUSION(S): Pregnancy is difficult to achieve for women >40 years of age and especially so for a woman at the age of 44 years. In vitro fertilization cycles may not lead to the desired result, but the availability of a frozen embryo replacement cycle is associated with better prognosis. PMID- 18675413 TI - Second trimester amniocentesis in assisted conception versus spontaneously conceived twins. AB - OBJECTIVE: To compare the outcome of amniocentesis in twins conceived with assisted reproduction technology (ART) versus spontaneously conceived twins. DESIGN: Retrospective analysis of case records between 1993 and 2006. SETTING: University-affiliated tertiary center for fetal medicine. PATIENT(S): 167 ART twin pregnancies and 275 spontaneous twin pregnancies. INTERVENTION(S): Genetic amniocentesis. MAIN OUTCOME MEASURE(S): Comparison of pregnancy loss rate and perinatal outcome between the ART and spontaneous twin-pregnancy groups. RESULT(S): The fetal loss rate was similar between the two groups (4.2% vs. 4.0% in the ART twins and spontaneous twins, respectively), although the interval between amniocentesis to miscarriage was statistically significantly shorter in the ART twins than the spontaneous twins (6.2 and 20.1 days, respectively). In all cases, fetal loss refers to the loss of the entire pregnancy. The preterm delivery rate before 37 weeks was statistically significantly higher in the ART group (64.1%) compared with controls (49.5%). CONCLUSION(S): Amniocentesis in ART twins carries a fetal loss rate similar to spontaneous twins. However, ART twins have a statistically significantly increased risk of preterm delivery especially before 32 weeks' gestation. PMID- 18675416 TI - Fuzzy-rule emulated networks, based on reinforcement learning for nonlinear discrete-time controllers. AB - This article introduces an adaptive controller for a class of nonlinear discrete time systems, based on self adjustable networks called Multi-Input Fuzzy Rules Emulated Networks (MIFRENs), and its reinforcement learning algorithm. Because of the universal function approximation of MIFREN, the first MIFREN called MIFREN(c) is used to estimate a long-term cost function, which demonstrates as a performance index for the tuning procedure. Another network or MIFREN(a) is designed as a direct controller via the human knowledge through defined If-Then rules. The selection procedure for any system parameters, such as learning rates and some constant parameters, is represented by the proof of proposed theorems. The system's performance is demonstrated by computer simulations via selected nonlinear discrete-time systems, and comparison results with other controllers to validate theoretical development. PMID- 18675415 TI - Pregnancy outcome in preimplantation genetic diagnosis cycle by blastomere biopsy is related to both quality and quantity of embryos on day 3. AB - The aim of this study was to determine the impact of day 3 embryo quality and quantity on subsequent embryonic development and pregnancy outcome following embryo biopsy. The findings demonstrate that a significantly lower proportion of poor/fair-quality embryos develop further after biopsy, and that patients with <4 good embryos on day 3 may have poor outcome. Therefore, appropriate counseling needs to be given to them before therapy is initiated. PMID- 18675414 TI - Intrafollicular antimullerian hormone levels predict follicle responsiveness to follicle-stimulating hormone (FSH) in normoandrogenic ovulatory women undergoing gonadotropin releasing-hormone analog/recombinant human FSH therapy for in vitro fertilization and embryo transfer. AB - OBJECTIVE: To investigate the relationship between antimullerian hormone (AMH) and steroidogenesis in follicles of normoandrogenic ovulatory women undergoing follicle-stimulating hormone (FSH) therapy for in vitro fertilization/embryo transfer (IVF-ET). DESIGN: Prospective cohort. SETTING: Institutional/private practice. PATIENT(S): 26 normoandrogenic ovulatory women. All women received gonadotropin-releasing hormone (GnRH) analog and ovarian stimulation for IVF-ET. INTERVENTION(S): Follicle fluid was aspirated at oocyte retrieval from the first follicle of each ovary. MAIN OUTCOME MEASURE(S): Follicle fluid was assayed for AMH, estradiol (E(2)), progesterone, androstenedione, testosterone, dihydrotestosterone, insulin, and FSH. RESULT(S): Intrafollicular AMH levels positively and negatively correlated with E(2) and FSH concentrations in follicles, respectively, causing a positive relationship between follicle fluid AMH levels and E(2)/FSH ratios as a measure of follicle sensitivity to FSH. A positive relationship also existed in follicles between AMH levels and E(2)/androgen ratios as a marker of aromatase activity. CONCLUSION(S): The AMH levels in follicles of IVF patients positively correlate with follicle sensitivity to FSH. PMID- 18675417 TI - When should we stop resuscitation efforts after blunt traumatic arrest? PMID- 18675418 TI - Intimate partner and nonpartner violence against pregnant women in rural Haiti. AB - OBJECTIVE: To examine the association between violence experienced by pregnant Haitian women in the previous 6 months and pregnancy-related symptom distress. METHODS: A total of 200 women seeking prenatal care at community health dispensaries in the Artibonite Valley were interviewed. RESULTS: Over 4 in 10 women (44.0%) reported that they had experienced violence in the 6 months prior to interview; 77.8% of these women reported that the violence was perpetrated by an intimate partner. Those who experienced intimate partner violence reported significantly greater pregnancy-related symptom distress (beta=0.23, P=0.001). No significant differences between violence perpetrated by family members or others and reporting of symptoms were observed (beta=0.06, P=0.38). CONCLUSION: The findings indicate the need to integrate violence screening, resources, and primary prevention into prenatal care in rural Haiti. PMID- 18675419 TI - The relationship between nurse staffing and patient outcomes: a case study. AB - BACKGROUND: Research has shown a direct relationship between staffing levels and patient outcomes for specific nurse-sensitive indicators, with lower patient to nurse ratios (i.e. less patients per nurse) associated with better outcomes. OBJECTIVES: To explore the relationship between nurse staffing characteristics (the nursing hours worked by permanent and temporary staff and nurse hours per patient day) and patient outcomes: pressure sores, patient falls, upper gastrointestinal bleed, pneumonia, sepsis, shock and deep vein thrombosis. DESIGN: A case study using retrospective hospital data, at ward level. SETTING: A tertiary cardio-respiratory NHS Trust in England, comprising two hospitals. PARTICIPANTS: All patients, including day cases, who were admitted to either hospital as an in-patient over 12 months. METHODS: Data were extracted from corporate hospital systems. The clinical areas were categorised as lower dependency, i.e. wards, or critical care which included ICU and high dependency units. The relationship between nurse staffing characteristics and patient outcomes was assessed using either a Poisson or negative binomial regression model as appropriate. We sought to establish whether the outcomes were affected by the nurse hours per patient day, the permanent nurse hours worked as a percentage of the total hours, and the permanent nurse hours worked as a percentage of the permanent and bank hours combined. RESULTS: In the lower dependency category wards there was only a weak association demonstrated between nurse staffing and the majority of the outcomes. The results from the high dependency critical care areas showed few significant results with only the rate of sepsis being significantly reduced as the ratio of permanent staff hours increased. CONCLUSIONS: The study demonstrated the possibility of using existing hospital data to examine the relationship between nurse staffing and patient outcomes, however the associations found were weak and did not replicate reliably the findings from previous work. PMID- 18675420 TI - Establishing reference values for cervical spine range of motion in pre-pubescent children. AB - Medical professionals, physical therapists, product designers, and computational models all use cervical spine range of motion reference values. To support these functions, researchers have collected a plethora of data to determine the normal range of motion of the cervical spine of adult subjects. However, little to no data exists for subjects under the age of 14. This study utilized the cervical range of motion device, referenced with respect to the Frankfort Plane, to measure the active cervical spine range of motion in all three cardinal planes of the human body, for 106 subjects whose ages ranged from 8 to 10 years. The active range of motion for flexion, extension, lateral extension, and rotation was calculated as 66+/-13 degrees , 85+/-14 degrees , 58+/-8 degrees , and 77+/-7 degrees , respectively, using linear statistics. The observed data significantly differed from the published American Medical Association guidelines for adults but fell within the range of the reference values for 10 year olds. Stratifying and analyzing the range of motion data with respect to gender yielded no significant effect. Appendix A analyzes the data using angular statistics, and produces virtually identical results as those from linear statistics. PMID- 18675421 TI - Regional peak plantar pressures are highly sensitive to region boundary definitions. AB - Traditional pedobarographic analyses subsample pressure data over a number of discrete anatomical regions of interest (ROIs). To our knowledge, the sensitivity of these data to ROI boundary definitions has not been previously addressed. Eight subjects each performed 20 trials of self-paced walking; commercial software was used to define 10 ROIs for each of the 160 total peak pressure images, and regional peak pressures (RPPs) were extracted for each image (total: 1600 values). We then asked three specific questions regarding RPP sensitivity to ROI boundary definition: (1) Is the ROI centroid representative of the RPP location? (2) How frequently do RPPs lie on the ROI boundary? and (3) By how much do RPP values change if the ROI boundary is changed by one pixel (resolution: 5.08 x 7.62 mm)? We found that the RPP locations differed from the ROI centroid in 80% of the cases and that the RPPs lay on the ROI boundary with a probability of 65%. We also found that a single-pixel change in the ROI boundary caused a mean RPP change of 10.8%. The most sensitive region was the midfoot for which a single-pixel ROI change yielded a median 29.4% change in RPP. These results indicate that RPP data are biased by regionalization schemes, which delineate pressure fields based on anatomy rather than on the field's geometric properties, and ultimately that regionalization may constitute a poor method of quantifying complex pressure fields. RPP sensitivity should be considered when making statistical inferences regarding foot function. PMID- 18675422 TI - Patient-specific knee joint finite element model validation with high-accuracy kinematics from biplane dynamic Roentgen stereogrammetric analysis. AB - Little is known about in vivo menisci loads and displacements in the knee during strenuous activities. A new method that combines high-speed kinematics measured with biplane dynamic Roentgen stereogrammetric analysis (DRSA) and a subject specific finite element (FE) model for studying in vivo meniscal behavior is presented here. Further model calibration in a very controlled uniaxial low and high-rate compression loading condition is presented by comparing the model behavior against the measured high-accuracy menisci DRSA kinematics and direct tibio-femoral pressure measurement from a K-scan sensor. It is apparent that certain model aspects such as removing of the pressure sensor from the model can result in relatively large errors (14%) in contact parameters that are not reflected in the change of the measured meniscal kinematics. Changing mesh size to 1mm by 1mm elements increased the magnitude of all but one of the contact variables by up to 45%. This local validation using accurate localized patient specific geometry and meniscal kinematics was needed to enhance model fidelity at the level of contact between menisci and cartilage. PMID- 18675423 TI - Multi-residue determination of seven quinolones antibiotics in gilthead seabream using liquid chromatography-tandem mass spectrometry. AB - A sensitive and selective confirmatory analytical method for the multi-residue determination of seven quinolones (ciprofloxacin, enrofloxacin, sarafloxacin, danofloxacin, oxolinic acid, nalidixic acid and flumequine) in gilthead seabream (Sparus aurata) was developed. The sample pre-treatment involves extraction with 0.1 M NaOH and purification by solid-phase extraction (SPE) on Waters Oasis HLB cartridges followed by the determination of all compounds in a single LC electrospray ionization MS/MS run. Separation was achieved on a Perfectsil ODS-2, 5 microm, 250 mm x 4 mm, analytical column (MZ Analysentechnik) by gradient elution using a mixture of 0.2% (v/v) formic acid, methanol and acetonitrile within 30 min. Multiple reaction monitoring (MRM) was used for selective detection of each quinolone. Accuracy was evaluated through recovery studies at three different fortification levels. The mean recoveries are between 90 and 132% for the selected levels with RSD values lower than 20%. The method presents satisfactory results for linearity, precision and limits of quantification. The latter are much lower than the maximum residue limits (MRLs) established by the European Union for quinolones in fish tissues (6-8 microg/kg). PMID- 18675424 TI - The calcite/water interface II. Effect of added lattice ions on the charge properties and adsorption of sodium polyacrylate. AB - The origin of the surface potential of calcium carbonate in aqueous dispersions and the dissolution of calcite in systems containing excess Ca(2+) and CO(3)(2-) have been the subjects of this study. In addition, stabilization of calcite particles with an anionic polyelectrolyte (sodium polyacrylate (NaPA)) and the effect on surface potential and dissolution of calcite have been studied. Preferential dissolution of either Ca(2+) or CO(3)(2-) from the surface, which is governed by the partial pressure of CO(2) in solution and the pH of the solution, mainly determines the surface potential. Both lattice ions (Ca(2+) and CO(3)(2-)) adsorb onto the surface and thus alter the surface potential. NaPA adsorbs strongly onto the calcite surface regardless of background electrolyte concentration, and reverses the surface potential to negative values. Chelation of the surface due to NaPA can be partly prevented by adding Ca(2+) to the dispersion. PMID- 18675425 TI - Applicability of waste materials--bottom ash and deoiled soya--as adsorbents for the removal and recovery of a hazardous dye, brilliant green. AB - Deoiled soya, an agricultural waste material, and bottom ash, a waste of power plants, have been successfully used for the removal and recovery of the hazardous water-soluble dye brilliant green from water. To remove the dye from water, batch adsorption studies have been carried out by observing the effects of pH, concentration, amounts of adsorbents, size of adsorbent particles, etc. Attempts have also been made to monitor the adsorption process through Langmuir, Freundlich, Tempkin, and D-R adsorption isotherm models. Relevant thermodynamic parameters have also been calculated from these models. The adsorption process has been found endothermic and feasible at all the temperatures. The kinetics of the adsorption was also recorded and indicates pseudo-second-order kinetics in both cases. Kinetic operations also reveal the involvement of a film diffusion mechanism for the deoiled soya adsorption at all the temperatures, while bottom ash undergoes through a particle diffusion mechanism at only 30 degrees C and at higher temperatures a film diffusion mechanism operates. Bulk removal of the dye has been carried out through column studies for both adsorbents. Attempts have also been made to recover the dye from exhausted columns by eluting sulfuric acid of pH 3. PMID- 18675426 TI - Electrochemically produced responsive hydrogel films: influence of added salt on thickness and morphology. AB - We report on electrochemically prepared hydrogel layers of poly-N isopropylacrylamide (pNIPAm) and on the influence that the supporting electrolyte has on their thickness and morphology. Ions that are destabilizing in the Hofmeister sense increase the thickness. The effect correlates well with the ion's tendency to lower the lower critical solution temperature (LCST) of pNIPAm films. AFM micrographs show small-scale globules. When the films were produced in the presence of a destabilizing salt (such as ammonium sulfate) one also observes larger features, resembling wrinkles. We attribute the globules to nucleated growth of surface-attached microgels, whereas the wrinkles presumably are produced by the collapse of hydrogen bubbles underneath a well-crosslinked film. Adding a chain transfer agent to the reactant solution reduces the lateral heterogeneities. PMID- 18675427 TI - Probing of polyelectrolyte monolayers by zeta potential and wettability measurements. AB - Detection of the very first step of polyelectrolyte adsorption onto a solid support is of great importance for understanding mechanisms of solid surface modification. It was shown that streaming potential and contact angle measurements can be successfully used for polyelectrolyte (PE) adsorption characterization in a broad range of surface coverage. Cationic polyallylamine hydrochloride (PAH) was used for the formation of the layer. The electrokinetic characteristics of the substrate covered by the PAH layer were compared with contact angles measured under wet (captive air bubble/substrate in water) and dry (sessile water droplet/dried substrate) conditions. It has been demonstrated that contact angle values determined under both conditions are in good agreement. The observed rapid increase in the contact angle from zero for the bare mica surface to the value close to one characteristic of the PAH monolayer appears in the same PAH coverage range as zeta potential value changes due to adsorption. These results show that wettability can be as sensitive to the presence of small amounts of adsorbed species as electrokinetic measurements. PMID- 18675428 TI - Learning to label letters by sounds or names: a comparison of England and the United States. AB - Learning about letters is an important foundation for literacy development. Should children be taught to label letters by conventional names, such as /bi/ for b, or by sounds, such as /b/? We queried parents and teachers, finding that those in the United States stress letter names with young children, whereas those in England begin with sounds. Looking at 5- to 7-year-olds in the two countries, we found that U.S. children were better at providing the names of letters than were English children. English children outperformed U.S. children on letter sound tasks, and differences between children in the two countries declined with age. We further found that children use the first-learned set of labels to inform the learning of the second set. As a result, English and U.S. children made different types of errors in letter-name and letter-sound tasks. The children's invented spellings also differed in ways reflecting the labels they used for letters. PMID- 18675429 TI - Reflecting on imitation in autism: introduction to the special issue. PMID- 18675430 TI - How young children judge whether a word is one they know: a dual criterion account. AB - Word familiarity judgment may be important for word learning, yet little is known about how children make this judgment. We hypothesized that preschool-age children differ in the judgment criteria that they use and that this difference derives from individual differences in basic memory processes. Those who have superior phonological working memory, but who retrieve less semantic information than their peers, base the judgment on whether they recognize a word's sound form. Those who show the opposite memory profile base the judgment on whether they retrieve a word's meaning. The results of two studies of 3- and 4-year-olds were consistent with these claims. Among those performing poorly on one memory measure, judgment accuracy was directly related to performance on the other memory measure. These memory-judgment relations were also found to be highly specific. This is the first investigation to demonstrate the usefulness of an individual differences approach for identifying relations between linguistic judgment processes and basic memory processes during early childhood. PMID- 18675431 TI - "Don't try this at home": toddlers' imitation of new skills from people on video. AB - Imitation of people on educational television is a potential way for very young children to learn new skills. Although toddlers in previous studies exhibited a "video deficit" in learning, 24-month-olds in Study 1 successfully reproduced behaviors modeled by a person who was on video as well as they did those modeled by a person who was present in the room (even after a 24-h delay). Neither displaced filming context nor cuts between actions affected toddlers' imitation from video. Shortening the demonstration in Study 2 affected imitation in the video condition but not in the live condition. In Study 3, 24-month-olds who viewed the original longer videos on their family TV screens (with which they had a viewing history) imitated significantly less than those who viewed the videos on the laboratory monitor. Imitation of a live modeler was the same across settings (home or lab). Implications for toddlers' judgments of reliable information sources and for the design of educational television programs are discussed. PMID- 18675432 TI - Hydronephrosis index: a better physiological reference in antenatal ultrasound for assessment of fetal hydronephrosis. AB - OBJECTIVE: To establish a nomogram of fetal hydronephrosis index (HI) (anteroposterior diameter of renal pelvis divided by urinary bladder volume) at different gestational ages, to serve as a new reference for antenatal ultrasound examination, and to avoid overestimation of fetal hydronephrosis due to transient effect of a distended fetal bladder. STUDY DESIGN: 504 uncomplicated singleton pregnancies from 20 to 38 weeks' gestation were included. In each fetus, the maximum anteroposterior diameters of both renal pelves were measured on transverse view of fetal kidneys. Urinary bladder volume was calculated using the ovoid volume formula. HI was derived accordingly. RESULTS: Values of HI vary significantly at different trimesters of pregnancy. HI was much higher (mean = 0.1543) from 20 to 27 weeks' gestation, and its value decreased significantly (mean = 0.0253) from 28 to 38 weeks' gestation (P < .05, independent-sample t test). As gestational age increased, HI decreased (R(2) = 0.5921). CONCLUSIONS: HI is easy to be measured and can be used as a new physiological reference for assessment of fetal hydronephrosis by eliminating the confounding effect of a full fetal bladder. The change in values of HI throughout gestation supports the clinical importance of a nomogram for this new index. PMID- 18675433 TI - Hypotension in preterm infants with significant patent ductus arteriosus: effects of dopamine. AB - OBJECTIVE: To study the effects of dopamine on systemic arterial pressure (SAP) and systemic blood flow (SBF) (estimated with the superior vena cava [SVC] flow) in preterm infants with hypotension and patent ductus arteriosus (PDA). STUDY DESIGN: Clinical and echocardiographic variables were measured before and 2 hours after starting dopamine in premature infants <32 weeks gestational age with PDA and systemic hypotension. RESULTS: Seventeen premature infants were included (gestational age, 28+/-2 weeks; birth weight, 1030 +/- 400 g). A mean rate of 8 +/- 2 microg/kg/min of dopamine raised SAP from 30 +/- 3 to 41 +/- 5 mm Hg (P < .05), and the pulmonary artery pressures from 25 +/- 5 to 32 +/- 8 mm Hg (P < .05). The SVC flow increased by 30% (from 130 +/- 40 to 170 +/- 44 mL/kg/min; P < .05). The left ventricular output and the end-diastolic and mean left pulmonary artery blood flow velocities did not change despite the increase in pulmonary artery pressure. CONCLUSION: In preterm infants with hypotension and PDA, dopamine (<10 microg/kg/min) increases the systemic blood pressure and the systemic blood flow. Our results suggest that dopamine decreases left-to-right shunting across ductus arteriosus, caused by a rise in pulmonary vascular resistances. PMID- 18675434 TI - Body mass index, waist circumference, and cardiovascular risk factors in adolescents. AB - OBJECTIVE: To determine optimal threshold values for body mass index (BMI) and waist circumference (WC) for detecting cardiovascular disease (CVD) risk (as defined with >/=3 CVD risk factors [RFs]) in adolescents. STUDY DESIGN: The 1999 2004 National Health and Nutrition Examination Surveys (NHANES) cross-sectional data for 12- to 19-year-old adolescents (n = 2581) was analyzed. Main outcome measures were >/=3 age-adjusted CVD RFs (high- and low-density lipoprotein cholesterol level, triglyceride level, glucose level, insulin level, and systolic and diastolic blood pressure). The presence of >/=3 RFs was predicted from age- and sex-adjusted BMI and WC values with receiver operating characteristics analyses. RESULTS: The proportion of adolescents at risk for >/=3 RFs ranged from 17% to 19%. Both BMI and WC had good diagnostic accuracy, ranging from 0.73 to 0.83, and good sensitivity and specificity, ranging from 0.68 to 0.77. The BMI cutoff points ranged from 19.5 to 25.0 kg/m(2) for boys and from 19.4 to 27.0 kg/m(2) for girls, and WC cutoff points ranged from 66.8 to 87.5 cm for boys and from 71.5 to 87.2 cm for girls. CONCLUSIONS: Age-, sex-, and ethnicity/race specific threshold values for BMI and WC may have significant clinical usefulness in identifying adolescents and teenagers at risk for later CVD onset. PMID- 18675435 TI - A wounding-induced PPO from cowpea (Vigna unguiculata) seedlings. AB - Polyphenol oxidases (PPO) are induced in cowpea plants by wounding. The highest activity levels were detected 48h after this stimulus in both wounded and neighbor-to-wounded unifoliates of cowpea seedlings; the increase of activity was in the order of 13 to 15-fold, respectively, in comparison to control unifoliates. Multiple molecular forms of active PPO (Mrs 58, 73 and congruent with220kDa) were detected by partially denaturing SDS-PAGE. Wounding-induced cowpea PPO were extracted and purified through (NH4)2SO4 precipitation and ion exchange chromatography. The effects of substrate specificity, pH, thermal stability and sensitivity to various inhibitors - resorcinol, EDTA, sodium azide and tropolone - of partially purified soluble PPO were investigated. Purified wounding-induced cowpea PPO (wicPPO) showed the highest activities towards 4 methylcatechol (Km=9.86mM, Vmax=24.66 EU [DeltaAmin(-1)]) and catechol (Km=3.44mM, Vmax=6.64 EU [DeltaAmin(-1)]); no activity was observed towards l tyrosine, under the assay conditions used. The optimum pH for wound-induced cowpea PPO was 6.0 with 4-methylcatechol as substrate. The enzyme was optimally activated by 10 mM SDS and was highly stable even after 5 min at 80 degrees C. The most effective inhibitor was tropolone, whereas addition of 10mM of resorcinol, EDTA and sodium azide were able to reduce PPO activities by 40%, 15% and 100%, respectively. PMID- 18675436 TI - APOBEC3G-independent reduction in virion infectivity during long-term HIV-1 replication in terminally differentiated macrophages. AB - APOBEC3G (APO3G) is a cellular cytidine deaminase with potent antiviral activity. In the case of HIV, the antiviral activity of APO3G is counteracted by the viral Vif protein. Monocyte-derived macrophages (MDM) are terminally differentiated, non-dividing cells susceptible to HIV infection. Human MDM are known to express APO3G and HIV replication in these cells is dependent on Vif. Here we analyzed the correlation between HIV-1 replication and APO3G expression in MDM. Replication of wild type HIV-1 induced a gradual 4-5-fold reduction in APO3G expression. The efficiency of APO3G downregulation correlated with the efficiency of virus replication. Interestingly, despite downregulation of APO3G, the relative infectivity of viruses rapidly declined during the course of infection and was already reduced approximately 90% prior to peak virus production. Cell free virus preparations showed increased levels of a 41 kDa MA-CA processing intermediate. Sequence analysis around the MA-CA cleavage site and the protease and LTR regions did not reveal deaminase-induced hypermutation of the viral genome, suggesting that APO3G activity is not responsible for the incomplete Gag processing. Thus, the loss of infectivity of HIV-1 viruses produced from long term infected primary macrophages is due to an APO3G-independent mechanism. PMID- 18675437 TI - Host responses in the bursa of Fabricius of chickens infected with virulent Marek's disease virus. AB - The bursa of Fabricius serves as an important tissue in the process of Marek's disease virus (MDV) pathogenesis, since B cells of the bursa harbor the cytolytic phase of MDV replication cycle. In the present study, host responses associated with MDV infection in the bursa of Fabricius of chickens were investigated. The expression of MDV phosphoprotein (pp)38 antigen, MDV glycoprotein (gB) and MDV viral interleukin (vIL)-8 transcripts was at the highest at 4 days post-infection (d.p.i.) and then showed a declining trend. On the contrary, the expression of meq (MDV EcoRI Q) gene as well as the viral genome load increased gradually until day 14 post-infection. The changes in viral parameters were associated with significantly higher infiltration of macrophages and T cell subsets, particularly CD4+ T cells into the bursa of Fabricius. Of the genes examined, the expression of interferon (IFN)-alpha, IFN-gamma genes and inducible nitric oxide synthase (iNOS) was significantly up-regulated in response to MDV infection in the bursa of Fabricius. The results suggest a role for these cells and cytokines in MDV induced responses in the bursa of Fabricius. PMID- 18675438 TI - Human vergence eye movements to oblique disparity stimuli: evidence for an anisotropy favoring horizontal disparities. AB - Binocular disparities applied to large-field patterns elicit vergence eye movements at ultra-short latencies. We used the electromagnetic search coil technique to record the horizontal and vertical positions of both eyes while subjects briefly viewed (150 ms) large patterns that were identical at the two eyes except for a difference in position (binocular disparity) that was varied in direction from trial to trial. For accurate alignment with the stimuli, the horizontal and vertical disparity vergence responses (HDVRs, VDVRs) should vary as the sine and cosine, respectively, of the direction of the disparity stimulus vector. In a first experiment, using random-dots patterns (RDs) with a binocular disparity of 0.2 degrees , this was indeed the case. In a second experiment, using 1-D sine-wave gratings with a binocular phase difference (disparity) of 1/4 wavelength, it was not the case: HDVRs were maximal when the grating was vertical and showed little decrement until the grating was oriented more than approximately 65 degrees away from vertical, whereas VDVRs were maximal when the grating was horizontal and began to decrement roughly linearly when the grating was oriented away from the horizontal. We attribute these complex directional dependencies with gratings to the aperture problem, and the HDVR data strongly resemble the stereothresholds for 1-D gratings, which are minimal when the gratings are vertical and remain constant for orientations up to approximately 80 degrees away from the vertical when expressed as spatial phase disparities [Morgan, M. J., & Castet, E. (1997). The aperture problem in stereopsis. Vision Research, 37, 2737-2744.]. To explain this constancy of stereothresholds, Morgan and Castet (1997) postulated detectors sensitive to the phase disparity of the gratings seen by the two eyes (rather than their linear separation along some fixed axis, such as the horizontal). However, because (1) our VDVR data with gratings did not show this constancy and (2) the available evidence strongly suggests that there are no major differences in the disparity detectors mediating the initial HDVR and VDVR, we sought an alternative explanation for our data. We show that the dependence of the initial HDVR and VDVR on grating orientation can be successfully modeled by a bias in the number and/or efficacy of the detectors that favors horizontal disparities. PMID- 18675439 TI - Effects of carbon source on denitrification efficiency and microbial community structure in a saline wastewater treatment process. AB - Two different denitrifying reactors were monitored in order to evaluate the effects of carbon source on denitrification efficiency and microbial community structure under various saline conditions. Nitrogen removal performances were determined when salinity concentrations increase gradually in acetate- or methanol-fed denitrifying reactor. As a result, acetate-fed process attained high nitrate removal at 0-10% NaCl, while methanol was proven beneficial electron donors at 0-3% NaCl. A parallel analysis of T-RFLP and cloning in the acetate-fed sludge showed that a specialized microbial population (i.e. the genera Halomonas and Marinobacter) adapted to a high saline environment. Meanwhile, there were no major changes of bacterial populations in the methanol-fed reactor at 4% NaCl, although the relative abundances of the genera Azoarcus and Methylophaga increased when salinity concentration was at 1-3% NaCl, indicating that methanol utilizing populations in activated sludge was unable to adapt to a high saline environments (>4% NaCl). PMID- 18675440 TI - Nitrite effectively inhibits sulfide and methane production in a laboratory scale sewer reactor. AB - The production and emission of hydrogen sulfide and methane by anaerobic microoganisms in sewer systems is a well-documented problem. The effectiveness of nitrite in controlling sulfide and methane production was tested in a laboratory scale sewer reactor. Nitrite was continuously dosed in the reactor for 25 days at concentrations of 20-140mgN/L. No sulfide and methane accumulation was observed in the reactor in the presence of nitrite. A significant reduction was observed in the sulfate reduction and methane production capabilities of the biofilm. Nitrite also stimulated biological sulfide oxidation within the biofilm. The nitrite uptake rate of the reactor increased over the nitrite dosing period and nitrous oxide production was observed within the biofilm. When nitrite addition was stopped, sulfate reduction and methane production gradually resumed, and reached pre-nitrite addition levels after 2.5 months. The slow recovery suggests that nitrite can be applied intermittently for sulfide and methane control, which represents a key advantage over similar chemicals such as nitrate and oxygen. The study demonstrates nitrite addition as a promising and effective strategy for the management of sulfide and methane in sewers. Further investigation and optimization are still required before application in the field. PMID- 18675441 TI - Longitudinal development of chlorophyll and phytoplankton assemblages in a regulated large river (the Ebro River). AB - The distribution of chlorophyll and phytoplankton communities were compared to nutrient concentrations and hydrological parameters along the main stretch of the river Ebro. A progressive increase in planktonic chlorophyll was observed from the upper reaches to the middle section of the river. Chlorophyll reached a maximum (60-80 microg L(-1)) in the meandering section (downstream of the city of Zaragoza), where nutrient inputs (both N and P) and the residence time of the water are very high. In this meandering section phytoplankton assemblages consisted of large centric diatoms and Scenedesmus sp.pl. These longitudinal patterns were interrupted by the presence of three large reservoirs in the lower section of the river. In the section below the reservoirs, the shorter residence water time, the presence of the invasive zebra mussel, and the massive macrophyte development may explain the historical decrease in chlorophyll-a (from 20-45 microg L(-1) in the 1990s to the present 2-5 microg L(-1)). Phytoplankton densities were extremely poor in this section of the river, where large colonial Coelastrum sp.pl. and Pediastrum sp.pl. were the most characteristic taxa. PMID- 18675442 TI - The influence of small-scale circum-neutral pH change on Cu-bioavailability and toxicity to an estuarine bivalve (Austriella cf plicifera) in whole-sediment toxicity tests. AB - In whole-sediment toxicity tests employing metal-spiked sediment, pH is a crucial determinant of sediment geochemistry, sediment-porewater metal partitioning and metal bioavailability to benthic organisms. Small pH changes in metal-spiked sediment may therefore have important implications for toxicity to organisms. The present study evaluated the effect of small-scale pH changes (pH 6.6, 7.2, 7.6) in Cu-spiked sediment on metal bioavailability and subsequent organism response. Variation of pH produced distinct changes in sediment geochemistry. Increasing pH from 6.6 to 7.6 resulted in lower redox potential, greater acid volatile sulfide (AVS) formation, and lower partitioning of Cu, Fe and Mn to the porewater. The minor pH changes and associated variation in sediment geochemistry modified reburial rates and copper accumulation by the bivalve, Austriella cf plicifera. Bivalve reburial was slowest for the 1400 microg g(-1) Cu-spiked pH 6.6 sediment, in which porewater copper concentrations exceeded 50 microg L(-1). Bivalve reburial was higher for pH 7.2 Cu-spiked sediments at all concentrations of copper investigated than in pH 6.6 and 7.6 sediments, where reburial rates were similar. Higher levels of bivalve activity suggest that A. cf plicifera may have an optimal pH range in the vicinity of pH 7.2. Tissue Cu concentrations varied with pH and appear dependent on spiked Cu concentration and organism activity. Strong linear correlations of tissue Cu concentration with all three Cu phases (overlying water, porewater and particulate phases), however, limited the ability to accurately determine the source of Cu uptake. PMID- 18675443 TI - The environmental fate of the antiviral drug oseltamivir carboxylate in different waters. AB - Since the efficacy of oseltamivir carboxylate (OC) as the active metabolite of Tamiflu has been demonstrated against influenza viruses and even against the avian influenza virus (H5N1), millions of Tamiflu treatment courses are stockpiled worldwide. This was done not at least to follow the recommendations of the World Health Organization (WHO) to cope with a viral influenza pandemic. Concentrations up to 26-32 microg l(-1) OC in river catchment areas in the United States and in the United Kingdom had been predicted recently for a pandemic case, assuming an apparent persistence of the Tamiflu metabolite. Such concentrations may involve the risk of generation of antiviral resistance. But there is still a lack of data concerning the stability of OC in a surface water environment. Under this aspect these predictions have to be validated with concrete facts about the environmental fate of OC. In this article we summarized the results of three different daylight exposure experiments with OC in different waters under sterile and non-sterile conditions simulating shallow water processes at the latitude of approximately 52 degrees N. Using a river water solution containing 50 microg l( 1) OC under non-sterile conditions a half-life time of 17.8 days was observed. Direct photolysis plays no or only a negligible role for the decomposition of OC. Degradation of OC seems to occur as a combination of microbial metabolism and indirect photolysis. PMID- 18675444 TI - The response of terrestrial ecosystems to global climate change: towards an integrated approach. AB - Accumulating evidence points to an anthropogenic 'fingerprint' on the global climate change that has occurred in the last century. Climate change has, and will continue to have, profound effects on the structure and function of terrestrial ecosystems. As such, there is a critical need to continue to develop a sound scientific basis for national and international policies regulating carbon sequestration and greenhouse gas emissions. This paper reflects on the nature of current global change experiments, and provides recommendations for a unified multidisciplinary approach to future research in this dynamic field. These recommendations include: (1) better integration between experiments and models, and amongst experimental, monitoring, and space-for-time studies; (2) stable and increased support for long-term studies and multi-factor experiments; (3) explicit inclusion of biodiversity, disturbance, and extreme events in experiments and models; (4) consideration of timing vs intensity of global change factors in experiments and models; (5) evaluation of potential thresholds or ecosystem 'tipping points'; and (6) increased support for model-model and model experiment comparisons. These recommendations, which reflect discussions within the TERACC international network of global change scientists, will facilitate the unraveling of the complex direct and indirect effects of global climate change on terrestrial ecosystems and their components. PMID- 18675445 TI - Feasibility of an easy-to-use risk score in the prevention of venous thromboembolism and placental vascular complications in pregnant women. A rebuttal. PMID- 18675446 TI - A phase II evaluation of weekly docetaxel in the treatment of recurrent or persistent endometrial carcinoma: a study by the Gynecologic Oncology Group. AB - OBJECTIVE: A phase II study was conducted to evaluate the anti-tumor activity and adverse effects of weekly docetaxel in patients with previously treated endometrial cancer. METHODS: Eligible patients were to have measurable disease with no more than one prior chemotherapy regimen. Docetaxel 36 mg/m(2) was administered intravenously over 1 h on days 1, 8 and 15. Cycles were repeated every 28 days until progression of disease or adverse effects prohibited further therapy. RESULTS: Twenty-seven patients were entered onto this study, of whom 26 were eligible and evaluable. All patients had received prior platinum with twenty (76.9%) having received prior treatment with paclitaxel. There were two (7.7%) partial responses, eight patients (30.8%) with stable disease, and fourteen patients (53.8%) with increasing disease. The most frequently reported adverse events were leucopenia, neutropenia, gastrointestinal, constitutional and peripheral neuropathy. Grade 3 and 4 adverse events were uncommon. CONCLUSIONS: Docetaxel, at the dose and schedule tested, has modest activity in this patient population. PMID- 18675447 TI - Electrical activation induces reactive oxygen species in porcine embryos. AB - The objectives were to determine factors affecting generation of reactive oxygen species (ROS) in porcine embryos after electrical activation of oocytes, and the effects of an antioxidant and chemical agent on ROS generation. Greater ROS were induced by electrical activation compared to IVF (mean+/-S.E.M., 14.6+/-0.8 vs. 9.2+/-0.4, P<0.05). Furthermore, ROS generation in embryos after electrical activation was significantly increased by higher intensity and longer duration electrical pulses and by higher exogenous Ca(2+) concentrations. Cleavage rate and blastocyst formation rate were not directly related to the level of ROS. Supplementation of the IVC medium with 0.5mM glutathione (GSH) reduced ROS (9.2+/ 0.4 vs. 14.7+/-0.9, P<0.05). Treatment with the chemical activation agent, 6 dimethylaminopurine (6-DMAP) for 3h did not induce further ROS generation in combination with electrical activation, but it improved blastocyst formation rate (53.8+/-1.1 vs. 23.7+/-3.5, P<0.05). We concluded that generation of ROS should be considered for optimizing electrical activation and that supplementing an antioxidant or combining electrical and chemical activation induced lower ROS generation in electrically activated porcine embryos. PMID- 18675448 TI - The effect of different zwitterionic buffers and PBS used for out-of-incubator procedures during standard in vitro embryo production on development, morphology and gene expression of bovine embryos. AB - The effect of the zwitterionic buffers HEPES, TES and MOPS and of PBS used for out-of-incubator procedures during standard in vitro embryo production on bovine oocytes and embryo development, morphology and on the expression patterns of eight selected genes: Fgf-4, Lama1, Ube2a, Gsta4, Il6, Sod1, Prss11 and Hspb1, was evaluated. All buffers were prepared at a concentration of 10 mM in TALP medium, with the exception of PBS. The total time of oocyte/embryo exposure to each buffer was approximately 41 min. The cleavage rates and number of embryos that developed to > or =8 cells at day 4 were no different among the buffers tested, however, more blastocysts developed at day 7, 8 and 9 in HEPES and MOPS treatments than in PBS and TES (P<0.05). No difference between buffers in total and apoptotic cell number was found. Except for Hspb1 and Ube2a genes, the levels of expression of the six remaining transcripts were higher in in vivo than in in vitro embryos irrespective of buffer used (P<0.05). In addition, higher expression of Hspb1 and lower expression of Ube2a and Lama1 were observed in PBS and TES than in MOPS and HEPES treatments (P<0.05). Expression of Fgf-4 and Gsta4 in the in vitro embryos was lower in PBS than in the remaining three buffers (P<0.05) and the level of expression of the Il6 gene was not affected by any buffer tested but was lower in in vitro than in in vivo derived embryos. Expression of both Sod1 and Prss11 genes in MOPS were at the level of the in vivo embryos. These results showed that the choice of buffer and short exposure time of approximately 41 min, affects mRNA expression of in vitro produced bovine embryos. PMID- 18675449 TI - Preliminary assessment of somatic cell nuclear transfer in the dromedary (Camelus dromedarius). AB - Somatic cloning may enable the maintenance/expansion of the population of camels with the highest potential for milk production or the best racing performances. However, there have been no reports of embryonic or somatic nuclear transfer in camels. The aim of this study was to produce dromedary embryos by nuclear transfer using in vitro matured oocytes and two somatic cells from two sources (adult fibroblasts or granulosa cells). A total of 58 adult females were superstimulated by a single dose of eCG (3500 IU). Ten days later, their ovaries were collected postmortem. Cumulus-oocytes-complexes (COCs) were aspirated from stimulated follicles and were matured in vitro for 30 h. Fibroblasts (from live adult male) and granulosa cells (from slaughtered adult females) were used as donor karyoplasts and injected into mature enucleated dromedary oocytes. The cleavage rate was significantly higher (P<0.05) for embryos reconstructed with fibroblasts (59%) versus those with granulosa cells (45%). However, there was no difference between the two groups in the proportion of cloned embryos reaching the blastocyst stage (fibroblasts: 14% vs. granulosa cells: 15%) or those that hatched (fibroblasts: 10% vs. granulosa cells: 12%). The viability of reconstructed dromedary embryos from the two sources of donor cells (fibroblasts; n=5 vs. granulosa cells; n=7) was examined by transferring them to synchronized recipients. Two females (fibroblasts: 1/5; 20%, granulosa cells: 1/7; 14%) were confirmed pregnant by ultrasonography at 15 and 25 days following transfer. Later, the pregnancies were followed by pregnancy empirical-symptoms. These two pregnancies were lost between 25 and 60 days following transfer, respectively. In conclusion, the present study shows for the first time that the development of dromedary NT embryos derived from either adult fibroblasts or granulosa cells can occur in vitro and the transfer of these cloned embryos to recipients can result in pregnancies. PMID- 18675450 TI - Uterine blood flow during the first 3 weeks of pregnancy in dairy cows. AB - Transrectal color Doppler sonography was used to compare changes in uterine blood flow between cyclic and early pregnant lactating dairy cows. Examinations were carried out in 53 multiparous lactating cows on days 3, 6, 9, 11, 13, 15, 18 and 21 (day 0=estrus). Fourteen cows were examined during the estrous cycle and thirty-nine cows after insemination with frozen/thawed sperm. Uterine blood flow was reflected by the time-averaged maximum velocity (TAMV) and the pulsatility index (PI) in the uterine artery ipsilateral to the corpus luteum. Twenty-one cows that were not pregnant on day 25 were excluded from the study. There was high inter-individual variability in PI- (CV: 22-62%) and TAMV-values (CV: 22 42%) at all examinations. In cyclic cows, TAMV values increased between days 13 and 18 and PI values decreased between days 15 and 21 (P<0.05). In pregnant cows (n=18), TAMV values increased from days 9 to 11 and then decreased to minimal values by day 18 (P<0.05). The PI values decreased between days 3 and 11 and then increased to maximum levels on day 18 (P<0.05). On day 21, both variables reached (P<0.05) values that did not differ (P>0.05) from those on day 11. The changes in TAMV were correlated with estrogen and progesterone concentrations (r=0.69 and 0.70, respectively; P<0.05) in cycling cows, but not in pregnant cows (P>0.05). The PI values did not correlate with steroid hormone levels (P>0.05). Differences in uterine blood flow between cycling and early pregnant cows were observed only on day 18 (P<0.05). The results show that in pregnant cows changes in uterine blood supply can be detected already in the second week after insemination; these changes do not occur in the second week of the cycle. PMID- 18675451 TI - Analysis of the expression of putatively imprinted genes in bovine peri implantation embryos. AB - The application of assisted reproductive technologies (ART) has been shown to induce changes in the methylation of the embryonic genome, leading to aberrant gene expression, including that of imprinted genes. Aberrant methylation and gene expression has been linked to the large offspring syndrome (LOS) in bovine embryos resulting in increased embryonic morbidity and mortality. In the bovine, limited numbers of imprinted genes have been studied and studies have primarily been restricted to pre-implantation stages. This study reports original data on the expression pattern of 8 putatively imprinted genes (Ata3, Dlk1, Gnas, Grb10, Magel2, Mest-1, Ndn and Sgce) in bovine peri-implantation embryos. Two embryonic developmental stages were examined, Day 14 and Day 21. The gene expression pattern of single embryos was recorded for in vivo, in vitro produced (IVP) and parthenogenetic embryos. The IVP embryos allow us to estimate the effect of in vitro procedures and the analysis of parthenogenetic embryos provides provisional information on maternal genomic imprinting. Among the 8 genes investigated, only Mest-1 showed differential expression in Day 21 parthenogenetic embryos compared to in vivo and IVP counterparts, indicating maternal imprinting of this gene. In addition, our expression analysis of single embryos revealed a more heterogeneous gene expression in IVP than in in vivo developed embryos, adding further to the hypothesis of transcriptional dysregulation induced by in vitro procedures, either by in vitro maturation, fertilization or culture. In conclusion, effects of genomic imprinting and of in vitro procedures for embryo production may influence the success of bovine embryo implantation. PMID- 18675452 TI - Retrograde ejaculation occurs in the dog, but can be prevented by pre-treatment with phenylpropanolamine: a urodynamic study. AB - Retrograde ejaculation is partial or total propulsion of semen from the posterior urethra into the urinary bladder; it is well characterized (and relatively common) in humans, with only a few reports in animals. Our objectives were to determine whether retrograde flow of semen occurred during ejaculation in mature dogs with normal fertility, and to determine the effects of phenylpropanolamine on this phenomenon (dose-titration, switch-back study). Retrograde ejaculation and urethral pressure profile measurements were evaluated (double-blind) in six dogs after 5 days of oral treatment with phenylpropanolamine (0, 2, 4, or 8 mg/kg); all dogs received all treatments (at 2-week intervals). The number of sperm in the urine was determined before and after each manual sperm collection. Urethral pressure profiles were obtained three times during each procedure. In the absence of phenylpropanolamine, sperm were present in the bladder after semen collection in all dogs (number varied significantly among individuals). The mean (+/-S.D.) number of sperm in the bladder was 17.0+/-5.0, 18.5+/-1.2, 5.1+/-5.0, and 4.8+/-0.1 x 10(6) sperm for 0, 2, 4, and 8 mg/kg, respectively (no significant difference between dogs given 4 or 8 mg/kg, but both were significantly lower than those given 0 or 2mg/kg). This reduction was significantly correlated to the increase in mean urethral pressure at the level of the sphincter (39 cm versus 59 cm H(2)O in placebo-treated dogs versus those given 8 mg/kg). In conclusion, we confirmed that retrograde ejaculation occurred during the ejaculatory process in normal dogs, and we demonstrated that phenylpropanolamine (4 or 8 mg/kg once daily for 5 days before collection) increased urethral pressure and reduced the number of sperm voided into the bladder during ejaculation. PMID- 18675453 TI - Biocompatibility of silica coated NaYF(4) upconversion fluorescent nanocrystals. AB - Here we report the synthesis of uniform silica coated hexagonal-phase NaYF(4) nanocrystals with strong NIR-to-visible upconversion fluorescence and its cytotoxicity and biodistribution in a rat model. The silica coated NaYF(4) nanocrystals were incubated with rat skeletal myoblasts and bone marrow-derived mesenchymal stem cells and cytotoxicity was assessed by using MTS and LDH assay. Healthy rats were injected intravenously with the nanocrystals so as to further investigate their biocompatibility and tissue distribution. The results from this study revealed that the silica coated NaYF(4) upconversion nanocrystals displayed good in vitro and in vivo biocompatibility, demonstrating their potential applications in both cellular and animal imaging systems. PMID- 18675454 TI - The biocompatibility of mesoporous silicates. AB - Micro- and nano-mesoporous silicate particles are considered potential drug delivery systems because of their ordered pore structures, large surface areas and the ease with which they can be chemically modified. However, few cytotoxicity or biocompatibility studies have been reported, especially when silicates are administered in the quantities necessary to deliver low-potency drugs. The biocompatibility of mesoporous silicates of particle sizes approximately 150 nm, approximately 800 nm and approximately 4 microm and pore sizes of 3 nm, 7 nm and 16 nm, respectively, is examined here. In vitro, mesoporous silicates showed a significant degree of toxicity at high concentrations with mesothelial cells. Following subcutaneous injection of silicates in rats, the amount of residual material decreased progressively over 3 months, with good biocompatibility on histology at all time points. In contrast, intra-peritoneal and intra-venous injections in mice resulted in death or euthanasia. No toxicity was seen with subcutaneous injection of the same particles in mice. Microscopic analysis of the lung tissue of the mice indicated that death may be due to thrombosis. Although local tissue reaction to mesoporous silicates was benign, they caused severe systemic toxicity. This toxicity might be mitigated by modification of the materials. PMID- 18675455 TI - Placental oxidative stress alters expression of murine osteogenic genes and impairs fetal skeletal formation. AB - Fetal and placental developments rely on an intricate balance of nutrients, growth factors, and signaling pathways at precise times in gestation. Disruptions to this balance may result in disease that manifests in adulthood, a situation termed "developmental origins of health and disease". Diet, exercise, and certain chemical exposures during pregnancy increase oxidative stress (OS), and may alter trajectory of fetal osteogenic regulation in a manner that increases risk of adult bone dysfunction. The present study used gestational methylnitrosourea (MNU), a known inducer of OS, in C57BL/6 mice with or without dietary antioxidant quercetin (Q) supplementation. Several key placental genes that influence placental development and fetal osteogenesis (Hgf, Kitl, IFNalpha4, Ifrd, and IL 1beta) were altered by MNU, and largely normalized by Q. MNU treatment also resulted in small fetuses with disproportionately shortened limbs and distal limb malformations, and caused placental endothelial and trophoblast damage. Q was again protective against these fetal and placental pathologies. An unanticipated finding with Q supplementation was increased interdigital webbing, perhaps due to dose-related effects on apoptosis required for digital sculpting, or pro-oxidant effects of Q that caused a maturational delay. These results suggest that elevated OS may alter normal placental osteogenic signaling and fetal skeletal formation. PMID- 18675456 TI - The distribution of angiopoietin-1, angiopoietin-2 and their receptors tie-1 and tie-2 in the very early human placenta. AB - Angiopoietins are integral to vasculogenesis and angiogenesis, which play crucial roles in the growth and development of the placenta. The current study assessed expression of angiopoietins (Ang-1 and Ang-2) and their receptors (Tie-1 and Tie 2) during development of the early human placenta. First-trimester placental tissues were obtained from women undergoing curettage during normal pregnancies. The use of immunohistochemistry (IHC) showed that Ang-1 was primarily localized to syncytiotrophoblasts where it displayed moderate immunoreactivity, whereas weak immunoreactivity for Ang-1 was observed in endothelial cells and angiogenic cell cords (ACC). Strong immunoreactivity for Ang-2 was also found predominantly in syncytiotrophoblasts with lower immunostaining levels evident in cytotrophoblasts. Moderate immunoreactivity for Ang-2 was observed in endothelial cells, ACC and Hofbauer cells. By contrast, the trophoblastic shell, as well as endothelial cells and ACC exhibited strong staining intensity for Tie-1 with the strongest immunoreactivity for Tie-2 observed in cytotrophoblasts, ACC and endothelial cells. Western blotting of tissue extracts confirmed the IHC results. Previous studies focused on VEGF and its receptors in controlling vasculogenesis and angiogenesis in human placenta. However, the specific localization patterns of angiopoietins and their receptors revealed by the current study emphasize the importance of these molecules in placental vascular development. Functional studies aimed at identifying the molecular mechanisms of actions of these factors and receptors may prove essential in elucidating the pathophysiology of placental disorders such as intrauterine growth restriction and pre-eclampsia. PMID- 18675457 TI - Cross-linking of FcepsilonRI causes Ca2+ mobilization via a sphingosine kinase pathway in a clathrin-dependent manner. AB - Clathrin-coated pits are now recognized to be involved in cell signaling in addition to receptor down-regulation. Here we tried to identify signaling pathways that might be dependent on clathrin. Our initial data with pharmacological inhibitors of formation of clathrin-coated pits or lipid-rafts indicated that Ca(2+) response evoked by cross-linking of the high affinity receptors for IgE (FcepsilonRI) was dependent on clathrin. To confirm this finding, we created clathrin-knockdown cells by transfecting the mast cell line RBL-2H3 with a shRNA-clathrin heavy chain construct. In these cells, the FcepsilonRI-mediated Ca(2+) response was almost completely abolished, which was accompanied by the inhibition of sphingosine 1-phosphate (S1P) production with no changes in inositol 1,4,5-trisphosphate (IP(3)) production. This suggests that the Ca(2+) signaling pathway via a sphingosine kinase (SK) is dependent on clathrin. Furthermore, antigen-induced tyrosine phosphorylation of p85 and p110 subunits of PI3K was almost completely inhibited in clathrin-knockdown cells. In contrast, antigen-induced tyrosine phosphorylation of phospholipase Cgamma was not affected by clathrin-knockdown and tyrosine phosphorylation of Syk and degranulation were partially inhibited in clathrin-knockdown cells. The present study identifies the SK/Ca(2+) pathway to be dependent on clathrin. PMID- 18675458 TI - Neonatal manipulation of oxytocin influences the partner preference in mandarin voles (Microtus mandarinus). AB - Neonatal manipulation of oxytocin (OT) has long-term effects on behavior and physiology. The objective of this research was to determine if neonatal exposure to OT can affect partner preferences and to characterize the mechanisms underlying social behavior such as neural activities of relevant brain regions in socially monogamous mandarin voles (Microtus mandarinus). After receiving a subcutaneous injection of isotonic saline (SAL) or OT within 24h of birth, mandarin vole at 60 days of age was paired with an unfamiliar opposite sex for 24h, followed immediately by an examination of their partner preference and 30 days later by the second examination of their partner preference and Fos expression in some brain regions. The results indicated that (1) while 24h cohabitation was insufficient for both female and male SAL-treated mandarin voles to form partner preference, neonatal exposure to OT significantly facilitate female, but not male mandarin voles to form partner preference within 24h of cohabitation; (2) the maintenance of partner preference in females was suppressed by neonatal OT treatment, while neonatal OT-treated males showed significant partner preference as neonatal SAL-treated males and females; in addition, the tendency of aggression to the strangers was impaired in both females and males, and neonatal OT-treated males showed significantly higher mounting behavior to the partner; (3) in comparison with saline-treated females, OT-treated females showed a significant decrease in Fos expression in all brain regions examined in response to partner preference. Relative to saline treatment, neonatal OT treatment induced to males a significant decrease of Fos expression in the bed nucleus of the stria terminalis (BNST), the hypothalamic paraventricular nucleus (PVN) and the mediodorsal thalamic nucleus (MD) as well as a significant increase in the medial preoptic area (MPOA), the dorsal part of the lateral septal nucleus (LSD) and the central amygdaloid nucleus (CeA). These results demonstrate that neonatal OT exposure has different effects on the formation and maintenance of partner preference in mandarin voles and the latter effects possibly via OT induced changes of the neural activities of relevant brain regions. PMID- 18675459 TI - IL-23 and IL-12 responses in activated human T cells retrovirally transduced with IL-23 receptor variants. AB - Interleukin-23 (IL-23) is a regulator of cellular immune responses involved in controlling infections and autoimmune diseases. Effects of IL-23 on T cells are mediated via a receptor complex consisting of an IL-12Rbeta1 and a specific IL 23R chain. The R381Q and P310L variants of the IL-23R were recently reported to be associated with autoimmune diseases, suggesting they have an effect on IL-23R function. To investigate this matter, these variants and a newly identified variant, Y173H, were retrovirally transduced into human T cell blasts and functionally characterized by measuring the IL-23-induced signal transduction pathway (i.e., STAT1, STAT3 and STAT4 phosphorylation), and IFN-gamma and IL-10 production. No differences were detected between the genetic variants and wild type in the function of the IL-23R-chain. Furthermore, while comparing IFN-gamma and IL-10 production in response to IL-23 and IL-12, we found IL-23 to be a more potent IL-10 inducer, and IL-12 a more potent IFN-gamma inducer. In addition, IL 23 also exerted a minor IL-12-like effect by inducing IL-23R-independent, IL 12Rbeta1-dependent STAT4 phosphorylation and IFN-gamma production. In conclusion, the reported association between R381Q and P310L variants of the IL-23R and autoimmune diseases does not depend on differences in functional activity between wild-type and R381Q and P310L variants of the IL-23R. PMID- 18675460 TI - Metal specificity in DNA damage prevention by sulfur antioxidants. AB - Metals such as CuI and FeII generate hydroxyl radical (.OH) by reducing endogenous hydrogen peroxide (H2O2). Because antioxidants can ameliorate metal mediated oxidative damage, we have quantified the ability of glutathione, a primary intracellular antioxidant, and other biological sulfur-containing compounds to inhibit metal-mediated DNA damage caused hydroxyl radical. In the CuI/H2O2 system, six sulfur compounds, including both reduced and oxidized glutathione, inhibited DNA damage with IC50 values ranging from 3.4 to 12.4 microM. Glutathione and 3-carboxypropyl disulfide also demonstrated significant antioxidant activity with FeII and H2O2. Additional gel electrophoresis and UV vis spectroscopy studies confirm that antioxidant activity for sulfur compounds in the CuI system is attributed to metal coordination, a previously unexplored mechanism. The antioxidant mechanism for sulfur compounds in the FeII system, however, is unlike that of CuI. Our results demonstrate that glutathione and other sulfur compounds are potent antioxidants capable of preventing metal mediated oxidative DNA damage at well below their biological concentrations. This novel metal-binding antioxidant mechanism may play a significant role in the antioxidant behavior of these sulfur compounds and help refine understanding of glutathione function in vivo. PMID- 18675461 TI - The risk for cancer among patients with schizoaffective disorders. AB - BACKGROUND: Several epidemiological studies explored the risk for cancer among both persons with schizophrenia and their first-degree relatives, and among patients with bipolar disorder. No studies have yet explored the risk among persons with schizoaffective disorders. METHOD: Linkage analysis was conducted based on the psychiatric and the cancer national databases. Standardized incidence ratios (SIR) for aggregated cancer sites were calculated by comparing the incidence rates among patients in the psychiatric case register with schizoaffective disorders with the incidence rates in the Jewish-Israeli general population. RESULTS: No significant alteration in cancer risk was found for both genders: males, SIR=1.11, 95% CI (0.48-1.73) and females, SIR=1.38, 95% CI (0.96 1.80). LIMITATIONS: Our sample was derived from patients with a history of psychiatric hospitalization. Putative factors such as diet, smoking and medications were not investigated. CONCLUSIONS: Our study showed no significant increase in the risk for cancer in schizoaffective disorders. Those results appear to be positioned between the schizophrenia findings that show a lower risk for cancer and the bipolar disorder findings that show an increased risk. PMID- 18675463 TI - Mutations in the hemochromatosis gene and the clinical outcome of multiple sclerosis. AB - Multiple sclerosis (MS) is a common inflammatory disease of the central nervous system unsurpassed for its variability in disease outcome. Given a possible role for dysregulation of iron metabolism in MS disease pathogenesis, we investigated whether or not mutations in the HFE gene influence the prognosis of the disease. A cohort of sporadic MS cases, taken from opposite extremes of the putative distribution of long-term outcome using the most stringent clinical criteria to date, was used to determine the role of HFE on MS disease severity. This approach increases the effective sample size by some 40-fold. Genotyping the two sets of MS patients (112 benign and 51 malignant) provided no evidence to suggest that mutations in HFE have any outcome modifying activity, although small effects cannot be ruled out. The frequency of HFE mutations was not different in MS compared to the general population. PMID- 18675462 TI - Granzyme B: evidence for a role in the origin of myasthenia gravis. AB - PURPOSE OF RESEARCH: Although the pathogenesis of myasthenia gravis (MG) as an antibody mediated disorder of acetylcholine receptors (AChRs) at neuromuscular junctions is well understood, the origin of the autoimmune response is unclear. The thymus is intimately involved in initiation of the autoimmune response; the antigen, AChR, is present in the thymus, but how the autoimmune response is triggered is not known. Granzyme B (GrB), a proteolytic enzyme present in cytolytic T cells and natural killer (NK) cells, selectively cleaves many potential autoantigens (but few non-autoantigens), generating novel fragments that trigger autoreactive responses. This protease has been strongly implicated in the pathogenesis of several autoimmune diseases including lupus, rheumatoid arthritis, dermatomyositis, and others. In the studies described in this manuscript, we examined the ability of GrB to cleave the AChR subunits, and performed biochemical, immunohistochemical and molecular studies on thymus glands from myasthenic patients and controls to assess GrB expression. MAIN RESULTS: GrB efficiently and specifically cleaves subunits of AChR, especially the epsilon subunit. GrB is present in thymus glands from myasthenia patients, but is absent in control thymuses. CONCLUSIONS: Our results provide evidence supporting a potential role for GrB in the process of initiation of MG, and are consistent with the concept of an immunodominant epsilon epitope. PMID- 18675464 TI - The MGTX experience: challenges in planning and executing an international, multicenter clinical trial. AB - We present our experience planning and launching a multinational, NIH/NINDS funded study of thymectomy in myasthenia gravis. We highlight the additional steps required for international sites and analyze and contrast the time investment required to bring U.S. and non-U.S. sites into full regulatory compliance. Results show the mean time for non-U.S. centers to achieve regulatory approval was significantly longer (mean 13.4+/0.96 [corrected] months) than for U.S. sites (9.67+/0.74 [corrected] months; p=0.003, [corrected] t-test). The delay for non-U.S. sites was mainly attributable to Federalwide Assurance certification and State Department clearance. PMID- 18675465 TI - A double mutation of MBP(83-99) peptide induces IL-4 responses and antagonizes IFN-gamma responses. AB - A number of treatment options are available to multiple sclerosis patients, however this needs to be improved. Herein, we designed and synthesized a number of peptides by mutating principal TCR contact residues based on MBP(83-99) peptide epitope. Immunization of SJL/J mice with MBP(83-99) and mutant [A(91)]MBP(83-99), [E(91)]MBP(83-99), [F(91)]MBP(83-99), [Y(91)]MBP(83-99), and [R(91), A(96)]MBP(83-99) peptides, induced IFN-gamma, and only [R(91), A(96)]MBP(83-99) mutant peptide was able to induce IL-4 secretion by T cells. T cells against the native MBP(83-99) peptide cross-reacted with all peptides except [Y(91)]MBP(83-99) and [R(91),A(96)]MBP(83-99). The double mutant [R(91), A(96)]MBP(83-99) was able to antagonize IFN-gamma production in vitro by T cells against the native MBP(83-99) peptide. Antibodies generated to [R(91), A(96)]MBP(83-99) did not cross-react with whole MBP protein. Molecular modeling between peptide analogs and H2 I-A(s) demonstrated novel interactions. The [R(91), A(96)]MBP(83-99) double mutant peptide analog is the most promising for further therapeutic studies. PMID- 18675466 TI - Remembering John Newsom-Davis' contribution to human imaging in Oxford. AB - John Newsom-Davis played a crucial role in supporting areas of scientific exploration beyond his own research interests. In particular, he was one of the key players in establishing human neuroimaging in Oxford. Here, we celebrate the role that he played in this endeavour, both in the early days of pulling together funding, and solving practical challenges, and in the following years, when we all appreciated his ongoing encouragement and support. PMID- 18675467 TI - Molecular form follows function: (un)snaring the SNAREs. AB - Exocytotic release of transmitters is mediated by the ternary SNARE complex. The form of this complex is consistent with its function in the positioning of vesicles to the plasma membrane and their fusion to it. Recent advances in single molecule techniques, however, bring an additional layer of complexity to this process, implicating that there might be various modes of operation. For example, the binary syntaxin-synaptobrevin 2 complex, in addition to the ternary complex containing SNAP25, might enable vesicular docking. Single-molecule techniques allow direct measurements of the distance/extension, rupture force, spontaneous dissociation times and interaction energy for SNARE protein-protein interactions. These measurements are complementary to results and conclusions drawn from other techniques. Consequently, single-molecule techniques promise tremendous opportunities for in vitro investigations of SNARE proteins to improve our understanding of their role in exocytosis. PMID- 18675469 TI - The HPA axis in major depression: classical theories and new developments. AB - Studies over the last 40 years have demonstrated that hyperactivity of the hypothalamic-pituitary-adrenal axis is one of the most consistent biological findings in major depression psychiatry, but the mechanisms underlying this abnormality are still unclear. PMID- 18675470 TI - Modification of chromate toxicity by sulphate in duckweeds (Lemnaceae). AB - Two duckweed species, Spirodela polyrhiza and Lemna minor, were used to measure the toxicity of chromate (100 microM) at three levels of sulphate (13 microM, low sulphate=LS; 410 microM, normal sulphate=NS; 10,000 microM, high sulphate=HS). Growth rates calculated on the basis of dry weight, chlorophyll and carotenoid content were all reduced by chromate. This inhibition was the strongest under LS conditions and the weakest under HS conditions. Thus, sulphate decreases chromate toxicity-which conforms with its influence on chromate uptake reported previously (Kaszycki, P., Gabrys, H., Appenroth, K.-J., Jaglarz, A., Sedziwy, S., Walczak, T., Koloczek, H., 2005. Exogenously applied sulphate as a tool to investigate transport and reduction of chromate in the duckweed Spirodela polyrhiza. Plant Cell Environ. 28, 260-269). The three levels of sulphate were also applied during pre-cultivation of both species for 2 weeks and the consequences for chromate toxicity were tested thereafter. When S. polyrhiza was pre-cultivated in NS medium, the growth inhibition by chromate was approximately 80% of the control (no chromate) in the subsequently applied LS medium, and approximately 50% in HS. L. minor showed similar relationships but a lower overall chromate sensitivity. In comparison to the plants pre-treated in NS medium, those pre-treated in LS were more sensitive whereas those pre-treated in HS were less sensitive toward chromate. The present data demonstrate that chromate is taken up into cells of the two duckweed species by sulphate transporter(s). The rather weak influence of sulphate on chromate toxicity indicates that chromate binds to the transporters much stronger than sulphate. Moreover, the relative effects of sulphate on the chromate toxicity remain very similar regardless of pre-treatment. This confirms the conclusion from uptake experiments that pre-treatment with different levels of sulphate changes the number of sulphate transporters but their affinity remains unchanged. In summary, the influence of sulphate on the toxicity of chromate reflects mainly its influence on chromate uptake, with a negligible impact of other physiological processes. PMID- 18675468 TI - Neuronal calcium mishandling and the pathogenesis of Alzheimer's disease. AB - Perturbed neuronal Ca(2+) homeostasis is implicated in age-related cognitive impairment and Alzheimer's disease (AD). With advancing age, neurons encounter increased oxidative stress and impaired energy metabolism, which compromise the function of proteins that control membrane excitability and subcellular Ca(2+) dynamics. Toxic forms of amyloid beta-peptide (Abeta) can induce Ca(2+) influx into neurons by inducing membrane-associated oxidative stress or by forming an oligomeric pore in the membrane, thereby rendering neurons vulnerable to excitotoxicity and apoptosis. AD-causing mutations in the beta-amyloid precursor protein and presenilins can compromise these normal proteins in the plasma membrane and endoplasmic reticulum, respectively. Emerging knowledge of the actions of Ca(2+) upstream and downstream of Abeta provides opportunities to develop novel preventative and therapeutic interventions for AD. PMID- 18675471 TI - Endocrine effects of contaminated sediments on the freshwater snail Potamopyrgus antipodarum in vivo and in the cell bioassays in vitro. AB - Lake Pilnok located in the black coal-mining region Ostrava-Karvina, Czech Republic, contains sediments highly contaminated with powdered waste coal. Moreover, population of the endangered species of narrow-clawed crayfish Pontastacus leptodactylus with high proportion of intersex individuals (18%) was observed at this site. These findings motivated our work that aimed to evaluate contamination, endocrine disruptive potency using in vitro assays and in vivo effects of contaminated sediments on reproduction of sediment-dwelling invertebrates. Chemical analyses revealed low concentrations of persistent chlorinated compounds and heavy metals but concentrations of polycyclic aromatic hydrocarbons (PAH) were high (sum of 16 PAHs 10 microg/g dw). Organic extracts from sediments caused significant in vitro AhR-mediated activity in the bioassay with H4IIE-luc cells, estrogenicity in MVLN cells and anti-androgenicity in recombinant yeast assay, and these effects could be attributed to non-persistent compounds derived from the waste coal. We have also observed significant in vivo effects of the sediments in laboratory experiments with the Prosobranchian euryhaline mud snail Potamopyrgus antipodarum. Sediments from Lake Pilnok as well as organic extracts of the sediments (externally added to the control sediment) significantly affected fecundity during 8 weeks of exposure. The effects were stimulations of fecundity at lower concentrations at the beginning of the experiment followed by inhibitions of fecundity and general toxicity. Our study indicates presence of chemicals that affected endocrine balance in invertebrates, and emphasizes the need for integrated approaches combining in vitro and in vivo bioassays with identification of chemicals to elucidate ecotoxicogical impacts of contaminated sediment samples. PMID- 18675472 TI - High dose norepinephrine-induced apoptosis in cultured rat cardiac fibroblast. AB - BACKGROUND: Prolonged sympathetic activation is damaging to the heart. Recent findings suggest that norepinephrine (NE) may contribute to the apoptotic cardiac cell loss. This study investigated high doses NE apoptotic effect on cardiac fibroblasts (CF) culture and compared the anti-apoptotic effect of alpha and beta (selective and non-selective) adrenergic receptor antagonists. METHODS AND RESULTS: Rat CF were cultured in the presence of NE (1 to 100 microM) for 48 h. bax and bcl(XL) genes expression were measured by real-time quantitative PCR method. Cell viability percentage, apoptotic cell percentage and caspase 3 activity was measured by MTT assay, flow cytometric method and caspase 3 flurogenic assay kit respectively. NE (100 microM) increased bax gene expression by 1.96+/-0.96 fold while bcl(XL) gene decreased by 0.53+/-0.15 fold when compared with the control group (p<0.01). The apoptotic cell percentage increased significantly from 5.09+/-2.94% in control group to 31.48+/-6.35% (p<0.01) with NE and the caspase 3 activity increased from 1432.2+/-658.8 in the control group to 5162+/-2028.6 (OD/microg protein, p<0.05). Addition of carvedilol (non selective beta blocker with alpha-blockade activity), doxazosin (alpha blocker), metoprolol (beta1 selective blocker) and propranolol (non-selective beta blocker) were not capable of inhibiting the apoptotic effect of high dose NE on CF. CONCLUSION: High dose NE has cytotoxic and apoptotic effect on CF. The process involved the activation of caspase 3, up-regulation of pro-apoptotic gene bax and down-regulation of anti-apoptotic gene bcl(XL). Both non-selective and selective adrenergic receptor antagonists tested were not capable of inhibiting the apoptotic effect of high dose NE on CF. PMID- 18675473 TI - Transcatheter closure of perimembranous ventricular septal defect in a patient with abnormal inferior vena cava return. AB - An 8-year-old boy with a 3.8 mm perimembranous ventricular septal defect (PmVSD) and abnormal inferior vena cava return was attempted cardiac catheterization to occlude the VSD. Through the right jugular vein and right femoral artery approach, an 8 mm Amplatzer VSD occluder was successfully deployed under fluoroscopic and echocardiographic guidance. After a 2 year follow-up, the patient is symptom-free. Our report offers a new perspective to percutaneous treatment of PmVSD with abnormal inferior vena cava return. PMID- 18675474 TI - Impact of previous myocardial infarction on the incremental value of myocardial contrast to two-dimensional supine bicycle stress echocardiography in evaluation of coronary artery disease. AB - BACKGROUND: If compared to two-dimensional echocardiography (2DE), myocardial contrast echocardiography (MCE) improves detection of coronary artery disease (CAD) during pharmacological stress, but data on MCE vs. 2DE during supine bicycle stress is limited. Although previous myocardial infarction (MI) influences sensitivity of 2DE, its effect on MCE has not been evaluated. OBJECTIVES: The study sought to determine the incremental benefit of MCE over 2DE for evaluation of CAD during supine bicycle stress and to assess the impact of previous MI on diagnostic values of both methods. METHODS: We studied 103 consecutive patients scheduled for coronary angiography. Prior to coronary angiography, all patients underwent supine bicycle stress. 2DE and MCE were performed during this stress test. The diagnosis of obstructive CAD (> or =50% stenosis) was based on the presence of inducible wall motion and perfusion abnormalities. RESULTS: Quantitative coronary angiography revealed > or =50% stenosis in 53 of 77 patients without previous MI and in 21 of 26 patients with previous MI. If compared to 2DE, MCE was more sensitive (68% vs. 86%; p<0.001) and more accurate (73% vs. 86%; p < 0.001) to detect > or =50% stenosis. In patients without previous MI, 2DE and MCE yielded sensitivity of 65% and 85% (p < 0.01) and accuracy of 71% and 85% (p < 0.01), whereas in patients with previous MI sensitivity was 79% and 90% (p=NS) and accuracy 79% and 88% (p = NS), respectively. CONCLUSIONS: MCE enhances sensitivity and accuracy of 2DE in detection of obstructive CAD during supine bicycle stress. The incremental benefit of MCE is especially present in patients without previous MI. PMID- 18675475 TI - Current management of unprotected left main coronary artery disease: run-in survey of the RITMO (Registro Italiano sul Trattamento del tronco coMune non protettO) study. AB - The optimal approach for a significant unprotected left main coronary stenosis (ULM) is debated, in light of the recent progresses of percutaneous coronary intervention (PCI). However, coronary artery bypass grafting (CABG) is still considered the first choice treatment. Randomized trials comparing PCI and CABG are ongoing, yet patient selection will considerably limit their clinical applicability. We thus designed a prospective multicenter registry which will include patients with ULM disease independently from the subsequent medical, interventional or surgical treatment: the RITMO Study (Registro Italiano sul Trattamento del tronco coMune non protettO). During the RITMO run-in phase, we conducted a systematic survey of Italian catheterization laboratories to define current management strategies for ULM. A total of 240 Italian catheterization laboratory were sent an email questionnaire on current practices for ULM, with 45 (19%) detailed replies, for a total of 61,370 annual coronary angiographies. Data provided from responders showed a 5% (95% interval: 2-16) prevalence of ULM, with 50% (9-99) of ULM treated surgically and 10% (0-81) treated percutaneously. In conclusion, treatment of ULM in Italy remains prevalently surgical, even if PCI is performed in a sizable portion of patients with ULM. PMID- 18675476 TI - Culprit only versus multivessel coronary revascularization in patients presenting with acute ST elevation myocardial infarction: unending debate. AB - The optimal percutaneous interventional strategy for dealing with significant non culprit lesions in patients with multivessel disease with acute myocardial infarction (AMI) at presentation remains to be controversial. For the time being, the current guidelines recommended that primary percutaneous coronary intervention (PCI) for non-culprit lesions should be limited to the infarct related artery. We believe that decisions about PCI of the non-infarct vessel(s) should be individualized and guided by objective evidence of significant residual ischemia except in patients with multivessel disease showing hemodynamic compromise. Further large, randomized trials will help us solve this dilemma. PMID- 18675477 TI - Granulocyte-colony stimulating factor or granulocyte-colony stimulating factor associated to stem cell intracoronary infusion effects in non ischemic refractory heart failure. AB - We prospectively studied bone marrow stem cell (BMSC) therapy in 23 patients with non ischemic refractory heart failure(HF) in comparison with a HF control group with 17 patients. BMSC patients randomly underwent granulocyte-colony stimulating factor (G-CSF) administration (14 patients) or G-CSF associated to BMSC intracoronary infusion (eight patients). After the first month all BMSC patients received G-CSF with one-month interval between each one. CD34+ cell peaks (per mm(3)) in BMSC patients were 19+/-12 and in normal control 60+/-20 (p=0.003). In BMSC patients after the 1st G-CSF left ventricular(LV) ejection fraction (EF) increased from 21.4+/-4.7% to 23.6+/-7.7%(p=.048), peak VO(2) (ml/kg/min) from 9.9+/-2.4 to 11.6+/-3 (p=.04), functional class and quality of life improved whereas in the HF control group LVEF, RFEF and functional class were unchanged. Both BMSC subgroups presented improvement of LV function evaluated by DTI velocities. Evaluations after the first month in BMSC patients showed improvements in LVEF (p=.001), right VEF (p=0.01), DTI velocities (p=.009), peak VO(2) (p=0.04), functional class (p<0.001) and quality of life (p<0.001). In conclusion, CD34+ mobilization is impaired in HF. Stem cell therapy can improve HF. Randomized trials should be developed to confirm our results. PMID- 18675478 TI - The cult of the amateur in agriculture threatens food security. AB - The incorporation of science and technology into agriculture has led to enormous growth in crop yields, providing food security in many countries. From the 1950s onwards there has been increasing interference in agricultural policy by a few scientists who are marginal to agriculture and from a variety of unqualified groups. These groups and individuals have used fear and anxiety and have greatly exaggerated minor problems to persuade an unqualified public of supposed dangers in food and to try and change agricultural policy. Fear and emotion do not lead to good policy, and the cult of the amateur that has developed could have serious repercussions on vital food security and future agriculture in developing countries; it must be soundly rejected. PMID- 18675479 TI - Affinity chromatography approaches to overcome the challenges of purifying plasmid DNA. AB - The diversity of biomolecules present in plasmid DNA (pDNA)-containing extracts and the structural and chemical similarities between pDNA and impurities are some of the main challenges of improving or establishing novel purification procedures. In view of the unequalled specificity of affinity purification, this technique has recently begun to be applied in downstream processing of plasmids. This paper discusses the progress and importance of affinity chromatography (AC) for the purification of pDNA-based therapeutic products. Several affinity approaches have already been successfully developed for a variety of applications, and we will focus here on highlighting their possible contributions to the pDNA purification challenge. Diverse affinity applications and their advantages and disadvantages are discussed, as well as the most significant results and improvements in the challenging task of purifying plasmids. PMID- 18675480 TI - Metabolic footprinting in microbiology: methods and applications in functional genomics and biotechnology. AB - Metabolomics embraces several strategies that aim to quantify cell metabolites in order to increase our understanding of how metabolite levels and interactions influence phenotypes. Metabolic footprinting represents a niche within metabolomics, because it focuses on the analysis of extracellular metabolites. Although metabolic footprinting represents only a fraction of the entire metabolome, it provides important information for functional genomics and strain characterization, and it can also provide scientists with a key understanding of cell communication mechanisms, metabolic engineering and industrial biotechnological processes. Due to the tight and convoluted relationship between intracellular metabolism and metabolic footprinting, metabolic footprinting can provide precious information about the intracellular metabolic status. Hereby, we state that integrative information from metabolic footprinting can assist in further interpretation of metabolic networks. PMID- 18675481 TI - Cybrid human embryos--warranting opportunities to augment embryonic stem cell research. AB - The recent vote in the British Parliament allows scientists in principle to create hybrid embryos by transferring human somatic cell nuclei into animal oocytes. This vote opens a fascinating new area of research with the central aim of generating interspecific lines of embryonic stem cells (ESCs) that could potentially be used to understand development, differentiation, gene expression and genomic compatibility. It will also promote human cell therapies, as well as the pharmaceutical industry's search for new drug targets. If this approach is to be successful, many biological questions need to be answered and, in addition, some moral and ethical aspects must be taken into account. PMID- 18675482 TI - Developing therapeutic proteins by engineering ligand-receptor interactions. AB - Ligand-receptor interactions govern myriad cell signaling pathways that regulate homeostasis and ensure that cells respond properly to stimuli. Growth factors, cytokines and other regulatory elements use these interactions to mediate cell responses, including proliferation, migration, angiogenesis, immune responses and cell death. Proteins that inhibit these processes have potential as therapeutics for cancer and autoimmune disorders, whereas proteins that stimulate these processes offer promise in regenerative medicine. Although much of the focus in this area over the past decade has been on monoclonal antibodies, recently there has been increased interest in the use of non-antibody proteins as therapeutic agents. Here, we review recent advances and accomplishments in the use of rational and combinatorial protein engineering approaches to developing ligands and receptors as agonists and antagonists against clinically important targets. PMID- 18675483 TI - Engineering microbial consortia: a new frontier in synthetic biology. AB - Microbial consortia are ubiquitous in nature and are implicated in processes of great importance to humans, from environmental remediation and wastewater treatment to assistance in food digestion. Synthetic biologists are honing their ability to program the behavior of individual microbial populations, forcing the microbes to focus on specific applications, such as the production of drugs and fuels. Given that microbial consortia can perform even more complicated tasks and endure more changeable environments than monocultures can, they represent an important new frontier for synthetic biology. Here, we review recent efforts to engineer synthetic microbial consortia, and we suggest future applications. PMID- 18675484 TI - Evaluation of antibacterial effects of a combination of Coptidis Rhizoma, Mume Fructus, and Schizandrae Fructus against Salmonella. AB - The antibacterial potential of a preparation of medicinal herbs for the treatment of salmonellosis was evaluated. The preparation contained a combination of methanolic extracts from Mume Fructus, Coptidis Rhizoma and Schizandrae Fructus, and is designated NP532. NP532 was tested against 26 different Salmonella stains. All these strains were affected by NP532 even though many of the salmonella strains were resistant to multiple antibiotics. The MIC values of this combination against the Salmonella strains varied from 0.49 to 7.8 mg/ml. The in vivo antibacterial activity of NP532 was examined using a S. Gallinarum infection chicken model. The chickens were initially infected with S. Gallinarum, and NP532 was then administered. This preparation was found to have major effects on mortality. The histopathological observations of the NP532 treated animals did not show any of the clinical signs and rarely showed histological damage associated with the disease. On the other hand, the untreated controls showed the clinical signs, e.g. congestion and necrotic changes in the liver, kidney, and spleen. This suggests that NP532 might be an effective treatment for salmonellosis. PMID- 18675485 TI - A standardized conjugation protocol to asses antibiotic resistance transfer between lactococcal species. AB - Optimal conditions and a standardized method for conjugation between two model lactococcal strains, Lactococcus lactis SH4174 (pAMbeta1-containing, erythromycin resistant donor) and L. lactis Bu2-60 (plasmid-free, erythromycin sensitive recipient), were developed and tested in a inter-laboratory experiments involving five laboratories from different countries. The ultimate goal of the study was to assess the microbial potential of antibiotic resistance transfer among Lactic Acid Bacteria (LAB). The influence of culture age (various OD values) and ratios of donor and recipient cultures as well as filter, solid and liquid mating techniques, were examined in order to optimize the conjugation protocol. In the result of these studies, we concluded that the donor-to-recipient ratio appear to be important; the most efficient technique for conjugation was filter mating and the optimal conditions for gene transfer were observed when late logarithmic cultures of both donor and recipient were used. Comparison of conjugal transfer frequencies between five partner laboratories showed that results are sufficiently intra-laboratory repeatable and inter-laboratory comparable. This is the first study of this kind, in which a standardized protocol of conjugal mating for testing antibiotic resistance dissemination among LAB was established and validated. PMID- 18675486 TI - Inactivation model equations and their associated parameter values obtained under static acid stress conditions cannot be used directly for predicting inactivation under dynamic conditions. AB - Organic acids (e.g., lactic acid, acetic acid and citric acid) are popular preservatives. In this study, the Listeria innocua inactivation is investigated under dynamic conditions of pH and undissociated lactic acid ([LaH]). A combined primary (Weibull-type) and secondary model developed for the L. innocua inactivation under static conditions [Janssen, M., Geeraerd, A.H., Cappuyns, A., Garcia-Gonzalez, L., Schockaert, G., Van Houteghem, N., Vereecken, K.M., Debevere, J., Devlieghere, F., Van Impe, J.F., 2007. Individual and combined effects of pH and lactic acid concentration on L. innocua inactivation: development of a predictive model and assessment of experimental variability. Applied and Environmental Microbiology 73(5), 1601-1611] was applied to predict the microbial inactivation under dynamic conditions. Because of its non autonomous character, two approaches were proposed for the application of the Weibull-type model to dynamic conditions. The results quantitatively indicated that the L. innocua cell population was able to develop an induced acid stress resistance under dynamic conditions of pH and [LaH]. From a modeling point of view, it needs to be stressed that (i) inactivation model equations and associated parameter values, derived under static conditions, may not be suitable for use as such under dynamic conditions, and (ii) non-autonomous dynamic models reveal additional technical intricacies in comparison with autonomous models. PMID- 18675488 TI - Tools to study DNA repair: what's in the box? AB - Our understanding of the DNA repair mechanisms that preserve genome integrity has increased greatly in recent years. To follow the DNA repair process, researchers have developed sophisticated techniques including live cell imaging, local damage induction and refined biochemical assays. These techniques have helped to elucidate the 'orchestration' of DNA repair mechanisms (i.e. the order of factor assembly around the lesion, the identification of new functions of known factors and the discovery of novel key regulators involved in DNA repair). We will discuss the uses and the limitations of these methods and their applications in the study of DNA repair. PMID- 18675489 TI - Low duplicability and network fragility of cancer genes. AB - We identified genomic and network properties of approximately 600 genes mutated in different cancer types. These genes tend not to duplicate but, unlike most human singletons, they encode central hubs of highly interconnected modules within the protein-protein interaction network (PIN). We find that cancer genes are fragile components of the human gene repertoire, sensitive to dosage modification. Furthermore, other nodes of the human PIN with similar properties are rare and probably enriched in candidate cancer genes. PMID- 18675487 TI - Whole chromosome instability and cancer: a complex relationship. AB - Although chromosome mis-segregation is a hallmark of cancer cells, its genetic basis and role in malignant transformation remain poorly understood. In recent years, several mouse models have been generated that harbor gene defects that perturb high-fidelity chromosome segregation. Analysis of these models has revealed that whole chromosome instability (W-CIN) can cause, inhibit or have no effect on tumorigenesis. Here we propose that the effect of W-CIN on tumor development depends on the particular W-CIN gene that is defective, including its other cellular functions, the extent or nature of the gene defect, the affected tissue or cell type and the context of other cancer gene mutations. PMID- 18675490 TI - Ectopic expression of prelamin A in early Xenopus embryos induces apoptosis. AB - Lamin proteins are components of metazoan cell nuclei. During evolution, two classes of lamin proteins evolved, A- and B-type lamins. B-type lamins are expressed in nearly all cell types and in all developmental stages and are thought to be indispensable for cellular survival. In contrast, A-type lamins have a more restricted expression pattern. They are expressed in differentiated cells and appear late in embryogenesis. In the earliest steps of mammalian development, A-type lamins are present in oocytes, pronuclei and during the first cleavage stages of the developing embryo. But latest after the 16-cell stage, A type lamin proteins are not any longer detectable in embryonic cells. Amphibian oocytes and early embryos do not express lamin A. Moreover, extracts of Xenopus oocytes and eggs have the ability to selectively remove A-type lamins from somatic nuclei. This observation and the restricted expression pattern suggest that the presence of lamin A might interfere with developmental processes in the early phase of embryogenesis. To test this, we ectopically expressed lamin A during early embryonic development of Xenopus laevis by microinjection of synthetic mRNA. Here, we show that introducing mature lamin A does not interfere with normal development. However, expression of prelamin A or lamin A variants that cannot be fully processed cause severe disturbances and lead to apoptosis during gastrulation. The toxic effect is due to lack of the conversion of prenylated prelamin A to its mature form. Remarkably, even a cytoplasmic prelamin A variant that is excluded from the nucleus drives embryos into apoptosis. PMID- 18675491 TI - Antiamoebic coumarins from the root bark of Adina cordifolia and their new thiosemicarbazone derivatives. AB - In continuation of our search for potential antiamoebic agents from folklore Indian medicinal plants, we found that the benzene and ethyl acetate extracts from the root bark of Adina cordifolia exhibited strong antiamoebic activity with IC(50) values of 2.92 and 2.50 microg/ml, respectively. Bioassay-guided fractionation of benzene and ethyl acetate extracts led to the isolation of 7 hydroxycoumarin (umbelliferone 1) and 7-beta-D-glucosylcoumarin (skimmin 2), respectively. Umbelliferone 1 was converted into 7-acetoxycoumarin 1a, which on treatment with aluminium chloride afforded 7-hydroxy-8-acetylcoumarin 2a. A new series of thiosemicarbazones 3a-e of 7-hydroxy-8-acetylcoumarin with different thiosemicarbazides were synthesized. Umbelliferone was also converted into its methoxy derivative (7-methoxycoumarin 4). Subsequently, all the compounds were assessed for antiamoebic activity against HM1:IMMS strain of Entamoeba histolytica. Umbelliferone and skimmin were found to possess a very good activity with IC(50) values of 6.38 and 4.35 microM/ml, respectively. The activity drastically increased on converting compound 2a into its thiosemicarbazone derivatives 3a-e with IC(50) values ranging between 1.06 and 4.46 microM/ml. Compounds 3b,c and e with IC(50) values of 1.49, 1.56 and 1.06 microM/ml, respectively, exhibited even higher antiamoebic activity than the standard drug metronidazole (IC(50)=2.62 microg/ml). The activity of 7-methoxycoumarin (IC(50)=8.92 microM/ml) was less than umbelliferone. Compounds 3b, c and e were tested for toxicity using H9c2 cardiac myoblasts cell line. The compounds exhibit >80% viability at 3.125-200 microg/ml. It is apparent from these results that umbelliferone and skimmin may be a useful lead for the development of new antiamoebic drugs. PMID- 18675492 TI - Radionuclides in terrestrial ecosystems of the zone of Kyshtym accident in the Urals. AB - It was shown that along the Eastern Ural Radioactive Trace central axis, about 100 km in length, decrease of the (90)Sr and (137)Cs deposition densities in soil samples may be described as an exponential function. At the western and eastern periphery of the trace, (90)Sr contents in soils approached to the background level due to global fallout. (90)Sr and (137)Cs concentrations in seeds of some herbaceous plants have been determined. The radionuclide concentrations and the resulting dose loads upon plant seeds showed an excess over the background level of about two or three orders of magnitude. PMID- 18675493 TI - White board to aid indexing of clinical photographs. PMID- 18675494 TI - Screw and "washer" fixation for onlay rib grafts to the mandible. PMID- 18675495 TI - Trigeminal neuralgia: assessment and management by oral and maxillofacial surgeons in the United Kingdom. AB - We aimed to find out how oral and maxillofacial surgeons in the United Kingdom currently assess and manage patients with suspected trigeminal neuralgia, and compared this with best practice. We undertook a postal survey of 297 consultant oral and maxillofacial surgeons in the UK (of whom 178 (60%) replied) asking questions about examinations and routine investigations of new patients with symptoms of trigeminal neuralgia, and their subsequent management. Clinical examination including cranial nerve function was made by almost all respondents (92%). Magnetic resonance imaging gives the most helpful diagnostic information and was requested by most surgeons (55%). Current publications have suggested that medical management of trigeminal neuralgia is appropriate for most patients, and this was reflected in current practice. Most surgeons (93%) used carbamazepine as their first choice of drug. PMID- 18675496 TI - Comparison of cage designs for transforaminal lumbar interbody fusion: a biomechanical study. AB - BACKGROUND: Prior biomechanical studies of transforaminal lumbar interbody fusion were primarily focused on various posterior instrumentation options, comparison with other fusion techniques, and cage positioning inside disc space. Few studies investigated the biomechanics of various cage designs in terms of construct stability. METHODS: Twelve lumbar motion segments were used in this study. The experimental procedure has two steps: multidirectional flexibility test and cyclic test. In the multidirectional flexibility test, all twelve specimens were tested following intact and five different cages (straight or banana shaped). The straight cages had biconvex or flat profile. In the cyclic test, the twelve specimens were randomly divided into two groups for biconvex and flat cages. Three thousand cycles in axial torsion, lateral bending and flexion extension were applied sequentially and cage migration was measured. FINDINGS: On average, the cage and posterior fixation reduced the range of motion of the intact condition by 40%, 69% and 75% in axial torsion, lateral bending and flexion extension, respectively. There was no statistical difference in construct stability among all five cages. The cage migration (biconvex vs flat) under cyclic loading was less than 0.2mm and no statistical difference was found. INTERPRETATION: The experimental results suggest that the geometry of cages, including shape (banana or straight), length, and surface profile (biconvex or flat), does not affect construct stability when the cages are used in conjunction with posterior fixation. With posterior fixation and surface serration, cage migration was minimal under cyclic loading for both biconvex and flat cages. PMID- 18675497 TI - Compensatory movements of transradial prosthesis users during common tasks. AB - BACKGROUND: Recent studies have documented motions of the upper limbs of healthy subjects during activities of daily living. The aim of this study was to investigate compensatory motions of the upper extremity and torso during tasks for transradial prosthesis users and to determine if bracing simulates prosthesis use. METHODS: Seven transradial myoelectric prosthesis users and 10 non-amputee volunteers performed four common tasks. Bracing was used to simulate the use of a transradial prosthesis by the non-amputee subjects. Range of motion of the glenohumeral (shoulder) joint, elbow joint and torso were calculated from optical motion analysis data. The motions between the non-braced, braced and transradial prosthesis user groups were statistically compared. Degree of asymmetry between the affected and unaffected arm was computed for the bilateral tasks. FINDINGS: Myoelectric transradial prosthesis users compensate for lack of wrist and forearm movement differently depending on the task. Compensatory motion in torso bending occurs while opening a door. For the box lift task, prosthesis users rely more on the sound arm and torso bending. While drinking from a cup, decreasing flexion of the glenohumeral joint and increasing elbow flexion was shown while using a prosthesis. While turning a steering wheel, prosthesis users are similar to non amputee subjects. INTERPRETATION: By looking at the compensatory motions caused by limiting forearm and wrist movement, a greater understanding of the problems with transradial prosthetic design can be developed. Although bracing intact subjects showed similar mechanisms of compensation in most tasks, the magnitude of compensation was greater for prosthesis users. PMID- 18675499 TI - Characterization of extractable and non-extractable polycyclic aromatic hydrocarbons in soils and sediments from the Pearl River Delta, China. AB - Formation of bound residues of pollutants in soils and sediments is an important process to control the fate of pollutants in the environment. The most of bound residue is not solvent extractable. In this paper, we measured both extractable and non-extractable polycyclic aromatic hydrocarbons (PAHs) in different organic matter fractions of samples from the Pearl River Delta, China. Non-extractable PAHs concentration was 234.45-1424.57microg/kg and accounted for 33.78-57.44% of total PAHs. 2-3 Ring PAHs were the dominant species and differed in concentration substantially between the samples. The atomic ratio of PAHs over organic-C in the fractions ordered as solvent soluble organic matter>humin>humic acids, matching the content of aliphatic moieties in the fractions of organic matter. The ratio of extractable and non-extractable PAHs may relate to the aging process of PAHs in soil and sediment. PMID- 18675498 TI - Effect of birch (Betula spp.) and associated rhizoidal bacteria on the degradation of soil polyaromatic hydrocarbons, PAH-induced changes in birch proteome and bacterial community. AB - Two birch clones originating from metal-contaminated sites were exposed for 3 months to soils (sand-peat ratio 1:1 or 4:1) spiked with a mixture of polyaromatic hydrocarbons (PAHs; anthracene, fluoranthene, phenanthrene, pyrene). PAH degradation differed between the two birch clones and also by the soil type. The statistically most significant elimination (p < or = 0.01), i.e. 88% of total PAHs, was observed in the more sandy soil planted with birch, the clearest positive effect being found with Betula pubescens clone on phenanthrene. PAHs and soil composition had rather small effects on birch protein complement. Three proteins with clonal differences were identified: ferritin-like protein, auxin induced protein and peroxidase. Differences in planted and non-planted soils were detected in bacterial communities by 16S rRNA T-RFLP, and the overall bacterial community structures were diverse. Even though both represent complex systems, trees and rhizoidal microbes in combination can provide interesting possibilities for bioremediation of PAH-polluted soils. PMID- 18675500 TI - Marital and cohabitation status as predictors of mortality: a 10-year follow-up of an Italian elderly cohort. AB - The relationship between mortality and marital status has long been recognized, but only a small number of investigations consider also the association with cohabitation status. Moreover, age and gender differences have not been sufficiently clarified. In addition, little is known on this matter about the Italian elderly population. The aim of this study is to examine differentials in survival with respect to marital status and cohabitation status in order to evaluate their possible predictive value on mortality of an Italian elderly cohort. This paper employs data from the Italian Longitudinal Study on Aging (ILSA), an extensive epidemiologic project on subjects aged 65-84 years. Of the 5376 individuals followed-up from 1992 to 2002, 1977 died, and 1492 were lost during follow-up period. The baseline interview was administered to 84% of the 5376 individuals and 65% of them underwent biological and instrumental examination. Relative risks of mortality for marital (married vs. non-married) and cohabitation (not living alone vs. living alone) categories are estimated through hazard ratios (HR), obtained by means of the Cox proportional hazards regression model, adjusting for age and several other potentially confounding variables. Non-married men (HR=1.25; 95% CI: 1.03-1.52) and those living alone (HR=1.42; 95% CI: 1.05-1.92) show a statistically significant increased mortality risk compared to their married or cohabiting counterparts. After age-adjustment, women's survival is influenced neither by marital status nor by cohabitation status. None of the other covariates significantly alters the observed differences in mortality, in either gender. Neither marital nor cohabitation status are independent predictors of mortality among Italian women 65+, while among men living alone is a predictor of mortality even stronger than not being married. These results suggest that Italian men benefit more than women from the protective effect of living with someone. PMID- 18675501 TI - Defining early morbidity of radical cystectomy for patients with bladder cancer using a standardized reporting methodology. AB - BACKGROUND: Reporting methodology is highly variable and nonstandardized, yet surgical outcomes are utilized in clinical trial design and evaluation of healthcare provider performance. OBJECTIVE: We sought to define the type, incidence, and severity of early postoperative morbidities following radical cystectomy (RC) using a standardized reporting methodology. DESIGN, SETTING, AND PARTICIPANTS: Between 1995 and 2005, 1142 consecutive RCs were entered into a prospective complication database and retrospectively reviewed for accuracy. All patients underwent RC/urinary diversion by high-volume fellowship-trained urologic oncologists. MEASUREMENTS: All complications within 90 d of surgery were analyzed and graded according to the Memorial Sloan-Kettering Cancer Center complication grading system. Complications were defined and stratified into 11 specific categories. Univariate and multivariate regression models were used to define predictors of complications. RESULTS AND LIMITATIONS: Sixty-four percent (735/1142) of patients experienced a complication within 90 d of surgery. Among patients experiencing a complication, 67% experienced a complication during the operative hospital admission and 58% following discharge. Overall, the highest grade of complication was grade 0 in 36% (n=407), grade 1-2 in 51% (n=582), and grade 3-5 in 13% (n=153). Gastrointestinal complications were most common (29%), followed by infectious complications (25%) and wound-related complications (15%). The 30-d mortality rate was 1.5%. CONCLUSIONS: Surgical morbidity following RC is significant and, when strict reporting guidelines are incorporated, higher than previously published. Accurate reporting of postoperative complications after RC is essential for counseling patients, combined modality treatment planning, clinical trial design, and assessment of surgical success. PMID- 18675502 TI - Editorial comment on: Defining early morbidity of radical cystectomy for patients with bladder cancer using a standardized reporting methodology. PMID- 18675503 TI - Editorial comment on: Defining early morbidity of radical cystectomy for patients with bladder cancer using a standardized reporting methodology. PMID- 18675504 TI - A minimally invasive approach in the treatment of vesicoureteral reflux in neurogenic bladder in children. PMID- 18675505 TI - Prostate-specific antigen: could it be a useful marker for bladder outlet obstruction? PMID- 18675506 TI - Vaccination with transforming growth factor-beta insensitive dendritic cells suppresses pulmonary metastases of renal carcinoma in mice. AB - Dendritic cells (DCs) have been widely used as cancer vaccines. However, their functional abilities have often been suppressed by tumor-secreted immunosuppressants such as transforming growth factor-beta (TGF-beta). We developed a new strategy using a TGF-beta insensitive DC as cancer vaccine. The effect of this vaccine was tested in a murine pulmonary metastases model of renal carcinoma (Renca). Tumor lysate-pulsed DCs (TP-DCs) were infected with retrovirus containing gene of dominant negative TGF-beta type II receptor (TbetaRIIDN) and thus made TGF-beta insensitive. Vaccination of the mice bearing Renca pulmonary metastases with the TbetaRIIDN TP-DC induced powerful tumor-specific cytotoxic T lymphocyte (CTL) responses, suppressed pulmonary metastases, and prolonged survival times. These results suggest TGF-beta-insensitive TP-DC vaccine can be used to enhance the antitumor efficacy of DC vaccine. PMID- 18675507 TI - Short hairpin RNA targeting c-FLIP sensitizes human cervical adenocarcinoma Hela cells to chemotherapy and radiotherapy. AB - c-FLIP inhibits caspase-8 activation and cell apoptosis mediated by death receptors. The present study aims at determining the effects of c-FLIP targeted vector-based short hairpin RNA (shRNA) on cell growth and evaluating its modulation of responsiveness to drugs and radiotherapy in cervical adenocarcinoma Hela cells. cFLIP expression of the cells transfected with shRNA against c-FLIP was significantly down-regulated after 72 h. c-FLIP silencing markedly suppressed cell proliferation and increased cell apoptosis. The activation of caspase-8 and caspase-3 was induced with shRNA targeting cFLIP with the passage of time after transfection. Furthermore, Vector-based shRNA against c-FLIP subsequently increased the sensitivity to cisplatin, iritican and Co60 radiotherapy by about 4 to 6-folds in Hela cells. Our data suggest that vector-based shRNA effectively inhibited c-FLIP expression, enhanced the expression level of caspase-8 and caspase-3 to induce cell apoptosis, probably with the higher efficacy in combination therapies with conventional chemotherapy and radiotherapy in cervical adenocarcinoma. PMID- 18675508 TI - Spatio-temporal dynamics of nutrients in the upper Han River basin, China. AB - The upper Han River basin with an area of approximately 95,000 km(2), is the water source area of the Middle Route of China's South to North Water Transfer Project. Thus, water quality in the basin's river network is of great importance. Nutrients including dissolved inorganic nitrogen (DIN), NO(3)(-)-N, NH(4)(+)-N, and dissolved phosphorus (DP) were analyzed in 41 sites during the period of 2005 2006. Cluster analysis (CA), analysis of variance (ANOVA) and general linear models (GLM) were performed to explore their spatio-temporal variations in the basin. The results revealed that the DIN, NO(3)(-)-N and NH(4)(+)-N increased over the 2 year study period, and their concentrations in the wet season was higher than those in the dry season. The seasonal variation in nitrogen was strongly associated with seasonal pattern of precipitation and there was a negative relationship between DP concentration and river flow. Cluster analysis indicated high nutrient contents in the urban and agricultural production areas. The research will help articulate water resource management strategy for the interbasin water transfer project. PMID- 18675509 TI - Aqueous heavy metals removal by adsorption on amine-functionalized mesoporous silica. AB - Amino functional mesoporous silica SBA-15 materials have been prepared to develop efficient adsorbents of heavy metals in wastewater. Functionalization with amino groups has been carried out by using two independent methods, grafting and co condensation. Three organic moieties have been selected to incorporate the active amino sites: aminopropyl (H(2)N-(CH(2))(3)-), [2-aminoethylamino]-propyl (H(2)N (CH(2))(2)-NH-(CH(2))(3)-) and [(2-aminoethylamino)-ethylamino]-propyl (H(2)N (CH(2))(2)-NH-(CH(2))(2)-NH-(CH(2))(3)-). Materials have been characterized by XRD, nitrogen sorption measurements and chemical analysis. We have found that all materials preserve the mesoscopic order and exhibit suitable textural properties and nitrogen contents to act as potential adsorbents. Metal removal from aqueous solution has been examined for Cu(II), Ni(II), Pb(II), Cd(II), and Zn(II); adsorption performances of materials prepared by the two functionalization methods have been compared. In addition, copper adsorption process has been thoroughly studied from both kinetic and equilibrium points of view for some selected materials. Aqueous Cu(II) adsorption rates show that the overall process is fast and the time evolution can be successfully reproduced with a pseudo second-order kinetic model. Whole copper adsorption isotherms have been obtained at 25 degrees C. Significant maximum adsorption capacities have been found with excellent behavior at low concentration. PMID- 18675510 TI - Potential use of cotton plant wastes for the removal of Remazol Black B reactive dye. AB - In this study, the potential use of cotton plant wastes - stalk (CS) and hull (CH) - as sorbents for the removal of Remazol Black B (RB5), a vinyl sulfone type reactive dye, was investigated. The results indicated that adsorption was strongly pH-dependent but slightly temperature-dependent for each sorbent-dye system. The Freundlich, Langmuir, Redlich-Peterson and Langmuir-Freundlich adsorption models were used for the mathematical description of adsorption equilibrium and isotherm constants were evaluated at 25 degrees C. All models except the Freundlich model were applicable for the description of dye adsorption by both sorbents in the concentration range studied. According to the Langmuir model, CS and CH sorbents exhibited the highest RB5 dye uptake capacities of 35.7 and 50.9 mg g(-1), respectively, at an initial pH value of 1.0. Simple mass transfer and kinetic models were applied to the experimental data to examine the mechanisms of adsorption and potential rate-controlling steps. It was found that both external mass transfer and intra-particle diffusion played an important role in the adsorption mechanisms of dye, and adsorption kinetics followed the pseudo second-order type kinetic model for each sorbent. Using the Langmuir model parameters, thermodynamic constant DeltaG degrees was also evaluated for each sorption system. PMID- 18675511 TI - Separation of zinc and nickel ions in a strong acid through liquid-liquid extraction. AB - Many solid wastes contain both zinc and nickel at the same time. For recycling or recovery of metals, it is essential to separate materials. Among those materials, zinc and nickel are very difficult to be separated because there is not so much difference in the chemical and physical properties. This paper focuses on the separation of zinc and nickel ions in a diluted aqua regia solution. Liquid liquid extraction by TBP, Cyanex 272 and Cyanex 301 was used and a distribution coefficient (D), a separation factor (S) and a relative purity (R) were induced to evaluate the degree of separation. All of the extractions were proportional to the concentration of the extractants, and zinc ions were extracted more easily than nickel ions. Among the extractants, Cyanex 301 showed the best characteristics regarding Zn/Ni separation. In particular, the extraction of zinc ions in the range of pHor=40 kg; 10mg for those weighing between 40 and 60 kg; 20mg for those weighing >60 kg. RESULTS: Sixty participants, 16 females and 44 males, had an average body surface area burn of 53% (S.D.=18) and average age of 11 years (S.D.=4). Imipramine subjects received an average daily dose of 1.00+/-0.29 mg/kg. Fluoxetine subjects received an average daily dose of 0.29+/-0.16 mg/kg. Between group differences were not detected. Fifty-five percent responded positively to placebo; 60% responded positively to imipramine; and 72% responded positively to fluoxetine. CONCLUSION: Within the parameters of this study design and sample, placebo was statistically as effective as either drug in treating symptoms of ASD. PMID- 18675521 TI - Facial emotion recognition and amygdala activation are associated with menstrual cycle phase. AB - Converging evidence has accumulated that menstrual cycle and thus hormonal levels can affect emotional behavior, in particular facial emotion recognition. Here we explored the association of ovarian hormone levels and amygdala activation during an explicit emotion recognition task in two groups of healthy young females: one group was measured while in their follicular phase (n=11) and the other during their luteal phase (n=11). Using a 3T scanner in combination with a protocol specifically optimized to reliably detect amygdala activation we found significantly stronger amygdala activation in females during their follicular phase. Also, emotion recognition performance was significantly better in the follicular phase. We observed significant negative correlations between progesterone levels and amygdala response to fearful, sad and neutral faces, further supporting a significant modulation of behavior and neural response by hormonal changes during the menstrual cycle. From an evolutionary point of view this significant influence of ovarian hormone level on emotion processing and an important neural correlate, the amygdala, may enable a higher social sensitivity in females during their follicular phase, thus facilitating socio-emotional behavior (and social interaction) which may possibly facilitate mating behavior as well. PMID- 18675520 TI - Novel mRNA isoforms of the sodium channels Na(v)1.2, Na(v)1.3 and Na(v)1.7 encode predicted two-domain, truncated proteins. AB - The expression of voltage-gated sodium channels is regulated at multiple levels, and in this study we addressed the potential for alternative splicing of the Na(v)1.2, Na(v)1.3, Na(v)1.6 and Na(v)1.7 mRNAs. We isolated novel mRNA isoforms of Na(v)1.2 and Na(v)1.3 from adult mouse and rat dorsal root ganglia (DRG), Na(v)1.3 and Na(v)1.7 from adult mouse brain, and Na(v)1.7 from neonatal rat brain. These alternatively spliced isoforms introduce an additional exon (Na(v)1.2 exon 17A and topologically equivalent Na(v)1.7 exon 16A) or exon pair (Na(v)1.3 exons 17A and 17B) that contain an in-frame stop codon and result in predicted two-domain, truncated proteins. The mouse and rat orthologous exon sequences are highly conserved (94-100% identities), as are the paralogous Na(v)1.2 and Na(v)1.3 exons (93% identity in mouse) to which the Na(v)1.7 exon has only 60% identity. Previously, Na(v)1.3 mRNA has been shown to be upregulated in rat DRG following peripheral nerve injury, unlike the downregulation of all other sodium channel transcripts. Here we show that the expression of Na(v)1.3 mRNA containing exons 17A and 17B is unchanged in mouse following peripheral nerve injury (axotomy), whereas total Na(v)1.3 mRNA expression is upregulated by 33% (P=0.003), suggesting differential regulation of the alternatively spliced transcripts. The alternatively spliced rodent exon sequences are highly conserved in both the human and chicken genomes, with 77-89% and 72-76% identities to mouse, respectively. The widespread conservation of these sequences strongly suggests an additional level of regulation in the expression of these channels, that is also tissue-specific. PMID- 18675522 TI - Fatal granulomatous amoebic meningoencephalitis due to Balamuthia mandrillaris. AB - Amoebic infections of the central nervous system (CNS) are very rare and usually fatal. A 33-year-old Bolivian male injured his elbow 18 months ago in an accident and, months later, developed multiple skin lesions. He was admitted with confusion, and brain images showed large multifocal lesions with mass effect. Review of the skin biopsy revealed the presence of amoebic trophozoites within a granulomatous inflammation. Despite treatment, he continued to deteriorate and died 17 days after admission. The brain was swollen with prominent uncal herniation, and the leptomeninges showed patchy exudate. Coronal sections revealed widespread hemorrhagic and necrotic lesions. Histology confirmed granulomatous amoebic meningoencephalitis (GAM) with the presence of amoebic trophozoites and occasional cysts. Post-mortem culture confirmed Balamuthia mandrillaris. GAM due to B. mandrillaris may occur in any age group, may or may not be associated with immunosuppression, and is present worldwide. It usually has a subacute and fatal course from hematogenous dissemination of chronic skin or lung lesions. PMID- 18675523 TI - Cloning, characterization, and gene expression analysis of a novel cadmium metallothionein gene in Tetrahymena pigmentosa. AB - A novel cadmium-inducible metallothionein (MT) gene (Tpig-MT1) was cloned and sequenced from the ciliate Tetrahymena pigmentosa. The number of deduced amino acids is 118. The polypeptide possesses CCC and CC clusters characteristic of typical Tetrahymena Cd-inducible MTs. The structure of Tpig-MT1 is different from the reported Cd-MT in T. pyriformis, T. thermophila and T. pigmentosa. Tpig-MT1 contains two intragenic tandem repeats with 72.9% identity described as Tpig-MT1 (repeat A1) and Tpig-MT1 (repeat A2). The transcriptional response of Tpig-MT1 gene to different heavy metals (Cd, Cu, Zn, Hg, Pb) and oxidative stress (H(2)O(2)) was measured using real-time quantitative PCR. The results showed that the gene was quickly induced (1 h) by the five heavy metals and the order of expression level was Hg>Pb>Cd>Cu>Zn. The induction effect of H(2)O(2) was 5-fold after about 15 min, but soon decreased to a non-significant level (30 min). The genetic diversity of Tetrahymena MT genes is discussed in relation to the unique structure of the Tpig-MT1 gene and other reported Cd-MT isoforms. PMID- 18675524 TI - Pathogenicity of highly pathogenic avian influenza viruses of H5N1 subtype isolated in Thailand for different poultry species. AB - Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype have caused several rounds of outbreaks in Thailand. In this study, we used 3 HPAI viruses isolated in Thailand in January 2004 from chicken, quail, and duck for genetic and pathogenetic studies. Sequence analysis of the entire genomes of these isolates revealed that they were genetically similar to each other. Chickens, quails, domestic ducks, and cross-bred ducks were inoculated with these isolates to evaluate their pathogenicity to different host species. A/chicken/Yamaguchi/7/04 (H5N1), an HPAI virus isolated in Japan, was also used in the chicken and quail studies for comparison. All four isolates were shown to be highly pathogenic to chickens and quails, with 100% mortality by 10(6) EID50 inoculants of the viruses. They caused sudden death in chickens and quails within 2-4 days after inoculation. The mean death times (MDT) of quails infected with the Thai isolates were shorter than those of chickens infected with the same isolates. Mortality against domestic and cross-bred ducks ranged from 50 to 75% by intranasal inoculation with the 10(6) EID50 viruses. Neurological symptoms were observed in most of the inoculated domestic ducks and appeared less severe in the cross-bred ducks. The MDTs of the ducks infected with the Thai isolates were 4.8-6 days post-inoculation. Most of the surviving ducks infected with the Thai isolates had sero-converted until 14 dpi. Our study illustrated the pathobiology of the Thai isolates against different poultry species and would provide useful information for improving control strategies against HPAI. PMID- 18675525 TI - Inhibition of chemically induced inflammation and pain by orally and topically administered leaf extract of Manihot esculenta Crantz in rodents. AB - The aqueous leaf extract of Manihot esculenta Crantz (MELE) is being used orally and topically in traditional African medicine for the treatment of inflammation and pain, and claimed to be safe. The anti-inflammatory effects of MELE (100-400 mg/kg, p.o. or 1-4%, w/w in petroleum jelly, topically) were tested against carrageenan-induced paw oedema in rats as well as against xylene-induced ear oedema in mice. The analgesic effect of MELE (100-400 mg/kg, p.o. or 1-4%, w/w in petroleum jelly, topically) was tested against acetic acid-induced (20 microl, 0.6%, v/v in normal saline, i.p.) and acetylcholine-induced (8.3 mg/kg, i.p.) mouse writhing models. At 100-400 mg/kg, p.o. and 1-4% (w/w), topically, MELE produced significant inhibitions of carrageenan-induced rat paw oedema and xylene induced ear swelling in mice. Effects produced by MELE were significantly higher than those produced by indomethacin (10 mg/kg, s.c. or 1%, w/w in petroleum jelly) in the anti-inflammatory models. For the analgesic effect, MELE (100-400 mg/kg, orally) and (1-4%, w/w, topically), like aspirin (100 mg/kg, i.p.) exhibited significant (P<0.05) inhibition of acetic acid- and acetylcholine induced mouse writhing tests, compared to untreated control. Effects produced by MELE were significantly lower than those produced by aspirin (100 mg/kg, i.p.) in the analgesic models, except for the topically administered extract on acetylcholine-induced pain. Acute oral administration up to 10 g/kg did not cause death within 14 days, but mortalities were produced in i.p. administered extract with LD(50) of 2.5+/-0.3 g/kg. Based on these, the extract may contain orally safe, topically and orally effective anti-inflammatory and analgesic principles, which justify its use in traditional African medicine. PMID- 18675526 TI - An interactive CD-ROM to inform patients about stem cell transplantation. AB - OBJECTIVE: Cancer patients receiving chemotherapy or a Stem Cell Transplantation (SCT) are in need of information about their disease, treatment options and side effects. Patient education usually has to be given within limited time. Under these circumstances, patients may find it difficult to completely understand and to retain the information given. METHODS: As a supplement to standard information methods we developed an interactive CD-ROM with information on SCT. This CD-ROM provides both medical information and more subjective patients' experiences. Part one provides information regarding the treatment course from diagnosis through to post-discharge care. The second part consists of interviews with former patients and describes their experiences. As the system is interactive, it can be utilised according to the patient's individual preferences. The CD-ROM comprises audio, video, animations, pictures, and text. Printing of certain sections is optional. The technical format of the CD-ROM makes it relatively simple to utilise the information and to make it suitable for other institutions or even other treatments. In this preliminary study the acceptability of the interactive CD-ROM by patients undergoing a SCT is described. RESULTS: Patients' overall evaluations of the interactive CD-ROM were highly positive. For example, 90.2% (N=51) found it interesting, clear, useful and valued getting information by means of a CD ROM. Most patients would recommend the interactive CD-ROM to other patients in the same situation. CONCLUSION: The content of the CD-ROM on SCT as well as the computer-based interactive method are well accepted by patients. PRACTICE IMPLICATIONS: Computer-based education may enhance patient education and thus the quality of patient care. We must now establish the program's effectiveness. Moreover, plans have been developed to disseminate the information on SCT over the Internet. Future development of comparable programs and their evaluation should be encouraged to promote the well-being of cancer patients. PMID- 18675527 TI - Metabolic traits affecting growth rates of pre-pubertal calves and their relationship with subsequent survival. AB - This study related growth and metabolic traits in Holstein-Friesian calves (n=45) to subsequent performance and longevity. Animals were measured at birth, 3, 6 and 9 months (weight, crown rump length (CRL), heart girth and height). Endocrine and metabolic traits were assessed at 6 months of age. These traits were not influenced by sire (n=5). Milk production and mortality records were obtained through 3 lactations. Seven heifers failed to achieve a lactation, 6 were culled after one, 17 after two and 15 (33%) survived >/=3 lactations. Birth weight and girth but not skeletal measures were highly positively correlated with the repeated size measurements at 3-9 months. Calves with higher cortisol at 6 months were smaller in weight and girth at 3 months, with reduced 3-6 months skeletal growth. GH secretion was related to height, weight and growth, whereas urea was most strongly related to CRL. Calves not achieving a single lactation had lower IGF-I. Calves with higher glucose or BHB were more likely to be culled after 2 lactations, an effect which was independent of yield. Size and metabolic measurements of growing calves may therefore prove useful in predicting longevity. PMID- 18675528 TI - Tobacco quitline use: enhancing benefit and increasing abstinence. AB - OBJECTIVE: This study sought to evaluate the impact on the quitline utilization and abstinence rates of increasing a free nicotine-replacement therapy (NRT) benefit from 4 weeks to 6 weeks. METHODS: Intake data were utilized to assess the number of callers to the quitline during the time period in which 4 weeks of free NRT was provided (January-November 2006) and the time period of the enhanced NRT benefit (December 2006-June 2007). Abstinence rates at 3 and 6 months were calculated for people utilizing the quitline program during both time periods. RESULTS: The mean number of intake calls to the quitline increased from 397 (range 326-509) prior to the enhanced NRT benefit to 712 (range 592-1227) during the 6-week NRT benefit period. The 6-month tobacco abstinence rates were significantly higher among people receiving the 6-week NRT benefit compared to those receiving the 4-week benefit (OR=1.51; 95% CI=1.07, 1.66). CONCLUSIONS: The findings suggest that the promotion of an expanded NRT benefit can increase quitline utilization and abstinence rates. PMID- 18675530 TI - Human papillomavirus vaccination coverage on YouTube. AB - BACKGROUND: A large percentage of Internet users regularly search for health related information. In recent years, participatory Internet sites such as YouTube have become increasingly popular, in part because individuals are able to both retrieve and post information. This study analyzed how human papillomavirus (HPV) vaccination was portrayed in videoclips and viewer-posted comments available on YouTube. METHODS: YouTube (www.youtube.com) was queried on February 8, 2008, using the search terms Gardasil, cervical cancer vaccination, and HPV vaccination to identify and download relevant videoclips. Videoclips were classified as either positively or negatively portraying HPV vaccination, and viewer-posted comments were enumerated. Data analyses were conducted immediately following videoclip retrieval. RESULTS: A total of 146 unique YouTube videoclips were located, using the three search keywords combined. Three quarters (n=109; 74.7%) of the videoclips portrayed HPV vaccination in a positive manner. One third (n=47; 32.2%) of the videoclips had generated at least one posted comment. CONCLUSIONS: These results demonstrate that there is a wide variety of information on YouTube regarding HPV vaccination and cervical cancer. As a result, public health and medical professionals need to be cognizant of the nature of the HPV-related information available, so that they are better equipped to respond to patients who acquire information posted on YouTube and other Internet sources. PMID- 18675529 TI - Comparison of cigarette and water-pipe smoking by Arab and non-Arab-American youth. AB - BACKGROUND: Water-pipe smoking is a rapidly growing form of tobacco use worldwide. Building on an earlier report of experimentation with cigarette and water-pipe smoking in a U.S. community sample of Arab-American youth aged 14-18 years, this article examines water-pipe smoking in more detail (e.g., smoking history, belief in harmfulness compared to cigarettes, family members in home who smoke water pipes) and compares the water-pipe-smoking behaviors of Arab-American youth with non-Arab-American youth in the same community. METHODS: A convenience sample of 1872 Arab-American and non-Arab-American high school students from the Midwest completed a 24-item tobacco survey. Data were collected in 2004-2005 and analyzed in 2007-2008. RESULTS: Arab-American youth reported lower percentages of ever cigarette smoking (20% vs 39%); current cigarette smoking (7% vs 22%); and regular cigarette smoking (3% vs 15%) than non-Arab-American youth. In contrast, Arab-American youth reported significantly higher percentages of ever water-pipe smoking (38% vs 21%) and current water-pipe smoking (17% vs 11%) than non-Arab American youth. Seventy-seven percent perceived water-pipe smoking to be as harmful as or more harmful than cigarette smoking. Logistic regression showed that youth were 11.0 times more likely to be currently smoking cigarettes if they currently smoked water pipes. Youth were also 11.0 times more likely to be current water-pipe smokers if they currently smoked cigarettes. If one or more family members smoked water pipes in the home, youth were 6.3 times more likely to be current water-pipe smokers. The effects of ethnicity were reduced as a result of the explanatory value of family smoking. CONCLUSIONS: Further research is needed to determine the percentages, patterns, and health risks of water-pipe smoking and its relationship to cigarette smoking among all youth. Additionally, youth tobacco prevention/cessation programs need to focus attention on water-pipe smoking in order to further dispel the myth that water-pipe smoking is a safe alternative to cigarette smoking. PMID- 18675531 TI - [Takotsubo syndrome: a young woman case]. AB - After induction of anaesthesia for lumbar disc herniation surgery, an ASA-1, 39 year-old woman presented an acute heart failure. A persistent hypotension with tachycardia was observed after a brief hypertension peak during orotracheal intubation. After electrocardiogram, echocardiogram and biologic dosages, the diagnosis seems to be coronary ischemia. Coronarography was normal; takotsubo syndrome diagnosis was made with typical ventriculographic aspect. Evolution at two days and one month was favorable. PMID- 18675533 TI - [Reputation and "electronic" consultations in progress go with the demands of the editorial board of La Presse Medicale]. PMID- 18675532 TI - Resveratrol, a natural phytoalexin, normalizes hyperglycemia in streptozotocin nicotinamide induced experimental diabetic rats. AB - Resveratrol is a polyphenolic phytoalexin produced in appreciable amounts as a secondary metabolite in grapevines in response to fungal infections. Based on the present knowledge, it appears to be a promising bioactive natural molecule with potential applications in phytotherapy or pharmacology. The present study was aimed to evaluate the antidiabetic properties of resveratrol in streptozotocin nicotinamide induced experimental diabetes in rats. The diabetic rats orally treated with resveratrol (5 mg kg(-)(1)b.w d(-)(1)) for 30 days resulted in significant (p<0.05) decrease in the levels of blood glucose, glycosylated hemoglobin, blood urea, serum uric acid, serum creatinine and diminished activities of pathophysiological enzymes such as aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP). The antihyperglycemic nature of resveratrol is also evidenced from the improvement in the levels of plasma insulin and hemoglobin. Further, the results are comparable with glyclazide, an oral standard drug. Thus, the present findings suggest that resveratrol may be considered as an effective therapeutic agent for the treatment of diabetes mellitus. PMID- 18675534 TI - Breathiness as a feminine voice characteristic: a perceptual approach. AB - The present study tested suggestions in the literature that vocal breathiness may influence the perception of femininity of the speaker. Two listener experiments were conducted using normal and breathy voice productions by normal females. Panels of students rated femininity of samples from seven biological female speakers, each producing a normal /a/ and a breathy /a/ at similar pitch and intensity. Two listening experiments were enacted. In experiment 1, the 14 samples were presented at random to the judges as if they were from different speakers. Judges rated feminity on a five-point rating scale with 1 (little feminine) and 5 (very feminine) as left and right extremes, respectively. In experiment 2, the normal and breathy samples of each of the seven speakers were presented pair wise and judges were required to indicate which of the two in their opinion sounded most feminine. In all seven participants, the breathy voice samples were judged to be more feminine than the natural voice samples. This was the case when the breathy and natural samples were presented randomly in experiment 1 and when the samples of each speaker were presented pair wise in experiment 2. Results demonstrate that breathiness indeed may contribute to the perception of femininity but replication in a study involving biological males and transgender clients is indicated. It is unclear which degree of breathiness is required or is the most suitable for facilitating the perception of femininity. PMID- 18675535 TI - Energy minimization in the nonlinear dynamic recurrent associative memory. AB - Chartier and his colleagues have recently proposed a nonlinear synchronous attractor neural network. In the Nonlinear Dynamic Recurrent Associative Memory (NDRAM), learning has been shown to converge to a set of real-valued attractors in single-layered neural networks and bidirectional associative memories. However, the transmission is highly nonlinear and its global stability has never been analytically proven. In this article, it is shown that NDRAM is an instance of the Cohen-Grossberg class of models and its energy function is defined. Analysis of the energy function shows that the transmission is stable in the entire domain of NDRAM. Numerical simulations further support this analysis. PMID- 18675536 TI - Deterministic neural dynamics transmitted through neural networks. AB - Precise spatiotemporal sequences of neuronal discharges (i.e., intervals between epochs repeating more often than expected by chance), have been observed in a large set of experimental electrophysiological recordings. Sensitivity to temporal information, by itself, does not demonstrate that dynamics embedded in spike trains can be transmitted through a neural network. This study analyzes how synaptic transmission through three archetypical types of neurons (regular spiking, thalamo-cortical and resonator), simulated by a simple spiking model, can affect the transmission of precise timings generated by a nonlinear deterministic system (i.e., the Zaslavskii mapping in the present study). The results show that cells with subthreshold oscillations (resonators) are very sensitive to stochastic inputs, and are not a good candidate for transmitting temporally coded information. Thalamo-cortical neurons may transmit very well temporal patterns in the absence of background activity, but jitter accumulates along the synaptic chain. Conversely, we observed that cortical regular-spiking neurons can propagate filtered temporal information in a reliable way through the network, and with high temporal accuracy. We discuss the results in the general framework of neural dynamics and brain theories. PMID- 18675537 TI - Re: The radiological fate of dermis-fat grafts in the human temporomandibular joint using magnetic resonance imaging. Dimotroulis G, Trost N, Morrison W. Int J Oral Maxillofac. Surg. 2008; 37: 249-254. PMID- 18675538 TI - Insulin resistance following continuous, chronic olanzapine treatment: an animal model. AB - Some atypical antipsychotics have been linked to an increased propensity for weight gain and metabolic disturbances, including type II diabetes. The objective of this study was to investigate an animal model to help understand the mechanisms underlying this phenomenon. Female, Sprague-Dawley rats were treated with olanzapine (2.0 or 7.5 mg/kg, via osmotic mini-pump) for 4 weeks, followed by the hyperinsulinemic/euglycemic and hyperglycemic clamp procedures to assess insulin sensitivity and secretion in vivo. Changes in body weight, visceral fat, food intake and locomotor activity were also assessed. Hepatic glucose production (R(A)) was increased in the hyperinsulinemic/euglycemic clamp for both treatment groups compared to control rats, while the high-dose olanzapine group had decreased peripheral glucose utilization (R(D)). No changes in insulin secretion were detected in the hyperglycemic clamp. Olanzapine did not change body weight or food intake, but did result in significant accumulation of visceral fat and decreases in locomotor activity. Like others, we found that a rodent model for antipsychotic-related weight gain per se is not tenable. However, chronic treatment with olanzapine was found to confer both hepatic and peripheral insulin resistance independent of weight gain, indicating a direct effect on glucose dysregulation. PMID- 18675539 TI - NHS-ester functionalized poly(PEGMA) brushes on silicon surface for covalent protein immobilization. AB - Poly(PEGMA) homopolymer brushes were developed by atom transfer radical polymerization (ATRP) on the initiator-modified silicon surface (Si-initiator). Through covalent binding, protein immobilization on the poly(PEGMA) films was enabled by further NHS-ester functionalization of the poly(PEGMA) chain ends. The formation of polymer brushes was confirmed by assessing the surface composition (XPS) and morphology (atomic force microscopy (AFM), scanning electronic microscopy (SEM)) of the modified silicon wafer. The binding performance of the NHS-ester functionalized surfaces with two proteins horseradish peroxidase (HRP) and chicken immunoglobulin (IgG) was monitored by direct observation. These results suggest that this method which incorporates the properties of polymer brush onto the binding surfaces may be a good strategy suitable for covalent protein immobilization. PMID- 18675540 TI - Transition metal ion binding studies of carnosine and histidine: biologically relevant antioxidants. AB - Carnosine and histidine are biologically interesting antioxidants. In order to probe whether they exert their antioxidant effect through metal ion chelation, the Cu(II) ion chelating abilities of these compounds were measured by UV-vis spectroscopy. Both of these compounds showed 1:1 complexations with Cu(II) ions as shown by their Job's plot. The binding constants for histidine and carnosine, as determined by Benesi-Hildebrand method, at pH 7.84+/-0.18, were found to be 71 and 1.1M(-1), respectively. The unexpectedly lower binding constant values of carnosine show the relatively minor role of the transition metal ion chelation in their antioxidant abilities. PMID- 18675541 TI - [Vestibular neuronitis in a teenager with sickle cell disease. Treatment is urgent]. AB - Vestibular syndrome is not frequently described in patients with sickle cell disease. We report the case of a teenager with sickle cell disease who had a vestibular syndrome with vertigo that successfully responded to exchange transfusion. We discuss guidelines and review the literature in view of this case report. Sensorineural disorders should be considered as stroke syndromes. They require urgent treatment consisting of exchange transfusion or maintaining optimal hydration associated with blood withdrawal. Treatment of vestibular syndrome in sickle cell disease is urgent. PMID- 18675542 TI - Low alcohol alternatives: a promising strategy for reducing alcohol related harm. AB - BACKGROUND: Less than 1% of the beer market in British Columbia comprises beers with an alcohol content below 4%, despite the success of low alcohol beers in other countries, e.g. Australia. A small experimental study is described in which male students were given either unmarked low alcohol beer (3.8%) or regular strength beer (5.3%) to investigate their enjoyment and subjective intoxication. METHODS: Thirty-four male students who reported drinking 5 or more beers in 1 day at least once in the last month volunteered for the study. In each drinking session, small groups of between 6 and 10 students consumed two servings of beer while playing dominoes. Each subject was his own control in the experiment by attending two group-drinking sessions, drinking a different beverage each time. The different beers were given in balanced order with half the subjects in each group drinking each type of beer. Standard measures of subjective intoxication and enjoyment were used. Blood alcohol levels were tested before, during and after drinking. RESULTS: Although significantly higher blood alcohol levels were obtained with the higher strength beer (means of 0.026 versus 0.033 mg/100 ml at the end of the study, p < 0.001), (i) most participants reported enjoying the two sessions equally or preferred the low alcohol beer session, (ii) most did not report feeling different between the two sessions and (iii) only about half correctly guessed which was the higher alcohol content beer. There was a preference, however, for the taste of the stronger beer. CONCLUSION: We conclude beer drinkers cannot readily distinguish low and regular strength beers and can enjoy socializing equally with either. We recommend taxation strategies to create incentives for the manufacture, marketing and consumption of low alcohol alternatives. PMID- 18675543 TI - Towards on-site pathogen detection using antibody-based sensors. AB - In this paper, the recent progress within biosensors for plant pathogen detection will be reviewed. Bio-recognition layers on sensors can be designed in various ways, however the most popular approach is to immobilise antibodies for specific capture of analytes. Focus will be put on antibody surface-immobilisation strategies as well as the use of antibodies in the widely used sensors, quartz crystal microbalance, surface plasmon resonance and cantilevers. We will describe the available data on antibody-based plant pathogen detection and furthermore use examples from detection of the pathogens Salmonella, Listeria monocytogenes, Streptococcus mutans, Bacillus cereus, Bacillus anthracis, Campylobacter and Escherichia coli. We will touch upon optimal assay design and further discuss the strengths and limitations of current sensor technologies for detection of viruses, bacteria and fungi. PMID- 18675544 TI - Phenolics composition in Erica sp. differentially exposed to metal pollution in the Iberian Southwestern Pyritic Belt. AB - The phenolic composition of different populations of Erica andevalensis,E. australis and E. arborea was analyzed according to the different degree of exposition to metals in soils. E. andevalensis populations, highly exposed to metal pollution, had the lowest total phenol content and the lowest antioxidant activity measured as trolox equivalents. The HPLC analysis of leaf extracts revealed that phenolic composition in all analyzed populations of E. australis and E. arborea was very similar, although wild populations were differentially exposed to metal pollutions. However, E. andevalensis showed a phenolic profile characterized by the absence of many phenolic compounds quantified in the other species, although two compounds derived from cinnamic and coumaric acids were exclusively observed in this species. PMID- 18675545 TI - Stereoselective synthesis and biological evaluation of 3,4 diaminocyclohexanecarboxylic acid derivatives as factor Xa inhibitors. AB - There have been few reports on synthetic methods for cis-1,2-diaminocyclohexane bearing a third ring substituent. Starting from 3-cyclohexenecarboxylic acid, we developed efficient methods for synthesizing the 3,4-diaminocyclohexanecarboxylic acid derivatives 2-5. We also evaluated their anti-Xa and anticoagulant activities. Among the compounds, acid 2a and amide 2b exhibited the most potent in vitro anti-fXa activity, indicating that the position and stereochemistry of a polar functional group on the cyclohexane ring greatly affected the in vitro anti fXa activity. PMID- 18675546 TI - Surgical management of Gynaecomastia: outcomes from our experience. AB - The study aims to assess the morbidity and outcomes associated with gynaecomastia surgery. Between 1998 and 2007, 748 males with a mean age 44.67 years (10-90) were referred to us with breast-related symptoms. From these only 65 patients (102 breasts), with a median age of 26 years (11-82) had an operation for gynaecomastia. We considered for the purpose of the study each operated breast as an individual case. Overall, 42 cases of grade I gynaecomastia, 40 with grade II and 20 with grade III were treated mainly with subcutaneous mastectomies, 22 with skin reduction. Acute major complications requiring intervention occurred in 12 cases. Twenty-three cases required a late corrective operation for unsatisfactory results. The surgical approach appears to be the most important determinant of good cosmesis with the circumareolar approach to give the better results. The majority of the patients can be managed conservatively. Surgical candidates should be made aware of the significant morbidity. PMID- 18675547 TI - Determination of femoral component size in unicompartmental knee replacement. AB - It is difficult to determine the ideal size of femoral component when a unicompartmental knee replacement (UKR) is implanted with a minimally invasive approach. The aim of the study was to identify the best method of determining femoral component size pre-operatively. This was done by assessing the accuracy with which it could be predicted by pre-operative templating, patient height, gender and tibial component size. One hundred patients who had had Oxford UKR were studied. Pre-operative radiographs were templated and patient height, gender and implanted tibial component size were recorded. From a review of post operative radiographs and the known size of component used the ideal size of component that should have been used was established. By relating patient height, gender and tibial component size to the ideal femoral component size the range of these various parameters that best predicted each component size was identified. The reliability of these predictions was assessed assuming that a component one size off ideal was acceptable and two sizes off was unacceptable. Templating was correct in 67% of cases, acceptable in 33% and was never unacceptable. Height alone was correct in 56%, acceptable in 38% and unacceptable in 6%. Height based on gender was correct in 75%, acceptable in 25% and was never unacceptable. Tibial size was correct in 56%, acceptable in 41% and unacceptable in 3%. We conclude that both templating and height based on gender are reliable methods of assessing femoral component size. However because errors can occur surgeons are advised to use both. PMID- 18675548 TI - The evaluation of biologically important trace metals in liver, kidney and breast tissue. AB - The levels of iron (Fe), copper (Cu), zinc (Zn) and potassium (K) have been measured in normal and malignant liver and kidney tissue using X-ray fluorescence. Zn was reduced in liver and kidney tumours by 63% and 26%, respectively. Fe, Cu and K were increased in kidney tumours by 150%, 8% and 90%, and reduced in liver tumours by 76%, 29% and 43%. A synchrotron microprobe was used to map calcium (Ca), Fe, Cu and Zn in breast tissue. The distributions were compared to reference images and higher concentrations of metals correlated with areas of cancer cells. The average increase of Ca, Fe, Cu and Zn concentrations in areas of malignancy were 70%, 43%, 53% and 87%. PMID- 18675549 TI - The history and principles of chemical dosimetry for 3-D radiation fields: gels, polymers and plastics. AB - Over recent decades, modern protocols of external beam radiotherapy have been developed that involve very steep dose gradients and are thus extremely sensitive to errors in treatment delivery. A recent credentialling study by the Radiological Physics Center at the MD Anderson Cancer Center (Texas, USA) has noted potentially significant inaccuracies in test treatments at a variety of institutions. 3-D radiation dosimetry (often referred to as "gel dosimetry") may have an important role in commissioning new treatment protocols, to help prevent this type of error. This article discusses the various techniques of 3-D radiation dosimetry, with a focus on the types of radiosensitive samples used and on the optical computed tomography readout technique. PMID- 18675550 TI - Protontherapy of eye tumours in the UK: a review of treatment at Clatterbridge. AB - The Scanditronix MC-60 PF cyclotron at Clatterbridge was commissioned in 1984 for fast neutrontherapy trials. It also produced a 60.0 MeV clinical beam suitable for treating ocular tumours with a maximum penetration of 31 mm (water) and a 0.9 mm fall-off. An additional treatment room was built with an ocular beamline constructed in-house. The first group of eye patients was treated in June 1989, making this the first hospital-based proton facility. More than 1700 eye patients have been treated by the only UK proton service. PMID- 18675551 TI - Bone marrow-derived cells and tumor growth: contribution of bone marrow-derived cells to tumor micro-environments with special focus on mesenchymal stem cells. AB - Research has provided evidence that tumor growth depends on the interaction of tumor cells with stromal cells, as already suggested in 1889 by Paget. Experimental and clinical studies have revealed that tumor stromal cells can be derived from bone marrow (BM)-derived progenitor cells, such as mesenchymal stem cells (MSCs), which can be mobilized into the circulation and incorporate into tumor micro-environments. Many observations indicate that, in the tumor micro environment, MSCs have several tumor growth promoting functions, including expression of growth factors, promotion of tumor vessel formation and creation of tumor stem cell niches. This review will discuss the currently known tumor growth promoting BM-derived cells and focus on the role of MSCs in modulating tumor micro-environments. In addition, we will discuss the potential of inhibiting BM derived cells and their utilization as cellular vehicles for selective delivery of cancer therapeutics as additional strategies in the treatment of cancer. PMID- 18675552 TI - Therapeutic down-regulation of central and peripheral B-cell-activating factor (BAFF) production in pediatric opsoclonus-myoclonus syndrome. AB - Opsoclonus-myoclonus syndrome (OMS) is an autoimmune, paraneoplastic, central nervous system disorder, characterized by cerebrospinal fluid (CSF) B-cell expansion and various putative autoantibodies. To investigate the role of B-cell activating factor (BAFF) in OMS and the effect of disease-modifying immunotherapies used to treat it, BAFF was measured by enzyme-linked immunoadsorbent assay in the CSF and serum of 161 children with OMS and 116 pediatric controls. The mean concentration of CSF BAFF and the CSF/serum BAFF ratio were significantly higher in untreated OMS compared to neurological controls. CSF and serum BAFF levels were significantly lower in children treated with ACTH or corticosteroids, as was the CSF/serum BAFF ratio. There was a strong, negative correlation between CSF or serum BAFF levels and ACTH dose. Monthly IVIg infusions had no net impact on BAFF levels, and the combination of IVIg with ACTH or steroids did not reduce or enhance their anti-BAFF effects. These data indicate that BAFF production is increased centrally, not peripherally, in OMS, implying astrocytic over production. The novel dose-related central and peripheral anti-BAFF properties of ACTH, especially, have implications for other BAFF-related autoimmune disorders, infectious diseases, and cancers. PMID- 18675553 TI - Multi-structure network shape analysis via normal surface momentum maps. AB - We present a shape analysis pipeline for the assessment of anatomical variations in subcortical networks in MR images. The shape analysis pipeline injects the global shape properties of the CFA subcortical template into the subcortical parcellations generated from FreeSurfer via large deformation diffeomorphic metric mapping (LDDMM). Examples are shown for this injection in several subcortical structures whose raw MR images were sampled from the database of Open Access Series of Imaging Studies (OASIS). The shape analysis is performed on random field representation of the template surface momentum maps that encode the shape variation of subcortical structure targets of each individual subject relative to the template. The momentum maps have the optimum property that they are supported only on the boundary of the subcortical structures with the direction normal to the subcortical nuclei boundary thereby reducing the dimension of shape variation significantly. A two-level statistical model was built on these momentum maps to assess anatomical connectivity among the subcortical structures on the basis of similar surface deformation (compression or expansion). Results in the study of healthy aging on the hippocampus-amygdala network indicate the anatomical connectivity between the basolateral complex of the amygdala and the subiculum of the hippocampus on the basis of shape compression in healthy elders relative to young adults. PMID- 18675554 TI - Morphologic analysis of the proximal ulna with special interest in elbow implant sizing and alignment. AB - A better understanding of the morphology of the proximal ulna should permit the development of ulnar component designs which have an improved fit to the native bone, thus leading to more accurate implant positioning. Computed-tomography (CT) scans of 31 cadaveric proximal ulnae were analyzed using computer aided design software to determine the shape of the medullary canal relative to the articular surface. The diameter, curvature, cross-sectional centroid position, and coronal and sagittal angulation of the ulnar canal were all calculated with respect to the center of the greater sigmoid notch. Posterior and lateral offsets increased distally from the articulation center, and the mean diameter of the canal was larger in males than in females (P < .05). The average valgus angulation was 8.0 +/- 4.0 degrees for males and 7.2 +/- 3.1 degrees for females (P = .6). Longer stemmed ulnar implants may require a modular design to meet anatomic constraints during implant positioning. PMID- 18675555 TI - Reduced interhemispheric hippocampal BOLD signal coupling related to early epilepsy onset. AB - In order to investigate the factors influencing interhemispheric hippocampal connectivity we used fMRI to assess the time course of hippocampal activity during a spatial memory task. Data were obtained from 14 patients with unilateral hippocampal sclerosis and focal unilateral medial temporal lobe epilepsy. Correlation coefficients between left and right hippocampal BOLD signal fluctuations were determined in each patient as a measure for interhemispheric hippocampal coupling. On the group level a non-parametric Spearman correlation analysis was performed to assess relations of hippocampal connectivity to age, age at epilepsy onset, disease duration, and task performance. In all except for one patient correlation coefficients between left and right hippocampal time courses reached significance. Interhippocampal connectivity exhibited the strongest correlation to age at onset, i.e. patients with later onset showed greater connectivity. It was negatively correlated with disease duration, but not with age. Our findings underline the malicious effect of epileptic seizures on brain structures inevitably engaged with core cognitive functions. The results suggest that a measure of interhemispheric interplay - even at a coarse time scale - might reflect the functional status of brain structures. PMID- 18675556 TI - Ketogenic diet treatment in adults with refractory epilepsy: a prospective pilot study. AB - PURPOSE: To assess the efficacy of ketogenic diet (KD) in adults with refractory epilepsy. METHODS: Eligible subjects were 18-45 years old with at least two monthly focal seizures (with or without secondary generalization) documented by 8 weeks' follow-up. Classic form of KD treatment (90% fat) was planned for 12 weeks: daily seizure diaries were kept and measurements of the urinary ketones were recorded. Blood studies were done monthly and resting energy expenditure (REE), substrate utilization; body composition and quality of life (QOL) were measured before and after intervention. RESULTS: Nine patients were enrolled (average age 28+/-6 years; seven women). Only two subjects concluded the study per protocol due to an early drop-out. The average length of KD treatment was 8+/ 4 weeks (two patients completed 12 weeks of KD; feelings of hunger and lack of efficacy resulted in withdrawal of the rest). The two patients who concluded the study had a more than 50% reduction in the frequency of the seizures. The others experienced no improvement. Adherence to the KD protocol (100%) was documented by constant ketonuria and increased fat utilization as indicated by the change in respiratory quotient (p<0.031). The KD increased the cholesterol levels (mainly LDL; p=0.0001). CONCLUSIONS: In our experience with relatively small adult population, adherence to KD is difficult. In patients who had compliance over 3 weeks (6/8), KD does not seem to have a significant effect. Yet, the significant reduction in the two patients who concluded the study per protocol may indicate that some patients may benefit from this diet. Significant increase in LDL levels and the unlikable dietary changes are additional impediments to its implementation among adults with refractory epilepsy. PMID- 18675557 TI - Preservation for future use of the autologous saphenous vein during femoro popliteal bypass surgery is inexpedient. AB - PURPOSE: To investigate the usefulness of greater saphenous vein preservation for future vascular reconstructions during femoro-popliteal bypass surgery. DESIGN: Post-hoc analysis of data acquired in a randomized multi-centre clinical trial comparing two different vascular prostheses (ClinicalTrials.gov ID: NCT 00523263). PATIENTS AND METHODS: The true frequency of ipsilateral saphenous vein use in subsequent femoro-popliteal and coronary bypass surgery was investigated through case-record analysis with a median follow-up of 60 months in 100 consecutive patients, that received a prosthetic femoro-popliteal bypass between 1996 and 2001. RESULTS: An ipsilateral secondary femoro-popliteal bypass was performed in 11 patients (11%) at a mean interval of 34 months (range 1-96). The ipsilateral saphenous vein was applied for these procedures in 8 cases (8%). The cumulative probability of receiving a subsequent bypass was 8% at 3 years and 10% at 5 years follow-up respectively. One patient (1%) underwent CABG at 8 years follow-up with the use of ipsilateral lower leg saphenous vein segments only. CONCLUSION: Preservation of the greater saphenous vein in supragenicular femoro popliteal bypass surgery is not a valid argument for application of prosthetic material. PMID- 18675558 TI - The role of superficial venous surgery in the management of venous ulcers: a systematic review. AB - BACKGROUND: The complicated natural history of venous ulcers requires the continued development and improvement of treatments to ensure the most effective management. Compression therapy or surgical correction of superficial venous incompetence (SVI) are currently the main methods employed for the treatment for venous ulceration (VU). This review compares and summates the healing and recurrence rates for each treatment modality used over the last thirty years. METHODS: Sixty-one articles investigating compression and superficial venous surgical treatments were obtained from a systematic search of electronic databases (Medline, Embase, The Cochrane Library, and Google Scholar) and then an expanded reference list review. Patient demographics, CEAP classification, patterns of venous insufficiency, type of intervention, length of follow up, healing and recurrence rates for venous ulceration was assessed. Inadequate data in seven reports led to their exclusion. Recent randomised controlled trials (RCTs) specifically comparing superficial surgery to compression therapy were reviewed and data from non-randomised and/or 'small' clinical studies prior to 2000 underwent summation analysis. RESULTS: Five RCTs since 2000 demonstrate a similar healing rate of VU with surgery and conservative compression treatments, but a reduction in ulcer recurrence rate with surgery. The effect of deep venous incompetence (DVI) on the ulcer healing is unclear, but sub-group analysis of long-term data from the ESCHAR trial suggests that although surgery results in a less impressive reduction in ulcer recurrence in patients with DVI, these patients appear to still benefit from surgery due to the haemodynamic and clinical benefits that result. The RCTs also highlight that a significant proportion of VU patients are unsuitable for surgical treatment. Summation of data from earlier studies (before 2000), included twenty-one studies employing conservative compression alone resulted in an overall healing rate of 65% (range 34-95%) and ulcer recurrence of 33% (range 0-100%). In thirty-one studies investigating superficial venous surgery, the overall rate of ulcer healing was 81% (range 40-100%) with a post-operative recurrence rate of 15% (range 0-55%). The duration of follow up care in the surgical studies was approximately twice as long as in the conservative studies, which would lend to more reliable recurrence data. CONCLUSIONS: Evidence from the current literature, would suggest that superficial venous surgery is associated with similar rates of ulcer healing to compression alone, but with less recurrence. The effects of post-operative compression and DVI on the efficacy of surgery are still unclear. PMID- 18675559 TI - Negative pressure wound therapy: a systematic review on effectiveness and safety. AB - OBJECTIVES: We reviewed the use of negative pressure wound treatment (NPWT) for problematic wounds. METHODS: A systematic literature review was undertaken to assess the effectiveness and safety of NPWT. Randomized controlled trials (RCTs) assessing NPWT were included. RESULTS: A total of 14 RCTs were included. Trials included patients with pressure wounds (2), post-traumatic wounds (3), diabetic foot ulcers (4) and miscellaneous chronic ulcers (5). In all trials NPWT was at least as effective and in some cases more effective than the control treatment. Most evidence supports the effectiveness of NPWT on chronic leg ulcers and posttraumatic ulcers. NPWT appears to be a safe treatment, and serious adverse events have been rarely reported. Only two trials were classified as high quality studies, whereas the remaining were classified as having poor internal validity. CONCLUSIONS: Reliable evidence on the effectiveness of NPWT is scarce. Tentative evidence indicates that the effectiveness of NPWT is at least as good as or better than current local treatment for wounds. The need for large high-quality randomised studies is apparent. PMID- 18675560 TI - Venous valve reconstruction in patients with secondary chronic venous insufficiency. AB - OBJECTIVES: To evaluate the durability of venous valve reconstruction (VVR) and its benefits in terms of symptom improvement, ulcer healing and symptom/ulcer recurrence among patients with secondary chronic venous insufficiency (SCVI) in whom superficial venous surgery and compression treatment had failed. METHODS: During a ten year period (1993-2004) 1800 patients with chronic venous insufficiency (CVI) were evaluated by colour duplex ultrasound (CDU) and ambulatory venous pressure measurement (AVP). Approximately two thirds of patients had SCVI. Initial treatment consisted of compression therapy for a 6 month period. In addition, superficial vein and perforator surgery was performed in those presenting with reflux in these venous systems. 121 patients who did not improve with this treatment were investigated by ascending venography, descending video venography, air plethysmography and measurement of post-ischaemic venous pressure gradient. Thirty two cases having venous reflux without obstruction were selected for VVR. RESULTS: The ulcer healing rate within three months was 68% (13/19 patients). VVR resulted in valvular competence and a clinical success rate of 47% and 40% after 3 and 7 years respectively. In 8/13 (54%) of patients with a healed leg ulcer, a median post-operative AVP reduction of 33 mm Hg (range 20-38) was recorded. The durability of clinical success was numerically longer in patients with haemodynamic improvement (n=10) median 24 months (12-108), when compared with that in those without haemodynamic improvement (n=22) median 18 months (6-108). Popliteal vein reconstruction was part of the VVR procedure in all patients with haemodynamic improvement (post-op. AVP reduction >or=20 mm Hg). VVR at the popliteal level alone or combined with inguinal reconstruction seemed to be the one significant factor associated with haemodynamic improvement (P=0.014, Chi squared). CONCLUSION: VVR may lead to ulcer healing, but when performed at the popliteal level, haemodynamic improvement can be obtained along with a longer recurrence-free period (durability). VVR should be considered in the treatment of patients with SCVI who do not respond to superficial venous surgery and compression treatment. PMID- 18675561 TI - Effect of varicose vein surgery on venous reflux scoring and plethysmographic assessment of venous function. AB - BACKGROUND: Colour duplex ultrasonography (CDU) is widely recommended before varicose vein surgery, combined with quantification of venous reflux by plethysmography where required. This study assessed venous haemodynamics before and after varicose vein surgery by venous outflow plethysmography (VOP), venous reflux plethysmography (VRP) and by adoption of a modified segmental venous reflux score (VRS). The effect of wearing one or two class I medical compression stockings was also assessed. The aim of the study was to identify parameters which reflect the outcome of treatment using medical compression stockings or surgical intervention. METHODS: 24 legs of 21 patients with superficial vein incompetence of clinical grade C(2-4a) (CEAP) were assessed before and a mean of 8 S.D. 4 months after superficial vein surgery. Investigations were CDU, as well as VOP and VRP using mercury in rubber gauges fitted either around the calf or the forefoot. Venous reflux was semi-quantitatively graded by CDU in relation to the actual vein diameter and transformed into a VRS with respect to the number of involved serial vein segments. The venous reflux rates were measured in standing patients after knee bending before and after application of one or two superimposed compression stockings (class I). RESULTS: According to VRP, one compression stocking reduced the maximum venous reflux rates (VR(max)) by about 30% which was comparable with the effect of surgery on VR(max). Two superimposed compression stockings were almost twice as effective and diminished VR(max) pre- and post operatively by around 60%. Varicose surgery reduced the maximum venous outflow rates significantly (pre-op: 166 S.D. 77 ml/min x 100 ml tissue, post op: 120 S.D. 34) and improved VRS (pre-op median 5.0 IQR: 4.5-5.5, post-op median 0.5 IQR: 0-1.0). Surgery had no effect on venous refilling time or venous reflux rates when measured without compression stockings. CONCLUSION: Venous reflux assessed by plethysmography was moderated by the use of compression stockings pre operatively but did not reflect the outcome of surgical treatment of superficial venous reflux. Increased venous volume and venous outflow were restored to the levels of normal contralateral limbs by surgery. The VRS decreased considerably following surgery, reflecting the effect of surgical treatment on the number of incompetent venous segments. Changes in this parameter did not correlate with any of the plethysmographic measurements. PMID- 18675562 TI - Hypertension in the very elderly--a call to improve blood pressure management of our PAD patients. PMID- 18675563 TI - It is possible to cause damage to a laser fibre during delivery of tumescent anaesthesia for endovenous laser ablation (EVLA). AB - AIMS: To establish a possible mechanism of damage to a laser fibre significant enough to cause a retained segment within a patient. METHODS: A 21 G needle was used to pierce a VARILASE 810 nm Laser Fibre inserted within a 4F sheath. A tiny pin source of light from the aiming beam emerged from the needle hole in the sheath. Using laser protection protocol, the generator was fired for one minute at 14 Watts (W) continuous wave. The sheath and fibre were then examined. In a control experiment, we were unable manually to break a fibre where the coating had been damaged prior to the laser being fired. RESULTS: The aiming beam was noted to be concentrated at the side of the catheter at the point of needle damage rather than at the fibre tip. When the fibre was removed from the sheath the distal length, from the point of damage to the tip, was retained within the sheath. Longer firing with the sheath surrounded by a wet towel or a pork loin resulted in complete severance of the sheath and fibre. CONCLUSION: There are no firm manufacturer's guidelines on whether Tumescent Anaesthesia should be delivered before or after the laser fibre has been inserted into the patient. Some units performing EVLA prefer to do this with the laser fibre in situ as it is easier to image on ultrasound than the sheath alone. The results of this in vitro experiment would suggest it is possible to cause sufficient needle damage to fracture a laser fibre when fired. In the interests of safety we would recommend administration of tumescent anaesthesia should always be carried out before introduction of the laser fibre. PMID- 18675564 TI - Widening of the inferior alveolar canal: a case report with atypical lymphocytic infiltration of the nerve. AB - Widening of the inferior alveolar (mandibular) canal is a rare radiological finding. It is most often associated with neurofibromatosis. Rarely, a malignant process such as lymphoma may cause ill-defined enlargement of the mandibular canal. We present a unique case of a 33-year-old male who gradually developed sensory loss of his left lower lip and cheek and a well-defined tube-like widening of his left mandibular canal. The histopathological findings of the lesion were unusual in that they indicated atypical lymphocytic infiltration of the nerve tissue. The differential diagnoses regarding the clinical, radiological, and histopathological findings are discussed. PMID- 18675565 TI - Relative expression of different Ikaros isoforms in childhood acute leukemia. AB - Ikaros is a zinc-finger transcriptional factor playing an essential role in lymphoid lineage commitment and differentiation. Animal models and analysis of human Ikaros in leukemic cells demonstrate deregulation of Ikaros expression. Short isoforms with a truncated DNA-binding domain suppress functions of Ikaros in a dominant-negative manner. Previous studies demonstrated that human leukemias are heterogeneous for Ikaros expression. We estimate the relative level of Ikaros mRNA transcripts in 80 childhood ALL cases in comparison with AML and healthy donor groups. We detected eight major isoforms and several minor mutant isoforms in most patients with acute lymphoblastic and myeloid leukemia and in healthy donors, but the relative level of expression varied. The relatively high level of Ik4A isoform, rarely mentioned in previous reports, was detected in all analyzed groups. The ratio between functional and all isoforms was used to determine functional activity of Ikaros. The ratio was significantly less in AML (p=0.027) and BCR-ABL positive ALL (p=0.0028) than in healthy bone marrow. We found a negative association between the Ikaros ratio and myeloid coexpression in B-cell ALL, the most prominent was for CD15. The Ikaros ratio positively correlates with CD5 and negatively with CD7 expression in T-ALL. We suggest that an anti proliferation and anti-activation effect of full-length Ikaros may be mediated through regulation of CD5 and CD7. PMID- 18675566 TI - Advances in surgical treatment of septic arthritis of the distal interphalangeal joint in cattle: a review. AB - This review describes the indications for surgical treatment of distal interphalangeal joint (DIJ) infection in cattle and presents an overview of former surgical methods that have been modified for current use, including arthrotomy, resection of the distal sesamoid bone, resection of the DIJ and digital amputation. The post-operative life span, reasons for culling, pros and cons of claw-preserving techniques versus digital amputation, and pain management in general are also discussed. It would appear that severe claw diseases that result in septic arthritis of the DIJ do not necessarily lead to a reduced life span if cows are adequately treated and thorough aftercare is provided. PMID- 18675567 TI - Predictive value of natriuretic peptides in dogs with mitral valve disease. AB - Natriuretic peptides are useful in diagnosing heart failure in dogs. However, their usefulness in detecting early stages of myxomatous mitral valve disease (MMVD) has been debated. This study evaluated N-terminal (NT) fragment pro-atrial natriuretic peptide (NT-proANP) and NT-pro-brain natriuretic peptide (NT-proBNP) in 39 Cavalier King Charles Spaniels (CKCS) with pre-clinical mitral valve regurgitation (MR), sixteen dogs with clinical signs of heart failure (HF) and thirteen healthy control dogs. Twenty seven CKCS and ten control dogs were re examined 4 years after the initial examination and the status of the dogs 5 years after the initial examination was determined by telephone calls to the owner. All dogs were evaluated by clinical examination and echocardiography. CKCS with severe MR had higher NT-proANP and NT-proBNP compared to controls and CKCS with less severe MR. Dogs with clinical signs of HF had markedly elevated NT-proANP and NT-proBNP. Plasma concentrations of the natriuretic peptides measured at re examination could predict progression in regurgitant jet size. PMID- 18675568 TI - Seroprevalence of caprine arthritis-encephalitis virus in goats in the Cariri region, Paraiba state, Brazil. AB - The seroprevalence of caprine arthritis-encephalitis virus (CAEV) in the Cariri Region of Paraiba State, Brazil, was determined in 60 goat herds using the agar gel immunodiffusion test. The overall seroprevalence was 8.2%, with seropositivity in 21/60 (35%) herds and 13/15 (86.6%) municipalities. Bucks had a significantly higher frequency of infection (28.3%) than does (5.9%), and bucks that originated in other states had a significantly higher frequency of infection (76.5%) than those from Paraiba State (9.3%). PMID- 18675569 TI - Lack of efficacy of alpha-lipoic acid in burning mouth syndrome: a double-blind, randomized, placebo-controlled study. AB - BACKGROUND: A systematic review from the Cochrane Collaboration stated that alpha lipoic acid (ALA) may help in the management of burning mouth syndrome (BMS). Because all of the data on ALA came from a single group, it has been stressed that its effectiveness should be reproduced in other populations. AIM: A double blind, randomized, placebo-controlled study, including two test groups (Group A and Group B) and one control group (Group C), was carried out to evaluate the efficacy of systemic ALA (400 mg) and ALA (400 mg) plus vitamins in the treatment of BMS. METHODS: Sixty-six patients (54 females and 12 males) were included in an 8-week trial. Symptoms were evaluated by using a visual analogue scale (VAS) and the McGill Pain Questionnaire (MPQ) at 0, 2, 4, 8 and 16 weeks. RESULTS: Fifty two patients (43 females and 9 males, aged 67.3+/-11.9 years) completed the study. All three groups had significant reductions in the VAS score and in the mixed affective/evaluative subscale of the MPQ; the responders' rate (at least 50% improvement in the VAS score) was about 30%. No significant differences were observed among the groups either in the response rate or in the mean latency of the therapeutic effect. CONCLUSIONS: The fairly high placebo effect observed is very similar to data obtained from patients affected by atypical facial pain. This study failed to support a role for ALA in the treatment of BMS, and further investigations are needed to identify the cause of BMS in order to develop efficacious therapies. PMID- 18675570 TI - Animal models of asthma and chronic obstructive pulmonary disease. PMID- 18675572 TI - Antinociceptive and side-effects of hydromorphone after subcutaneous administration in cats. AB - The subcutaneous (SC) route is often chosen for drug administration in cats because it is easier to perform than intravenous (IV) injection and is perceived as less painful than intramuscular (IM) injection. However, little is known of how the route of administration influences the pharmacodynamics of drugs. This study measured the changes in skin temperature and thermal threshold (TT) and recorded the side-effects after SC injection of 0.1mg/kg of hydromorphone in six cats. Time to peak TT was 105min. Skin temperature was elevated at 15min and between 45 and 360min. Five cats vomited and two exhibited marked dysphoria. Compared to previously published studies of IV and IM administration of hydromorphone, the SC route results in a slower onset of peak effect, a shorter duration of antinociception and is associated with more undesirable side-effects. As with IV and IM injections, SC administration of hydromorphone at 0.1mg/kg is associated with a significant elevation in skin temperature. Overall, the SC route appears to have the least utility. PMID- 18675571 TI - Characterization of the null murine sodium/myo-inositol cotransporter 1 (Smit1 or Slc5a3) phenotype: myo-inositol rescue is independent of expression of its cognate mitochondrial ribosomal protein subunit 6 (Mrps6) gene and of phosphatidylinositol levels in neonatal brain. AB - Ablation of the murine Slc5a3 gene results in severe myo-inositol (Ins) deficiency and congenital central apnea due to abnormal respiratory rhythmogenesis. The lethal knockout phenotype may be rescued by supplementing the maternal drinking water with 1% Ins. In order to test the hypothesis that Ins deficiency leads to inositide deficiencies, which are corrected by prenatal treatment, we measured the effects of Ins rescue on Ins, phosphatidylinositol (PtdIns) and myo-inositol polyphosphate levels in brains of E18.5 knockout fetuses. As the Slc5a3 gene structure is unique in the sodium/solute cotransporter (SLC5) family, and exon 1 is shared with the mitochondrial ribosomal protein subunit 6 (Mrps6) gene, we also sought to determine whether expression of its cognate Mrps6 gene is abnormal in knockout fetuses. The mean level of Ins was increased by 92% in brains of rescued Slc5a3 knockout fetuses (0.48 versus 0.25 nmol/mg), but was still greatly reduced in comparison to wildtype (6.97 nmol/mg). The PtdIns, InsP(5) and InsP(6) levels were normal without treatment. Mrps6 gene expression was unaffected in the E18.5 knockout fetuses. This enigmatic model is not associated with neonatal PtdIns deficiency and rescue of the phenotype may be accomplished without restoration of Ins. The biochemical mechanism that both uniformly leads to death and allows for Ins rescue remains unknown. In conclusion, in neonatal brain tissue, Mrps6 gene expression may not be contingent on function of its embedded Slc5a3 gene, while inositide deficiency may not be the mechanism of lethal apnea in null Slc5a3 mice. PMID- 18675573 TI - [The first ray of the hand and the foot in the primates (II). Functional anatomy]. AB - To study the articular amplitude of the first ray of both hands and feet using passive mobilisation, either on fresh human cadavers or on living primates after anesthesia (namely eight pongidae, 15 cercopithecidae, two platirrhinii, eight lemuroidae, three daubentonidae, two loridae and two galagonidae). Plans slightly parasagittal and parafrontal have been chosen to obtain the maximum dorsal and palmar or plantar inclinations and the maximum lateral and medial inclinations. The pronosupination of the first ray has been evaluated in relation of these plans and alike for the hand of the simian primates on the plan of the concavity and the convexity of trapezial surface. The results have been compared with these obtained in the man. The pronosupination of the first ray, so denominated by analogy with the pronosupination of the fore-arm is the movement which allows the pulp of the thumb or of the hallux to look now forward or now backward in order to be opposed at the other fingers during the pollici- or the hallucidigital pinch. It can be considered like "effective" unless if there is rotation around the longitudinal axis of its metacarpus (or metatarsus). The prosimian primates had great and sensibly similar circular amplitude of their first metacarpus and first metatarsus, which reached more than 90 degrees in all the directions. It allowed a pronosupination of 90 degrees, but this was only "apparent" because it has been obtained by the alone combined action of the two orthogonal axes at the basis of the ray by simple effect of cardan joint. The anthropoids had only limited circular amplitude, not exceeding 45 degrees. It was associated with an "effective" pronosupination according to a longitudinal axis. The rotation reached 90 degrees. It was produced around the point of anchorage made up by the medial ligaments for the trapezometacarpal articulation and the lateral ligaments for the cuneometatarsal. This movement was possible because of the peculiar relief of the articular surfaces: modified cardan for the trapezometacarpal and spiraled trochoid for the cuneometacarpal joint. The man was the sole primate, which had practically no mobility at the level of the first ray of his foot. However, the morphology of his cuneometatarsal articulation was similar to that of the other anthropomorphous primates. The muscles which was inserted on the first metacarpus (or metatarsus) played an active role in the circular amplitude and in the apparent circular amplitude, but very discreet in the effective circumduction. The transversal fascicle of the short abductor of the thumb and of the hallux had a little pronator effect. At the level of the foot, the tendon of the fibular longus had an action of pronation and the abductor longus muscle an action of supination. The metacarpophalangeal and the metatarsophalangeal articulations of all the primates were condylar with laterality movements, which were able to reach 40 degrees and which were then unsteady. The metacarpophalangeal articulation of the man was the single exception. He was the sole one with a really steady thumb. PMID- 18675574 TI - Are inflammation and ossification on separate tracks in ankylosing spondylitis? PMID- 18675575 TI - Analysis of stresses in complete upper dentures with flat teeth at differing inclinations. AB - This study measures the effect of the inclination of the occlusal surfaces of flat teeth, on the magnitude of exerted stresses in a model of the complete upper denture (CUD), in order to estimate which inclination could be more beneficial for the stressed denture. Three groups of dentures were produced, with three different inclinations of their posterior flat teeth on the buccolingual direction: group 1 with an inward inclination of flat posterior teeth on the buccolingual direction (anti-Monson arrangement), group 2 with flat occlusion (no inclination) and group 3 with outward inclination (Monson arrangement). Using commercial edentulous molds and standardized procedures, six identical CUDs were produced for each group. Two rosette strain gauges were cemented onto the midline of each denture specimen, for measuring the stress magnitudes. The use of flat teeth with outward inclination may prolong the lifetime of the denture, by reducing the detrimental stresses (tensile principal and maximum shear stress). When flat posterior teeth with inward inclination are used, the reinforcement of CUD is necessary, to withstand the significant increase of the developed stresses (p<0.05). PMID- 18675577 TI - "Convenient space" or "a tight squeeze": insider views on the community pharmacy. AB - This paper presents research aimed at identifying the extent to which pharmacy spaces are aligned to good professional practice, enhance a professional's sense of self and meet the demands of the public. Findings from a novel, qualitative, mixed-methods approach employing biographic and photographic techniques indicate that UK pharmacy spaces are less accessible than intended by the Department of Health's pharmacy contract. Pharmacists escape to the dispensary to preserve their professional self-identity and to avoid the expectations of a demanding public. Recent innovations such as consultation rooms lack clarity of intent amid the multiple functions that a busy community pharmacy demands. PMID- 18675579 TI - Modelling of image-catheter motion for 3-D IVUS. AB - Three-dimensional intravascular ultrasound (IVUS) allows to visualize and obtain volumetric measurements of coronary lesions through an exploration of the cross sections and longitudinal views of arteries. However, the visualization and subsequent morpho-geometric measurements in IVUS longitudinal cuts are subject to distortion caused by periodic image/vessel motion around the IVUS catheter. Usually, to overcome the image motion artifact ECG-gating and image-gated approaches are proposed, leading to slowing the pullback acquisition or disregarding part of IVUS data. In this paper, we argue that the image motion is due to 3-D vessel geometry as well as cardiac dynamics, and propose a dynamic model based on the tracking of an elliptical vessel approximation to recover the rigid transformation and align IVUS images without loosing any IVUS data. We report an extensive validation with synthetic simulated data and in vivo IVUS sequences of 30 patients achieving an average reduction of the image artifact of 97% in synthetic data and 79% in real-data. Our study shows that IVUS alignment improves longitudinal analysis of the IVUS data and is a necessary step towards accurate reconstruction and volumetric measurements of 3-D IVUS. PMID- 18675578 TI - Effects of biventricular pacing and scar size in a computational model of the failing heart with left bundle branch block. AB - OBJECTIVES: To study the impact of biventricular pacing (BiV) and scar size on left ventricular (LV) regional and global function using a detailed finite element model of three-dimensional electromechanics in the failing canine heart. BACKGROUND: Cardiac resynchronization therapy (CRT) clinical trials have demonstrated that up to 30% of patients may be classified as non-responders. The presence of a scar appears to contribute to those that do not respond to CRT. A recent study in patients with myocardial scar showed that LV dyssynchrony was the sole independent predictor of reverse remodeling, and not scar location or size. METHODS: Two activation sequences were simulated: left bundle branch block (LBBB) and acute simultaneous BiV (with leads in the left and right ventricle) in hearts with chronic scars of various sizes. The dependence of regional function (mean fiber ejection strain, variance of fiber isovolumic strain and fraction of tissue stretched during ejection) and global function (left ventricular dP/dt(max), ejection fraction, stroke work) on scar size and pacing protocol was tested. RESULTS: Global function and regional function averaged over the whole LV during LBBB and BiV decreased as a function of scar size. In the non-scarred regions, however, regional function was largely independent of scar size for a fixed pacing site. CONCLUSIONS: The model results suggest that uniformity of mechanical contraction in non-scarred regions in the failing heart during biventricular pacing is independent of scar size for a fixed pacing site. PMID- 18675580 TI - Primary oral melanoma: population-based incidence. AB - Primary oral melanoma (POM) is a rare form of melanoma, generally diagnosed at a late stage and has poor prognosis. Epidemiological information has scarcely been reported in the literature over the past 40 years. Analyzing the Cancer Incidence in Five Continents volume IX (CI5-IX) database we found a total of 124,436 oral cavity cancers reported from all population-based cancer registries and 319 cases of POM reported by 67 cancer registries. To our knowledge this is the largest number of POM cases analyzed and accounts for 0.26% of all oral cavity cancers. ASR(W) for POM do not exceed 0.01 per 100,000 persons-year in all regions. The most common anatomic sites were palate (47%) and gum (27.6%) and a detailed oral mucosa examination and biopsies of nodular or macular lesions in these sites, especially on elderly patients, is advised to be routinely performed. PMID- 18675581 TI - Hypoxia-inducible factor 1 alpha in oral squamous cell carcinoma and its relation to prognosis. AB - The aim of this study was to investigate the correlation between the expression of hypoxia-inducible factor 1 alpha (HIF-1 alpha) and proliferative activity in tumor cells, lymph node metastasis, as well as prognosis in patients with oral squamous cell carcinoma (OSCC). Fifty-seven biopsy specimens of OSCC were investigated for the expression of HIF-1 alpha and proliferating cell nuclear antigen (PCNA) by immunohistochemistry. None of the patients had received any prior treatments. The percentage of HIF-1 alpha immunopositive area (PHIA) was calculated using computer-assisted image analysis for quantitative assessment of HIF-1 alpha expression. The PCNA labeling index (LI) was evaluated as a proliferation marker. We found that the mean PHIA in all stages was 12.1% in the poor prognosis patients, and it was 6.4% in the good prognosis patients. There was a significant difference of PHIA between poor prognosis and good prognosis patients (P=0.0065). Furthermore, the mean PHIA in the patients who had no metastatic lymph nodes was 7.5%, while it was 11.7% in the patients who had metastatic lymph nodes. There was also a significant difference of PHIA between patients who had no metastatic lymph nodes and those who had metastatic lymph nodes (P=0.0487). On the other hand, significant correlation between PHIA and PCNA LI was not observed. These results provide the clinical data indicating that HIF-1 alpha may play an important role in lymph node metastasis and prognosis in patients with OSCC. PMID- 18675583 TI - Using a modified technology acceptance model in hospitals. AB - PURPOSE: The use of information technology in the health care sector and especially in hospitals offers great potential for improving the quality of services provided and the efficiency and effectiveness of the personnel, but also for reducing the organizational expenses. However, the main question that arises according to the literature is whether hospital personnel are willing to use state of the art information technology while performing their tasks. This study attempts to address this issue by developing and testing a modified technology acceptance model taking into consideration other relevant models found in the literature. METHOD: The original TAM has been extended to include some exogenous variables in order to examine HIS acceptance by Greek hospital personnel. Correlation, explanatory and confirmation factor analysis was performed to test the reliability and validity of the measurement model. The structural equation modeling technique has also been used to evaluate the causal model. RESULTS: The results indicate that perceived usefulness, ease of use, social influence, attitude, facilitating conditions and self-efficacy significantly affect hospital personnel behavioral intention. Training has a strong indirect impact on behavioral intention through the mediators of facilitating condition and ease of use. Furthermore, the existence of significant positive effects between self efficacy and social influence, perceived usefulness and anxiety, and facilitating conditions and social influence is also supported. CONCLUSIONS: The proposed model can explain 87% of the variance of behavioral intention indicating that the core constructs of the technology acceptance models have a strong and statistically significant influence on hospital personnel usage intention. PMID- 18675582 TI - Determination of reactions between free radicals and selected Chilean wines and transition metals by ESR and UV-vis technique. AB - Four different types of Chilean wines (Cabernet Sauvignon, Merlot, Carmenere and Syrah) were selected and examined in their free radical scavenging capacities by electron spin resonance (ESR) and spectrophotometric methods. The free radical scavenging properties were evaluated against 2,2-diphenyl-1-picrylhydrazyl (DPPH*) radical, 2,6-di-tert-butyl-alpha-(3,5-di-tert-butyl-4-oxo-2,5 cyclohexadien-1-ylidene)-p-tolyloxy (Galvinoxyl) radical and hydroxyl radical (HO*). The possible effect on these scavenging properties of added transition metals to these wines was evaluated. Among the wines evaluated, Cabernet Sauvignon was the one with the highest activity against all radicals tested. The presence of added copper or iron to wines resulted in a reduced free radical scavenging capacity for all type of wines studied. The formation of redox inactive complexes between polyphenols of wine and transition metals is the possible cause of this reduction in antioxidant activity. PMID- 18675584 TI - A commercial ELISA detects high levels of human H5 antibody but cross-reacts with influenza A antibodies. AB - BACKGROUND AND OBJECTIVES: Commercial serological assays to determine influenza A H5N1 infection are available, although the accuracy and reproducibility of these are not reported in detail. This study aimed to assess the validity of a commercial ELISA H5 hemagglutinin (HA) antibody kit. STUDY DESIGN: A commercial ELISA for detection of antibodies towards influenza A H5 HA was evaluated using human sera from vaccinated individuals. The ELISA was used to screen 304 sera with elevated influenza A complement fixation titres collected between the period 1995-2007. RESULTS AND CONCLUSIONS: The ELISA was found to be accurate for sera with high levels of anti-H5 antibodies, and would be useful in clinical settings where a rapid result is required. Thirteen of the stored sera were positive using the ELISA, but were confirmed as negative for H5N1 exposure using further serological tests. Absorption studies suggested that antibodies towards seasonal H3N2 and H1N1 influenza may cross-react with H5 antigen, giving false positive results with the ELISA. PMID- 18675585 TI - Analysis of the internal transcribed spacer 2 (ITS2) region of scuticociliates and related taxa (Ciliophora, Oligohymenophorea) to infer their evolution and phylogeny. AB - The ITS2 (ITS--internal transcribed spacer) region of the rDNA in 11 representative scuticociliates and two ambiguously related genera was analyzed. In common with other eukaryotes, the putative ITS2 folding pattern consists of a closed loop with four helices supported by minimum free energy and compensatory base changes (CBCs), although two of these helices are variable and sometimes absent. Three topologies were obtained on the basis of traditional primary sequence analysis, "string" strategy of secondary structure and analysis of the combined data. It was found that the secondary structure information could help to improve alignment and utilize appropriately phylogenetic strategies. The proposed phylogenies, though differing between sequence- and structure-based results, provide consistent support for high-level clades: the systematically questionable genera Dexiotrichides and Cardiostomatella always cluster together in a clade basal to the scuticociliates s.s., whereas Pleuronema branches from other uronematids at a deep level, and is hence a divergent taxon. Within the well-supported monophyletic philasterids, a sister relationship exists between Orchitophrya and Mesanophrys, while Uronema shows a close relationship with the group including Paranophrys and Parauronema. The positions of Metanophrys, Pseudocohnilembus and Anophryoides among the philasterids remain poorly resolved. Our findings firmly support the proposed evolutionary scenario inferred previously both from morphological and molecular data. PMID- 18675586 TI - Agricultural pesticides and risk of childhood cancers. AB - Agricultural pesticide applications have the potential for significant drift beyond the target spray area and may result in exposure to non-farming residents in surrounding communities. Using geographic information system (GIS) methods, 1778 childhood cancer cases and 1802 controls born in Texas between 1990 and 1998 were assigned probable agricultural pesticide exposure based on proximity of birth residence to crop fields. Multivariate modeling was used to estimate odds ratios and 95% confidence intervals for selected cancers. For most childhood cancers, we found no evidence of elevated risk associated with residential proximity at birth to cropland. There was an overall pattern of increased risk for germ-cell tumors but the odds ratios were based on few number of exposed cases. There was also some indication of increased risk for non-Hodgkin lymphoma (NHL) and Burkitt lymphoma, but point estimates were imprecise and not statistically significant. Previous studies have assessed pesticide exposure primarily based on parental occupational history or household use, while our focus was on agricultural pesticides and so may represent a different array of chemical agents occurring at lower doses. PMID- 18675587 TI - Regulation of autoimmune arthritis by self-heat-shock proteins. AB - Heat-shock proteins (hsps) are highly conserved and immunogenic, and they are generally perceived to be attractive initiators or targets of a pathogenic immune response, and as such, have been implicated in the pathogenesis of autoimmune arthritis. However, studies in animal models and arthritis patients have unraveled the disease-regulating attributes of self-hsp65. We propose that the self-hsp65 induces a protective and beneficial immune response because of its ubiquitous distribution, stress inducibility and participation in tolerogenic processes. By contrast, the foreign hsp65 that does not influence the above processes and that resides admixed with microbial ligands for innate receptors generates an inflammatory pathogenic response. The regulatory properties of self hsps need be fully explored and might be used for therapeutic purposes. PMID- 18675588 TI - MHC class I assembly: out and about. AB - The assembly of major histocompatibility complex (MHC) class I molecules with peptides is orchestrated by several assembly factors including the transporter associated with antigen processing (TAP) and tapasin, the endoplasmic reticulum (ER) oxido-reductases ERp57 and protein disulfide isomerase (PDI), the lectin chaperones calnexin and calreticulin, and the ER aminopeptidase (ERAAP). Typically, MHC class I molecules present endogenous antigens to cytotoxic T lymphocytes (CTLs). However, the initiation of CD8(+) T-cell responses against many pathogens and tumors also requires the presentation of exogenous antigens by MHC class I molecules. We discuss recent developments relating to interactions and mechanisms of function of the various assembly factors and pathways by which exogenous antigens access MHC class I molecules. PMID- 18675589 TI - Malaria vaccines: immunity, models and monoclonal antibodies. AB - Although experts in the field have agreed on the malaria vaccine technology roadmap that should be followed (http://www.malariavaccineroadmap.net/), the path towards an effective malaria vaccine remains littered with intellectual and practical pot-holes. The animal models that are currently available are problematic, and current understanding of the exact mechanisms and targets of protective immune responses is incomplete. However, recent technological advances might help overcome some of these hurdles. PMID- 18675590 TI - Will new antischistosomal drugs finally emerge? AB - It has been often observed that the chemotherapeutic armamentarium against an important disease such as schistosomiasis consists of just one drug, praziquantel. Thus, development of drug resistance is an impending danger, with serious implications for the health protection of many millions of people. This rational and legitimate concern might now begin to be relieved by the recent proposal of a new class of compounds that could represent a novel source of drugs against schistosomiasis. PMID- 18675591 TI - Concurrent detection of secreted products from human lymphocytes by microengraving: cytokines and antigen-reactive antibodies. AB - Cell surface determinants, cytokines and antibodies secreted by hematopoietic cells are used to classify their lineage and function. Currently available techniques are unable to elucidate multiple secreted proteins while also assigning phenotypic surface-displayed markers to the individual living cells. Here, a soft lithographic method, microengraving, was adapted for the multiplexed interrogation of populations of individual human peripheral blood mononuclear cells for secreted cytokines (IFN-gamma and IL-6), antigen-specific antibodies, and lineage-specific surface-expressed markers. Application of the method to a clinical sample from a recent-onset Type 1 diabetic subject with a positive titer of anti-insulin antibodies showed that approximately 0.58% of circulating CD19(+) B cells secreted proinsulin-reactive antibodies of the IgG isotype and 2-3% of circulating cells secreted IL-6. These data demonstrate the utility of microengraving for interrogating multiple phenotypes of single human cells concurrently and for detecting rare populations of cells by their secreted products. PMID- 18675593 TI - Multidisciplinary training in obstetric critical care. AB - Once identified, the critically ill obstetric patient will often fare well. Unfortunately, several recent publications show that many such patients are not identified and come to harm. Training for those caring for pregnant women has changed in recent years. Medical training has contracted with a shorter working week, reducing experience of the critically ill. Midwifery training has also changed. Previously, midwives trained in general nursing prior to entering midwifery. Now they enter training directly, meaning their experience of the critically ill, especially those with non-obstetric problems, is limited. Other medical staff (anaesthetists, intensivists) may have experience of critical care in non-pregnant patients, but may be less aware of physiological changes of pregnancy that can affect how these patients are managed. The best way to address these issues is with multidisciplinary training, as highlighted in 'Safer Childbirth' and 'Saving Mothers Lives'. Such training allows each team member to understand his/her role and that of each of the other disciplines involved. PMID- 18675592 TI - Gene gun-mediated DNA vaccination enhances antigen-specific immunotherapy at a late preclinical stage of type 1 diabetes in nonobese diabetic mice. AB - Type 1 diabetes (T1D) is characterized by the T cell mediated destruction of the insulin-producing beta cells. Antigen-specific immunotherapies are used to selectively tolerize beta cell-specific pathogenic T cells either directly, or indirectly through the induction of immunoregulatory T cells. A key concern of antigen-specific immunotherapy is exacerbating autoimmunity. We compared the T cell reactivity and efficacy induced by plasmid DNA (pDNA) encoding glutamic acid decarboxylase 65 (GAD65) administered via intramuscular versus gene gun vaccination in NOD mice at a late preclinical stage of T1D. Whereas intramuscular injection of pGAD65 promoted a predominant type 1 CD4(+) T cell response and failed to suppress ongoing beta cell autoimmunity, gene gun vaccination preferentially induced IL-4 secreting CD4(+) T cells and significantly delayed the onset of diabetes. These findings demonstrate that gene gun delivery of autoantigen-encoding pDNA preferentially elicits immunoregulatory T cells and offers a safe, effective mode of pDNA vaccination for the treatment of T1D and other autoimmune diseases. PMID- 18675594 TI - Underlying medical conditions. AB - More than half of maternal deaths in the UK are due to pre-existing medical conditions, and medical emergencies often mimic acute obstetric conditions. An acute flare of systemic lupus erythaematosus, a thyroid storm or a phaeochromocytoma have many of the signs and symptoms of imminent eclampsia. Similarly, severe postpartum haemorrhage can result in diagnostic difficulties of medical conditions such as Addison's syndrome or acute renal failure. An acute collapse can be due to a pulmonary embolus, myocardial infarction, tachyarrhythmia or myasthenia gravis. These conditions are rare; however, unless they are considered in a differential diagnosis, they will not be diagnosed, to the detriment of the woman and her infant. This chapter deals with acute medical conditions occurring uncommonly in pregnant women. An obstetrician should know the initial steps to take in the emergency management of these cases. PMID- 18675595 TI - Outreach in obstetric critical care. AB - The present chapter considers the evolving role of critical care outreach in the general hospital setting and applied to obstetric patients, the mechanics of transferring critically ill obstetric patients to critical care and radiology areas, the scoring systems in use in critical care, and the difficulties in applying these scoring systems to obstetric patients. PMID- 18675596 TI - First experience of a modified device for excision of the endocervix in a reverse cone during laparoscopic supracervical hysterectomy. AB - The objective of this study was to evaluate the performance and safety of a modified unipolar electrode developed for excision of the endocervix in a reverse cone pattern during laparoscopic supracervical hysterectomy (LSH) to reduce the occurrence of persistent vaginal bleeding after the procedure. The study is a prospective pilot trial performed at a Norwegian university teaching hospital. Excision of the endocervix was performed with the unipolar electrode immediately after the amputation of the cervix during LSH in 23 consecutive women referred to the department for hysterectomy on the basis of a benign condition. The mean duration of the inverse conization was 61.9 seconds (SD 24.7). The excision of the endocervix in a reverse cone pattern was reported to be without technical problems in 22 procedures (96%). The results of this pilot study indicate that the modified device is safe, technically easy to use, and prolongs the duration of the LSH procedure insignificantly. Long-term effects of the device will be evaluated in a prospective randomized trial. PMID- 18675597 TI - Endometrial aspiration before or after saline infusion sonography and the effect on specimen quality: a randomized study. AB - STUDY OBJECTIVE: Saline infusion sonography (SIS) and endometrial aspiration can both be performed with the same catheter in 1 session. The objective of this study was to evaluate the effect of the order of investigations (aspiration-SIS vs SIS-aspiration) on the quality of the endometrial sample. DESIGN: Single-blind randomized controlled trial (Canadian Task Force classification I). SETTING: Academic teaching hospital. PATIENTS: A total of 113 women with abnormal uterine bleeding who visited the outpatient clinic were included in the study. INTERVENTIONS: The included women were randomly allocated either to SIS and subsequent endometrial aspiration, or to the reverse order. Both examinations were performed in 1 session with the same catheter. All aspiration specimens were sent to the same pathologist, who evaluated the quality of the samples. MEASUREMENTS AND MAIN RESULTS: The quality of the endometrial aspiration specimen was significantly better in the group of women who had aspiration before SIS, compared with women who were allocated to the reverse order (p=.04). Blood and mucus staining and the presence of endometrial epithelium had a significant influence on the sample quality. The difference between both groups was most pronounced in premenopausal women (p=.005). CONCLUSION: This study shows that the proportion of adequate specimens is higher when endometrial aspiration is done first with subsequent SIS. Aspiration should be performed before SIS to decrease the chance of obtaining an insufficient endometrial aspiration specimen. PMID- 18675598 TI - Determination of venlafaxine in human plasma by high-performance liquid chromatography using cloud-point extraction and spectrofluorimetric detection. AB - A new straightforward method based on cloud-point extraction (CPE) has been developed, optimized and validated for the determination of venlafaxine in human plasma by reversed-phase high-performance liquid chromatography with fluorescence detection. The non-ionic surfactant Triton X-114 (polyethylene glycol tert octylphenyl ether) was chosen as the extract solvent. Separation was obtained using a reversed-phase Diamonsil column (C(18), 250 mm x 4.6 mm I.D., 5 microm) and a mobile phase composed of acetonitrile-phosphate buffer solution (pH 3.0) triethylamine (33.5:66.5:0.4). Fluorescence detection was used (lambda(ex) 276 nm, lambda(em) 598 nm). Maprotiline was used as the internal standard. Under the optimum conditions, the linear range of venlafaxine in human plasma was 10-800 ng mL(-1) (r(2)=0.9995). The limit of detection (LOD) was less than 2 ng mL(-1) (S/N=3) and the limit of quantification (LOQ) was less than 10 ng mL(-1) (S/N=10). The method was successfully applied for the evaluation of pharmacokinetic profiles of venlafaxine capsules in nine healthy volunteers. PMID- 18675599 TI - Identification of quorum sensing signal molecules and oligolignols associated with watermark disease in willow (Salix sp.). AB - The bacterium Brenneria salicis is the causal agent of watermark disease in willow. This work shows the importance of in situ studies and high-resolution separation of biological samples with ultrahigh performance liquid chromatography combined with ion trap mass spectrometry to unambiguously identify molecular compounds associated with this disease. Approximately 40 oligolignols accumulated in wood sap of watermark diseased willow, and are indicative for degradation of the xylem cell wall, of which 15 were structurally assigned based on an earlier study. Many bacteria are known to produce and release quorum sensing signal molecules that switch on the expression of specific, sometimes pathogenic functions. Two quorum sensing signal molecules, N-(3-oxohexanoyl)-l-homoserine lactone and N-(hexanoyl)-l-homoserine lactone, were present in 4/1 ratios in diseased wood and in high-density in vitro cultures of B. salicis at 0.13-1.2 microM concentrations, and absent in healthy wood and in low-density in vitro cultures of B. salicis. Although it is not a proof, it can be an indication for involvement of quorum sensing in B. salicis pathogenesis. Cyclic dipeptides were present at high concentrations in high-density in vitro cultures of B. salicis, but not in situ, and were found not to be involved in quorum sensing signaling, therefore, the attribution of quorum signal properties to cyclic dipeptides isolated from in vitro cultures of pathogenic bacteria should be reconsidered. PMID- 18675601 TI - Preparation of chitosan-gelatin hybrid scaffolds with well-organized microstructures for hepatic tissue engineering. AB - The structural organization of natural liver is instrumental in the multifunctionality of hepatocytes, and mimicking these specific architectures in tissue-engineered scaffold plays an important role in the engineering of an implantable liver equivalent in vitro. To achieve this goal, we have developed a novel fabrication process to create chitosan-gelatin hybrid scaffolds with well organized architectures and highly porous structures by combining rapid prototyping, microreplication and freeze-drying techniques. The scaffolds obtained not only have analogous configurations of portal vein, central vein, flow-channel network and hepatic chambers, but also have high (>90%) porosity, with the mean pore size of 100microm. Swelling and degradation studies showed that the scaffold has excellent properties of hydrophilicity and biodegradability. A hepatocyte culture experiment was conducted to evaluate the efficiency of the well-defined chitosan-gelatin scaffold in facilitating hepatocyte growth in the inner layer of the scaffold in vitro. Scanning electron microscopy and histological analysis showed that hepatocytes could form large colonies in the predefined hepatic chambers, and these cavities could the completely filled with hepatocytes during 7 day culture. Albumin secretion and urea synthesis further indicated that the well-organized scaffolds were more suitable for hepatocyte culture. PMID- 18675600 TI - Determination of rivaroxaban--a novel, oral, direct Factor Xa inhibitor--in human plasma by high-performance liquid chromatography-tandem mass spectrometry. AB - A high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method allowing the sensitive and specific quantification of rivaroxaban (BAY 59 7939), a Factor Xa inhibitor in advanced development for the prevention and treatment of thromboembolic disorders, in human plasma is described. After precipitation of plasma proteins with methanol containing the internal standard followed by centrifugation, the plasma supernatant was injected directly onto the HPLC-MS/MS system. Concentrations could be determined between 0.50 and 500 microg/L. Inter-assay precision was < or = 7.4% and inter-assay accuracy was between 96.3 and 102.9% throughout the entire working range. The method was applied successfully in several clinical studies, which allowed an accurate determination of rivaroxaban pharmacokinetics in human plasma. PMID- 18675602 TI - Appendicitis in pre-school children: a continuing clinical challenge. A retrospective study. AB - OBJECTIVE: The purpose of this study was to examine the presenting symptoms, signs perforation rates and outcomes of appendicitis in pre-school children. In addition, we sought to determine the reasons of delay in the diagnosis of those with complicated appendicitis. MATERIAL AND METHODS: This is a retrospective study of all children of 5 years or less who underwent appendectomy for appendicitis in King Khalid University Hospital, Riyadh, Saudi Arabia between January 2001 and December 2007. RESULT: During the 7-year period, 106 patients 5 years of age or less required appendectomy for appendicitis. There were 66 boys and 40 girls. Sixty-four children (60.3%) had complicated appendicitis, 38 (35.3%) had acute appendicitis and 4 (3.7%) had normal appendix. Although classic symptoms were present in the majority of patients, atypical symptoms were found in some children. The duration of symptoms in patients with complicated appendicitis was more (5.2 vs 2.1 days). A diagnosis other than appendicitis was suspected in 47 patients (73.4%). Sixty (56.6%) patients had perforation at the time of surgery. Complicated appendicitis was associated with a longer hospital stay and more post-operative complications. CONCLUSION: The accurate diagnosis of appendicitis in pre-school children is still a clinical challenge. A high index of suspicion of appendicitis is necessary in pre-school children in view of the atypical presentation and high incidence of complicated appendicitis. Primary care physician, emergency room physician and young surgeons have a crucial role in recognizing the symptoms and signs of early appendicitis in pre-school children. PMID- 18675603 TI - Strategy for management of skin necrosis in neonates with lower extremity vascular birthmarks. PMID- 18675604 TI - Reconstruction of the retroauricular fold by 'nonpedicled' superficial mastoid fascia: details of anatomy and surgical technique. AB - During ear reconstruction, it is often difficult to achieve a good projection of the new ear. This is due to the retraction of retroauricular tissues, and the use of a cartilage wedge and superficial temporal fascia is frequently needed to elevate the ear framework. The aim of this article is to identify, by cadaver dissection, the anatomical structures of the retroauricular and mastoid regions and also to demonstrate the possibility of increasing the size of the superficial mastoid fascia by defibrating and cutting up its structure. The cartilage wedge is then inserted into a three-dimensional fascial environment. This allows reconstruction of the retroauricular fold with 'nonpedicled' fascia and to have a firm, stable ear elevation without sacrificing the temporal fascia. Nine ear reconstructions were performed using this technique and no major complications occurred; however, one procedure was complicated by a limited necrosis of the skin graft. In all cases we obtained a good and persistent elevation of the ear, as well as a well-defined fold, with the possibility of wearing glasses. Symmetry was satisfactory and aesthetic results were pleasant for all treated patients. This simple procedure allows one to obtain a good projection of the ear without involving the use of the superficial temporal fascia, which in addition is very useful in case of eventual complications such as cartilage extrusion. PMID- 18675606 TI - Antihelix, antehelix or anthelix? Comments on 'Upping the Anti?'. PMID- 18675605 TI - Preoperative CT angiography reduces surgery time in perforator flap reconstruction. AB - The use of perforator flaps in breast reconstructions has increased considerably in the past decade. A disadvantage of the perforator flap is difficult dissection, which results in a longer procedure. During spring 2006, we introduced CT angiography (CTA) as part of the diagnostic work-up in perforator flap reconstructions to visualise each perforator more accurately. The main objectives were to reduce surgery time and the number of complications. A chart review was conducted 1 year after CTA introduction to investigate if these objectives were met. MATERIALS AND METHODS: Patients with a deep inferior epigastric perforator (DIEP) flap who underwent preoperative analysis through CTA were retrospectively evaluated. The population .05). CONCLUSION: Prolonged latencies in inguinal hernia may be a result of nerve trap caused by hernia sac. GFN motor functions showed no causative role in other inguinoscrotal pathologies. It can be also suggested that clinical features of other inguinoscrotal pathologies were not affected by GFN motor functions. Electrophysiological studies in younger age groups with large number of patients are needed to support our suggestions. PMID- 18675651 TI - Polyorchidism in a child with imperforate anus. AB - Polyorchidism is a rare diagnosis. When recovered, it is frequently found in combination with other urologic pathologies. We report the case of a 14-month-old child with imperforate anus who was found to have polyorchia during repair of his inguinal hernia. Although cryptorchidism is not an uncommon finding in patients with imperforate anus, polyorchidism has never been reported. This is an unusual presentation of a rare entity. PMID- 18675650 TI - An investigation on the mechanism of contralateral manifestations after unilateral herniorrhaphy in children based on laparoscopic evaluation. AB - BACKGROUND/PURPOSE: No straightforward theoretical descriptions explaining the discrepant incidences of metachronous contralateral hernia and positive contralateral patent processus vaginalis (cPPV) exist. The indications for contralateral groin repair are still not clearly defined. METHODS: During laparoscopic herniorrhaphy, the intraoperative appearances of the internal inguinal rings (IIRs) were characterized and classified in 211 children with symptomatic unilateral inguinal hernias. A total of 125 boys and 86 girls with 130 right-sided and 81 left-sided indirect inguinal hernias were examined. The morphological appearances of the IIRs in patients with cPPV were compared before and after the ligation of the symptomatic IIRs. RESULTS: Of 211 children, 108 (51%) had positive cPPV findings. The cPPVs had significantly smaller orifices and shorter lengths than those on the side of the symptomatic hernia. Among these children, 24 (22%) patients exhibited morphological changes in their cPPVs, characterized by an upward movement of the overlying peritoneal veil after the closure of the symptomatic hernia. CONCLUSION: Metachronous contralateral hernia might occur because of morphological changes to the cPPV resulting from closure of the symptomatic IIRs. The predicted incidence of contralateral hernia in this series was 11.2%. PMID- 18675652 TI - Sigmoid fixation associated with rectopexy using a laparoscopic approach could prevent relapse of rectal prolapse in pediatric patients with spinal dysraphia. AB - Recurrent rectal prolapse, resistant to medical treatment, is an indication for surgical treatment. Patients with spinal dysraphia frequently have already been treated by sclerotherapy or other surgical techniques, but unsuccessfully. METHODS: We present 2 patients, who underwent laparoscopic rectopexy, with spinal dysraphia and complete rectal prolapse relapse after conservative treatment. In these patients, we performed, as an additional technique, fixation of the rectosigmoid to avoid recurrence by invagination or prolapse of the anterior wall. RESULTS: Follow-up at 14 and 11 months, respectively, did not find any recurrence. CONCLUSION: We suggest that laparoscopic rectopexy with sigmoid fixation should be considered as an alternative for the treatment for patients with spinal dysraphia and rectal prolapse to avoid recurrence. PMID- 18675653 TI - Failure of appendectomy to resolve appendiceal intussusception. AB - Appendiceal intussusception is a rare entity that primarily affects children. The optimal surgical management is not known. We describe a child with appendiceal intussusception treated by simple appendectomy who suffered recurrent intussusception of the cecum at the base of the appendiceal stump. He was cured by resection of a rim of cecum around the appendiceal staple line. Partial cecectomy with appendectomy may be the most effective operation for appendiceal intussusception. PMID- 18675654 TI - Congenital thickening and shortening of the round ligament of the liver causing gastric outlet obstruction. AB - This report describes a case of congenital thickening and shortening of the round ligament of the liver, a rare cause of gastric outlet obstruction. There are few published cases in the literature and these were generally diagnosed at a late stage and were mostly among adults. The patient underwent exploratory laparotomy, which showed gastric compression in the distal antrum, close to the pylorus, caused by the round ligament, which was shortened and thickened. The round ligament was suture ligated and divided, and the patient's symptoms resolved. Comments on this abnormality and its rarity are presented. PMID- 18675655 TI - Laparoscopic-assisted correction of a retrocaval ureter. AB - Retrocaval ureter has been traditionally corrected surgically through an open retroperitoneal procedure; however, in recent years, the correction has been accomplished using laparoscopy. The great advantages of this procedure remain limited because of the prolonged time it takes to complete the intracorporeal suture of the ureter, besides the need for a highly experienced surgeon. We present a case in which the ureter was freed using laparoscopy, and the ureteral anastomosis was carried out extracorporeally, which notably diminished the duration of the surgery. This technique would have limited use in obese patients, patients with ureters of limited length, patients with very proximal obstructions of the ureter, and patients with risk of devascularization of both ureteral segments. However, the technique does appear effective and reproducible in adequately selected patients. PMID- 18675656 TI - Perinatal testicular torsion: preoperative radiological findings and the argument for urgent surgical exploration. AB - Perinatal testicular torsion is an infrequent event, the management of which has been controversial. Occurrence is rare, estimated at 1 in 7500 newborns (Kaplan, G. W., Silber, I.: Neonatal torsion--to pex or not? In: Urologic surgery in neonates and young infants. Edited by King, L.R. Philadelphia: W.B. Saunders Co., 1988; Chapter 20, pp. 386-395). The frequency of bilateral perinatal torsion is up to 22% (J Urol. 2005;174:1579). Here, we describe two cases of bilateral asynchronous perinatal torsion, in which the only presenting abnormality on exam after birth was a unilateral scrotal mass. These cases illustrate that contralateral perinatal torsion may be present even when physical exam findings suggest unilateral involvement. PMID- 18675657 TI - Mother-to-daughter occurrence of biliary atresia: a case report. AB - We herein report a case of mother-to-daughter occurrence of biliary atresia (BA). The mother underwent a Kasai hepatic portoenterostomy at the age of 64 days with a diagnosis of type III BA. Her postoperative course was uneventful, with a good bile discharge, and she became pregnant at the age of 20 years. At 21 weeks of gestational age, fetal ultrasonographic examination of the baby revealed a cystic mass at the hepatic hilum. The baby was born via vaginal delivery at 41 weeks of gestation with a birth weight of 3269 g. Because of an increasing direct bilirubin level and the absence of bile excretion, a laparotomy was performed at the age of 29 days. Intraoperative cholangiography revealed the diagnosis of BA I cyst, and a hepaticojejunostomy was performed with a good bile discharge. To our knowledge, these patients are the first occurrence of BA in a transmission pattern of mother to daughter. PMID- 18675658 TI - Submucosal hematoma presenting as small bowel obturator obstruction in a patient on low-molecular-weight heparin. AB - Recent studies have shown the efficacy of low-molecular-weight heparin (LMWH) in the treatment of venous thromboembolic disease in children. Compared to unfractionated heparin and coumadin, LMWH has more predictable pharmacokinetics and a reported lower incidence of osteoporosis and heparin-induced thrombocytopenia in children. The overall incidence of severe hemorrhage on LMWH in children is low. To date, there is a single report of a small bowel obstruction in a child secondary to a hematoma while on LMWH. We report the second case of a child, on enoxaparin (Lovenox) therapy, who underwent bowel resection secondary to a completely obstructing small bowel wall hematoma. PMID- 18675659 TI - Laparoscopic excision of a newborn rectal duplication cyst. AB - Congenital rectal duplication cyst is a rare entity treated with surgical excision. Without treatment, a rectal duplication cyst may cause a variety of complications, most notably, transforming into a malignancy. We report on a 7 week-old girl who was found to have a rectal duplication cyst. The rectal duplication cyst was successfully excised laparoscopically. Rectal duplication cysts are rare alimentary tract anomalies generally discovered during childhood. Complications include symptoms arising from the cyst and the possibility of malignant degeneration. They are typically managed by surgical excision. PMID- 18675660 TI - Living donor liver transplantation with hyperreduced left lateral segments. AB - Liver transplantation is now an established technique to treat children with end stage liver disease. Implantation of left lateral segment grafts (Couidaud's segments II and III) can be a problem in small infants because of a large-for size graft. Reduced left lateral segmental liver transplantation has been recently introduced for small infants to mitigate the problem of large-for-size graft. Further reduction of the left lateral segment graft increases the possibility of supplying an adequate hyperreduced left lateral segment graft as an alternative surgical technique. We report 3 cases of our experience of transplantation using hyperreduced left lateral segment grafts from living donors. PMID- 18675661 TI - The perceived cosmetic value of laparoscopic pyloromyotomy. PMID- 18675662 TI - Smaller scars--what is the big deal: a survey ofthe perceived value of laparoscopic pyloromyotomy. PMID- 18675664 TI - Re: orchidopexy for undescended testis in England: is it evidence based? PMID- 18675666 TI - The resurgence of therapeutic ultrasound--a 21st century phenomenon. PMID- 18675667 TI - Teaming up for tobacco control. PMID- 18675668 TI - Progress towards revalidation of the UK medical profession. PMID- 18675669 TI - The Lords of despair. PMID- 18675670 TI - Is it time to screen and treat H pylori to prevent gastric cancer? PMID- 18675671 TI - Influenza vaccine for community-acquired pneumonia. PMID- 18675672 TI - Supervised injecting facilities: time for scale-up? PMID- 18675673 TI - Lessons from a multicentre paediatric HIV trial. PMID- 18675674 TI - The challenges of technological innovation in HIV. PMID- 18675675 TI - The untold stories of the Sichuan earthquake. PMID- 18675676 TI - Brian Greenwood: making milestones in malaria research. PMID- 18675677 TI - Countdown to 2015 for maternal, newborn, and child survival. PMID- 18675678 TI - MDGs, Countdown to 2015, and "concern" for Africa. PMID- 18675679 TI - Countdown to 2015 for maternal, newborn, and child survival. PMID- 18675680 TI - Basal insulin glargine vs prandial insulin lispro in type 2 diabetes. PMID- 18675681 TI - Steps to enhance Peru's progress on maternal mortality. PMID- 18675682 TI - Basal insulin glargine vs prandial insulin lispro in type 2 diabetes. PMID- 18675683 TI - Basal insulin glargine vs prandial insulin lispro in type 2 diabetes. PMID- 18675685 TI - Mass screening for neuroblastoma at 6 months of age. PMID- 18675687 TI - Ethics of a 6-month delay. PMID- 18675688 TI - International aid in disaster zones: help or headache? PMID- 18675690 TI - Influenza vaccination and risk of community-acquired pneumonia in immunocompetent elderly people: a population-based, nested case-control study. AB - BACKGROUND: Pneumonia is a common complication of influenza infection in elderly individuals and could therefore potentially be prevented by influenza vaccination. In studies with data from administrative sources, vaccinated elderly people had a reduced risk of admission for pneumonia compared with unvaccinated seniors; however, these findings could have been biased by underlying differences in health between the groups. Furthermore, since most individuals with pneumonia are not treated in hospital, such studies should include both outpatient and inpatient events. We therefore assessed whether influenza vaccination is associated with a reduced risk of community-acquired pneumonia in immunocompetent elderly people after controlling for health status indicators. METHODS: We did a population-based, nested case-control study in immunocompetent elderly people aged 65-94 years (cases and controls) enrolled in Group Health (a health maintenance organisation) during the 2000, 2001, and 2002 preinfluenza periods and influenza seasons. Cases were individuals with an episode of outpatient or inpatient community-acquired pneumonia (validated by review of medical records or chest radiograph reports). We randomly selected two age-matched and sex-matched controls for each case. The exposure of interest was influenza vaccination. We reviewed medical records to define potential confounders, including smoking history, presence and severity of lung and heart disease, and frailty indicators. FINDINGS: 1173 cases and 2346 controls were included in the study. After we adjusted for the presence and severity of comorbidities, as defined by chart review, influenza vaccination was not associated with a reduced risk of community acquired pneumonia (odds ratio 0.92, 95% CI 0.77-1.10) during the influenza season. INTERPRETATION: The effect of influenza vaccination on the risk of pneumonia in elderly people during influenza seasons might be less than previously estimated. FUNDING: Group Health Center for Health Studies internal funds and Group Health Community Foundation fellowship grant. PMID- 18675691 TI - Meniere's disease. AB - Meniere's disease is a chronic illness that affects a substantial number of patients every year worldwide. The disease is characterised by intermittent episodes of vertigo lasting from minutes to hours, with fluctuating sensorineural hearing loss, tinnitus, and aural pressure. Although there is currently no cure, more than 85% of patients with Meniere's disease are helped by either changes in lifestyle and medical treatment, or minimally invasive surgical procedures such as intratympanic steroid therapy, intratympanic gentamicin therapy, and endolymphatic sac surgery. Vestibular neurectomy has a very high rate of vertigo control and is available for patients with good hearing who have failed all other treatments. Labyrinthectomy is undertaken as a last resort and is best reserved for patients with unilateral disease and deafness. PMID- 18675689 TI - Effect of eradication of Helicobacter pylori on incidence of metachronous gastric carcinoma after endoscopic resection of early gastric cancer: an open-label, randomised controlled trial. AB - BACKGROUND: The relation between Helicobacter pylori infection and gastric cancer has been proven in epidemiological studies and animal experiments. Our aim was to investigate the prophylactic effect of H pylori eradication on the development of metachronous gastric carcinoma after endoscopic resection for early gastric cancer. METHODS: In this multi-centre, open-label, randomised controlled trial, 544 patients with early gastric cancer, either newly diagnosed and planning to have endoscopic treatment or in post-resection follow-up after endoscopic treatment, were randomly assigned to receive an H pylori eradication regimen (n=272) or control (n=272). Randomisation was done by a computer-generated randomisation list and was stratified by whether the patient was newly diagnosed or post-resection. Patients in the eradication group received lansoprazole 30 mg twice daily, amoxicillin 750 mg twice daily, and clarithromycin 200 mg twice daily for a week; those in the control group received standard care, but no treatment for H pylori. Patients were examined endoscopically at 6, 12, 24, and 36 months after allocation. The primary endpoint was diagnosis of new carcinoma at another site in the stomach. Analyses were by intention to treat. This trial is registered with the UMIN Clinical Trials Registry, number UMIN000001169. FINDINGS: At 3-year follow-up, metachronous gastric carcinoma had developed in nine patients in the eradication group and 24 in the control group. In the full intention-to-treat population, including all patients irrespective of length of follow-up (272 patients in each group), the odds ratio for metachronous gastric carcinoma was 0.353 (95% CI 0.161-0.775; p=0.009); in the modified intention-to treat population, including patients with at least one post-randomisation assessment of tumour status and adjusting for loss to follow-up (255 patients in the eradication group, 250 in the control group), the hazard ratio for metachronous gastric carcinoma was 0.339 (95% CI 0.157-0.729; p=0.003). In the eradication group, 19 (7%) patients had diarrhoea and 32 (12%) had soft stools. INTERPRETATION: Prophylactic eradication of H pylori after endoscopic resection of early gastric cancer should be used to prevent the development of metachronous gastric carcinoma. FUNDING: Hiroshima Cancer Seminar Foundation. PMID- 18675692 TI - Cystic fibrosis: impaired bicarbonate secretion and mucoviscidosis. AB - For more than 20 years, the abnormally thick mucus (mucoviscidosis) in cystic fibrosis has been widely shown to be linked to a genetic defect in the cystic fibrosis transmembrane conductance regulator Cl(-) channel. The defect is widely thought to cause mucus to become dehydrated as a result of basic defects in Cl(-) dependent fluid transport. However, this widely held explanation is inconsistent with the known physiological properties and functions of organs affected by cystic fibrosis. During the process of releasing highly condensed mucins from intracellular granules, Ca(2+) and H(+) cations must be removed to enable the mucins to expand by as much as 1000 times, forming extracellular mucus-gel networks. Over the past few years, that HCO(3)(-) transport is also defective in patients with cystic fibrosis has become apparent. I propose that HCO(3)(-) is crucial to normal mucin expansion because it forms complexes with these cations. Thus, because HCO(3)(-) secretion is defective in cystic fibrosis, mucins in organs affected by cystic fibrosis tend to remain aggregated, poorly solubilised, and less transportable. If the hypothesis is valid, pathogenesis in cystic fibrosis could be due as much to defective transport of HCO(3)(-) as to defective Cl(-) transport. PMID- 18675693 TI - A young man who kept falling over. PMID- 18675694 TI - Time course of morphologic effects on different retinal compartments after ranibizumab therapy in age-related macular degeneration. AB - PURPOSE: To analyze the effect of ranibizumab therapy on retinal and subretinal compartments in age-related macular degeneration and to compare the time course of compartment specific effects to visual function. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Fourteen patients with changes in 3 major compartments owing to neovascular age-related macular degeneration. METHODS: Standard treatment with 3 monthly doses of intravitreal ranibizumab was performed. Eyes were examined at baseline and weeks 1, 4, and 12 using a standardized protocol. Manual segmentation was applied to all 128 B-scans contained in a macular raster scan (MRS). MAIN OUTCOME MEASURES: Morphology and time course of different retinal and subretinal compartments. RESULTS: High definition optical coherence tomography and manual segmentation allowed for precise identification of volumes within individual compartments. All morphologic parameters responded positively to therapy, but demonstrated a specific time course. Subretinal fluid was identified as the most relevant factor for visual function, whereas changes in retinal and subpigment epithelial volumes did not correlate with the time course of functional rehabilitation. CONCLUSION: Analysis of MRS identified a characteristic impact of therapy on retinal and subretinal morphology. PMID- 18675695 TI - Mandating comprehensive eye examinations for children: where is the evidence? PMID- 18675697 TI - Cataract surgery in uveitis. PMID- 18675698 TI - Cataract surgery complication rates. PMID- 18675701 TI - Cost-effectiveness. PMID- 18675702 TI - DSEK in eyes with double glaucoma tubes. PMID- 18675704 TI - Intravitreal triamcinolone. PMID- 18675705 TI - Rehabilitation and intraocular telescopes. PMID- 18675707 TI - Depth of focus. PMID- 18675710 TI - Eye drop compliance in religious fasting. PMID- 18675696 TI - Optical coherence tomography measurements and analysis methods in optical coherence tomography studies of diabetic macular edema. AB - OBJECTIVE: To evaluate optical coherence tomography (OCT) measurements and methods of analysis of OCT data in studies of diabetic macular edema (DME). DESIGN: Associations of pairs of OCT variables and results of 3 analysis methods using data from 2 studies of DME. PARTICIPANTS: Two hundred sixty-three subjects from a study of modified Early Treatment of Diabetic Retinopathy Study (mETDRS) versus modified macular grid (MMG) photocoagulation for DME and 96 subjects from a study of diurnal variation of DME. METHODS: Correlations were calculated for pairs of OCT variables at baseline and for changes in the variables over time. Distribution of OCT measurement changes, predictive factors for OCT measurement changes, and treatment group outcomes were compared when 3 measures of change in macular thickness were analyzed: absolute change in retinal thickness, relative change in retinal thickness, and relative change in retinal thickening. MAIN OUTCOME MEASURES: Concordance of results using different OCT variables and analysis methods. RESULTS: Center point thickness correlated highly with central subfield mean thickness (CSMT) at baseline (0.98-0.99). The distributions of changes in CSMT were approximately normally distributed for absolute change in retinal thickness and relative change in retinal thickness, but not for relative change in retinal thickening. Macular thinning in the mETDRS group was significantly greater than in the MMG group when absolute change in retinal thickness was used, but not when relative change in thickness and relative change in thickening were used. Relative change in macular thickening provides unstable data in eyes with mild degrees of baseline thickening, unlike the situation with absolute or relative change in retinal thickness. CONCLUSIONS: Central subfield mean thickness is the preferred OCT measurement for the central macula because of its higher reproducibility and correlation with other measurements of the central macula. Total macular volume may be preferred when the central macula is less important. Absolute change in retinal thickness is the preferred analysis method in studies involving eyes with mild macular thickening. Relative change in thickening may be preferable when retinal thickening is more severe. PMID- 18675713 TI - Anatomy and pathomechanics of the thumb. AB - The prehensile thumb provides the human mind an outlet for coordinated activity through its fine motions of prehension, opposition, and circumduction. A comprehensive understanding of the anatomy and biomechanics of the thumb provides a foundation on which functional disorders may be recognized and effectively treated. PMID- 18675711 TI - Early androgen exposure modulates spatial cognition in congenital adrenal hyperplasia (CAH). AB - Major questions remain about the exact role of hormones in cognition. Furthermore, the extent to which early perturbation in steroid function affects human brain development continues to be a wide open area of research. Congenital adrenal hyperplasia (CAH), a genetic disorder of steroid dysfunction characterized in part by in utero over-production of testosterone, was used as a natural model for addressing this question. Here, CAH (n=54, mean age=17.53, 31 female) patients were compared to healthy age- and sex-matched individuals (n=55, mean age=19.02, 22 female) on a virtual equivalent of the Morris Water Maze task [Morris, R., 1984. Developments of a water-maze procedure for studying spatial learning in the rat. J. Neurosci. Methods 11, 47-60], an established measure of sex differences in spatial cognition in rodents. Findings revealed that females with CAH with the most severe form of the disease and expected highest level of in utero exposure to androgens were found to perform similarly to both healthy males and CAH males, whereas strong sex differences were apparent in milder forms of the disorder and in controls. Moreover, advanced bone age, an indicator of long-term childhood exposure to testosterone was correlated with improved performance. The results indicate that individuals exposed to both excess androgens prenatally and prolonged exposure during childhood may manifest long lasting changes in cognitive function. Such finding suggests a pivotal role of hormonal function on brain development in humans, mirroring results from the animal literature. PMID- 18675714 TI - Physical diagnosis and radiographic examination of the thumb. AB - Arthritis of the thumb joints is a common problem and remains a significant cause of morbidity in the adult population. Careful physical examination is critical in the assessment of these patients, given the large differential diagnosis of conditions affecting the thumb and the radial side of the wrist. Because treatment should be specifically directed at the area of pathology, adequate diagnosis is vital. Plain radiograph evaluation remains the diagnostic modality of choice in the evaluation of patients with degenerative conditions about the hand and wrist. PMID- 18675715 TI - Treatment of thumb metacarpophalangeal and interphalangeal joint arthritis. AB - Degenerative joint disease affecting the thumb metacarpophalangeal and interphalangeal joints is a debilitating condition, which can significantly restrict activities of daily living. Conservative measures to address symptoms include oral anti-inflammatory medications, activity modification and splinting, and intraarticular corticosteroid injections. Surgical interventions include arthroscopic synovectomy, arthroplasty, and finally arthrodesis of the affected articulations. Although short-term results from synovectomy and arthroplasty seem promising, long-term data are not yet available. Arthrodesis of the metacarpophalangeal and interphalangeal joints yields a stable yet functional thumb with reliably good pain relief. PMID- 18675716 TI - Early treatment of degenerative arthritis of the thumb carpometacarpal joint. AB - Degenerative arthritis of the thumb carpometacarpal (CMC) joint is a common disorder that may affect anyone but most frequently affects the postmenopausal female population. Because of its high prevalence, the management of the condition has been a popular topic among hand surgeons and therapists worldwide. There are many decisions to consider when devising the appropriate treatment plan for each patient. In particular, early stages of thumb CMC joint arthritis may be treated nonoperatively or with less invasive surgical techniques to relieve symptoms, restore function and strength, stop the progression of the disease, and even potentially reverse the process. This article explores treatment options at the disposal of primary care physicians and hand surgeons for early thumb CMC arthritis. PMID- 18675717 TI - Treatment of advanced carpometacarpal joint disease: carpometacarpal arthroplasty with ligament interposition. AB - Basal joint arthritis is a common condition, primarily affecting postmenopausal women. Persistent pain and functional impairment despite conservative treatment are indications for operative intervention. Ligament reconstruction and tendon interposition (LRTI) arthroplasty is one of the most popular and time-tested operations to treat metacarpal instability and basal joint arthritis. LRTI incorporates three fundamental principles that address the underlying anatomic pathology: (1) trapezium excision, either partial or complete, to eliminate eburnated bone and the source of pain; (2) anterior oblique ligament reconstruction for carpometacarpal joint stability; and (3) tendon interposition to minimize axial shortening and prevent bony impingement. PMID- 18675718 TI - Treatment of advanced carpometacarpal joint disease: trapeziectomy and hematoma arthroplasty. AB - Some surgical treatment options of painful basal joint arthritis do not require complex arthroplasty or suspensionplasty techniques. Simple trapeziectomy with temporary pinning of a slightly overdistracted thumb metacarpal can provide reliable pain relief, good motion, and functional stability. Recent literature supports alternative options when compared with formal ligament reconstruction or suspensionplasty procedures. Simple trapeziectomy is associated with less morbidity due to shorter operative times and the lack of need for graft harvest and ligament reconstruction. PMID- 18675719 TI - Treatment of advanced CMC joint disease: trapeziectomy and implant arthroplasty (silastic-metal-synthetic allograft). AB - Osteoarthritis of the basal joint of the thumb is a common and frequently debilitating condition, most often affecting middle-aged women. Nonoperative treatment with activity modification, splinting, oral anti-inflammatory medication, and intraarticular steroid injection frequently leads to acceptable control of symptoms. If nonoperative treatment fails, many surgical techniques have been described for management of symptomatic advanced degenerative joint changes. This article reviews the literature related to various arthroplasty options for advanced disease. Treatment decisions must clearly be tempered by the surgeon's experience, the patient's goals and expectations, and the extent of degenerative disease. PMID- 18675720 TI - Treatment of advanced carpometacarpal joint disease: arthrodesis. AB - Thumb carpometacarpal arthritis is a common condition that may result in debilitating pain and loss of acceptable hand function. If patients continue to be symptomatic after a trial of nonoperative management, many surgical options exist in the treatment of carpometacarpal arthritis. For isolated trapeziometacarpal arthritis, arthrodesis is a viable option to create a pain free, strong, and stable thumb. PMID- 18675721 TI - Carpometacarpal joint disease: addressing the metacarpophalangeal joint deformity. AB - The successful surgical treatment of arthrosis of the carpometacarpal articulation of the thumb requires a thorough understanding and evaluation of the intercalated axis of the first ray. A hyperextension/adduction deformity commonly occurs at the metacarpophalangeal joint of the thumb with advanced stages of carpometacarpal arthrosis. Failure to recognize and treat the metacarpophalangeal deformity may result in continued pain and poor outcomes. Additionally, the stability of the ligament reconstruction may become compromised, resulting in recurrence of deformity and longitudinal collapse. This article presents an orderly means of clinical and radiographic evaluation of this deformity and recommends surgical treatments to correct hyperextension and maximize functional outcomes. A treatment algorithm is provided. PMID- 18675722 TI - Treatment of scaphotrapezio-trapezoid arthritis. AB - Arthritis of the scaphotrapeziotrapezoid (STT) joint presents with deep thenar eminence and thumb basilar pain and is often coexistent with carpometacarpal arthritis of the thumb. Conservative treatment includes splinting and corticosteroid injections. Operative treatment consists primarily of fusion of the STT joint, although alternatives include trapeziectomy, fibrous arthroplasty, and prosthetic replacements. When STT arthritis is coexistent with carpometacarpal arthritis, excision of the trapezium and proximal 2 mm of trapezoid has been recommended. Complications of surgery include pericarpal arthrosis, superficial radial nerve injury, and nonunion. PMID- 18675723 TI - The rheumatoid thumb. AB - Rheumatoid arthritis of the thumb is a common source of disability. Obtaining an understanding of the underlying biologic and physical manifestations of rheumatoid arthritis is essential in the choice of treatment of the disease. In the early stages of the disease, conservative and less invasive measures can be used. In the more advanced stages, arthrodesis and arthroplasty are often used. Isolated interphalangeal involvement is best managed with arthrodesis. Metacarpophalangeal involvement in low-demand patients can be treated with arthroplasty, whereas arthrodesis can be used in more active patients. Patients who have carpometacarpal joint damage are best treated with trapezium resection arthroplasty. PMID- 18675724 TI - A Zn-surprise in Ehlers-Danlos syndrome. PMID- 18675725 TI - B cell positive cross-match not due to anti-HLA Class I antibodies and first kidney graft outcome. AB - The effect of B cell cross-match (XM) was investigated in 680 first deceased donor kidney transplants in a single centre from 1990 to 1999: 74 transplants presented a B-positive XM (Group 1) 606 had a B-negative XM (Group 2). The absence in Group 1 of weak/low-titre anti-HLA Class I antibodies was assured blocking anti-Class I reactivity by treating B cells with non-cytotoxic anti beta2 microglobulin (alphabeta2 M) serum before XM. Graft survivals up to 5 years were not significantly different; some differences were nevertheless observed: HLA-A,B,DR mismatches influenced graft outcome in Group 1: patients with 0-2 mismatches had better survival than patients with 3-4. When analysed according DR mismatch, patients with 1 mismatch had worse graft survival than well matched patients (p<0.05). No significant difference depending on HLA match was observed in Group 2. Early acute rejection rate was similar in the Groups except the rejection episodes after one year: Group 1 had significantly more. 61/74 patients of Group 1 were retrospectively analysed for anti-HLA-DR,DQ reactivity: only 11/61 had anti-HLA-DR or DQ antibodies (3/11 were donor specific); graft survival and rejections were not significantly different in the patients with and without anti-HLA Class II antibodies. Anti-donor B cell reactivity, at XM, once excluded the presence of weak/low-titre anti-HLA Class I antibodies, did not influence first kidney graft survival. PMID- 18675727 TI - An overview of restraint and blood collection techniques in exotic pet practice. AB - As more data about blood test results become available, blood sampling becomes increasingly important as a diagnostic tool. The increased availability of biochemistry analyzers that sample small volumes of blood has made it possible to get diagnostic profiles and hematology results from pets that have only small volumes of blood to contribute. This article describes techniques for restraint, phlebotomy, and sample preparation of several of the more popular exotic animals in the pet trade. The species covered include birds, lizards, chelonians, snakes, frogs, salamanders, fish, rabbits, ferrets, guinea pigs, mice, rats, hamsters, gerbils, hedgehogs, sugar gliders, and giant spiders (tarantulas). PMID- 18675728 TI - Hematologic disorders of fish. AB - Hematology can be a useful tool for monitoring health status, detecting illness, and following the progress of disease and response to therapy. Despite advances in fish medicine in recent years, interpretation of fish hematology often is hampered by a lack of meaningful reference values and the bewildering diversity of fish species. A multitude of intrinsic and extrinsic factors cause normal and abnormal variation in hematologic data. This article provides an overview of some of the hematologic abnormalities in fish induced by infectious agents and environmental, husbandry, and nutritional issues. PMID- 18675729 TI - Amphibian hematology. AB - Amphibians are a diverse class of animals with a unique life cycle. Intrinsic and extrinsic factors contribute to the wide variability in normal hematologic parameters. Reference values are scarce, and normal hematology of many species is poorly understood. Challenges include analytic obstacles posed by nucleated red blood cells and thrombocytes and potential difficulty with obtaining blood samples of adequate volume and without lymph contamination. Despite these limitations, it is possible to obtain hematologic data that may be useful in assessing an animal's current health, progression of disease, or response to therapy. In this article, amphibian blood sample collection and handling guidelines, hematologic tests, cell morphology and function, hematopoiesis, interpretation of results, and disorders and diseases are described. PMID- 18675730 TI - Reptile hematology. AB - The basic principles of hematology used in mammalian medicine can be applied to reptiles. Available phlebotomy sites vary with the taxa and may be chosen based on the experience of the handler or phlebotomist, health status or temperament of the patient, and species. Methods of analysis and the cytologic appearance of erythrocytes, leukocytes, and thrombocytes are different from those of mammals and also vary by taxa. Many causes for abnormalities of the reptilian hemogram are similar to those for mammals, although additional factors, such as venipuncture site, season, hibernation status, captivity status, and environmental factors, can affect measured values. Interpretation of hematologic results can be frustrating, although new case reports and research studies help to elucidate possible causes that are often unique to reptiles. PMID- 18675731 TI - Avian hematology and related disorders. AB - Hematology is an essential component of veterinary practice. The interpretation of avian blood cells provides many challenges. Practitioners must be able to recognize normal morphology and function of cells to interpret changes in those cells. This article describes the normal morphology of avian erythrocytes, leukocytes, and thrombocytes. Changes observed in erythrocytes and leukocytes during disease and major differential diagnoses are discussed. A brief overview of avian blood parasites is also presented. PMID- 18675732 TI - Clinical hematology of rodent species. AB - Pet rodents, such as rats, guinea pigs, and chinchillas, differ from more traditional companion animal species in many aspects of their hematologic parameters. Animals within this order have much diversity in size, anatomy, methods of restraint, and blood collection technique. Appropriate sample collection is often the most challenging aspect of the diagnostic protocol, and inappropriate restraint may cause a stress response that interferes with blood test results. For many of these patients, sedation is required and can also affect results as well. In most cases, however, obtaining a standard database is necessary and very possible when providing medical care for this popular group of pets. PMID- 18675733 TI - Ferret hematology and related disorders. AB - This article summarizes the general topic of ferret hematology, including discussion of restraint for phlebotomy and phlebotomy sites, red and white blood cell morphology, interpretation of the hemogram, and normal and abnormal factors affecting the hemogram. In addition, the apparent lack of blood groups and techniques for bone marrow aspirates and blood transfusion are addressed. There is much still to be learned about the ferret and its diseases. We often depend on tests as simple as the complete blood cell count to help guide us in our diagnosis and treatments of this patient. PMID- 18675734 TI - Rabbit hematology. AB - Using laboratory animal medicine as an established resource, companion animal veterinarians have access to many physiologic and basic science studies that we can now merge with our clinical impressions. By working with reference laboratories, companion animal veterinarians are poised to accelerate our knowledge of the normal rabbit rapidly. The aim of this article is to discuss normal hematopoiesis and infectious and metabolic diseases that specifically target the hemolymphatic system. Additionally, photographic representation of cell types is provided. PMID- 18675735 TI - Normal hematology and hematologic disorders in potbellied pigs. AB - Potbellied pigs have become a notable portion of small animal and farm animal practitioners' caseload. Relatively little information is readily accessible for the veterinary practitioner in regard to normal hematologic values or alterations of the hemogram in response to disease, however. This article is a review of blood collection techniques in swine adaptable to potbellied pigs in addition to collection artifacts observed in the swine hemogram. Alterations of the hemogram in disease states that may be encountered in potbellied pig medicine are reviewed. PMID- 18675736 TI - Flow cytometry applications for exotic animals. AB - This article describes applications of flow cytometry in hematology. It includes a basic description of how flow cytometers work and their use in enumerating cell populations based on phenotypic markers and measurement of cell functions. Challenges presented by limitations of reagents for exotic animals are described in addition to sources of cross-reactive antibodies. PMID- 18675737 TI - Intraneural catheterization of the sciatic nerve in humans: a pilot study. AB - BACKGROUND AND OBJECTIVES: Imaging studies in humans have shown that intraneural injection of local anesthetic may be relatively frequent. The incidence of intraneural catheterization is unknown. We speculated that early neural blockade after the injection of a small dose of local anesthetic might be a common finding produced by unintended intraneural catheterization. We investigated the clinical effect of a small dose of local anesthetic injected through a stimulating sciatic catheter, and also the placement of the tip of the catheters by computed tomography (CT) scan. METHODS: In this descriptive study, a dose of 3 to 5 mL of 1.5% mepivacaine was injected through a stimulating sciatic catheter in 45 patients undergoing hallux valgus repair. Patients with early neural blockade at the foot, within 5 minutes after injection, were suspected to have an intraneural placement of the catheter. A CT scan of the thighs was performed in 10 additional patients to assess catheter tip placement. RESULTS: Nine of 45 patients had early neural blockade with the small dose of local anesthetic. Seven patients underwent surgery without further supplementation. There was a statistically significant lower intensity of stimulating current through the catheter in the group of patients with early neural blockade. Of 10 CT scans, 3 showed clear intraneural placement of the catheter. No postoperative sequelae were observed. CONCLUSIONS: Intraneural catheterization of the sciatic nerve may be a frequent finding and was not followed by nerve injury under the conditions of our small pilot study. PMID- 18675738 TI - The sensitivity of motor response to needle nerve stimulation during ultrasound guided interscalene catheter placement. AB - BACKGROUND AND OBJECTIVES: Neurostimulation during single shot interscalene block has a significant false negative motor response rate. Compared with tangential needle approaches for single shot block, interscalene catheter (ISC) placement commonly involves Tuohy needles inserted longitudinally to the brachial plexus. This study aimed to determine the sensitivity of neurostimulation during ultrasound-guided ISC needle placement, and the feasibility of an ultrasound guided ISC needle endpoint. METHODS: One hundred fifty-five consecutive nonstimulating ISCs were placed with the needle tip position confirmed by the sonographic spread of 5 mL dextrose 5%. Catheter advancement was then blind 2 to 3 cm past the needle tip. A 0.8 mA electrical stimulus at 2 Hz was applied throughout. When a satisfactory image was obtained, neurostimulation was ignored and the minimum motor response amplitude noted. If imaging was equivocal, a brief appropriate motor response at 0.8 mA was sought. A sustained response at <0.5 mA was only sought if imaging was suboptimal. Prior to surgery conducted under general anesthesia, 30 mL ropivacaine 0.5% was administered through the ISC. Catheter success was defined as a recovery room numerical rating pain score of or =75 years). METHODS: Patients with CKD but not on dialysis, receiving darbepoetin alfa every other week (Q2W), and with stable hemoglobin (Hb) levels between 11 and 13 g/dL, inclusive, were enrolled in this 33-week, multicenter, open-label, single-arm study. The study was carried out at 36 US centers and consisted of a 24-week QM darbepoetin alfa dose-titration period followed by an 8-week evaluation period. Hb levels were measured Q2W. Study results were stratified according to patient age (<65, 65-74, and > or =75 years). RESULTS: A total of 152 patients (79 women, 73 men) were enrolled; 55 patients (36%) were <65 years of age, 46 (30%) were 65 to 74 years of age, and 51 (34%) were > or =75 years of age. In patients who received > or =1 dose of darbepoetin alfa, Hb levels > or =11 g/dL were maintained in 76%, 80%, and 71% of patients aged <65, 65 to 74, and > or =75 years, respectively. For patients who completed the study, the proportions who maintained Hb levels > or =11 g/dL were 83%, 88%, and 85%, respectively, for the 3 age groups. The safety profile of QM darbepoetin alfa in this study was consistent with that expected in patients with CKD not receiving dialysis. CONCLUSIONS: Darbepoetin alfa administered QM maintained Hb levels > or =11 g/dL in patients with CKD (not on dialysis) aged <65, 65 to 74, and > or =75 years. This treatment regimen may help optimize anemia management for older community dwelling and long-term care patients. PMID- 18675765 TI - Methodology of an ongoing, randomized controlled trial to prevent falls through enhanced pharmaceutical care. AB - BACKGROUND: Falls are the leading cause of both fatal and nonfatal injuries among adults aged > or =65 years in the United States. Past research suggests that individuals taking multiple medications are at increased risk of falls. Central nervous system-active drugs in particular have been associated with increased risk. OBJECTIVE: The goal of this research was to describe the design of a study evaluating the effectiveness of a community pharmacy-based falls prevention program. Also presented are the algorithms used to identify high-risk patients based on their prescription profile records and to deliver the experimental intervention. METHODS: The study is a randomized controlled trial. The target population was community-dwelling older adults (aged > or =65 years) at high risk for future falls because: (1) they had experienced > or =1 fall within the 12 month period preceding study enrollment; (2) they were currently using > or =4 chronic prescription medications; and (3) they were taking > or =1 of the high risk medications targeted by the intervention. Participants were recruited using pharmacy prescription profile records. Individuals in the intervention group received a face-to-face medication consultation provided by a community pharmacy resident. Identification of drug therapy problems and therapeutic recommendations was guided by a series of algorithms developed for this study. All participants were followed up for 24 months. The primary study end points were: (1) time to first fall; and (2) proportion of participants who experienced > or =1 fall during the first year of follow-up. RESULTS: Participant enrollment began in September 2005 and was completed in August 2007. A total of 186 individuals were enrolled in the study (mean [SD] age, 74.8 [6.9] years; 132 women, 54 men), and 67 have completed the first year of follow-up. CONCLUSIONS: The study is using a rigorous randomized controlled research design, which will enhance the internal validity of its findings. Results of the study, which will be reported after the completion of follow-up data collection activities, will enable us to assess the effects of the intervention on both medication use and the incidence of falls. If the intervention is found to be effective, it will provide a resource for community pharmacists working with older adults at high risk of medication related falls. PMID- 18675766 TI - A before and after study of medical students' and house staff members' knowledge of ACOVE quality of pharmacologic care standards on an acute care for elders unit. AB - BACKGROUND: The Assessing Care of Vulnerable Elders (ACOVE) comprehensive set of quality assessment tools for ill older persons is a standard designed to measure overall care delivered to vulnerable elders (ie, those aged > or =65 years) at the level of a health care system or plan. OBJECTIVE: The goal of this research was to quantify the pretest and posttest results of medical students and house staff participating in a pharmacotherapist-led educational intervention that focused on the ACOVE quality of pharmacologic care standards. METHODS: This was a before and after study assessing the knowledge ofACOVE standards following exposure to an educational intervention led by a pharmacotherapist. It was conducted at the 29-bed Acute Care for Elders (ACE) unit of Maimonides Medical Center, a 705-bed, independent teaching hospital located in Brooklyn, New York. Participants included all medical students and house staff completing a rotation on the ACE unit from August 2004 through May 2005 who completed both the pre-and posttests. A pharmacotherapist provided a 1-hour active learning session reviewing the evidence supporting the quality indicators and reviewed case-based questions with the medical students and house staff. Educational interventions also occurred daily through pharmacotherapeutic consultations and during work rounds. Medical students and house staff were administered the same 15-question, patient-specific, case-based, multiple-choice pre-and posttest to assess knowledge of the standards before and after receiving the intervention. RESULTS: A total of 54 medical students and house staff (median age, 28.58 years; 40 men, 14 women) completed the study. Significantly higher median scores were achieved on the multiple-choice test after the intervention than before (median scores, 14/15 [93.3%] vs 12/15 [80.0%], respectively; P = 0.001). CONCLUSION: A pharmacotherapist-led educational intervention improved the scores of medical students and house staff on a test evaluating knowledge of evidence-based recommendations for pharmacotherapy in the elderly. PMID- 18675767 TI - Mirtazapine associated with profound hyponatremia: two case reports. AB - BACKGROUND: Depression is a common problem in the elderly, and the recognition and appropriate management of this illness are important aspects of geriatric medicine. Selective serotonin reuptake inhibitors have been associated with hyponatremia, but the association of mirtazapine with hyponatremia is less well documented. OBJECTIVE: The goal of this research was to highlight and characterize the association between mirtazapine and hyponatremia. We present here 2 case reports as well as the results of a literature review. CASE SUMMARIES: Two patients, a 61-year-old man and a 79-year-old woman, developed profound hyponatremia (sodium levels, 112 and 113 mEq/L, respectively) 7 and 10 days after commencement of mirtazapine for symptoms of depression. Investigations excluded other causes, and cessation of mirtazapine was associated with recovery of sodium levels to > or =132 mEq/L after 7 and 10 days, respectively. CONCLUSIONS: The likelihood of mirtazapine use causing hyponatremia in these 2 cases was "probable" according to criteria of the Naranjo Adverse Drug Reaction Probability Scale (score, 6). A review of published cases found that mirtazapine associated hyponatremia occurred in patients aged >60 years, after a mean of 6.5 days and with doses as low as 7.5 mg daily. The mean sodium nadir was 117.2 mEq/L, but after stopping mirtazapine, the mean time to recovery was 11 days. Clinicians should be aware of the possibility of this reaction in elderly patients and should monitor sodium levels in high-risk patients if symptoms suggestive of hyponatremia develop. PMID- 18675768 TI - Adverse effects of propafenone after long-term therapy with the addition of citalopram. AB - BACKGROUND: Propafenone, a class IC antiarrhythmic, and citalopram, a selective serotonin reuptake inhibitor (SSRI), are widely used in older patients. Although a potential interaction between propafenone and SSRIs has been noted, a MEDLINE search revealed no published reports of an interaction between propafenone and citalopram. OBJECTIVE: The goal of this article was to describe a potential drug drug interaction between propafenone and citalopram, which caused symptoms of propafenone adverse effects. CASE SUMMARY: An 80-year-old white female, followed up at the Memory Clinic for mild cognitive impairment, had been taking propafenone 900 mg/d for >10 years for paroxysmal atrial fibrillation without adverse effects. Three months after starting citalopram, she experienced episodes of chest tightness and dizziness. The episodes became more frequent in the following months, causing several falls and requiring visits to the emergency department, but no acute coronary event was diagnosed. She was started on amlodipine 2.5 mg orally once daily, a nitroglycerin patch (0.4 mg/h), and warfarin 5 mg orally once daily. After one fall, the patient became delirious. Amlodipine and the nitroglycerin patch were discontinued and propafenone decreased to 450 mg/d; citalopram was continued at 20 mg/d. The patient recovered well, both cognitively and physically, and did not have any further symptoms in 1 year of follow-up. Results of coronary investigations were negative. CONCLUSIONS: This is the first report of a possible interaction between propafenone and citalopram, which caused propafenone adverse effects (eg, dizziness, falls) and mimicked coronary artery disease. PMID- 18675769 TI - Cardiovascular risk factors and dementia. AB - BACKGROUND: Dementias, such as Alzheimer's disease (AD) and vascular dementia, are disorders of aging populations and represent a significant economic burden. Evidence is accumulating to suggest that cardiovascular disease (CVD) risk factors may be instrumental in the development of dementia. OBJECTIVE: The goal of this review was to discuss the relationship between specific CVD risk factors and dementia and how current treatment strategies for dementia should focus on reducing CVD risks. METHODS: We conducted a review of the literature for the simultaneous presence of 2 major topics, cardiovascular risk factors and dementia (eg, AD). Special emphasis was placed on clinical outcome studies examining the effects of treatments of pharmacologically modifiable CVD risk factors on dementia and cognitive impairment. RESULTS: Lifestyle risk factors for CVD, such as obesity, lack of exercise, smoking, and certain psychosocial factors, have been associated with an increased risk of cognitive decline and dementia. Some evidence suggests that effectively managing these factors may prevent cognitive decline/dementia. Randomized, placebo-controlled trials of antihypertensive medications have found that such therapy may reduce the risk of cognitive decline, and limited data suggest a benefit for patients with AD. Some small open label and randomized clinical trials of statins have observed positive effects on cognitive function; larger studies of statins in patients with AD are ongoing. Although more research is needed, current evidence indicates an association between CVD risk factors--such as hypertension, dyslipidemia, and diabetes mellitus--and cognitive decline/dementia. CONCLUSIONS: From a clinical perspective, these data further support the rationale for physicians to provide effective management of CVD risk factors and for patients to be compliant with such recommendations to possibly prevent cognitive decline/dementia. PMID- 18675770 TI - Use of computer decision support interventions to improve medication prescribing in older adults: a systematic review. AB - BACKGROUND: Older adults take multiple medications and are at high risk for adverse drug effects. OBJECTIVE: This systematic review was conducted to describe the impact of computer decision support (CDS) interventions designed to improve the quality of medication prescribing in older adults. METHODS: PubMed and EMBASE databases were searched from January 1980 through July 2007 (English-language only); studies were eligible if they described a CDS intervention intended to improve medication prescribing in adults aged > or =60 years. Studies were retained if they were observational or experimental in design and reported > or =1 process or clinical outcome measurement related to medication prescribing. In the main analysis, study characteristics and major outcome results were extracted. A combination of searches was performed using relevant medical subject headings: aged; drug therapy, computer-assisted; medication errors; medication errors/prevention and control; decision making, computer-assisted; decision support systems, clinical; and clinical pharmacy information systems. RESULTS: After review of study abstracts, 10 articles met the eligibility criteria. Of those 10 studies testing CDS interventions, 8 showed at least modest improvements (median number needed to treat, 33) in prescribing, as measured by minimizing drugs to avoid, optimizing drug dosage, or more generally improving prescribing choices in older adults (according to each study's intervention protocols). Findings for the impact of CDS interventions on clinical outcomes were mixed and were reported for only 2 studies. CONCLUSIONS: Various types of CDS interventions may be effective in improving medication prescribing in older adults, but few studies reported clinical outcomes related to changes in medication prescribing. Data from this study should help to guide refinement and testing of future CDS interventions that specifically target older adult populations that are taking multiple medications. PMID- 18675772 TI - Free sulfhydryl measurement as an indicator of antibody stability. AB - Monoclonal antibodies are a major subclass of biopharmaceuticals. They are structurally different from other biopharmaceuticals in size and quaternary structure. Here we demonstrate a correlation between chemical stability of antibodies and thermal stability. We show that overall thermal protein stability can be predicted based on the measurement of free sulfhydryl (-SH) content on applying mildly denaturing conditions. We propose that this method can be adapted to a high-throughput screening format and used either as an absolute measure of thermal stability or for ranking a panel of possible variants. PMID- 18675773 TI - Monitoring the (photo)genotoxicity of photosensitizer drugs: direct quantitation of single-strand breaks in deoxyribonucleic acid using an oligonucleotide chip. AB - Oligonucleotide chip-based assays can be a sample-thrifty, time-saving, routine tool for evaluation of chemical-induced DNA strand breaks. This article describes a novel approach using an oligonucleotide chip to determine photosensitizer induced DNA single-strand breaks. Surface coverage of fluorophore-labeled oligonucleotides on silicon dioxide chip surfaces was determined on alkaline phosphatase digestion. Fluorescence maxima (at 520 nm) of the solutions were converted to molar concentrations of the fluorescein-modified oligonucleotide by interpolation from a predetermined standard linear calibration curve. The photosensitizing activity of chlorpromazine and triflupromazine toward DNA single strand breaks was then studied at different drug doses and also as a function of photoirradiation time. Photoinduced single-strand breaks calculated using the method described here agreed with values predicted by theoretical extrapolation of the single-strand breaks obtained for plasmid DNAs from agarose gel electrophoresis, and thereby indirectly validated the chip-based assays. Under UV irradiation (>or=93.6 kJ/m2) chlorpromazine (>or=0.08 mM) was found to have significant photogenotoxicity. However, triflupromazine did not exhibit any (photo)genotoxicity over the concentration range studied (0.04-0.20mM). The method developed will be useful for quantitative screening of drug genotoxicity in terms of induction of breaks in DNA. PMID- 18675774 TI - Human thrombospondin's (TSP-1) C-terminal domain opens to interact with the CD-47 receptor: a molecular modeling study. AB - Thrombospondin-1 (TSP-1) interaction with the membranous receptor CD-47 involves the peptide RFYVVMWK (4N-1) located in its C-terminal domain. However, the available X-ray structure of TSP-1 describes this peptide as completely buried into a hydrophobic pocket, preventing any interaction. Where classical standard methods failed, an appropriate approach combining normal mode analysis and an adapted protocol of energy minimization identified the large amplitude motions responsible of the partial solvent exposure of 4N-1. In agreement, the obtained model of the open TSP-1 was further used for protein-protein docking experiments against a homology model generated for CD-47. Considering the multiple applications of the CD-47 receptor as a target, our results open new pharmacological perspectives for the design of TSP-1:CD-47 inhibitors and CD-47 antagonists. We also suggest a common opening mechanism for proteins sharing the same fold as TSP-1. This work also suggests the usefulness of our approach in other topics in which predictions of protein-protein interactions are of importance. PMID- 18675775 TI - Glial metabolism of quercetin reduces its neurotoxic potential. AB - The neuroprotective effects of flavonoids will ultimately depend on their interaction with both neuronal and glial cells. In this study, we show that the potential neurotoxic effects of quercetin are modified by glial cell interactions. Specifically, quercetin is rapidly conjugated to glutathione within glial cells to yield 2'-glutathionyl-quercetin, which is exported from cells but has significantly reduced neurotoxicity. In addition, quercetin underwent intracellular O-methylation to yield 3'-O-methyl-quercetin and 4'-O-methyl quercetin, although these were not exported from glia at the same rate as the glutathionyl adduct. The neurotoxic potential of both quercetin and 2' glutathionyl-quercetin paralleled their ability to modulate the pro-survival Akt/PKB and extracellular signal-regulated kinase (ERK) signalling pathways. These data were supported by co-culture investigation, where the neurotoxic effects of quercetin were significantly reduced when they were cultured alongside glial cells. We propose that glial cells act to protect neurons against the neurotoxic effects of quercetin and that 2'-glutathionyl-quercetin represents a novel quercetin metabolite. PMID- 18675776 TI - Role of SelS in lipopolysaccharide-induced inflammatory response in hepatoma HepG2 cells. AB - To investigate the role of SelS in bacterial lipopolysaccharide (LPS) induced inflammatory response, some parameters in LPS-stimulated HepG2 cells were comparatively studied fore-and-aft SelS silence. LPS induced the decreases of cytoplasmic glutathione peroxidase (GPx-1) mRNA expression and activity, and the increases of reactive oxygen species (ROS) level, intracellular and extracellular nitric oxide (NO) levels, inducible nitric oxide synthase (iNOS) mRNA expression and activity, and serum amyloid A1 (SAA1) mRNA expression and secreted protein level in hepatoma HepG2 cells. When SelS was suppressed by small interfering RNA (siRNA), those decreases and increases were further aggravated under LPS stimulation, respectively. In conclusion, the negative association between SelS and the LPS-induced production of ROS, NO and SAA1 demonstrated that SelS had an important role in influencing inflammatory response, and that role may be related with SelS as a central component of retro-translocation channel in endoplasmic reticulum-associated protein degradation (ERAD) and its anti-oxidative property. PMID- 18675777 TI - Characterization of the palytoxin effect on Ca2+-ATPase from sarcoplasmic reticulum (SERCA). AB - The effect of palytoxin was studied in a microsomal fraction enriched in longitudinal tubules of the sarcoplasmic reticulum membrane. Half-maximal effect of palytoxin on Ca(2+)-ATPase activity yielded an apparent inhibition constant of approx. 0.4 microM. The inhibition process exhibited the following characteristics: (i) the degree of inhibition was dependent on membrane protein concentration; (ii) no protection was observed when the ATP concentration was raised; (iii) dependence on Ca(2+) concentration with a decreased maximum catalytic rate; (iv) it occurred in the absence of Ca(2+) ionophoric activity. Likewise, the inhibition mechanism was linked to: (i) rapid enzyme phosphorylation from ATP in the presence of Ca(2+) but lower steady-state levels of phosphoenzyme; (ii) more drastic effect on phosphoenzyme levels when the toxin was added to the enzyme in the absence of Ca(2+); (iii) decreased phosphoenzyme levels at saturating Ca(2+) concentrations; (iv) no effect on kinetics of phosphoenzyme decomposition. The palytoxin effect is related with lock of the enzyme in the Ca(2+)-free conformation so that progression of the catalytic cycle is impeded. PMID- 18675778 TI - Cellular uptake and membrane-destabilising properties of alpha-peptide/beta peptoid chimeras: lessons for the design of new cell-penetrating peptides. AB - Novel peptidomimetic backbone designs with stability towards proteases are of interest for several pharmaceutical applications including intracellular delivery. The present study concerns the cellular uptake and membrane destabilising effects of various cationic chimeras comprised of alternating N alkylated beta-alanine and alpha-amino acid residues. For comparison, homomeric peptides displaying octacationic functionalities as well as the Tat(47-57) sequence were included as reference compounds. Cellular uptake studies with fluorescently labelled compounds showed that guanidinylated chimeras were taken up four times more efficiently than Tat(47-57). After internalisation, the chimeras were localised primarily in vesicular compartments and diffusively in the cytoplasm. In murine NIH3T3 fibroblasts, the chimeras showed immediate plasma membrane permeabilising properties, which proved highly dependent on the chimera chain length, and were remarkably different from the effects induced by Tat(47 57). Finally, biophysical studies on model membranes showed that the chimeras in general increase the permeability of fluid phase and gel phase phosphatidylcholine (PC) vesicles without affecting membrane acyl chain packing, which suggests that they restrict lateral diffusion of the membrane lipids by interaction with phospholipid head groups. The alpha-peptide/beta-peptoid chimeras described herein exhibit promising cellular uptake properties, and thus represent proteolytically stable alternatives to currently known cell-penetrating peptides. PMID- 18675779 TI - Tight junction claudins and the kidney in sickness and in health. AB - The epithelial cell tight junction has several functions including the control of paracellular transport between epithelial cells. Renal paracellular transport has been long recognized to exhibit unique characteristics within different segments of the nephron, functions as an important component of normal renal physiology and has been speculated to contribute to renal related pathology if functioning abnormally. The discovery of a large family of tight junction associated 4 transmembrane spanning domain proteins named claudins has advanced our understanding on how the paracellular permeability properties of tight junctions are determined. In the kidney, claudins are expressed in a nephron-specific pattern and are major determinants of the paracellular permeability of tight junctions in different nephron segments. The combination of nephron segment claudin expression patterns, inherited renal diseases, and renal epithelial cell culture models is providing important clues about how tight junction claudin molecules function in different segments of the nephron under normal and pathological conditions. This review discusses early observations of renal tubule paracellular transport and more recent information on the discovery of the claudin family of tight junction associated membrane proteins and how they relate to normal renal function as well as diseases of the human kidney. PMID- 18675780 TI - Effect of sphingomyelinase treatment on ligand binding activity of human serotonin1A receptors. AB - The serotonin1A receptor is an important member of the G-protein coupled receptor family, and is involved in the generation and modulation of a variety of cognitive, behavioral, and developmental functions. We have monitored the ligand binding of the human serotonin1A receptor stably expressed in CHO cells (termed CHO-5-HT1AR) following treatment with sphingomyelinase (SMase), an enzyme that specifically catalyzes the hydrolysis of sphingomyelin into ceramide and phosphorylcholine. Our results show, for the first time, that the specific ligand binding activity of the serotonin1A receptor in membranes isolated from CHO-5 HT1AR cells is increased upon sphingomyelinase treatment. Saturation binding analysis reveals increase in binding affinity of the receptor under these conditions. This is accompanied by a reduction in membrane order, as monitored by fluorescence anisotropy of the membrane probe 1-[4-(trimethylammonio)phenyl]-6 phenyl-1,3,5-hexatriene (TMA-DPH) in intact cells. These results represent the first report on the effect of sphingomyelinase treatment on the ligand binding activity of this important neurotransmitter receptor. PMID- 18675781 TI - Specific adsorption of functionalized colloids at the surface of living cells: a quantitative kinetic analysis of the receptor-mediated binding. AB - This paper presents a statistical experimental study of the adsorption of colloids onto the plasma membrane of living cells mediated by specific ligand receptor interactions. The colloids consist of lipid multilamellar liposomes (spherulites) functionalized by Shiga toxin B-subunit (STxB), while cells are cervix carcinoma epithelial cells expressing the Shiga toxin receptor, the glycolipid globotriaosyl ceramide (Gb3). The specificity of the colloid adsorption is demonstrated using both confocal microscopy and flow cytometry, while a thorough cytometry study on living cells allows characterizing the kinetics of this specific adsorption. The final number of bound colloids and the characteristic adsorption time are shown to depend on bulk concentration, as expected for a thermodynamic equilibrium. However, the colloids appear to be irreversibly attached to the membrane. We interpret this apparent irreversibility as the result of a progressive recruitment of receptors. The methodology used here, whereby microscopic mechanisms are deduced from direct quantitative measurements on living cells, might allow the optimization of drug delivery systems or the quantification of virus infectivity. PMID- 18675782 TI - Spermine biphasically affects N-type calcium channel currents in adult dorsal root ganglion neurons of the rat. AB - Spermine (Spe) is a polyamine co-secreted with neurotransmitters. In this work its effects on N-type Ca2+ channel (CaV2.2) have been studied on adult sensory neurons of the rat by means of whole-cell patch-clamp. Spe exerted biphasic effects when added to the external solution: at 500 microM decreased N-type Ca2+ channel currents, reducing the maximum whole-cell conductance, shifting the activation curve to the right on the voltage axes and decreasing its slope; conversely, at lower concentration (500 nM) Spe induced completely opposite effects. In 62% of the neurons the inhibitory effects were accompanied by a slowing down of the activation kinetics relieved by a conditioning pre-pulse to +50 mV. The biphasic effects and their rapid onset and offset time course may be explained if multiple sites of action with a different affinity for Spe are present directly on the channel. The effects of Spe on HVA Ca2+ currents were strongly dependent on [Ca2+]ext, high [Ca2+] powerfully reducing Spe effects. This may be explained if we take into account that as Spe has four positive charges at physiological pH; it may compete with divalent cations for some negatively charged regulatory sites. In these experiments, Spe was effective at concentrations possibly reached in physiological conditions. PMID- 18675783 TI - Regulatory mechanism of TNFalpha autoregulation in HaCaT cells: the role of the transcription factor EGR-1. AB - Tumor necrosis factor alpha (TNFalpha) is a pro-inflammatory cytokine involved in innate immune response, as well as in the pathogenesis of many inflammatory diseases. Although several response elements in the TNFalpha promoter region are involved in the activation of gene transcription, few studies have examined the regulatory mechanism that controls TNFalpha autoregulation. In this study, we investigated the role of the Early Growth Response-1 (EGR-1) transcription factor in TNFalpha autoregulation in HaCaT human keratinocytes. The requirement for EGR 1 in TNFalpha autoregulation was confirmed using a construct harboring a point mutation in the EGR-1 binding site within the TNFalpha promoter and the introduction of EGR-1 siRNA. Inhibition of the ERK or JNK pathway suppressed TNFalpha-induced EGR-1 expression, resulting in the inhibition of TNFalpha induced TNFalpha promoter activation. These results reveal that the ERK and JNK MAPK pathways contribute to the autoregulation of TNFalpha synthesis via EGR-1 induction in HaCaT keratinocytes. PMID- 18675784 TI - A1762T/G1764A mutations of hepatitis B virus, associated with the increased risk of hepatocellular carcinoma, reduce basal core promoter activities. AB - Hepatitis B virus (HBV) infection is a major global health problem that causes over one million deaths annually. A1762T and G1764A mutations in the basal core promoter are often present in HBV patients but seldom in asymptomatic carriers, and are highly correlated with the increased risk of HBV-associated hepatocellular carcinoma (HCC). In this study, for the first time, we show that the basal core promoter activity of HBV strains isolated from asymptomatic carriers is decreased when 1762A is mutated to 1762T or 1764G is mutated to 1764A by site directed mutagenesis. By contrast, the promoter activity of HBV strains isolated from HCC patients is increased when 1762T and 1764A are reversely mutated into 1762A and 1764G, respectively. 1764G contributes more promoter activity than 1762T. We also show that T1762A and G1764A double mutations synergize the reduction of the promoter activity. A mechanism of HBV evasion from host immunoresponse that may facilitate disease development is also discussed. PMID- 18675786 TI - Involvement of neuromedin S in the oxytocin release response to suckling stimulus. AB - We recently identified neuromedin S (NMS) from the rat hypothalamus as an endogenous ligand for the FM-4/TGR-1 receptor distinct from neuromedin U. In the present study, we examined the role of NMS in the oxytocin release response to suckling stimulation by rat pups. Intracerebroventricular (icv) injection of NMS induced cFos expression in the paraventricular nucleus and supraoptic nucleus. Double immunohistochemical analysis revealed induction of cFos expression in a proportion of oxytocinergic neurons in both nuclei. In addition, icv injection of NMS stimulated oxytocin release dose-dependently in intact rats, and increased milk secretion in lactating rats. On the other hand, icv injection of anti-NMS antiserum into lactating rats significantly suppressed suckling-induced milk ejection. These results suggest that, in the rat, endogenous NMS plays an important role in the oxytocin release response to the suckling stimulus. PMID- 18675785 TI - Ribosomal frameshifting in response to hypomodified tRNAs in Xenopus oocytes. AB - We used Xenopus oocytes as an intracellular system to study ribosomal frameshifting. Microinjection of oocytes with a construct encoding the naturally occurring UUU or AAC codon at the frameshift site demonstrated that the level of frameshifting was similar or lower than found normally in retroviral frameshifting in mammalian cells, suggesting that oocytes are a reliable system to study this event. Phenylalanine (Phe) or asparagine (Asn) tRNAs with and without the highly modified wyebutoxine (Y) or queuosine (Q) base, respectively, were microinjected to assess their ability to promote frameshifting. tRNAPhe+Y inhibited the level of frameshifting, while tRNAPhe-Y promoted frameshifting providing evidence that the hypomodified form does not act only to enhance frameshifting, but is an essential requirement. Both tRNAAsn+Q and tRNAAsn-Q were used indiscriminately in frameshifting, whether the frameshift site contained the wild-type AAC, or the mutant AAU codon, suggesting that Q base modification status does not influence this process. PMID- 18675787 TI - Alternative splicing generates a short BCL6 (BCL6S) isoform encoding a compact repressor. AB - BCL6 gene transcribes two main mRNA isoforms, variant 1 and variant 2, which have distinct transcription start sites. However, these two isoforms encode identical BCL6 protein. Recently, a third variant was sequenced from a human hippocampus cDNA library (DB465062). In this study, we cloned and sequenced a novel BCL6 spliced isoform (BCL6S) which lacks exon 7 that encodes the first two zinc fingers of BCL6. We found a splicing isoform, BCL6S, that excludes exon 7 but retains the last four of the six zinc fingers (ZFs) coding region of the BCL6 gene. BCL6S mRNA and protein were expressed in human cell lines and tissues that expressed BCL6, but accounted for a minor portion of BCL6 transcripts or protein. BCL6S protein was also detectable in BCL6 positive cells. BCL6S could form homodimers or heterodimers with BCL6 and could bind to classical BCL6-binding sites. Luciferase reporter assays demonstrated that BCL6S could effectively repress typical BCL6 target genes. BCL6S is a compact repressor that may have a functional role in normal and neoplastic germinal center B cells. PMID- 18675788 TI - The dihydrolipoamide dehydrogenase of Aeromonas caviae ST exhibits NADH-dependent tellurite reductase activity. AB - Potassium tellurite (K(2)TeO(3)) is extremely toxic for most forms of life and only a limited number of organisms are naturally resistant to the toxic effects of this compound. Crude extracts prepared from the environmental isolate Aeromonas caviae ST catalize the in vitro reduction of TeO32- in a NADH-dependent reaction. Upon fractionation by ionic exchange column chromatography three major polypeptides identified as the E1, E2, and E3 components of the pyruvate dehydrogenase (PDH) complex were identified in fractions exhibiting tellurite reducing activity. Tellurite reductase and pyruvate dehydrogenase activities co eluted from a Sephadex gel filtration column. To determine which component(s) of the PDH complex has tellurite reductase activity, the A. caviae ST structural genes encoding for E1 (aceE), E2 (aceF), and E3 (lpdA) were independently cloned and expressed in Escherichia coli and their gene products purified. Results indicated that tellurite reductase activity lies almost exclusively in the E3 component, dihydrolipoamide dehydrogenase. The E3 component of the PDH complex from E. coli, Zymomonas mobilis, Streptococcus pneumoniae, and Geobacillus stearothermophilus also showed NADH-dependent tellurite reductase in vitro suggesting that this enzymatic activity is widely distributed among microorganisms. PMID- 18675789 TI - Adipogenic genes on induction and stabilization of commitment to adipose conversion. AB - Most late events of adipose conversion are known, but those early events that lead to cell commitment, and important aspects of its mechanism remain unknown. We recently described that, in the absence of any other adipogenic factor, 4h incubation with staurosporine promotes commitment of 3T3-F442A cells to adipogenesis. This commitment consists of two stages; a first stage of 4h induction by staurosporine, and, in the absence of this drug, a second stage of stabilization which becomes completed after 40-48h from staurosporine treatment. Here, we demonstrate that pparg2 gene is expressed early after induction stage but before commitment is stabilized, whereas cebpa is highly expressed during the last part of stabilization stage. A decrease of dlk1 expression, whose down regulation is indispensable for adipogenesis, began to take place between 24 and 48h of St-Dex incubation started, reaching the lowest levels well into the end of stabilization stage. PMID- 18675790 TI - Recombinant human laminin isoforms can support the undifferentiated growth of human embryonic stem cells. AB - Human embryonic stem cells (hESCs) are thought to be a promising cell source for cell transplantation therapy. For such a clinical application, the hESCs should be manipulated using appropriate and qualified materials. In this study, we examined the efficacy of recombinant human laminin (rhLM) isoforms on the undifferentiated growth of hESCs. We first determined the major integrins expressed on the hESCs to reveal the preference of the hESCs for rhLMs, and found that the hESCs mainly expressed integrin alpha6beta1, which binds predominantly to laminin-111, -332 and -511/-521. When the hESCs were seeded onto rhLMs, the cells indeed adhered markedly to rhLM-332, and to rhLM-511 and rhLM-111 to a lesser extent. The hESCs proliferated on these three rhLMs for several passages while preserving their pluripotency. These results show that rhLM-111, -332, and 511 are good substrates to expand undifferentiated hESCs due to their high affinity to integrin alpha6beta1 expressed on hESCs. PMID- 18675792 TI - Cortical representation of rhythmic foot movements. AB - The cortex is involved in rhythmic hand movements. The cortical contribution to rhythmic motor patterns of the feet, however, has never been evaluated in humans. In this study we investigated EEG activity related to rhythmic stepping and tapping movements in 10 healthy subjects. Subjects performed self-paced fast bilateral anti-phase, in-phase and unilateral rhythmic foot movements as well as an isometric cocontraction of the calf muscles, while being seated as relaxed as possible. Surface EMG from the anterior tibial muscles was recorded in parallel with a 64 channel EEG. Power spectra, corticomuscular coherence and corticomuscular delay were calculated. All subjects showed corticomuscular coherence at the stepping frequencies in the central midline region that extended further to the frontal mesial area. The magnitude and the topography of this coherence were equal for the right and left anterior tibial muscle and all movement conditions. During cocontraction there was coherence in the 15-30 Hz range which was refined to the central midline area. EEG-EMG delays were significant in 9 subjects with values between 14 and 26 ms, EMG-EEG feedback was only found in 6 subjects with delays between 25 and 40 ms. We conclude that rhythmic motor patterns of the feet are represented in the cortex, transmitted to the muscles with delays compatible with fast corticospinal transmission and fed back to the cortex. A similar cortical contribution may be important also for gait control in humans. PMID- 18675791 TI - Intracerebroventricular administration of Shiga toxin type 2 altered the expression levels of neuronal nitric oxide synthase and glial fibrillary acidic protein in rat brains. AB - Shiga toxin (Stx) from enterohemorrhagic Escherichia coli (STEC) is the main cause of hemorrhagic colitis which may derive into Hemolytic Uremic Syndrome (HUS) and acute encephalopathy, one of the major risk factors for infant death caused by the toxin. We have previously demonstrated that intracerebroventricular administration of Stx2 causes neuronal death and glial cell damage in rat brains. In the present work, we observed that the intracerebroventricular administration of Stx2 increased the expression of glial fibrillary acidic protein (GFAP) leading to astrogliosis. Confocal microscopy showed reactive astrocytes in contact with Stx2-containing neurons. Immunocolocalization of increased GFAP and Stx2 in astrocytes was also observed. This insult in the brain was correlated with changes in the expression and activity of neuronal nitric oxide synthase (nNOS) by using the NADPH-diaphorase histochemical technique (NADPH-d HT). A significant decrease in NOS/NADPH-d-positive neurons and NOS/NADPH-d activity was observed in cerebral cortex and striatum, whereas an opposite effect was found in the hypothalamic paraventricular nucleus. We concluded that the i.c.v. administration of Stx2 promotes a typical pattern of brain injury showing reactive astrocytes and an alteration in the number and activity of nNOS/NADPH-d. According to the functional state of nNOS/NADPH-d and to brain cell morphology data, it could be inferred that the i.c.v. administration of Stx2 leads to either a neurodegenerative or a neuroprotective mechanism in the affected brain areas. The present animal model resembles the encephalopathy developed in Hemolytic Uremic Syndrome (HUS) patients by STEC intoxication. PMID- 18675793 TI - Serum leptin levels are associated with cognitive function in older adults. AB - Recent work suggests that leptin, a circulating adipokinine hormone, might contribute to age-related cognitive decline. The present study investigated the relationship between serum leptin levels and cognitive function in older adults. Thirty-five older adults (73.69+/-6.62 years of age) without significant neurologic or psychiatric history completed a fasting blood draw and a brief neuropsychological test battery. Partial correlations adjusting for demographic and medical conditions showed that higher leptin levels were associated with poorer performance on Trail Making Test B (r = .46, p = .01). These findings indicate that serum leptin levels are negatively associated with speeded executive function in older adults without significant neurological or psychiatric conditions. The mechanisms for this relationship are unknown and require further examination. Such studies may provide key insight into the mechanisms of age-related cognitive decline and identify possible interventions. PMID- 18675794 TI - The glutamate-rich region of the larger lamprey neurofilament sidearm is essential for proper neurofilament architecture. AB - The carboxyl terminal "tail" domains of the heavy and middle molecular weight mammalian neurofilament (NF) proteins regulate inter-NF spacing and formation of organized networks. The C-terminal region of the larger of the two lamprey NF subunits (NF-180) resembles these mammalian proteins in that it consists of a proximal glutamate-rich region and a distal region containing multiple phosphorylation sites. To investigate the role of these two sidearm domains in the organization of lamprey NFs, we generated plasmids lacking the glutamate-rich domain, the domain containing multiple phosphorylation sites, or both, and examined the impact of the resultant mutant proteins on the endogenous NF network in differentiated NB21/d1 neuroblastoma cells. We present evidence that, like mammalian NFs, the glutamate-rich region of NF-180 sidearm plays a critical role in NF architecture. PMID- 18675795 TI - Cross-frequency phase coupling of brain rhythms during the orienting response. AB - A critical function of the brain's orienting response is to evaluate novel environmental events in order to prepare for potential behavioral action. Here, measures of synchronization (power, coherence) and nonlinear cross-frequency phase coupling (m:n phase locking measured with bicoherence and cross bicoherence) were computed on 62-channel electroencephalographic (EEG) data during a paradigm in which unexpected, highly-deviant, novel sounds were randomly intermixed with frequent standard and infrequent target tones. Low frequency resolution analyses showed no significant changes in phase coupling for any stimulus type, though significant changes in power and synchrony did occur. High frequency resolution analyses, on the other hand, showed significant differences in phase coupling, but only for novel sounds compared to standard tones. Novel sounds elicited increased power and coherence in the delta band together with m:n phase locking (bicoherence) of delta:theta (1:3) and delta:alpha (1:4) rhythms in widespread fronto-central, right parietal, temporal, and occipital regions. Cross bicoherence revealed that globally synchronized delta oscillations were phase coupled to theta oscillations in central regions and to alpha oscillations in right parietal and posterior regions. These results suggest that globally synchronized low frequency oscillations with phase coupling to more localized higher frequency oscillations provide a neural mechanism for the orienting response. PMID- 18675796 TI - A non-ideal replacement for the Boyle van't Hoff equation. AB - A non-ideal osmotic equilibrium equation is proposed as a replacement for the Boyle van't Hoff equation to describe the equilibrium volume of a living cell as a function of external osmolality. Contrary to common understanding, the Boyle van't Hoff equation is only thermodynamically correct for ideal, dilute solutions. However, the Boyle van't Hoff equation is commonly used to determine the osmotically inactive fraction of the cell. This involves extrapolating to infinite osmolality, which violates the ideal, dilute solution constraint. It has been noted that the osmotically inactive fractions obtained from the Boyle van't Hoff equation for human erythrocytes are markedly larger than measured values of the dry volume fraction of the cell. Using the new osmotic equilibrium equation to analyze experimental osmotic equilibrium data reduces the inferred osmotically inactive fraction of human erythrocytes by approximately 20%. PMID- 18675798 TI - WITHDRAWN: Tanshinone IIA protects rat cardiomyocytes against apoptosis induced by angiotensin II in vitro. AB - This article has been withdrawn at the request of the authors. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 18675797 TI - Eye formation in the absence of retina. AB - Eye development is a complex process that involves the formation of the retina and the lens, collectively called the eyeball, as well as the formation of auxiliary eye structures such as the eyelid, lacrimal gland, cornea and conjunctiva. The developmental requirements for the formation of each individual structure are only partially understood. We have shown previously that the homeobox-containing gene Rx is a key component in eye formation, as retinal structures do not develop and retina-specific gene expression is not observed in Rx-deficient mice. In addition, Rx-/- embryos do not develop any lens structure, despite the fact that Rx is not expressed in the lens. This demonstrates that during normal mammalian development, retina-specific gene expression is necessary for lens formation. In this paper we show that lens formation can be restored in Rx-deficient embryos experimentally, by the elimination of beta-catenin expression in the head surface ectoderm. This suggests that beta-catenin is involved in lens specification either through Wnt signaling or through its function in cell adhesion. In contrast to lens formation, we demonstrate that the development of auxiliary eye structures does not depend on retina-specific gene expression or retinal morphogenesis. These results point to the existence of two separate developmental processes involved in the formation of the eye and its associated structures. One involved in the formation of the eyeball and the second involved in the formation of the auxiliary eye structures. PMID- 18675799 TI - The effect of dietary supplementation with limonene or myo-inositol on the induction of neoplasia and matrix metalloproteinase and plasminogen activator activities in accessory sex organs of male Lobund-Wistar rats. AB - Prostate cancer, the most prevalent non-cutaneous cancer in men, is associated with increased age. This suggests that dietary chemopreventive measures could be effective in delaying the onset or decreasing the severity of the disease. We utilized the Lobund-Wistar rat nitrosomethylurea induced, testosterone promoted (NMU-T) model of male sex accessory gland cancer to test the potential chemopreventive effects of myo-inositol and limonene on tumor incidence and associated protease activities. Tumors were found to arise in the seminal vesicles and dorsal and anterior prostate lobes. There were also some tumors that appeared to arise in both the seminal vesicles and anterior prostate, and in some cases the tissue of origin was not clear. The distribution of tumors as to site of origin in limonene or myo-inositol treated animals did not vary from that of the starch fed control animals, and the number of animals presenting with metastases did not vary significantly between treatment groups. There was a statistically significant delay in onset of tumors in myo-inositol, but not limonene fed rats, at 10 months post-induction of carcinogenesis; however, at 12 and 15 months this was not significant. The ventral prostate and seminal vesicles expressed pro-MMP-2 and plasminogen activator (PA) activities. Based on sensitivity to amiloride, the PA activities were predominately urokinase (uPA) in the ventral prostate and a mixture of tissue-type activator (tPA) and uPA in the seminal vesicles of non-treated rats. Sex accessory gland tumors, and metastases, expressed increased levels PA and pro- and active forms of MMP-2 and -9. The PA activities of the tumors were a mixture of uPA and tPA. There was no difference in the levels of these protease activities based on the tissue of tumor origin, nor in tumor vs metastasis. These studies indicate that MMP and PA activities play a role in sex accessory gland tumor biology and that dietary supplementation with myo-inositol can delay but not ultimately prevent the development of such tumors. PMID- 18675800 TI - Glycogen synthase kinase 3beta regulation of nuclear factor of activated T-cells isoform c1 in the vascular smooth muscle cell response to injury. AB - The migration and proliferation of vascular smooth muscle cells (vSMCs) are critical events in neointima formation during atherosclerosis and restenosis. The transcription factor nuclear factor of activated T-cells-isoform c1 (NFATc1) is regulated by atherogenic cytokines, and has been implicated in the migratory and proliferative responses of vSMCs through the regulation of gene expression. In T cells, calcineurin de-phosphorylates NFATc1, leading to its nuclear import, while glycogen synthase kinase 3 beta (GSK3beta) phosphorylates NFATc1 and promotes its nuclear export. However, the relationship between NFATc1 and GSK3beta has not been studied during SMC migration and proliferation. We investigated this by scrape wounding vSMCs in vitro, and studying wound repair. NFATc1 protein was transiently increased, reaching a peak at 8 h after wounding. Cell fractionation and immunocytochemistry revealed that NFATc1 accumulation in the nucleus was maximal at 4 h after injury, and this was coincident with a significant 9 fold increase in transcriptional activity. Silencing NFATc1 expression with siRNA or inhibition of NFAT with cyclosporin A (CsA) attenuated wound closure by vSMCs. Phospho-GSK3beta (inactive) increased to a peak at 30 min after injury, preceding the nuclear accumulation of NFATc1. Overexpression of a constitutively active mutant of GSK3beta delayed the nuclear accumulation of NFATc1, caused a 50% decrease in NFAT transcriptional activity, and attenuated vSMC wound repair. We conclude that NFATc1 promotes the vSMC response to injury, and that inhibition of GSK3beta is required for the activation of NFAT during wound repair. PMID- 18675801 TI - Asynchronous replication dynamics of imprinted and non-imprinted chromosome regions in early mouse embryos. AB - We have used interphase FISH to analyze the replication behavior of four imprinted chromosome regions (Snrpn, Zim1-Peg3, Dlk1-Gtl2, and Igf2r) and five non-imprinted regions in mouse one-cell to morula-stage embryos and embryonic fibroblasts. In general, imprinted chromosome regions showed the expected asynchronous pattern of replication throughout all analyzed stages of preimplantation development and in differentiated cells. The Dlk1-Gtl2 locus which is not expressed and Igf2r which is biallelically expressed in early embryos showed a relaxation of replication asynchrony at the morula stage. Asynchronous replication in zygotes and two-cell embryos was not specific to imprinted regions. Three non-imprinted loci (Emp1-Pbp2-Dyntl1, Hbb-b1-Hbb-b2-Hbb y, and Opa1) as well as one gene-free region on chromosome 7A1 switched from asynchronous replication in one- and two-cell embryos to synchronous replication in 4-cell embryos and later stages. Another gene-free region on chromosome 16C2 showed a more gradual transition from asynchronous to synchronous replication from two-cell to morula-stage embryos. We propose that replication asynchrony contributes to the striking asymmetry between the two parental genomes, which are epigenetically reprogrammed after fertilization into a diploid somatic genome. The switching of non-imprinted genes from asynchronous to synchronous replication may be associated with embryonic genome activation and restoration of transcriptional potential for somatic development. PMID- 18675802 TI - Are rodents an appropriate pre-clinical model for treating spinal cord injury? Examples from the respiratory system. AB - Because most studies of the effects of spinal cord injury (SCI) and resulting repair and treatments use rodent models, it is important to determine if these models are relevant to humans. In this review, we focus on alterations in respiratory function as a result of SCI. Several injury paradigms have been used in the rat to examine restoration of post-lesion respiratory function and potential benefits from repair strategies designed for humans. Unlike the corticospinal locomotor system, respiratory neural organization is well preserved between rodents and humans, and resembles the general organization of motor pathways in primates. These similarities justify the use of the rodent respiratory system as a model to analyze SCI and putative repair strategies. PMID- 18675804 TI - Reduced NGF secretion by Schwann cells under the high glucose condition decreases neurite outgrowth of DRG neurons. AB - BACKGROUND: Schwann cells (SCs) have been supposed to play prominent roles in axonal regeneration under various diseases. Here, to evaluate the direct interaction between SCs and dorsal root ganglion (DRG) neurons under a diabetic condition, the effects of Schwann cell-conditioned media on neurite outgrowth of DRG neurons were investigated. METHODS: Immortalized mouse Schwann cells (IMS) were cultured under 5.5 mM glucose (NG) or 30 mM glucose (HG) conditions for 4 days. IMS-conditioned media (IMS-media) were added to the culture media of neurons isolated from 8-week-old DDY mice. Neurons were cultured for 48 h with or without mouse recombinant NGF (mrNGF) or nerve growth factor (NGF) neutralizing antibody. The concentrations of NGF in IMS-media by ELISA and neurite outgrowth by a computed image analysis system were evaluated. RESULTS: Neurite outgrowth was significantly enhanced by IMS-media (IMS-media (-): 177+/-177 microm, IMS media (+): 1648+/-726). The neurite outgrowth cultured with IMS-media obtained under the HG condition was significantly reduced compared with that under the NG condition (NG: 1474+/-652, HG: 734+/-331). The NGF concentrations were significantly lower in IMS-media under the HG condition than in those under the NG condition. The accelerated neurite outgrowth by IMS-media was inhibited by NGF neutralizing antibody. CONCLUSIONS: These results suggest that SCs play important roles in neurite outgrowth of DRG neurons, and that the decreased NGF secretion by SCs under the diabetic condition would cause a defect of axonal regeneration, resulting in the development of diabetic neuropathy. PMID- 18675803 TI - GABA(A) receptor-mediated excitation in dissociated neurons from human hypothalamic hamartomas. AB - The cellular mechanisms underlying intrinsic epileptogenesis in human hypothalamic hamartoma (HH) are unknown. We previously reported that HH tissue is composed predominantly of GABAergic neurons, but how GABAergic-neuron-rich HH tissue is intrinsically epileptogenic is unclear. Here, we tested the hypotheses that some HH neurons exhibit immature features and that GABA excites these neurons via activation of GABA(A) receptors (GABA(A)Rs). Gramicidin-perforated and cell-attached patch-clamp recordings were performed using freshly-dissociated HH neurons to evaluate GABA(A)R-mediated currents, Cl(-) equilibrium potentials, and intracellular Cl(-) concentrations. Single-cell RT-PCR and immunocytochemical techniques were used to examine cation-Cl(-) co-transporter (NKCC1 and KCC2) gene and KCC2 protein expression and molecular markers of maturation. From a total of 93 acutely-dissociated HH neurons from 34 patients, 76% were small (soma: 6-9 microm) and 24% were large (soma: >20 microm) in size. Under gramicidin perforated patch recording conditions, GABA(A)R activation depolarized/excited large but hyperpolarized/inhibited small HH neurons in most cases. Compared to small HH neurons, large HH neurons exhibited more positive Cl(-) equilibrium potentials, higher intracellular Cl(-) concentrations, lower KCC2 expression, and an immature phenotype, consistent with GABA(A)R-mediated excitation. Taken collectively, we provide novel evidence for and mechanistic insights into HH epileptogenicity: GABA(A)R-mediated excitation. PMID- 18675805 TI - A single residue mutation in Hha preserving structure and binding to H-NS results in loss of H-NS mediated gene repression properties. AB - In this study, we report that a single mutation of cysteine 18 to isoleucine (C18I) in Escherichia coli Hha abolishes the repression of the hemolysin operon observed in the wild-type protein. The phenotype also includes a significant decrease in the growth rate of E. coli cells at low ionic strength. Other substitutions at this position (C18A, C18S) have no observable effects in E. coli growth or hemolysin repression. All mutants are stable and well folded and bind H NS in vitro with similar affinities suggesting that Cys 18 is not directly involved in H-NS binding but this position is essential for the activity of the H NS/Hha heterocomplexes in the regulation of gene expression. PMID- 18675806 TI - Denitration of L-type calcium channel. AB - Tyrosine nitration results in altered function of selective proteins, including human smooth muscle L-type calcium channel, hCa(v)1.2b. We report here that Ca(v)1.2 is also subject to "denitration". Cell lysates from activated macrophage like cell line, RAW264.7 cells, reversed peroxynitrite-induced nitration of the carboxy terminus of Ca(v)1.2 in a 1D gel assay. Tyrosine phosphorylation of the calcium channel by c-src kinase was blocked by nitration but reversed by pretreatment with RAW264.7 cell lysates. These findings indicate that denitration may be a physiological mechanism to restore cellular excitability during inflammation. PMID- 18675807 TI - Internal translation initiation on the foot-and-mouth disease virus IRES is affected by ribosomal stalk conformation. AB - A human cell line, in which expression of the ribosomal stalk proteins P1 and P2 has been suppressed by RNAi technology, has been used to test how the loss of these proteins affects IRES-dependent translation. Foot-and-mouth disease virus (FMDV) IRES-dependent translation from a bicistronic construct is about three fold higher in the P1/P2-depleted cells than in control cells in the presence of Lb protease. By contrast, no effect on Hepatitis C virus (HCV) IRES translation was observed. These results emphasize the functional heterogeneity of the IRES and they highlight a functional connection between the ribosomal stalk and picornavirus IRES-dependent translation. PMID- 18675808 TI - Only plant-type (GLYK) glycerate kinases produce d-glycerate 3-phosphate. AB - D-Glycerate kinases (GK) occur in three phylogenetically distinct classes. Class II GKs produce glycerate 2-phosphate, while both class I GK and class III GK (GLYK) are thought to produce glycerate 3-phosphate. We report on the identification of a bacterial-type class I GK in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 and of a plant-type GLYK in the filamentous cyanobacterium Nostoc sp. strain PCC 7120. The comparison with other prokaryotic and eukaryotic GKs of both classes shows that glycerate 3-phosphate is produced only by the GLYKs, but, in contrast to current thinking, not by any of the examined class I enzymes. PMID- 18675809 TI - Crystal structure of the IL-22/IL-22R1 complex and its implications for the IL-22 signaling mechanism. AB - Interleukin-22 (IL-22) is a member of the interleukin-10 cytokine family, which is involved in anti-microbial defenses, tissue damage protection and repair, and acute phase responses. Its signaling mechanism involves the sequential binding of IL-22 to interleukin-22 receptor 1 (IL-22R1), and of this dimer to interleukin-10 receptor 2 (IL-10R2) extracellular domain. We report a 1.9A crystal structure of the IL-22/IL-22R1 complex, revealing crucial interacting residues at the IL-22/IL 22R1 interface. Functional importance of key residues was confirmed by site directed mutagenesis and functional studies. Based on the X-ray structure of the binary complex, we discuss a molecular basis of the IL-22/IL-22R1 recognition by IL-10R2. PMID- 18675810 TI - Acceptor substrate binding revealed by crystal structure of human glucosamine-6 phosphate N-acetyltransferase 1. AB - Glucosamine-6-phosphate (GlcN6P) N-acetyltransferase 1 (GNA1) is a key enzyme in the pathway toward biosynthesis of UDP-N-acetylglucosamine, an important donor substrate for N-linked glycosylation. GNA1 catalyzes the formation of N acetylglucosamine-6-phosphate (GlcNAc6P) from acetyl-CoA (AcCoA) and the acceptor substrate GlcN6P. Here, we report crystal structures of human GNA1, including apo GNA1, the GNA1-GlcN6P complex and an E156A mutant. Our work showed that GlcN6P binds to GNA1 without the help of AcCoA binding. Structural analyses and mutagenesis studies have shed lights on the charge distribution in the GlcN6P binding pocket, and an important role for Glu156 in the substrate binding. Hence, these findings have broadened our knowledge of structural features required for the substrate affinity of GNA1. PMID- 18675811 TI - Nepmucin/CLM-9, an Ig domain-containing sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro. AB - Nepmucin/CLM-9 is an Ig domain-containing sialomucin expressed in vascular endothelial cells. Here we show that, like CD31, nepmucin was localized to interendothelial contacts and to vesicle-like structures along the cell border and underwent intracellular recycling. Functional analyses showed that nepmucin mediated homotypic and heterotypic cell adhesion via its Ig domain. Nepmucin expressing endothelial cells showed enhanced lymphocyte transendothelial migration (TEM), which was abrogated by anti-nepmucin mAbs that block either homophilic or heterophilic binding. Notably, the mAbs that inhibited homophilic binding blocked TEM without affecting lymphocyte adhesion. These results suggest that endothelial nepmucin promotes lymphocyte TEM using multiple adhesion pathways. PMID- 18675812 TI - Detection of higher-order G protein-coupled receptor oligomers by a combined BRET BiFC technique. AB - Despite some caveats, G protein-coupled receptor oligomerization is a phenomenon that is becoming largely accepted. Within these oligomers, however, stoichiometry remains to be elucidated. Here, by using bimolecular fluorescence complementation, we visualized adenosine A(2A) receptor homodimers in living cells, showing no apparent difference in the subcellular distribution when compared to the YFP-labelled adenosine A(2A) receptor protomer. Interestingly, the combination of bimolecular fluorescence complementation and bioluminescence resonance energy transfer techniques allowed us to detect the occurrence of adenosine A(2A) receptors oligomers containing more than two protomers. These results provide new insights into the molecular composition of G protein-coupled receptor oligomers. PMID- 18675813 TI - The alpha-kinases TRPM6 and TRPM7, but not eEF-2 kinase, phosphorylate the assembly domain of myosin IIA, IIB and IIC. AB - TRPM6 and TRPM7 encode channel-kinases. While these channels share electrophysiological properties and cellular functions, TRPM6 and TRPM7 are non redundant genes raising the possibility that the kinases have distinct substrates. Here, we demonstrate that TRPM6 and TRPM7 phosphorylate the assembly domain of myosin IIA, IIB and IIC on identical residues. Whereas phosphorylation of myosin IIA is restricted to the coiled-coil domain, TRPM6 and TRPM7 also phosphorylate the non-helical tails of myosin IIB and IIC. TRPM7 does not phosphorylate eukaryotic elongation factor-2 (eEF-2) and myosin II is a poor substrate for eEF-2 kinase. In conclusion, TRPM6 and TRPM7 share exogenous substrates among themselves but not with functionally distant alpha-kinases. PMID- 18675814 TI - 5-Hydroxytryptamine 2A receptor signaling cascade modulates adiponectin and plasminogen activator inhibitor 1 expression in adipose tissue. AB - Knowledge of the regulatory factors associated with down-regulation of adiponectin gene expression and up-regulation of PAI-1 gene expression is crucial to understand the pathophysiological basis of obesity and metabolic diseases, and could establish new treatment strategies for these conditions. We showed that expression of 5-HT(2A) receptors was up-regulated in hypertrophic 3T3-L1 adipocytes, which exhibited decreased expression of adiponectin and increased expression of PAI-1. 5-HT(2A) receptor antagonists and suppression of 5-HT(2A) receptor gene expression enhanced adiponectin expression. Activation of Gq negatively regulated adiponectin expression, and inhibition of mitogen-activated protein kinase reversed the Gq-induced effect. Moreover, the 5-HT(2A) receptor blockade reduced PAI-1 expression. These findings indicate that antagonism of 5 HT(2A) receptors in adipocytes could improve the obesity-linked decreases in adiponectin expression and increases in PAI-1 expression. PMID- 18675815 TI - Reversal of asynchrony between circular and longitudinal muscle contraction in nutcracker esophagus by atropine. AB - BACKGROUND & AIMS: Patients with high-amplitude esophageal contractions (nutcracker esophagus [NCE]) show asynchrony of circular muscle (CM) and longitudinal muscle (LM) contraction during peristalsis. The goal of our study was to determine if this asynchrony is related to an increase in the cholinergic receptor activity. METHODS: High-frequency intraluminal ultrasound images and pressures of the esophagus were recorded simultaneously in 10 normal subjects and 10 patients with NCE. Recordings were obtained at 2 cm above the lower esophageal sphincter under 2 study conditions in normal subjects (before and after 80 microgm/kg of edrophonium), and under 3 study conditions in the NCE patients (control, 5 microgm and 10 microgm/kg of atropine). RESULTS: In normal subjects, edrophonium induced an increase in the CM and LM contraction amplitude, an increase in the contraction duration, and asynchrony of LM and CM contraction during peristalsis. On the other hand, increased contraction amplitude, duration, and asynchrony of LM and CM contraction observed at the baseline in the NCE patients were reversed by atropine in a dose-dependent fashion. CONCLUSIONS: These data prove that the esophageal motor abnormalities in patients with nutcracker esophagus, including asynchrony of CM and LM contraction, are related to a hypercholinergic state. PMID- 18675816 TI - Molecular characterization of follicle stimulating hormone receptor (FSHR) gene in the Indian river buffalo (Bubalus bubalis). AB - Follicle stimulating hormone (FSH) plays a central role in regulation of ovarian function in mammals. The actions of follicle stimulating hormone are mediated through receptors present on the granulosa cells of the ovary. In the present study we have cloned and characterized the FSHR gene of buffalo. Sequence analysis indicated that the buffalo FSHR cDNA sequence comprised of an open reading frame of 2085bp encoding a 695 amino acid protein. Its nucleotide sequence showed more than 80% similarity to the homologous genes of mammalian species. At amino acid level buffalo FSHR exhibited a high percentage (84-96.7%) of identity with the corresponding mammalian homologs. This is the first isolation and characterization of FSHR cDNA from buffalo ovary. PMID- 18675817 TI - Genetic differences in coping strategies in response to prolonged and repeated restraint in Japanese quail divergently selected for long or short tonic immobility. AB - Exposure to fearful situations elicits behavioral and Hypothalamic-Pituitary Adrenal (HPA) axis responses characteristic of the coping response of individual animals to counteract environmental challenges. The aim of this study was to investigate behavioral and corticotropic responses concomitantly following prolonged or repeated restraint stress by placing two genotypes of Japanese quail divergently selected for long (LTI) or short (STI) duration of tonic immobility (TI) in a crush cage. In our study, STI quail exhibited higher corticosterone (CORT) levels than LTI quail in response to prolonged restraint. STI quail struggled sooner and much more than LTI quail, and struggling behavior in STI quail progressively decreased during the course of restraint whereas LTI quail displayed very little struggling behavior in the crush cage. LTI quail are thus more likely to adopt a passive behavior coping strategy upon exposure to threat whereas STI quail behave more as active copers. The corticosterone responses shown by LTI and STI quail under restraint stress suggest that adrenocortical correlates of coping behavior in these genotypes of quail may be different from the coping styles previously described in other species. Repeated restraint slightly decreased CORT responses to stress in all experimental groups, but more markedly in male STI quail, whereas adrenal sensitivity and maximum adrenal corticosterone response capacity did not change in any group. On the other hand, neither behavioral habituation nor sensitization processes occurred in the context of repeated restraint in female and male LTI quail and female STI quail, whereas the decreases observed in some behavioral responses were interpreted to be the result of a habituation process in male STI quail. PMID- 18675818 TI - A single exposure to immobilization causes long-lasting pituitary-adrenal and behavioral sensitization to mild stressors. AB - We have previously reported that a single exposure to immobilization (IMO) in rats causes a long-term desensitization of the hypothalamic-pituitary-adrenal (HPA) response to the same (homotypic) stressor. Since there are reports showing that a single exposure to other stressors causes sensitization of the HPA response to heterotypic stressors and increases anxiety-like behavior, we studied in the present work the long-term effects of IMO on behavioral and HPA response to mild superimposed stressors. In Experiments 1 and 2, adult male Sprague-Dawley rats were subjected to 2 h of IMO and then exposed for 5 min to the elevated plus maze (EPM) at 1, 3 or 7 days after IMO. Blood samples were taken at 15 min after initial exposure to the EPM. Increases in anxiety-like behavior and HPA responsiveness to the EPM were found at all times post-IMO. Changes in the resting levels of HPA hormones did not explain the enhanced HPA responsiveness to the EPM (Experiment 3). In Experiments 4 and 5, we studied the effects of a single exposure to a shorter session of IMO (1 h) on behavioral and HPA responses to a brief and mild session of foot-shocks done 10 days after IMO. Neither previous IMO nor exposure to shocks in control rats modified behavior in the EPM. However, a brief session of shocks in previously IMO-exposed rats dramatically increased anxiety in the EPM. HPA and freezing responses to shocks were similar in control and previous IMO groups. Therefore, a single exposure to IMO appears to induce long-lasting HPA and behavioral sensitization to mild superimposed stressors, although the two responses are likely to be at least partially independent. Long-term effects of IMO on the susceptibility to stress-induced endocrine and emotional disturbances may be relevant to the characterization of animal models of post-traumatic stress. PMID- 18675819 TI - An efficient culture method for generating large quantities of mature mouse splenic macrophages. AB - In this study, we established an efficient in vitro culture method for generating mature splenic macrophages (Sp-Mphi). Splenocytes were cultured in the presence of conditioned medium containing macrophage colony-stimulating factor (M-CSF) for 7 days post post-isolation and the generated Sp-Mphi were characterized phenotypically and functionally. Through this method, 9 x 10(6)/mouse Sp-Mphi were obtained in comparison to 2 x 10(5)/mouse when Mphi were cultured in regular medium. In addition, the purity of these cells was as high as 80% by day 5 and >90% by day 7 of culturing, confirmed with Mphi-specific markers. The increased Sp-Mphi yields, in the presence of M-CSF, point towards the existence of a precursor population in the spleen that can be influenced to differentiate into Sp-Mphi. Moreover, we compared the maturation of generated Sp-Mphi to conventional bone marrow-derived Mphi (BM-Mphi) in vitro. Interestingly, Sp-Mphi exhibited lower capacity to phagocytose dead cells after 3 days of maturation, but showed similar internalizing capacity after 5 and 7 of maturation to BM-Mphi cultured for the same time period. Importantly, Sp-Mphi upregulated the expression of several surface markers such as MOMA-2 and CD68 while downregulating SIGN-R1 after 7 days, indicating that these Sp-Mphi undergo further maturation in vitro due to culturing in M-CSF. Taken together, we describe and validate a method for generating Sp-Mphi in large quantities and high purity. These data should prove valuable in future studies characterizing the functions and maturation of Sp-Mphi. PMID- 18675820 TI - Gene expression and lysosomal content of silkworm peritracheal athrocytes. AB - To examine the function of silkworm Bombyx mori L. athrocytes (nephrocytes), we constructed cDNAs of larval peritracheal athrocytes that were anatomically isolated from surrounding tissues. Larval expression levels of genes encoding hemolymph proteins, such as arylphorin, the 30K proteins, and lysozyme, were lower in peritracheal athrocytes than in the fat body, whereas genes involved in protein degradation were highly expressed in athrocytes. Real time RT-PCR revealed that a member of the Hsp40/Dnaj protein family, DjA2 (also known as Rdj2, Dj3, Dnj3, Cpr3, and Hirip4), an endocytic gene, was highly expressed in the peritracheal athrocytes compared to the fat body. Homologs of the Drosophila ATG1, ATG5, ATG6, and ATG8 genes had high expression levels in the peritracheal athrocytes. Observations using laser confocal microscopy with lysosomal fluorescent probes showed that silkworm athrocytes, including pericardial cells, suboesophageal body, and peritracheal athrocytes, were rich in lysosomes, in contrast to other tissues. Peritracheal athrocytes had lysotracker-positive spots at all times from the fourth larval molt to the pupa. Of these, molting larval and pupal peritracheal athrocytes had larger spots. Starvation for 24h induced greater lysotracker staining, but the number of spots decreased. Silkworm peritracheal athrocytes are lysosome-rich tissues and may function in the degradation of proteins. PMID- 18675821 TI - Upregulation of the immune protein gene hemolin in the epidermis during the wandering larval stage of the Indian meal moth, Plodia interpunctella. AB - Expression of hemolin, which generates an immune protein, was up-regulated in wandering fifth instar larval stage of Plodia interpunctella. The mRNA level peaked in the middle of the wandering stage. Major expression was in the epidermis, rather than in the fat body or gut. To test a possible ecdysteroid effect on hemolin induction we treated with RH-5992, an ecdysteroid agonist, and KK-42, which inhibits ecdysteroid biosynthesis in both feeding and wandering fifth instar larvae. When feeding larvae were treated with RH-5992 the hemolin mRNA level was increased. When wandering larvae were treated with KK-42 its level was reduced. In addition, when KK-42-treated larvae were subsequently treated with RH-5992 the hemolin mRNA level was recovered. These results strongly suggest that ecdysteroid up-regulates the expression of hemolin mRNA. Hormonal and bacterial effects on hemolin induction were further analyzed at the tissue level. Major induction of hemolin mRNA was detected following both RH-5992 treatment and bacterial injection in the epidermis of both feeding and wandering larvae. Minor induction of hemolin was detected in the fat body following a bacterial injection, but not RH-5992 treatment. We infer that in P. interpunctella larvae, the epidermis is the major tissue for hemolin induction in naive insects and in insects manipulated with bacterial and hormonal treatments. PMID- 18675822 TI - Tyramine as a possible neurotransmitter/neuromodulator at the spermatheca of the African migratory locust, Locusta migratoria. AB - Tyramine-like immunoreactivity was identified in neurons of the VIIIth abdominal ganglion and in axons projecting to the spermatheca of adult females of Locusta migratoria. Tyramine-like immunoreactive processes were also found throughout all regions of the spermatheca and tyramine-like immunoreactive bipolar or multipolar neurons were present on the spermathecal sac. HPLC coupled with electrochemical detection revealed more tyramine than octopamine present in spermathecal tissue. Electrical stimulation of the ventral ovipositor nerve resulted in a significant increase in calcium-dependent release of tyramine from the spermatheca. Both tyramine and octopamine increase the frequency and basal tonus of spermathecal contractions in a dose-dependent manner, with octopamine having a lower threshold. When tyramine is applied along with a half maximal octopamine dose, there is an additive effect on contractions of the spermatheca with slight synergistic effects at lower doses of tyramine. High concentrations of tyramine (10(-4)M) stimulated increases in cyclic AMP levels of the spermatheca; an effect blocked by phentolamine. Phentolamine has a higher affinity (and thus a lower IC(50) value congruent with 5.6 x 10(-8)M) than yohimbine (IC(50) congruent with 1.1 x 10(-4)M) in reducing tyramine-induced spermathecal contractions. Taken together, these results suggest that tyramine may be a co-transmitter with octopamine at the spermatheca, with both neuroactive chemicals acting on an octopamine receptor. PMID- 18675823 TI - Hypertonia-associated protein Trak1 is a novel regulator of endosome-to-lysosome trafficking. AB - Hypertonia, which is characterized by stiff gait, abnormal posture, jerky movements, and tremor, is associated with a number of neurological disorders, including cerebral palsy, dystonia, Parkinson's disease, stroke, and spinal cord injury. Recently, a spontaneous mutation in the gene encoding trafficking protein, kinesin-binding 1 (Trak1), was identified as the genetic defect that causes hypertonia in mice. The subcellular localization and biological function of Trak1 remain unclear. Here we report that Trak1 interacts with hepatocyte growth-factor-regulated tyrosine kinase substrate (Hrs), an essential component of the endosomal sorting and trafficking machinery. Double-label immunofluorescence confocal studies show that the endogenous Trak1 protein partially colocalizes with Hrs on early endosomes. Like Hrs, both overexpression and small-interfering-RNA-mediated knockdown of Trak1 inhibit degradation of internalized epidermal growth factor receptors through a block in endosome-to lysosome trafficking. Our findings support a role for Trak1 in the regulation of Hrs-mediated endosomal sorting and have important implications for understanding hypertonia associated with neurological disorders. PMID- 18675824 TI - IL-22R, IL-10R2, and IL-22BP binding sites are topologically juxtaposed on adjacent and overlapping surfaces of IL-22. AB - Interleukin (IL) 22 is a type II cytokine that is produced by immune cells and acts on nonimmune cells to regulate local tissue inflammation. As a product of the recently identified T helper 17 lineage of CD4(+) effector lymphocytes, IL-22 plays a critical role in mucosal immunity as well as in dysregulated inflammation observed in autoimmune diseases. We used comprehensive mutagenesis combined with mammalian cell expression, ELISA cell-based, and structural methods to evaluate how IL-22 interacts with its cell surface receptor, IL-22R/IL-10R2, and with secreted IL-22 binding protein. This study identifies those amino acid side chains of IL-22 that are individually important for optimal binding to IL-22R, considerably expands the definition of IL-22 surface required for binding to IL 10R2, and demonstrates how IL-22 binding protein prevents IL-22R from binding to IL-22. The IL-22R and IL-10R2 binding sites are juxtaposed on adjacent IL-22 surfaces contributed mostly by helices A, D, and F and loop AB. Our results also provide a model for how IL-19, IL-20, IL-24, and IL-26 which are other IL-10-like cytokines, interact with their respective cell surface receptors. PMID- 18675825 TI - mGlu5 receptor deletion does not confer seizure protection to mice. AB - Metabotropic glutamate mGlu5 receptors have been implicated in the regulation of seizures and have been suggested as a target against which discovery of novel anticonvulsants may be possible. However, the experimental literature is not consistent in reporting anticonvulsant efficacy of mGlu5 receptor antagonists. Additional assessment of this target was approached in the present study by comparing convulsions in wild-type (WT) and mGlu5 receptor null (knockout or KO) mice. Chemically induced seizures induced by a variety of mechanisms including pentylenetetrazole, N-methyl-d-aspartic acid (NMDA), cocaine, kainic acid, aminophylline, 4-aminopyridine, strychnine, and nicotine did not differentially increase clonic, clonic/tonic, or lethality in WT vs. mGlu5 receptor KO mice. The mGlu5 receptor antagonist 3-[(2-Methyl-1,3-thiazol-4-yl) ethynyl]-pyridine (MTEP) did not significantly prevent seizures induced by NMDA; in contrast, the uncompetitive NMDA receptor antagonist, dizocilpine, significantly prevented NMDA induced seizures and lethality in both WT and KO mice. The present findings do not support the idea that mGlu5 receptors play as important a role in seizure control as previously speculated. PMID- 18675826 TI - The phosphatidylinositol 3-kinase-mediated production of interferon-beta is critical for the lipopolysaccharide inhibition of osteoclastogenesis. AB - AIMS: This study was performed to define molecular mechanisms underlying lipopolysaccharide (LPS)-induced inhibition of osteoclastogenesis. MAIN METHODS: The LPS-dependent signaling pathways leading to the inhibition of osteoclastogenesis were examined using chemical inhibitors and neutralizing antibodies. KEY FINDINGS: Lipopolysaccharide (LPS) has been shown to induce massive bone loss in vivo by stimulating osteoclast differentiation and activity. However, the direct effect of LPS on osteoclastogenesis in vitro is complex. Based on the context of the differentiation state of precursors, LPS has been shown to either augment or inhibit osteoclast differentiation. When added to receptor activator of NF-kappaB ligand (RANKL)-primed precursors, LPS enhances osteoclast differentiation via the production of TNF-alpha. On the other hand, LPS inhibits osteoclastogenesis from early precursors like bone marrow macrophages by hitherto unknown mechanism. In the present study, we investigated the mechanism by which LPS inhibits osteoclastogenesis. We have identified that the phosphatidylinositol 3-kinase (PI 3-kinase) dependent production of IFN-beta and resultant inhibition of c-Fos expression upon LPS stimulation of bone marrow macrophages are responsible for the LPS-induced inhibition of osteoclastogenesis. Inhibition of PI 3-kinase, neutralization of IFN-beta, and overexpression of c Fos respectively prevented the LPS-induced inhibition of osteoclast differentiation. SIGNIFICANCE: Our results provide a molecular understanding of the differentiation stage-specific dual effect of LPS on osteoclastogenesis. PMID- 18675827 TI - The tumor suppressor homolog in fission yeast, myh1(+), displays a strong interaction with the checkpoint gene rad1(+). AB - The DNA glycosylase MutY is strongly conserved in evolution, and homologs are found in most eukaryotes and prokaryotes examined. This protein is implicated in repair of oxidative DNA damage, in particular adenine mispaired opposite 7,8 dihydro-8-oxoguanine. Previous investigations in Escherichia coli, fission yeast, and mammalian cells show an association of mutations in MutY homologs with a mutator phenotype and carcinogenesis. Eukaryotic MutY homologs physically associate with several proteins with a role in replication, DNA repair, and checkpoint signaling, specifically the trimeric 9-1-1 complex. In a genetic investigation of the fission yeast MutY homolog, myh1(+), we show that the myh1 mutation confers a moderately increased UV sensitivity alone and in combination with mutations in several DNA repair genes. The myh1 rad1, and to a lesser degree myh1 rad9, double mutants display a synthetic interaction resulting in enhanced sensitivity to DNA damaging agents and hydroxyurea. UV irradiation of myh1 rad1 double mutants results in severe chromosome segregation defects and visible DNA fragmentation, and a failure to activate the checkpoint. Additionally, myh1 rad1 double mutants exhibit morphological defects in the absence of DNA damaging agents. We also found a moderate suppression of the slow growth and UV sensitivity of rhp51 mutants by the myh1 mutation. Our results implicate fission yeast Myh1 in repair of a wider range of DNA damage than previously thought, and functionally link it to the checkpoint pathway. PMID- 18675828 TI - UGT1A1 and UGT1A9 functional variants, meat intake, and colon cancer, among Caucasians and African-Americans. AB - Glucuronidation by the UDP-glucuronosyltransferase enzymes (UGTs) is one of the primary detoxification pathways of dietary heterocyclic amines (HCAs) and polycyclic aromatic hydrocarbons (PAHs). In a population-based case-control study of 537 cases and 866 controls, we investigated whether colon cancer was associated with genetic variations in UGT1A1 and UGT1A9 genes and we determined if those variations modify the association between colon cancer and dietary HCA and PAH exposure. We measured functional UGT1A1 polymorphisms at positions -53 (28; A(TA)6TAA to A(TA)7TAA), -3156 (G>A), -3279 (T>G) and the UGT1A9-275(T>A) polymorphism, and found no association with colon cancer overall. However, when stratified by race, the UGT1A1-3279 GG/TG intermediate/low activity genotypes were associated with an increased risk of colon cancer (odds ratio (OR)=1.5, 95% confidence interval (CI)=1.1-2.0) in Caucasians. This finding is also supported by haplotype analyses where the UGT1A1-3279G-allele-bearing haplotype is overrepresented in case group. Overall, UGT1A1-53 and -3156 genotypes modified the association between dietary benzo(a)pyrene (BaP) and colon cancer (P for interaction=0.02 and 0.03, respectively). The strongest association was observed for those with <7.7 ng/day BaP exposure and the low activity genotypes, for both UGT1A1 28/28 (OR=1.8, 95% CI=1.1-2.9) and -3156AA (OR=1.7, 95% CI=1.0-3.0), compared to >or=7.7 ng/day and combined high/intermediate genotypes. These data support a hypothesis that UGTs modify the association between meat-derived PAH exposure and colon cancer by their role in the elimination of dietary carcinogens. PMID- 18675829 TI - Effects of serotonin in failing cardiac ventricle: signalling mechanisms and potential therapeutic implications. AB - Previously, cardioexcitation by serotonin (5-hydroxytryptamine, 5-HT) was believed to be confined to atria in mammals including man, and mediated through 5 HT(4) receptors in pig and man, but 5-HT(2A) receptors in rat. Recent studies, reviewed here, demonstrate that functional 5-HT(4) receptors can be revealed in porcine and human ventricular myocardium during phosphodiesterase inhibition, and that 5-HT(4) receptor mRNA is increased in human heart failure. In rats, functional 5-HT(4) and 5-HT(2A) receptors appear in the cardiac ventricle during heart failure and mediate inotropic responses through different mechanisms. 5 HT(2A) receptor signalling resembles that from alpha(1)-adrenoceptors and causes inotropic effects through increased myosin light chain phosphorylation, resulting in Ca(2+) sensitisation. 5-HT(4) receptor signalling resembles that from beta adrenoceptors and causes inotropic effects through a pathway involving cAMP and PKA-mediated phosphorylation of proteins involved in Ca(2+) handling, resulting in enhanced contractility through increased Ca(2+) availability. Cyclic AMP generated through 5-HT(4) receptor stimulation seems more efficiently coupled to increased contractility than cAMP generated through beta-adrenoceptor stimulation. Increasing contractility through cAMP is considered less energy efficient than Ca(2+) sensitisation and this may be one reason why beta adrenoceptor antagonism is beneficial in heart failure patients. Treatment of heart failure rats with the 5-HT(4) antagonist SB207266 (piboserod) resulted in potentially beneficial effects, although small. Further studies are needed to clarify if such treatment will be useful for patients with heart failure. PMID- 18675830 TI - Calcium modulation of 5-HT3 receptor binding and function. AB - Calcium modulates the 5-HT3 receptor response by reducing peak current amplitude and increasing rates of activation, deactivation and desensitisation, but the binding site(s) and mechanism(s) of this modulation are unknown. Here we study residues that may be involved in calcium binding in two partially overlapping regions of the extracellular domain (E213-E215-E218 and D204-E218-V219). The modulatory effects of calcium were assessed by radioligand binding and whole-cell patch-clamp. Comparisons of [3H]granisetron binding showed an increase in Kd in 10mM calcium that was abolished by the substitutions E213Q, E215Q, D204N and V219L. E218Q mutant receptors displayed no specific binding or function, and immunofluorescence showed that they did not reach the cell surface. E213Q increased inherent rates of desensitisation, but the relative effects of calcium on these rates, and on the reduction in current amplitude, were similar to wild type receptors. Current responses and calcium-mediated effects at E215Q mutant receptors were indistinguishable from wild type. D204N and V219L mutants were non functional. A calcium impermeable mutant (E277A/S297R) revealed no changes in peak amplitude or kinetics with increased calcium. Our results are consistent with residues D204, E218 and V219 participating in receptor assembly, structure and/or trafficking to the plasma membrane, and we speculate that this might rely upon the stabilising effect of bound calcium. E213, E215, D204 and V219 may contribute to a calcium binding site that is responsible for the calcium-mediated effects on ligand binding. However, the major site for calcium-dependent modulation of the 5-HT3 current is located within the ion channel or cell interior. PMID- 18675831 TI - Blockade of group II metabotropic glutamate receptors produces hyper-locomotion in cocaine pre-exposed rats by interactions with dopamine receptors. AB - It was previously reported that blockade of group II metabotropic glutamate receptors (mGluRs) produces hyper-locomotion in rats previously exposed to amphetamine, indicating that group II mGluRs are well positioned to modulate the expression of behavioral sensitization by amphetamine. The present study further examined the locomotor activating effects of specific blockade of these receptors after cocaine pre-exposures. First, rats were pre-exposed to seven daily injections of cocaine (15mg/kg, IP). When challenged the next day with an injection of either saline or the group II mGluR antagonist LY341495 (0.5, 1.0 or 2.5mg/kg, IP), they produced hyper-locomotor activity, measured by infrared beam interruptions, to LY341495 compared to saline in a dose-dependent manner. Second, rats were pre-exposed to either saline or seven daily injections of cocaine (15mg/kg, IP). Three weeks later, when they were challenged with an injection of either saline or LY341495 (1.0mg/kg, IP), only rats pre-exposed to cocaine produced hyper-locomotor activity to LY341495 compared to saline. These effects, however, were not present when dopamine D1 (SCH23390; 5 or 10microg/kg), but not D2 (eticlopride; 10 or 50microg/kg), receptor antagonist was pre-injected, indicating that this cocaine-induced hyper-locomotor activity to LY341495 may be mediated in dopamine D1 receptor-dependent manner. These results suggest that group II mGluRs may be adapted to interact with dopaminergic neuronal signaling in mediating the sensitized locomotor activity produced by repeated cocaine pre exposures. PMID- 18675833 TI - Blinks of the eye predict blinks of the mind. AB - The Attentional Blink (AB)--a deficit in reporting the second of two target stimuli presented in close succession in a rapid sequence of distracters--has been related to individual processing limitations of working memory. Given the known role of dopamine (DA) in working memory processes, the present experiment tested the hypothesis that DA, and in particular the DA/D1 subsystem, plays a role in the AB. We present evidence that the spontaneous eyeblink rate (EBR), a functional marker of central dopaminergic function, reliably predicts the size of AB. Thus, in line with our hypothesis, these data point to a modulatory role for DA in the AB. PMID- 18675832 TI - Glutamate transmission in addiction. AB - Cortico-striatal glutamate transmission has been implicated in both the initiation and expression of addiction related behaviors, such as locomotor sensitization and drug-seeking. While glutamate transmission onto dopamine cells in the ventral tegmental area undergoes transient plasticity important for establishing addiction-related behaviors, glutamatergic plasticity in the nucleus accumbens is critical for the expression of these behaviors. This information points to the value of exploring pharmacotherapeutic manipulation of glutamate plasticity in treating drug addiction. PMID- 18675834 TI - Alliesthesia to food cues: heterogeneity across stimuli and sensory modalities. AB - Negative alliesthesia to olfactory and visual stimuli was assessed in 29 normal weight women who, on alternate days, were either fasting or in a postprandial state after an ad libitum lunch. The participants were alternatively exposed to food and non-food pictures and odorants, and then rated for their hedonic appreciation (liking) and their desire to ingest (wanting) the evoked foods. While negative alliesthesia was observed only for food stimuli, it did not equally affect all food categories in either sensory modality. The stimuli representing foods eaten in typical local main dishes or having high energy density (e.g., pizza, bacon, beef, cheese) evoked clear negative alliesthesia, whereas this was not the case for those less consumed within a customary meal or associated with desserts (i.e., fruits). Furthermore, the visual food stimuli triggered a more negative shift in liking than did the food odours. Finally, the shift in wanting between pre- and post-meal state was more important than the shift in liking. These results suggest that alliesthesia may be influenced by both metabolic and non-metabolic factors. PMID- 18675835 TI - Why do heavy littermates grow better than lighter ones? A study in wild and domestic European rabbits. AB - Birth mass can vary considerably among mammalian littermates. Heavier pups often show higher growth rates than their lighter siblings, which might positively affect fitness-relevant parameters during later life. Such a correlation between birth mass and pre-weaning growth within litters was confirmed by our study of wild-type and domestic European rabbits (Oryctolagus cuniculus) living in a semi natural environment and under laboratory housing conditions, respectively. Our study indicates that at least two main mechanisms account for this relationship in our study species: heavier pups had a higher milk intake and also showed a more efficient conversion of milk into body mass. Furthermore, our study suggests that the better milk conversion by heavy pups was driven by three synergistic mechanisms: heavier pups had comparatively more huddling partners in the nest, they did not need to perform large amounts of proactive behavior in order to reach and remain in a central position within the litter huddle, and they could maintain a comparatively higher body temperature most probably due to their more favorable surface area to volume ratio. In conclusion, our study of European rabbits provides strong evidence that both under natural conditions and in the laboratory, within-litter differences in birth mass are maintained and may even increase during pups' early postnatal development. PMID- 18675836 TI - Shaping the light/dark pattern for circadian adaptation to night shift work. AB - This is the second in a series of simulated night shift studies designed to achieve and subsequently maintain a compromise circadian phase position between complete entrainment to the daytime sleep period and no phase shift at all. We predict that this compromise will yield improved night shift alertness and daytime sleep, while still permitting adequate late night sleep and daytime wakefulness on days off. Our goal is to delay the dim light melatonin onset (DLMO) from its baseline phase of approximately 21:00 to our target of approximately 3:00. Healthy young subjects (n=31) underwent three night shifts followed by two days off. Two experimental groups received intermittent bright light pulses during night shifts (total durations of 75 and 120 min per night shift), wore dark sunglasses when outside, slept in dark bedrooms at scheduled times after night shifts and on days off, and received outdoor light exposure upon awakening from sleep. A control group remained in dim room light during night shifts, wore lighter sunglasses, and had unrestricted sleep and outdoor light exposure. After the days off, the DLMO of the experimental groups was approximately 00:00-1:00, not quite at the target of 3:00, but in a good position to reach the target after subsequent night shifts with bright light. The DLMO of the control group changed little from baseline. Experimental subjects performed better than control subjects during night shifts on a reaction time task. Subsequent studies will reveal whether the target phase is achieved and maintained through more alternations of night shifts and days off. PMID- 18675837 TI - Social context rather than behavioral output or winning modulates post-conflict testosterone responses in Japanese quail (Coturnix japonica). AB - Testosterone regulates the expression of sexual and aggressive behavior in male vertebrates and treatments with testosterone may promote territorial aggression and winning in dyadic contests. Conversely, individual testosterone levels respond to sexual or aggressive interactions and the social environment. Post conflict testosterone in winner males though appears to be more complex than simply reflecting conflict outcome. Expression and degree of post-conflict testosterone responses may adapt to additional modulators such as repeated winning experience, audience presence, opponent's fighting ability, and self assessment. We present simulated intrusion experiments with male Japanese quail using mirror-elicited aggression and fights with real opponents ('direct challenge'). We recorded agonistic behavior and measured immunoreactive testosterone metabolites (TM) non-invasively from individual droppings. Frequencies of initiated agonistic behavior were similar whether towards the mirror or in direct challenge tests, although some of the males were behaviorally non-responsive to the mirror ('mirror submissives'). However, there was no TM response to the mirror test in both, mirror fighters and mirror submissives, thus independently of behavioral output. After direct challenges TM levels were elevated in all males (focal males winning or conflict unresolved after 30 min), hence independently of conflict outcome. Thus, in male quail a combination of physical stimuli and the individual perception of own and opponent's fighting ability explained the expression of post-conflict TM responses rather than behavioral performance, conflict outcome, or any of these factors alone. In sum, our results emphasize that the degree of androgen responsiveness to agonistic behavior is fine-tuned by components related with social context and environment. PMID- 18675838 TI - Databases applicable to quantitative hazard/risk assessment--towards a predictive systems toxicology. AB - The Workshop on The Power of Aggregated Toxicity Data addressed the requirement for distributed databases to support quantitative hazard and risk assessment. The authors have conceived and constructed with federal support several databases that have been used in hazard identification and risk assessment. The first of these databases, the EPA Gene-Tox Database was developed for the EPA Office of Toxic Substances by the Oak Ridge National Laboratory, and is currently hosted by the National Library of Medicine. This public resource is based on the collaborative evaluation, by government, academia, and industry, of short-term tests for the detection of mutagens and presumptive carcinogens. The two-phased evaluation process resulted in more than 50 peer-reviewed publications on test system performance and a qualitative database on thousands of chemicals. Subsequently, the graphic and quantitative EPA/IARC Genetic Activity Profile (GAP) Database was developed in collaboration with the International Agency for Research on Cancer (IARC). A chemical database driven by consideration of the lowest effective dose, GAP has served IARC for many years in support of hazard classification of potential human carcinogens. The Toxicological Activity Profile (TAP) prototype database was patterned after GAP and utilized acute, subchronic, and chronic data from the Office of Air Quality Planning and Standards. TAP demonstrated the flexibility of the GAP format for air toxics, water pollutants and other environmental agents. The GAP format was also applied to developmental toxicants and was modified to represent quantitative results from the rodent carcinogen bioassay. More recently, the authors have constructed: 1) the NIEHS Genetic Alterations in Cancer (GAC) Database which quantifies specific mutations found in cancers induced by environmental agents, and 2) the NIEHS Chemical Effects in Biological Systems (CEBS) Knowledgebase that integrates genomic and other biological data including dose-response studies in toxicology and pathology. Each of the public databases has been discussed in prior publications. They will be briefly described in the present report from the perspective of aggregating datasets to augment the data and information contained within them. PMID- 18675839 TI - Increase of the cytotoxic effect of Bothrops jararacussu venom on mouse extensor digitorum longus and soleus by potassium channel blockers and by Na(+)/K(+) ATPase inhibition. AB - We investigated the myotoxicity of Bothrops jararacussu crude venom and other cytolytic agents on mouse isolated extensor digitorum longus (EDL) and soleus (SOL) muscles, which present distinct properties: EDL is a fast-twitch, white muscle with predominantly glycolytic fibers, while SOL is slow-twitch, red muscle with predominantly oxidative fibers. Muscles were exposed to B. jararacussu crude venom (25 microg/ml) and other crotaline venoms (Agkistrodon contortrix laticinctus; Crotalus viridis viridis; Crotalus durissus terrificus) at the same concentration. Basal creatine kinase (CK) release to bathing solution was 0.43+/ 0.06 for EDL and 0.29+/-0.06 for SOL (U g(-)(1) h(-)(1), n=36 for each muscle). Sixty minutes after exposure to B. jararacussu venom, EDL presented higher increase in the rate of CK release than SOL, respectively, 13.2+/-1.5 and 2.9+/ 0.7 U g(-)(1)h(-)(1), n=10-12. Muscle denervation, despite decreasing CK content, did not affect sensitivities to B. jararacussu venom. Ouabain and potassium channel blockers (TEA; clotrimazole; glibenclamide) increased the rate of CK release by B. jararacussu in EDL and SOL muscles, decreasing and almost abolishing the different sensitivity. When we exposed EDL or SOL muscles to Naja naja, Apis mellifera venoms (25 microg/ml), or Triton X-100 (0.01%), they showed similar rate of CK release. Our present data suggest that a mechanism involving intracellular calcium regulation or potassium channels may participate in the different sensitivity of EDL and SOL to B. jararacussu venom. PMID- 18675840 TI - Long-wave sensitivity in the masked greenling (Hexagrammos octogrammus), a shallow-water marine fish. AB - Microspectrophotometry (MSP) revealed that surprisingly for a "fully marine" species, in summer, photoreceptors of the nearshore scorpaeniform fish known as the masked greenling, Hexagrammos octogrammus, contained exclusively, or presumably, porphyropsin with a small admixture of rhodopsin. As a result of this, the lambda(max) of the spectral sensitivity of the photoreceptors were significantly shifted to longer wavelengths as compared to the lambda(max) typical of marine shallow-water fishes, showing about 530 nm for rods and single cones, and 570/625 nm for double-cone members. These unique spectral shifts would permit a cone-driven wavelength discrimination in spite of high-density orange corneal filters which block light at lower wavelengths. PMID- 18675841 TI - Radial frequency adaptation suggests polar-based coding of local shape cues. AB - The study of shape processing in the human visual system has frequently employed radial frequency (RF) patterns as conveniently manipulable stimuli. This study uses an adaptation paradigm to investigate how local shape information is sampled in the processing of RF contour shapes. Experiment 1 measured thresholds for detecting a fixed mean radius RF contour following adaptation to RF patterns which, in separate conditions, varied in mean radius and radial frequency. Results reveal that, adaptation is strongly tuned for RF over a range of pattern radii, but is not tuned for the number of cycles of radial modulation per visual degree of contour length; a characteristic that changes with both radius and radial frequency. Experiment 2 manipulated the polar angle separation on the fronto-parallel plane between curvature features on a fixed RF by foreshortening the pattern appearance (consistent with a rotation in depth) and shows that RF shape processing is tuned for fronto-parallel separation angles between curvature features. Results were near identical when a stereo rotation cue was added to the perspective modified RF. In the second part of Experiment 2 we showed that RF shape adaptation is also tuned for the polar angular extent of the curvature represented by the lobe at that angle. Collectively, our results indicate that the polar angle at which local curvature features appear, in addition to the angular extent of the curvature feature at that location, are both critical parameters for coding specific RF shapes. PMID- 18675842 TI - A mechanistic inter-species comparison of spatial contrast sensitivity. AB - The validity of the Rovamo-Barten modulation transfer function model for describing spatial contrast sensitivity in vertebrates was examined using published data for the human, macaque, cat, goldfish, pigeon and rat. Under photopic conditions, the model adequately described overall contrast sensitivity for changes in both stimulus luminance and stimulus size for each member of this diverse range of species. From this examination, optical, retinal and post retinal neural processes subserving contrast sensitivity were quantified. An important retinal process is lateral inhibition and values of its associated point spread function (PSF) were obtained for each species. Some auxiliary contrast sensitivity data obtained from the owl monkey were included for these calculations. Modeled values of the lateral inhibition PSF were found to correlate well with ganglion cell receptive field surround size measurements obtained directly from electrophysiology. The range of vertebrates studied was then further extended to include the squirrel monkey, tree shrew, rabbit, chicken and eagle. To a first approximation, modeled estimates of lateral inhibition PSF width were found to be inversely proportional to the square root of ganglion cell density. This finding is consistent with a receptive field surround diameter that changes in direct proportion to the distance between ganglion cells for central vision. For the main species examined, contrast sensitivity is considerably less than that for the human. Although this is due in part to a reduction in the performance of both optical and retinal mechanisms, the model indicates that poor cortical detection efficiency plays a significant role. PMID- 18675843 TI - The expression of locomotor sensitization to apomorphine is dependent on time interval between injection and testing. AB - This study examined the onset of locomotor sensitization induced by apomorphine as a function of the temporal delay between drug injection and testing. In experiment 1, rats received three daily administrations of 2.0 mg/kg apomorphine or vehicle either immediately (0 min) or 20 min before being placed into the test environment for 20 min test sessions. Apomorphine given immediately before testing induced a stimulant effect during the first session and sensitization by the second session. However, when testing was delayed 20 min, apomorphine induced stimulant effects only after the third injection. In experiment 2, separate groups received a single 2.0 (mg/kg) apomorphine/vehicle injection immediately before being placed into the test environment for 60 min. In this experiment, apomorphine induced a stimulant effect at 0-20 and 20-40 min. However, the 20-40 interval increase in locomotion was relative to the low level of activity in the vehicle group and was not greater than the 0-20 min locomotion of the vehicle group. Thus, sensitization depends both on peak drug concentration and habituation state of the control group. The variable post-injection delays could be a useful method to study sensitization because it can avoid ceiling effects and changing baselines in the control groups. PMID- 18675844 TI - Effectiveness of a nutrition program in reducing symptoms of respiratory morbidity in children: a randomized field trial. AB - OBJECTIVES: To assess the effectiveness of home visits advising mothers about breast feeding and weaning in reducing symptoms of respiratory morbidity at the age of 12 months. METHODS: A randomized field trial was conducted with mothers who gave birth within the public health system in the Brazilian city of Sao Leopoldo (2001/2002). The intervention group received dietary advice during the first year based on a Brazilian national health policy for primary care, which is based on WHO guidelines. Morbidity data was assessed in 397 children at 12 months. RESULTS: A total of 23.3% of the children in the intervention group and 39.7% of the controls had one or more of the following symptoms during the month preceding assessment: cough, stuffy nose, runny nose, or breathlessness. The risk of respiratory morbidity symptoms was 41% lower for the intervention group (RR: 0.59; 95% CI: 0.43-0.81). The number of families needed to be visited to avoid one children presenting symptoms of respiratory morbidity (Number Needed to Treat) was 6.1. The intervention-group status was also associated with a longer duration of exclusive (RR:1.59; 95% CI: 1.21-2.07; p=0.001) and total breastfeeding (RR: 1.25; 95% CI:1.02-1.55; p=0.032) later introduction of solid foods (RR:1.11; 95% CI: 1.02-1.21; p=0.023), and a lower proportion of current medication use (RR:0.56; 95% CI:0.34-0.91;p=0.016). CONCLUSION: A nutrition education program during the first year of life has a positive impact on reducing respiratory symptoms in infants. PMID- 18675845 TI - Long-term effectiveness of interventions promoting physical activity: a systematic review. AB - BACKGROUND: This systematic review aims to evaluate the long-term effectiveness of physical activity interventions targeted at healthy adults and to identify effective intervention components. METHODS: The systematic search in Pubmed, Embase, the Cochrane Library and Psycinfo identified literature published in English and German between January 2001 and June 2007. We included randomised controlled trials investigating physical activity interventions for healthy adults with a minimum study duration of 12 months. Two researchers independently assessed publications according to pre-defined inclusion criteria and methodological quality was rated according to the SIGN criteria. Study characteristics and outcome measures were extracted, and pooled effect estimates with 95% confidence intervals calculated. RESULTS: Of 5508 identified publications 25 studies met the inclusion criteria. There was substantial heterogeneity in study quality, intervention strategies and intervention effects. Compared to no-intervention and minimal-intervention control, gains in weekly energy expenditure and physical fitness of up to 975 kcal and 11% were achieved, respectively. Booster interventions were used in 16 studies. CONCLUSIONS: There is evidence for long-term increases in physical activity behaviour and physical fitness. To improve uptake of physical activity additional tailored exercise prescription strategies seem promising. Booster interventions such as phone, mail or internet can help to facilitate long-term effectiveness. PMID- 18675846 TI - Functional neuroimaging of reward processing and decision-making: a review of aberrant motivational and affective processing in addiction and mood disorders. AB - The adequate integration of reward- and decision-related information provided by the environment is critical for behavioral success and subjective well being in everyday life. Functional neuroimaging research has already presented a comprehensive picture on affective and motivational processing in the healthy human brain and has recently also turned its interest to the assessment of impaired brain function in psychiatric patients. This article presents an overview on neuroimaging studies dealing with reward processing and decision making by combining most recent findings from fundamental and clinical research. It provides an outline on the neural mechanisms guiding context-adequate reward processing and decision-making processes in the healthy brain, and also addresses pathophysiological alterations in the brain's reward system that have been observed in substance abuse and mood disorders, two highly prevalent classes of psychiatric disorders. The overall goal is to critically evaluate the specificity of neurophysiological alterations identified in these psychiatric disorders and associated symptoms, and to make suggestions concerning future research. PMID- 18675847 TI - Association between the interleukin-1beta polymorphisms and Alzheimer's disease: a systematic review and meta-analysis. AB - The pro-inflammatory cytokine interleukin(IL)-1beta is a main component in inflammatory pathways and is overexpressed in the brain of Alzheimer's disease (AD) patients. Several studies report associations between IL-1beta polymorphisms and AD, but findings from different studies are controversial. Our aim was to verify the correlation between the single nucleotide polymorphisms (SNPs) of the IL-1beta, at sites -511 and +3953, and AD by meta-analysis. Computerized bibliographic searches of PUBMED and AlzGene database (http://www.alzgene.org) were supplemented with manual searches of reference lists. There is evidence for association between IL-1beta +3953 SNP and AD, with an OR=1.60 (95% C.I.: 1.16 2.22; Z=2.83 p=0.005) for TT genotype. No significant difference in genotype distribution of the IL-1beta -511 SNP in AD was obtained, but high between-study heterogeneity was found. To reduce heterogeneity, subgroup analyses were performed using, as stratifying variables, characteristics of the population under study (age, gender, type of AD diagnosis, Mini Mental State Examination of the controls) and characteristics related to the study design (statistical power of individual studies). The frequency of the IL-1beta -511 TT genotype resulted significantly higher than other genotypes only when the Caucasian studies with the highest statistical power were included in the subgroup analysis (OR=1.32; 95% C.I.: 1.03-1.69; p=0.03), with no evidence of between-study heterogeneity. Our data support an association between the TT genotype of IL-1beta +3953 SNP and AD, and suggest a possible association of the -511 TT genotype. Unreplicability of the results seems to be due mainly to the lack of statistical power of the individual studies. PMID- 18675848 TI - CD83 on murine APC does not function as a costimulatory receptor for T cells. AB - The transmembrane glycoprotein CD83 is rapidly upregulated on murine and human DC upon maturation and therefore a costimulatory function for T cell activation has been suggested. Studies employing human APC indeed showed that CD83 expression was positively correlated to the stimulatory capacity of the APC. Murine APC that were CD83 deficient however, did not display a reduced capacity to activate T cells. To elucidate this contradiction, we thoroughly compared the stimulatory capacity of CD83-overexpressing and CD83-deficient APC. Here we show that CD83 expression levels on APC did not affect the capacity of the APC to activate CD8(+) T cells. CD83 expression levels did not significantly affect CD4(+) T cell activation in vivo, but a weak positive correlation of CD83 expression with CD4(+) T cell activation was observed in vitro under suboptimal stimulation conditions. As CD83 expression also positively correlated with MHC-II but not with MHC-I expression, this differential stimulation specifically of CD4(+) T cells could be explained by a higher density of MHC-II peptide complexes on the APC surface. Taken together, our results strongly suggest that CD83 does not deliver crucial costimulatory signals to murine T cells. PMID- 18675849 TI - BEI Resources: supporting antiviral research. AB - The Biodefense and Emerging Infections Research Resources Repository (BEI Resources) provides unique, quality-assured reagents to the scientific community for use in basic research and product development involving biodefense and emerging infectious diseases. These include microorganisms (up to Biosafety Level 3) on the National Institute of Allergy and Infectious Diseases (NIAID) and Centers for Disease Control and Prevention (CDC) lists of Category A, B and C priority pathogens. In addition to live microorganisms, related products such as polyclonal antisera, monoclonal antibodies, isolated nucleic acid preparations, overlapping peptide arrays, purified proteins, and assay kits are also available. Many of these materials have direct or indirect applications in antiviral research. These reagents are available free of charge to all registered investigators, regardless of funding source or affiliation. Acquisition of new reagents for the repository is one of the critically necessary and challenging tasks for BEI Resources. Therefore, investigators are encouraged to deposit relevant items, so as to provide access to materials, relief from the burden of distribution, protection of intellectual property rights, and secure storage. In addition, BEI Resources has the capability of contracting for the preparation of specific reagents. If there is a resource needed to advance a specific research area, contact an NIAID program officer or use the "suggest a reagent" option on the BEI Resources homepage, (www.beiresources.org). PMID- 18675850 TI - Treatment of Venezuelan equine encephalitis virus infection with (-)-carbodine. AB - Venezuelan equine encephalitis virus (VEEV) may cause encephalitis in humans, for which no FDA-approved antiviral treatment is available. Carbocyclic cytosine (carbodine) has broad-spectrum activity but toxicity has limited its utility. It was anticipated that one of the enantiomers of carbodine would show enhanced activity and reduced toxicity. The activity of the d-(-) enantiomer of carbodine [(-)-carbodine] was evaluated by infectious cell culture assay and was found to have a 50% effective concentration (EC50) of 0.2 microg/ml against the TC-83 vaccine strain of VEEV in Vero cells, while the l-(+) enantiomer had no activity. Virus titer inhibition correlated with intracellular cytidine triphosphate reduction after treatment with (-)-carbodine, as determined by HPLC analysis. Pre treatment with 200 mg/(kgd) resulted in significant improvement in survival, virus load in the brain, weight change, and mean day-to-death in a mouse model of TC-83 VEEV disease. A single dose of (-)-carbodine resulted in a slight extension of mean time to death in mice infected with wild-type VEEV. Post-virus exposure treatment with (-)-carbodine was effective in significantly improving disease parameters in mice infected with TC-83 VEEV when treatment was initiated as late as 4 days post-virus installation (dpi). It is remarkable that (-)-carbodine is effective when initiated after the establishment of brain infection. PMID- 18675851 TI - Interplay between glutamate and serotonin within the dorsal periaqueductal gray modulates anxiety-related behavior of rats exposed to the elevated plus-maze. AB - Several neurotransmitters, including glutamate and serotonin, modulate defensive behaviors related to anxiety in the rat dorsal periaqueductal gray (PAG). Although both glutamate N-methyl-d-aspartic acid (NMDA) and serotonin type 1-A (5 HT(1A)) receptors have been shown to interfere with these subtle responses, such as inhibitory avoidance, a possible interaction between them remains to be examined. To address this issue, the present study investigated whether the activation or the blockage of 5-HT(1A) receptors located in the dorsal PAG would interact with NMDA function in animals exposed to the elevated plus-maze task. The effect of the NMDA (25 pmol) was evaluated in rats pretreated with the 5 HT(1A) receptor antagonist WAY-100135 (2.0 or 5.0 nmol). In addition, the effect of the NMDA (100 pmol) was evaluated in rats pretreated with the 5-HT(1A) receptor agonist 8-OH-DPAT (2.0 or 8.0 nmol). Intra-dorsal PAG injection of NMDA (25 pmol) increased inhibitory avoidance behavior. This anxiogenic-like effect of the NMDA was counteracted by the pretreatment with WAY-100135 (5.0 nmol). Although 100 pmol of NMDA failed to increase inhibitory avoidance in the vehicle pretreated group, in rats pretreated with 8-OH-DPAT this NMDA dose produced an anxiogenic-like effect. These results suggest that 5-HT(1A) and NMDA receptors interact in the dorsal PAG to modulate the anxiety-related behavior. PMID- 18675852 TI - Rodent models of serial reaction time tasks and their implementation in neurobiological research. AB - Since the late eighties of the last century, when the so-called serial reaction time (SRT) task was first presented, analysis of sequential learning and performance has developed into a prosperous area of neurocognitive research in human subjects and non-human primates. In more recent years, rodent implementations of SRT tasks have become available, which will be summarized here and discussed with respect to their advantages, but also to their methodological and theoretical drawbacks. Then, a review of new neurobiological findings with such rodent tasks will be provided, that will be embedded into the existing theoretical framework, originally obtained from the work with primate species. Among others, this review will show that rodent SRT tasks allow actualizing and thus, studying many psychological features of human sequential tasks, for example, cognitive complexity of sequences. Since these rodent tasks can be used for a number of neuronal analyses, it is assumed that they will provide important insights for the neural mechanisms underlying such cognitive functions. PMID- 18675853 TI - Acid extrusion from the intraerythrocytic malaria parasite is not via a Na(+)/H(+) exchanger. AB - The intraerythrocytic malaria parasite, Plasmodium falciparum maintains an intracellular pH (pH(i)) of around 7.3. If subjected to an experimentally imposed acidification the parasite extrudes H(+), thereby undergoing a pH(i) recovery. In a recent study, Bennett et al. [Bennett TN, Patel J, Ferdig MT, Roepe PD. P. falciparum Na(+)/H(+) exchanger activity and quinine resistance. Mol Biochem Parasitol 2007;153:48-58] used the H(+) ionophore nigericin, in conjunction with an acidic medium, to acidify the parasite cytosol, and then used bovine serum albumin (BSA) to scavenge the nigericin from the parasite membrane. The ensuing Na(+)-dependent pH(i) recovery, seen following an increase in the extracellular pH, was attributed to a plasma membrane Na(+)/H(+) exchanger. This is at odds with previous reports that the primary H(+) extrusion mechanism in the parasite is a plasma membrane V-type H(+)-ATPase. Here we present evidence that the Na(+) dependent efflux of H(+) from parasites acidified using nigericin/BSA is attributable to Na(+)/H(+) exchange via residual nigericin remaining in the parasite plasma membrane, rather than to endogenous transporter activity. PMID- 18675854 TI - Effect of exenatide on gastric emptying and relationship to postprandial glycemia in type 2 diabetes. AB - OBJECTIVES: To evaluate the effect of exenatide on gastric emptying (GE) in type 2 diabetes using scintigraphy. METHODS: Seventeen subjects with type 2 diabetes participated in a randomized, single-blind, 3-period, crossover study. In each 5 day period, 5 or 10 microg exenatide or placebo was administered subcutaneously BID. Oral antidiabetic treatments were continued. The presence of cardiac autonomic neuropathy was assessed during screening. On day 5, after the morning dose, subjects consumed a 450-kcal breakfast containing (99m)Tc-labeled eggs and (111)In-labeled water, and GE was measured by scintigraphy. Plasma glucose and insulin, perceptions of appetite, and plasma exenatide were also quantified. RESULTS: Exenatide slowed GE of both solid and liquid meal components [solid (T(50)(90% confidence interval [CI]); placebo, 60(50-70) min; 5 microg exenatide, 111(94-132) min; 10 microg exenatide, 169(143-201) min; both P<0.01); liquid (T(50)(90% CI), placebo, 34(25-46) min; 5 microg exenatide, 87(65-117) min; 10 microg exenatide, 114(85-154) min; both P<0.01)]. GE was not different between subjects with cardiac autonomic neuropathy (n=7), compared with those without (n=10) (P>/=0.68). Exenatide reduced postprandial glucose (area under the curve [AUC((0-6 h))]) by 69-76% and peak insulin (C(max)) by 84-86% compared with placebo. There was an inverse relationship between the postprandial rise in glucose (AUC((0-3 h))) and GE (solid T(50), r=-0.49, P<0.001). CONCLUSIONS: Exenatide slows GE substantially in type 2 diabetes, which could be an important mechanism contributing to the beneficial effect of exenatide on postprandial glycemia. PMID- 18675855 TI - Visualizing chromatin dynamics in intact cells. AB - Chromatin and associated regulatory proteins regulate gene expression in the natural environment of the intact cell nucleus. Specific combinations of DNA binding transcription factors and recruited coregulatory proteins alter the conformation of chromatin at promoters and enhancers of target genes to stimulate or repress transcription. The dynamic nature of the regulatory proteins active in these processes allows the cell to modulate gene expression very rapidly, an important feature in many physiological processes. Live cell imaging and photobleaching studies of fluorescently-tagged proteins reveal that many transcription factors and other chromatin-associated proteins rapidly move through the nucleoplasm. Transcription factors also transiently interact with specific regulatory sequences in chromatin, suggesting that gene activation does not require the formation of stable long-lived regulatory complexes on the chromatin. In this review we discuss how dynamic interactions allow transcriptional regulatory proteins find their targets within the nucleus, alter target chromatin structure, and modulate physiological gene expression. PMID- 18675856 TI - Novel identification and differentiation of Brucella melitensis, B. abortus, B. suis, B. ovis, B. canis, and B. neotomae suitable for both conventional and real time PCR systems. AB - We describe the development of a novel PCR assay for the rapid detection of members of the Brucella genus, and the differentiation between six recognized Brucella species. The assay has proven to be highly specific with the additional advantage of being suitable for use with both conventional and real-time PCR. PMID- 18675857 TI - Protective effect of isoflavones from Trifolium pratense on dopaminergic neurons. AB - In the present study, protective effect of five isoflavones (formononetin, daidzein, pratensein, calycosin and irilone) from Trifolium pratense on lipopolysaccharide-induced dopaminergic neurodegeneration was studied for the first time. The results showed that all five isoflavones attenuated LPS-induced decrease in dopamine uptake and the number of dopaminergic neurons in a dose dependent manner in rat mesencephalic neuron-glia cultures. Moreover, they also significantly inhibited LPS-induced activation of microglia and production of tumor necrosis factor-alpha, nitric oxide and superoxide in mesencephalic neuron glia cultures and microglia-enriched cultures. In addition, the rank order of protective potency of five isoflavones was: pratensein>daidzein>calycosin>formononetin>irilone. This study suggested that all five isoflavones protected dopaminergic neurons against LPS-induced injury through inhibition of microglia activation and proinflammatory factors generation. PMID- 18675858 TI - A chemical lipid modification of recombinant preS antigen to study the mechanism of HBV attachment to the host cell. AB - Surface antigen preS of Hepatitis B virus plays fundamental roles in mediating receptor recognition and virus internalization. Myristoylation at N-terminal Gly(2) residue of preS is essential for viral attachment and infectivity. A number of myristoylated proteins have been shown to undergo a conformational change (myristoyl switch) that alters their affinity to cell membrane. However, there is little knowledge about what effect this fatty acylation contributes in virus-host cell interaction. Here we demonstrated a new method for lipid modification of recombinant preS protein at N-terminal residue 2 with alkylating chemicals. Modified preS was able to inhibit HBV penetrating into HepG2 cells with an increased efficiency compared to unmodified form. Flow cytometric analysis indicated that lipid modification enhanced the binding affinity of preS to hepatocytes, but not resulting from hydrophobic interaction. CD analysis further revealed a conformational change of modified preS in the presence of membrane mimetics. These findings imply that the conformation transition induced by fatty acylation is important for efficient attachment of virus to cell receptors, and this method of chemical lipid modification provides a basis for designing therapeutic inhibitors to Hepatitis B virus. PMID- 18675859 TI - Role of RNA chaperones in virus replication. AB - RNA molecules are functionally diverse in part due to their extreme structural flexibility that allows rapid regulation by refolding. RNA folding could be a difficult process as often molecules adopt a spatial conformation that is very stable but not biologically functional, named a kinetic trap. RNA chaperones are non-specific RNA binding proteins that help RNA folding by resolving misfolded structures or preventing their formation. There is a large number of viruses whose genome is RNA that allows some evolutionary advantages, such as rapid genome mutation. On the other hand, regions of the viral RNA genomes can adopt different structural conformations, some of them lacking functional relevance and acting as misfolded intermediates. In fact, for an efficient replication, they often require RNA chaperone activities. There is a growing list of RNA chaperones encoded by viruses involved in different steps of the viral cycle. Also, cellular RNA chaperones have been involved in replication of RNA viruses. This review briefly describes RNA chaperone activities and is focused in the roles that viral or cellular nucleic acid chaperones have in RNA virus replication, particularly in those viruses that require discontinuous RNA synthesis. PMID- 18675860 TI - Allopurinol, an inhibitor of purine catabolism, enhances susceptibility of tobacco to Tobacco mosaic virus. AB - Tobacco (cv. Xanthi nn) plants were watered with allopurinol [4-hydroxypyrazolo (3,4-d) pyrimidine, HPP], a xanthine oxidoreductase (XOR) inhibitor, to investigate its effects on infection by Tobacco mosaic virus engineered to express the green fluorescent protein (TMV.GFP). TMV.GFP infection was monitored by examination of inoculated leaves under UV light, by confocal scanning laser microscopy and by epifluorescence microscopy. Susceptibility to TMV.GFP was enhanced in HPP-treated plants. This was seen as a statistically significant increase in numbers of infection sites per leaf and in the number of infected cells per infection site. Two hypotheses are discussed to explain the enhanced susceptibility. The inhibition exerted by HPP against XOR activity could provoke either (i) an increased adenine and guanine nucleotide pool, which could facilitate viral RNA synthesis or (ii) it could cause changes in IAA/auxin levels, which has been proposed to influence TMV susceptibility in tobacco. PMID- 18675861 TI - Divergent picornavirus IRES elements. AB - Internal ribosome entry site (IRES) elements were first identified about 20 years ago within the 5' untranslated region of picornavirus RNAs. They direct a cap independent mechanism of translation initiation on the viral RNA. Within the picornavirus family it is now known that there are four classes of IRES element which vary in size (450-270 nt), they also have different, complex, secondary structures and distinct requirements for cellular proteins to allow them to function. This review describes the features of each class of picornavirus IRES element but focuses on the characteristics of the most recently described group, initially identified within the porcine teschovirus-1 RNA, which has strong similarities to the IRES elements from within the genomes of hepatitis C virus and the pestiviruses which are members of the flavivirus family. The selection of the initiation codon by these distinct IRES elements is also discussed. PMID- 18675862 TI - Polyhydroxyethylaspartamide-spermine copolymers: efficient vectors for gene delivery. AB - Aim of this paper was that to prepare biocompatible, polyaspartamide based copolymers containing spermine or spermine/hydrophobic side chains able to condense nucleic acids and to transfect mammalian cells. Copolymers were prepared starting from alpha,beta-poly-(N-2-hydroxyethyl)-D,L-aspartamide (PHEA) and exploiting the reactive hydroxyl groups in the polymeric side chains by subsequent activation reactions to obtain PHEA-Spermine (PHEA-Spm) and PHEA Spermine-Butyramide (PHEA-Spm-C(4)). Molecular, physico-chemical and biological characterization of copolymers and interpolyelectrolyte complexes with plasmid DNA was performed. Experimental results evidenced that these copolymers are able to form complexes with plasmid DNA already at low polycation/DNA weight ratio ranging from 0.75/1 to 2/1. Interpolyelectrolyte complexes with decreased size were obtained when increasing the polycation/DNA weight ratio, until nanosized dimensions were reached. Copolymers as well as complexes were not haemolytic and non toxic in vitro. In vitro cell transfection with PHEA derivatives showed good biocompatibility and high transfection efficiency (luciferase) in cancer cells in comparison with commercially available, but toxic transfection agents. PMID- 18675863 TI - Ghrelin induces proliferation in human aortic endothelial cells via ERK1/2 and PI3K/Akt activation. AB - The direct ghrelin (Ghr) involvement in cardiovascular (CV) system homeostasis has been suggested by the expression of its receptor in CV tissues and by evidence that ghrelin mediates CV activities in animals and in humans. Moreover, low Ghr plasma levels have been reported in pathological conditions characterized by high cardiovascular risk. In the present study, we investigated Ghr effect on proliferation of human aortic endothelial cell (HAEC) and involved transduction pathways. Our results indicate that ghrelin elicited proliferation in a dose dependent manner (EC(50) about of 5nmol/L) in cultured HAEC, and that this effect was inhibited by the receptor antagonist (D-Lys3)-GHRP-6. Western blot experiments documented an activation of external receptor activated kinases (ERK1/2) and Akt in a dose-dependent fashion, as well as involvement of the cAMP pathway in ERK1/2 phosphorylation. Experiments conducted with appropriate pharmacological inhibitors to investigate Ghr-induced HAEC proliferation confirmed the involvement of ERK1/2 and I3P/Akt pathways, as well as the role of AMP cyclase/PKA pathway in ERK1/2 phosphorylation. Our results indicate that Ghr promotes HAEC proliferation, and thus may be a protective factor against vascular damage. The low ghrelin serum levels reported in insulin-resistant states may not be able to effectively counteract endothelial cell injury. PMID- 18675864 TI - Synthesis and preliminary evaluation of [(18)F]-fluoro-(2S)-Exaprolol for imaging cerebral beta-adrenergic receptors with PET. AB - Cerebral beta-adrenergic receptors (beta-ARs) are of interest in several disorders including Parkinson's disease, Alzheimer's disease and in particular major depressive disorder. Development of a positron emission tomography (PET) ligand for imaging beta-ARs would allow the quantification of these receptors in the living human brain so as to better understand both the pathophysiology of depression and how to improve treatment. Currently there are no radioligands suitable for this purpose. In an attempt to achieve this goal, we prepared [(18)F]-labeled (2S)-1-(1-fluoropropan-2-ylamino)-3-(2-cyclohexylphenoxy)propan-2 ol (fluoro-Exaprolol; (2S)-1). Radiolabeling with fluorine-18 was accomplished via preparation of a precursor containing a tosyl leaving group (10), and utilizes the 2-oxazolidinone group to simultaneously protect both the amine and hydroxy groups. The oxazolidinone was readily removed with lithium aluminum hydride following a nucleophilic [(18)F]-fluoride for tosyl displacement to prepare [(18)F]-(2S)-1 in 31% radiochemical yield (uncorrected for decay), with >98% radiochemical purity in <1h. The specific activity of the formulated product was 927 mCi/micromol and the log P (pH 7.4) was 2.97. Preliminary biological evaluations in conscious rats indicated that [(18)F]-(2S)-1 had good brain uptake for imaging (0.8-1.3% injected dose/gram (% ID/g) of wet tissue, 5 min post injection of the radiotracer) with a slow washout (>0.5% ID/g at 60 min post injection) in all brain regions. Pharmacological challenges indicate that the binding is largely non-specific, as administration of Propranolol, authentic (2S) 1, or WAY 100635 prior to injection of [(18)F]-(2S)-1 did not block uptake of the radiotracer. These results indicate that [(18)F]-(2S)-1 is not a suitable candidate for PET imaging of cerebral beta-ARs. PMID- 18675866 TI - Efficient immune responses against Intimin and EspB of enterohaemorragic Escherichia coli after intranasal vaccination using the TLR2/6 agonist MALP-2 as adjuvant. AB - Mucosal vaccine formulations based on purified recombinant C280 gamma-Intimin and EspB (Escherichia coli secreted protein B) from enterohaemorragic E. coli co administered with a pegylated derivative of the TLR2/6 agonist MALP-2 (macrophage activating lipopeptide) as adjuvant were evaluated in BALB/c mice. After intranasal vaccination, strong humoral and cellular immune responses were observed against C280 gamma-Intimin and EspB. Sera of immunized mice inhibit bacterial haemolytic activity in vitro. Antigen-specific T-cell proliferation, IL 4, IL-2 and IFN-gamma producing cells, and secretory IgA were mostly detected in animals receiving MALP-2 as adjuvant. These results suggest that C280 gamma Intimin and EspB are good candidate antigens to be incorporated into mucosal vaccines against this important pathogen. PMID- 18675867 TI - Evaluation of recombinant Onchocerca volvulus activation associated protein-1 (ASP-1) as a potent Th1-biased adjuvant with a panel of protein or peptide-based antigens and commercial inactivated vaccines. AB - Alum, the only adjuvant approved for clinical applications, can induce strong humoral (Th2) but weak cellular (Th1) immune responses. It is necessary to develop safe and effective adjuvants capable of inducing both humoral and cellular immune responses. We previously showed that activation-associated protein-1 (ASP-1) derived from Onchocerca volvulus has potent adjuvant activity. In this study, we have further evaluated the adjuvanticity of recombinant ASP-1 using a panel of recombinant proteins or synthetic peptide-based antigens, including ovalbumin (OVA), synthetic HIV peptide (HIV-p), recombinant HIV gp41 (rgp41) and HBV HBsAg, as well as three commercially available inactivated vaccines against haemorrhagic fever with renal syndrome (HFRS), Influenza and Rabies. Our results indicate that ASP-1 induced significantly higher IgG1 (Th2 associated) and IgG2a (Th1-associated) responses than alum adjuvant against OVA antigen, HIV-p, and rgp41. Consistently, it induced similar level of IgG1 responses as alum but higher level of IgG2a and IFN-gamma-producing T cell responses than alum adjuvant against HBsAg. Further, ASP-1 improved both IgG1 and IgG2a responses to three commercial inactivated vaccines when used separately or in combination. In conclusion, the recombinant ASP-1, unlike alum adjuvant, is able to induce both Th1 and Th2-associated humoral responses and Th1 cellular responses, suggesting that it can be further developed as a promising adjuvant for subunit-based and inactivated vaccines. PMID- 18675868 TI - Immunogenicity assay of the Leishmune vaccine against canine visceral leishmaniasis in Brazil. AB - Leishmune is the industrialized version of the FML-saponin vaccine which has been shown to develop 92-95% protection in vaccinated dogs and 76-80% vaccine efficacy against field canine visceral leishmaniasis (CVL) in Brazil. Leishmune has been proven to be safe and tolerable and a transmission-blocking vaccine which renders vaccinated dogs non-infectious to sand fly vectors. In the present investigation, 550 healthy seronegative dogs of endemic and epidemic areas of Brazil were monitored for Leishmune-induced immunogenicity during a 2-year trial. Another group of 588 untreated exposed dogs was also studied in parallel. Both groups were seronegative on day 0. The strong immunogenicity induced by Leishmune vaccine was demonstrated by the 98% of FML-seroconversion, increase in absorbencies, the 82.7% DTH positive reactions and increase in skin test size diameters, the average increase in CD8+ total lymphocytes population in blood (27.1%), expected for QS21 saponin-containing vaccine, the sustained proportions of CD4+ T cells, and the average increased proportions of CD21+ B lymphocytes (42.3%). The Leishmune-induced protection against CVL is demonstrated by the results: 98.8% asymptomatic dogs (at the end of first year) and 99% healthy survivors (at the end of the second year) among vaccinated dogs, compared to the 79.4% asymptomatic and 61% survivor dogs (p<0.001) monitored in the untreated exposed cohort. In spite of the low vaccine coverage, it was possible to detect a 66.1% (p<0.005) reduction in Belo Horizonte and an 80.2% (p<0.005) reduction in Aracatuba of the incidence of CVL among vaccinated dogs, when compared to the global incidence of CVL of each town, respectively. Our preliminary results support the potential use of Leishmune to prevent CVL epidemics. PMID- 18675869 TI - Evaluation of Brucella melitensis B115 as rough-phenotype vaccine against B. melitensis and B. ovis infections. AB - Brucella melitensis strain Rev1 is used as vaccine for the prophylaxis of brucellosis in sheep and goats. Because of its smooth phenotype, however, it induces antibodies directed to the O-polysaccharide (O-PS) of the lipopolysaccharide (LPS), thus unabling to distinguish between vaccinated and infected animals. It has been speculated that alternative vaccines could be live, attenuated Brucella rough strains, which are devoid of the O-PS. B. melitensis B115 is a natural, attenuated, rough strain. The O-PS is not exposed at the surface but is present in the cytoplasm. We tested the protective activity of B115 against B. melitensis and B. ovis infections in mice, in comparison with that of Rev1. The residual virulence and the humoral response following B115 vaccination were also evaluated. Vaccination with B115 conferred significant protective immunity against B. melitensis 16M and B. ovis challenge strains, equivalent to that provided by Rev1. No interfering antibodies to O-PS were detected, while the B115 vaccination was monitored by a specific B115-based complement fixation test. These promising features suggest further evaluation of B. melitensis B115 as vaccine for target animal hosts. PMID- 18675870 TI - Safety and immunogenicity of a DTaP-IPV(Vero) (serum-free) combination vaccine in comparison to DTaP-IPV(Mkc) when administered simultaneously with Haemophilus influenzae type B conjugate vaccine (PRP-T) in children at 2, 3.5, 5 and 16 months of age. AB - In a phase III, double blind, randomized, noninferiority, multi-centre clinical trial, 817 infants were included and randomly assigned to vaccination with DTaP IPV(Vero) (N=410) or DTaP-IPV(Mkc) (N=407) vaccines (Statens Serum Institut (SSI), Denmark) in the right thigh. All infants were vaccinated with Act-HIB (Sanofi Pasteur, France) in the left thigh at the same time. The vaccination schedule was 2, 3.5, 5 and 16 months and serum samples were obtained at 6, 16 and 17 months. The primary objective was to demonstrate noninferiority of DTaP IPV(Vero) to DTaP-IPV(Mkc) as regards immunological protection against polio virus types 1, 2 and 3. Furthermore, the immunogenicity of all vaccine antigens and the safety profile of the vaccines were assessed. The study demonstrated that DTaP-IPV(Vero) was noninferior to DTaP-IPV(Mkc). All antibody concentrations/titres remained at an acceptable level from the end of the primary vaccination series (i.e. 2, 3.5 and 5 months) until the time of the booster vaccination at 16 months. A good booster response was, furthermore, demonstrated for all antigens. No vaccine-related serious adverse events and no injection site granulomas or swelling of the entire thigh occurred. The frequencies of local injection site erythema and swelling as well as systemic adverse events such as fever, irritability, somnolence and decreased appetite were low and acceptable in both treatment groups. In conclusion, DTaP-IPV(Vero) is immunogenic and safe for primary vaccination and for booster vaccination of healthy children. PMID- 18675871 TI - Induction of multispecific Th-1 type immune response against HCV in mice by protein immunization using CpG and Montanide ISA 720 as adjuvants. AB - Recent studies demonstrate that Th1-type immune responses against a broad spectrum of hepatitis C virus (HCV) gene products are crucial to the resolution of acute HCV infection. We investigated new vaccine approaches to augment the strength of HCV-specific Th1-type immune responses. ELISPOT assay revealed that single or multiple protein immunization using both CpG ODN and Montanide ISA 720 as adjuvants induced much stronger IFN-gamma-producing Th1 responses against core, NS3 and NS5b targets than did the formulation without these adjuvants. Protein vaccination using CpG ODN and Montanide ISA 720 as adjuvants also greatly enhanced humoral responses to HCV core, E1/E2 and NS3. When specific IgG isotypes were assayed, protein immunization using CpG ODN and Montanide ISA 720 as adjuvants produced higher titers of IgG2a dominant antibodies than did protein immunization alone, indicating a more Th1-biased pathway. This increase in IgG2a is consistent with the induction of Th1 cells secreting IFN-gamma demonstrated by ELISPOT assay. In conclusion, protein immunization using CpG ODN and Montanide ISA 720 as adjuvants greatly enhanced cellular (Th1 type) as well as humoral immune responses against HCV in Balb/c mice. The use of adjuvants appears critical to the induction of Th1 immune responses during HCV vaccination with recombinant proteins. PMID- 18675872 TI - Protective Th1 immune responses against chronic toxoplasmosis induced by a protein-protein vaccine combination but not by its DNA-protein counterpart. AB - Vaccine-induced protection against toxoplasmosis is correlated with cellular immune responses to Toxoplasma gondii, both in animals and man. The goal of the current study was to evaluate whether the combination of a recombinant protein and a plasmid DNA vaccine could offer an advantage over the protein mixture, and protect outbred mice against infection with T. gondii. To this purpose, the chimeric protein rEC2, encoding antigenic fragments of surface-associated proteins MIC2, MIC3 and SAG1, was combined with pGRA7 plasmid DNA or rGRA7 protein. High levels of antibodies were elicited by both vaccine formulations. The protein-DNA vaccine elicited a polarized Th1/Th2 immune response, characterized by IFN-gamma and IL-10, and afforded low protection (24%) against brain cyst formation. In contrast, the protein-protein vaccine elicited a Th1 focused immune response, characterized by IFN-gamma and IL-2 production, conferring a strong protection (79%) against brain cyst formation in chronic toxoplasmosis. We show here that GERBU adjuvanted protein vaccines confer better protection against toxoplasmosis than the protein-DNA heterologous vaccine. PMID- 18675873 TI - Toxoplasma gondii protease inhibitor-1 (TgPI-1) is a novel vaccine candidate against toxoplasmosis. AB - The Toxoplasma gondii serin protease inhibitor-1 (TgPI-1) is a dense granule antigen that showed to specifically inhibit trypsin, chymotrypsin and neutrophil elastase, suggesting a possible modulatory role during the parasite invasion process and on the development of the innate immune response. To study the immune protective value of TgPI-1, C3H/HeN mice were immunized with a recombinant form of the antigen rTgPI-1 combined with alum. All immunized mice produced specific anti-rTgPI-1 immunoglobulins, with high IgG antibody titers and a mixed IgG(1)/IgG(2a) response, with predominance of IgG(1) production. The cellular immune response was associated with the production of IFN-gamma and IL-10 cytokines. Vaccinated mice displayed significant protection against an oral challenge either after a lethal infection with Me49 cysts (90% survival vs. 50%) and also after a non-lethal infection (58% reduction in brain parasite load) compared to the non-vaccinated control group. In conclusion, rTgPI-1 elicits a strong specific immune response providing partial protection against both T. gondii acute and chronic infection, so it would be a good candidate in a vaccine against toxoplasmosis, which could be combined with other relevant parasite antigens. PMID- 18675874 TI - Inhibitory effect of state independent ponto-geniculo-occipital waves on seizure occurrence induced by local application of penicillin into the temporal lobe amygdala. AB - PURPOSE: In order to explore the possible inhibitory role of the phasic phenomena of REM sleep ponto-geniculo-occipital (PGO) waves over epilepsy, seizure activity produced by topic administration of Na-penicillin (PCN) has been analyzed during sustained PGO waves irrespective of current state. PGO waves were induced by the injection of carbachol in the peribrachial area. METHODS: The development of acute experimental epilepsy was compared among nine chronically implanted, adult, male cats, by means of polygraphic 23 h recordings. Our protocol consisted of sets of 4 trials: carbachol; PCN; carbachol followed by PCN and finally PCN followed by carbachol. Each cat received one single set and all trials were carried out with a seven days interval, in order to compare the epileptic activity both in the presence of PGOs and without them. RESULTS: PGO waves 1) exert an inhibitory influence over number and duration of the Generalized Convulsive Seizures (GCSs) and 2) spike frequency; 3) increase the latency of GCSs; and 4) restores sleep alterations produced by experimental epilepsy. CONCLUSIONS: PGO waves exhibit an inhibitory influence over seizures induced by PCN. These data support the hypothesis that this phasic phenomenon of REM sleep have a depressant effect on epilepsy, inhibit seizures and normalize sleep architecture changes induced by seizures. We suggest that one possible function of PGO activity is to protect the brain from intense changes in neuronal excitability; namely convulsive activity. PMID- 18675875 TI - Neuroprotective effects of Yi-Gan San against beta amyloid-induced cytotoxicity on rat cortical neurons. AB - INTRODUCTION: Recent clinical studies have demonstrated that Yi-Gan San (YGS, Yokukan-San in Japanese), a Chinese herbal medicine, alleviates various dementia related symptoms. However, Chinese herbal medicines have rarely been investigated scientifically and the underlying mechanism of YGS remains uncertain. In this study, we investigated the effect of YGS on beta amyloid protein (Abeta)-induced cytotoxicity in a primary culture of rat cortical neurons. METHODS: Cortical neurons prepared from rat embryos were exposed to Abeta in the presence or absence of YGS. The protective effect of YGS was measured as the % of control (unexposed neurons) by using MTT assay and LDH assay. RESULTS: Abeta significantly decreased the number of surviving cortical neurons at a dose of 20 microM and higher. In the presence of 20 microM Abeta, YGS concentrations of 10( 5) g/L (W/V) and higher significantly increased the number of viable neurons. CONCLUSION: Our study demonstrated a neuroprotective effect of Yi-Gan San against Abeta-induced cytotoxicity. Since according to traditional herbal medicine beliefs, YGS most likely exerts its clinical effects not through a single constituent but as a mixture of several herbal ingredients, the true mechanism of this neuroprotective action remains unclear. However, our results suggest that this Chinese herbal medicine might be a valuable treatment for clinical symptoms associated with dementia having fewer side effects and possible additional neuroprotective effects in the elderly. PMID- 18675876 TI - Adhesio interthalamica in individuals at high-risk for developing psychosis and patients with psychotic disorders. AB - Abnormal neurodevelopment in midline structures such as the adhesio interthalamica (AI) has been reported in psychotic disorders, but it is unknown whether individuals at risk for the disorder share the AI findings observed in patients with florid psychosis. Magnetic resonance imaging of 162 patients with first-episode psychosis (FEP), 89 patients with chronic schizophrenia, 135 individuals at ultra high-risk (UHR) of psychosis (of whom 39 later developed psychosis), and 87 healthy controls were used to investigate the length and prevalence of the AI. The relation of the AI length to lateral ventricular enlargement was also explored. The patients with FEP and chronic schizophrenia as well as UHR individuals had a shorter AI than the controls, but there was no difference in the AI findings between the UHR individuals who did and did not subsequently develop psychosis. There was a negative correlation between the AI length and lateral ventricular volume in all the diagnostic groups. The absence of the AI was more common in the chronic schizophrenia patients when compared with all other groups. These results support the notion that the AI absence or shorter length could be a neurodevelopmental marker related to vulnerability to psychopathology, but also suggest that schizophrenia patients may manifest progressive brain changes related to ongoing atrophy of the AI after the onset. PMID- 18675877 TI - Early effects of caffeinated and decaffeinated coffee on subjective state and gender differences. AB - OBJECTIVES: Some previous studies have demonstrated an early effect of caffeine administration on subjective state, but none of them has explored its existence after the administration of decaffeinated coffee, or the possible differences depending on the gender and circadian typology of the subjects. The aim of the present work is to investigate the early effects (10-30 min post-consumption) of a single low dose of caffeine (100 mg) and decaffeinated coffee on sleepiness, subjective activation and affect using a realistic design. The influence of gender and circadian typology is also explored. METHODS: A randomized double blind informed placebo controlled procedure was applied to 688 healthy undergraduate volunteers, mean age 22.03+/-2.21 years, 238 men and 450 women. Measures were recorded before and after beverage consumption (10, 20 and 30 min), in two experimental sessions: morning 11:00-13:00 h or afternoon 16:00-18:00 h. RESULTS: Caffeine administration induced arousing effects (lesser somnolence and greater activation) in all post-consumption records, while the effects of decaffeinated drink were only apparent at 10 min. Caffeine effects were greater in men, and the decaffeinated beverage produced greater effects in women. Circadian typology only showed effects for time of day (morning/afternoon) related with rhythmic expression. CONCLUSION: Future works should study more accurately the early effect of coffee beverages and the influence of gender, using other parameters which have proven to be sensitive to their administration. The effect of several caffeine doses should also be studied. PMID- 18675878 TI - Induction of characteristic hepatocyte proliferative lesion with dietary exposure of Wistar Hannover rats to tocotrienol for 1 year. AB - Tocotrienol is an antioxidant which has found commercial application as a food additive and health supplement all over the world. Since there have been no reports regarding toxicological effects of long-term exposure, we performed a 52 week chronic study using Wistar Hannover rats of both sexes given the compound at doses of 0, 0.08, 0.4 or 2% in powdered basal diet. Since 6 animals in the 2% male group died of hemorrhage of several organs by week 50, the maximum dose level was changed to 1% in both sexes for the last 2 weeks. Decrease of body weight gain was observed in the 2% males from week 5 and females from week 10, this persisting to the end of the study. With the high dose, prolongation of prothrombin time and increase of serum ALT in males, and increase of serum ALP in both sexes were observed with statistical significance. In male and female rats receiving 0.4% or less, there were no toxicological changes in any of the parameters examined. At necropsy, multiple cyst-like nodules on the liver surface were macroscopically pronounced in both sexes receiving 2%. On histopathological examination, hepatocellular nodules were evident with distortion of hepatic cords and compression of the surrounding tissue, almost all including areas of spongiosis hepatis. The constituent hepatocytes were immunohistochemically stained with proliferation cell nuclear antigen at high rates. Nevertheless, they did not exhibit overt atypia and the basic lobular architecture remained intact. Additionally, they were consistently negative for glutathione S-transferase placental form (GST-P). Accordingly, we propose the newly categorized but previously used name 'nodular hepatocellular hyperplasia', which may not necessarily have a neoplastic or regenerative nature. However, quantitative GST-P analysis of the liver sections overall showed numbers of GST-P foci in the high dose females to be significantly elevated as compared to the control value. Based on the present data demonstrating nodular liver lesions only at the high dose of both sexes, we conclude that the no-observed-adverse-effect level (NOAEL) is 0.4% (303 mg/kg/day for males, and 472 mg/kg/day for females). PMID- 18675879 TI - Stereochemistry of the carboxylation reaction catalyzed by the ATP-dependent phosphoenolpyruvate carboxykinases from Saccharomyces cerevisiae and Anaerobiospirillum succiniciproducens. AB - The stereochemistry of CO(2) addition to phosphoenolpyruvate (PEP) to yield oxaloacetate catalyzed by ATP-dependent Saccharomyces cerevisiae and Anaerobiospirillum succiniciproducens PEP carboxykinases was determined using (Z) 3-fluorophosphoenolpyruvate ((Z)-F-PEP) as a substrate analog. A. succiniciproducens and S. cerevisiae PEP carboxykinases utilized (Z)-F-PEP with 1/14 and 1/47 the respective K(m) values for PEP. On the other hand, in the bacterial and yeast enzymes k(cat) was reduced to 1/67 and 1/48 the value with PEP, respectively. The binding affinity of pyridoxylphosphate-labeled S. cerevisiae and A. succiniciproducens PEP carboxykinases for PEP and (Z)-F-PEP was checked and found to be of similar magnitude for both substrates, suggesting that the lowered K(m) values for the fluorine-containing PEP analog are due to kinetic effects. The lowered k(cat) values when using (Z)-F-PEP as substrate suggest that the electron withdrawing effect of fluorine affects the nucleophilic attack of the double bond of (Z)-F-PEP to CO(2). For the stereochemical analyses, the carboxylation of (Z)-F-PEP was coupled to malate dehydrogenase to yield 3 fluoromalate, which was analyzed by (19)F NMR. The fluoromalate obtained was identified as (2R, 3R)-3-fluoromalate for both the A. succiniciproducens and S. cerevisiae PEP carboxykinases, thus indicating that CO(2) addition to (Z)-F-PEP, and hence PEP, takes place through the 2-si face of the double bond. These results, together with previously published data [Rose, I.A. et al. J. Biol. Chem. 244 (1969) 6130-6133; Hwang, S.H. and Nowak, T. Biochemistry 25 (1986) 5590 5595] indicate that PEP carboxykinases, no matter their nucleotide specificity, catalyze the carboxylation of PEP from the 2-si face of the double bond. PMID- 18675880 TI - Homeostatically regulated synchronized oscillations induced by short-term tetrodotoxin treatment in cultured neuronal network. AB - Homeostatic plasticity plays a critical role in the stability of neuronal activities. Here, with high-density hippocampal networks cultured on multi electrode arrays (MEAs), the transformation of spontaneous neuronal firing patterns induced by 1microM tetrodotoxin was clarified. Once tetrodotoxin was washed out after a 4-h treatment, spontaneous activities rose significantly with spike rate increasing approximately three times, and synchronized burst oscillations appeared throughout the network, with the cross-correlation coefficient between the active sites rising from 0.06+/-0.03 to 0.27+/-0.05. The long-term recording showed that the oscillations lasted for more than 4h before the network recovered. These results suggest that short-term treatment by tetrodotoxin may induce the homeostatically enhanced neuronal excitability, and that the spontaneous synchronized oscillations should be an indicator of homeostatic plasticity in cultured neuronal network. Furthermore, the non invasive and long-term recording with MEAs as a novel sensing system is identified to be appropriate for pharmacological investigations of neuronal plasticity at the network level. PMID- 18675881 TI - Regulation of mRNAs encoding MMP-9 and MMP-2, and their inhibitors TIMP-1 and TIMP-2 by androgens in the rat ventral prostate. AB - The relative activities of matrix metalloproteinases (MMPs) and their natural inhibitors (tissue inhibitors of matrix metalloproteinases, TIMPs) determine the extent of matrix degradation in any tissue. Their identification and characterization is key towards understanding remodeling of the prostate in the context of both castration induced atrophy and tumor invasion and metastasis. Although the expression of MMPs and TIMPs in prostate tumors has been reported, their regulation by androgens has not been studied. Here, we show that androgen ablation by castration increases the steady state mRNA levels of MMP-9, MMP-2, TIMP-1 and TIMP-2. Blockade of the androgen receptor using flutamide, however, has differential effects on the steady state mRNA expressions of these genes. We also show that both castration and flutamide treatment cause enhanced expression of a high molecular weight gelatinolytic activity in the rat ventral prostate (RVP). Actinomycin D does not affect the increase in steady state mRNA levels of MMP-9 and TIMP-1. Furthermore we show that actinomycin D alone enhances the steady state mRNA and protein levels of these genes. Using RNA gel shift assay with 3'-UTR of TIMP-1, we show that an RNA binding protein is induced following castration. Taken together our data suggest that the induction of MMP-2, MMP-9, TIMP-1 and TIMP-2 mRNAs post-castration could be at least in part due to post transcriptional stabilization. PMID- 18675882 TI - Neuroprotective effect of prenyloxycoumarins from edible vegetables. AB - The present study is designed to investigate the effect of some natural prenyloxyphenylpropanoids as neuroprotective agents against NMDA-induced toxicity in mixed cortical cell cultures containing both neurons and astrocytes. Excitotoxicity was induced by exposure of cultures to NMDA (100microM) at room temperature in a HEPES-buffered salt solution followed by incubation at 37 degrees C for the following 24h in MEM-Eagle's supplemented with 15.8mM NaHCO(3) and 25mM glucose. Tested compounds were mixed with NMDA. Neuronal injury was measured in all experiments by examination of cultures with phase-contrast microscopy at 20x, 18-20h after the insult while neuronal damage was quantitatively assessed by counting dead neurons stained with trypan blue and by measuring lactate dehydrogenase (LDH) released in the medium. Results showed that only natural prenyloxyphenylpropanoids containing a coumarin nucleus, namely 7 isopentenyloxycoumarin and auraptene, both found in nature from plants belonging to the genus Citrus and other of the family of Rutaceae, including edible ones, exerted a good dose-dependent manner protective effect against NMDA-induced neurotoxicity in particular at concentrations ranging from 1 to 10microM. PMID- 18675883 TI - Nitric oxide-mediated neuronal apoptosis in rats with recurrent febrile seizures through endoplasmic reticulum stress pathway. AB - Nitric oxide (NO), as a neurotransmitter, exerts various physiological and pathological effects on the brain. Excess NO is toxic to neurons and may cause neuronal apoptosis. However, the cascade of NO-mediated apoptosis is not fully understood. We utilized a recurrent febrile seizures (FS) rat model and found that plasma NO was increased, neuronal apoptosis was evident, the expression of glucose-regulated protein78 (GRP78, a well-established marker of ER stress) was elevated, and caspase-12 (an ER stress-specific proapoptosis molecule) was activated in the hippocampus in a time-dependent manner after recurrent FS. Administration of sodium nitroprusside (SNP, an NO donor) enhanced neuronal apoptosis, down-regulated the expression of GRP78, and increased that of caspase 12 in FS+SNP groups compared with FS groups. In contrast, treatment with N(G) nitrol-l-arginine methyl ester (l-NAME, a competitive NO synthase inhibitor) inhibited neuronal apoptosis, up-regulated the expression of GRP78, and decreased that of caspase-12 in FS+l-NAME groups compared with FS groups. These results suggest that NO mediates neuronal apoptosis caused by recurrent FS, and that the ER stress pathway is involved in NO-mediated neuronal apoptosis. PMID- 18675884 TI - Contrasting genetic effects of major histocompatibility complex on ischemic peripheral nerve and spinal cord injury in female rats. AB - We have recently shown that the major histocompatibility complex (MHC) exerts a regulatory influence on the development of neuropathic pain-like behaviors after partial sciatic nerve injury in male rats. In the present study, we assessed the role of the MHC in peripheral nerve injury-induced pain as well as central pain following spinal cord injury in female rats using the following inbred strains: Dark Agouti (DA; RT1(av1)), Piebald Virol Glaxo (PVG; RT1(c)) and in the MHC congenic strain PVG-RT1(av1). In line with our previous observation in male rats, PVG-RT1(c) displayed more severe allodynia compared to the strains carrying the RT1(av1) haplotype (PVG-RT1(av1) and DA-RT1(av1)) following sciatic nerve injury in female rats. However, the MHC did not seem to impact the development of allodynia following spinal cord injury since the two congenic strains PVG-RT1(c) and PVG-RT1(av) did not differ after spinal cord injury. Interestingly, the DA RT1(av1) strain displayed significantly more severe allodynia than both PVG strains and this difference was not explained by the extent of spinal cord injury. These results suggest that there are differences in the genetic mechanisms for neuropathic pain development following peripheral or central nervous system injury, both in regarding to the role of the MHC complex as well as non-MHC genes. PMID- 18675885 TI - Immunosuppressive networks in the tumour environment and their effect in dendritic cells. AB - The failure of the immune system to provide protection against tumour cells is an important immunological problem. It is now evident that inadequate function of the host immune system is one of the main mechanisms by which tumours escape from immune control, as well as an important factor that limits the success of cancer immunotherapy. In recent years, it has become increasingly clear that defects in dendritic cells have a crucial role in non-responsiveness to tumours. This article focuses on the functional consequences and recently described mechanisms of the dendritic-cell defects in cancer. PMID- 18675887 TI - Brain nociceptive imaging in rats using (18)f-fluorodeoxyglucose small-animal positron emission tomography. AB - Preclinical exploration of pain processing in the brain as well as evaluating pain-relief drugs in small animals embodies the potential biophysical effects in humans. However, it is difficult to measure nociception-related cerebral metabolic changes in vivo, especially in unanesthetized animals. The present study used (18)F-fluorodeoxyglucose small-animal positron emission tomography to produce cerebral metabolic maps associated with formalin-induced nociception. Anesthesia was not applied during the uptake period so as to reduce possible confounding effects on pain processing in the brain. The formalin stimulation at the hind paw of rats resulted in significant metabolic increases in the bilateral cingulate cortex, motor cortex, primary somatosensory cortex, secondary somatosensory cortex, insular cortex, visual cortex, caudate putamen, hippocampus, periaqueductal gray, amygdala, thalamus, and hypothalamus. Among the measured areas, clear lateralization was only evident in the primary somatosensory cortex and hypothalamus. In addition, pretreatment with lidocaine (4 mg/kg, i.v.) and morphine (10 mg/kg, i.v.) significantly suppressed formalin induced cerebral metabolic increases in these areas. The present protocol allowed identification of the brain areas involved in pain processing, and should be useful in further evaluations of the effects of new drugs and preclinical therapies for pain. PMID- 18675889 TI - A simple method for phylogenomic inference using the information of gene content of genomes. AB - Many studies have been contributed to the inferences of phylogenies. Some studies are based on a single-gene (family), and some are based on entire genome data. In this paper, we propose a total loss genome distance approach based on gene content information to inferring phylogenies. Through various simulations, we demonstrate and evaluate the proposed approach. We compare it with some other approaches built upon gene content or extended gene content. Overall, the proposed approach performs equally well as the other methods do and is more efficient than some of the methods. We apply our approach to 34 microbial complete genomes from COG. The reconstructed tree agrees with the results from other approaches and the tree supports the concept of universal trees. PMID- 18675886 TI - Extended fear conditioning reveals a role for both N-methyl-D-aspartic acid and non-N-methyl-D-aspartic acid receptors in the amygdala in the acquisition of conditioned fear. AB - Pavlovian conditioning is a useful tool for elucidating the neural mechanisms involved with learning and memory, especially in regard to the stimuli associated with aversive events. The amygdala has been repeatedly implicated as playing a significant role in the acquisition and expression of fear. If the amygdala is critical for the acquisition of fear, then it should contribute to this processes regardless of the parameters used to induce or evaluate conditioned fear. A series of experiments using reversible inactivation techniques evaluated the role of the amygdala in the acquisition of conditioned fear when training was conducted over several days in rats. Fear-potentiated startle was used to evaluate the acquisition of conditioned fear. Pretraining infusions of N-methyl-d aspartic acid (NMDA) or non-NMDA receptor antagonists alone into the amygdala interfered with the acquisition of fear early in training, but not later. Pretraining infusions of a cocktail consisting of both an NMDA and non-NMDA antagonist interfered with the acquisition of conditioned fear across all days of training. Taken together these results suggest the amygdala may potentially be critical for the acquisition of conditioned fear regardless of the parameters utilized. PMID- 18675888 TI - Dynamic seizure-related changes in extracellular signal-regulated kinase activation in a mouse model of temporal lobe epilepsy. AB - Extracellular signal-regulated kinase (ERK) is highly sensitive to regulation by neuronal activity and is critically involved in several forms of synaptic plasticity. These features suggested that alterations in ERK signaling might occur in epilepsy. Previous studies have described increased ERK phosphorylation immediately after the induction of severe seizures, but patterns of ERK activation in epileptic animals during the chronic period have not been determined. Thus, the localization and abundance of phosphorylated extracellular signal-regulated kinase (pERK) were examined in a pilocarpine model of recurrent seizures in C57BL/6 mice during the seizure-free period and at short intervals after spontaneous seizures. Immunolabeling of pERK in control animals revealed an abundance of distinctly-labeled neurons within the hippocampal formation. However, in pilocarpine-treated mice during the seizure-free period, the numbers of pERK-labeled neurons were substantially decreased throughout much of the hippocampal formation. Double labeling with a general neuronal marker suggested that the decrease in pERK-labeled neurons was not due primarily to cell loss. The decreased ERK phosphorylation in seizure-prone animals was interpreted as a compensatory response to increased neuronal excitability within the network. Nevertheless, striking increases in pERK labeling occurred at the time of spontaneous seizures and were evident in large populations of neurons at very short intervals (as early as 2 min) after detection of a behavioral seizure. These findings suggest that increased pERK labeling could be one of the earliest immunohistochemical indicators of neurons that are activated at the time of a spontaneous seizure. PMID- 18675890 TI - Heme-binding to the nuclear receptor retinoid X receptor alpha (RXRalpha) leads to the inhibition of the transcriptional activity. AB - Heme acts as a ligand for transcription factors and regulates the expression of several genes. The nuclear receptor retinoid X receptor alpha (RXRalpha) plays important roles in various nuclear receptor-dependent signaling pathways. We here show that heme binds to RXRalpha and impairs its DNA-binding activity. Deletion and mutation studies of RXRalpha revealed that the binding region of hemin corresponded to the ligand binding domain of mouse RXRalpha and cysteine 374 was involved in the binding. The DNA-binding activity using the DR-1 consensus sequence of RXRalpha in electrophoretic mobility shift assays was inhibited by heme. The reporter assay also showed a decrease of RXRalpha-dependent transcriptional activity. It was reported that hemin enhanced the adipocyte differentiation of mouse 3T3-L1 cells, where the functions of several nuclear receptors including RXRalpha and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) are activated. However, the inductions of adipogenic factor mRNAs including PPAR-gamma, fatty acid binding protein-4 and glucose transporter-4 were markedly repressed by heme during adipocyte differentiation. These results suggest that heme causes the impairment of RXRalpha-dependent signal pathways and inhibits the adipocyte differentiation of 3T3-L1 cells. PMID- 18675891 TI - Simultaneous measurement of liquid-phase and solid-phase transformation kinetics in rotating disc and channel flow cell dissolution devices. AB - Solvent-mediated solid-phase transformations may occur during dissolution tests which complicates the evaluation of dissolution rates in cases of metastable drugs. The purpose of this study was to determine the effects of solvent-mediated transformations of theophylline anhydrate (TP (A)) on the intrinsic dissolution rate in simulated gastric fluid at pH 1.2. A combined method set-up for simultaneous measurement of the dissolved quantity of drug and the solid form composition was constructed from in situ Raman spectroscopy and UV-vis spectrophotometry. Transformation kinetics in the traditional USP rotating disc (RD) dissolution apparatus was compared with the recently introduced channel flow cell (CFC). Solid-phase data, supported by scanning electron micrographs taken off-line, explained the changes in the intrinsic dissolution rates due to hydrate formation. Kinetic modelling showed that first order kinetics fitted the data in CFC, but the conversion in RD was strongly S-shaped. These differences were related to dissimilar hydrodynamic conditions and diffusion characteristics in the two dissolution testing devices. In situ solid-phase measurement during dissolution testing can largely improve the understanding of the dissolution results of metastable drugs. This information is valuable in drug candidate selection as well as in explaining and controlling the behaviour of drug substances in the final drug products. PMID- 18675892 TI - Application of molecular topology to the prediction of the antimalarial activity of a group of uracil-based acyclic and deoxyuridine compounds. AB - A topological-mathematical model has been arranged to search for new derivatives of deoxyuridine and related compounds acting as antimalarials against Plasmodium falciparum. By using linear discriminant and multilinear regression analysis a model with two functions was capable to predict adequately the IC(50) for each compound of the training and test series. After carrying out a virtual screening based upon such a model, new structures potentially active against P. falciparum are proposed. PMID- 18675893 TI - Theoretical comparison of hydrodynamic diffusion layer models used for dissolution simulation in drug discovery and development. AB - The effective hydrodynamic diffusion layer thickness (h(eff)) of a drug particle dissolving into an agitated fluid is of great importance for oral absorption simulation. The purpose of the present study was to: (1) introduce a h(eff) estimation method based on the fluid dynamic theory (FD model), and (2) compare the FD model with the non-FD-based approximation models previously reported by Hintz and Johnson (HJ model) and Wang and Flanagan (WF model). In the FD model, the relative velocity of a particle suspended in an agitated fluid was estimated from the terminal slip velocity and the microeddy effect. For small particles (particle radius (r(p))ca. 15microm). One of the merits of the FD model is that it provides the a priori theoretical estimation of h(eff) from particle radius, drug density, agitation strength, fluid viscosity, and diffusion coefficient. The hydrodynamic conditions in the gastrointestinal (GI) tract differ among human and animals, the GI sites, and fasted/fed conditions, etc. Therefore, the FD model could provide a more comprehensive and sophisticated simulation of oral absorption. PMID- 18675894 TI - Pharmacological properties of purinergic receptors and their effects on proliferation and induction of neuronal differentiation of P19 embryonal carcinoma cells. AB - We have used P19 embryonal carcinoma cells as in vitro model for early neurogenesis to study ionotropic P2X and metabotropic P2Y receptor-induced Ca(2+) transients and their participation in induction of proliferation and differentiation. In embryonic P19 cells, P2Y(1), P2Y(2) and P2X(4) receptors or P2X-heteromultimers with similar P2X(4) pharmacology were responsible for ATP and ATP analogue-induced Ca(2+) transients. In neuronal-differentiated cells, P2Y(2,) P2Y(6), P2X(2) and possibly P2X(2)/P2X(6) heteromeric receptors were the major mediators of the elevations in intracellular free calcium concentration [Ca(2+)](i). We have collected evidence for the involvement of metabotropic purinergic receptors in proliferation induction of undifferentiated and neural progenitor cells by using a BrdU-incorporation assay. ATP-, UTP-, ADP-, 2-MeS-ATP and ADP-betaS-induced proliferation in P19 cells was mediated by P2Y(1) and P2Y(2) receptors as judged from pharmacological profiles of receptor responses. ATP-provoked acceleration of neuronal differentiation, determined by analysis of nestin and neuron-specific enolase gene and protein expression, also resulted from P2Y(1) and P2Y(2) receptor activation. Proliferation- and differentiation induction involved the activation of inositol-trisphosphate sensitive intracellular Ca(2+) stores. PMID- 18675895 TI - Identification and characterization of Inc766, an inclusion membrane protein in Chlamydophila abortus-infected cells. AB - We have identified the gene product of locus 766 in the transmembrane head region (TMH/Inc-region) in the Chlamydophila abortus genome by using mass spectrometry and a monoclonal antibody that reacted with the inclusion membrane. The identified protein at 32 kDa, termed Inc766, formed highly stable oligomers when solubilized in the absence of beta-mercaptoethanol. These oligomers were resistant to SDS, to heat denaturation and to 8M urea, but very sensitive to beta mercaptoethanol, consistent with conformations resulting from protein-protein interactions stabilized through disulphide bonds. Mass spectrometry analysis of immunoprecipitated infected cell lysates indicated that a dimer at 56 kDa was the most prominent form in solution. Cross-linking with DSP provided supporting evidence for the formation of oligomers in situ. Inc766 was expressed at 20-24h post infection and its localization pattern in the extra-inclusion space was common in all C. abortus strains tested. Taken together, Inc766 displays unique biochemical and cellular features not encountered in other Incs from other Chlamydiaceae species. Future studies of the particular characteristics especially the interactive properties of Inc766 should contribute to our understanding of the relationship of the different chlamydial species with their respective hosts. PMID- 18675896 TI - Alternative splicing and expression of human and mouse NFAT genes. AB - Four members of the nuclear factor of activated T cells (NFAT) family (NFATC1, NFATC2, NFATC3, and NFATC4) are Ca(2+)-regulated transcription factors that regulate several processes in vertebrates, including the development and function of the immune, cardiovascular, musculoskeletal, and nervous systems. Here we describe the structures and alternative splicing of the human and mouse NFAT genes, including novel splice variants for NFATC1, NFATC2, NFATC3, and NFATC4, and show the expression of different NFAT mRNAs in various mouse and human tissues and brain regions by RT-PCR. Our results show that alternatively spliced NFAT mRNAs are expressed differentially and could contribute to the diversity of functions of the NFAT proteins. Since NFAT family members are Ca(2+)-regulated and have critical roles in neuronal gene transcription in response to electrical activity, we describe the expression of NFATC1, NFATC2, NFATC3, and NFATC4 mRNAs in the adult mouse brain and in the adult human hippocampus using in situ hybridization and show that all NFAT mRNAs are expressed in the neurons of the mouse brain with specific patterns for each NFAT. PMID- 18675897 TI - Comparison of detection and signal amplification methods for DNA microarrays. AB - One of the factors limiting the use of DNA microarray technology for the detection of pathogenic organisms from clinical and environmental matrices has been inadequate assay sensitivity. To assess the effectiveness of post hybridization secondary detection steps to enhance the sensitivity of DNA microarray-based pathogen detection, we evaluated a panel of 11 commercial and novel hybridization detection and signal amplification methods (direct labeling, indirect aminoallyl labeling, antibody, DNA dendrimers, viral particles, internally fluorescent nanoparticles, tyramide signal amplification, resonance light scattering nanoparticles and quantum dots) using a multiplex PCR and spotted long oligonucleotide microarray for Vibrio cholerae. Quantitative parameters such as sensitivity, signal intensity, background, assay complexity, time and cost were assessed and provide comparative criteria to be considered for DNA microarray experimental design. While the most important parameter is likely to vary based on the assay, when weighted equally, the findings suggest that recognition element- and dye-functionalized viral particles provide the most attractive option for microarray detection and signal amplification. PMID- 18675898 TI - Association of cocaine- and amphetamine-regulated transcript and neuropeptide Y in the forebrain and pituitary of the catfish, Clarias batrachus: a double immunofluorescent labeling study. AB - Cocaine- and amphetamine-regulated transcript (CART) and neuropeptide Y (NPY) are involved in the regulation of food intake, body weight, pituitary hormones, and reproduction. While CART and NPY occupy overlapping fields in the brain of mammals, little is known about the interaction between these peptide-containing systems in other vertebrates. We explored neuroanatomical associations between CART and NPY in the olfactory system, forebrain and pituitary of the catfish, Clarias batrachus, using double immunofluorescence method. NPY-containing fascicles from olfactory receptor neurons innervated the olfactory glomeruli and mitral cell layer in close association with CART-containing terminal fields. Distinct CART- or NPY-containing fibers were seen in the medial olfactory tract. In the dorsal telencephalon, CART- and NPY-immunoreactive axons were closely associated in area dorsalis telencephali/pars lateralis dorsalis (Dld), and posterioris (Dlp). In the ventral telencephalon, while most of the cells of nucleus entopeduncularis (NE) showed the presence of CART as well as NPY, a few cells with only NPY-immunoreactivity were observed. Similarly, a CART and NPY colocalized cell population was prominent in the preoptic area (POA); and a small population of cells with NPY-immunoreactivity was also evident. Other areas where CART and NPY were colocalized included fibers in the tuberal area, inferior lobe, neurohypophysis, proximal pars distalis and pars intermedia of the pituitary. No association between CART and NPY was observed in the thalamus and habenular ganglion. These results suggest that CART- and NPY-peptidergic systems may interact in NE, POA, tuberal area, certain telencephalic areas and pituitary and jointly process information relating to reproduction, feeding and neuroendocrine regulation. PMID- 18675899 TI - Antioxidant defense in hepatic ischemia-reperfusion injury is regulated by damage associated molecular pattern signal molecules. AB - Hepatic ischemia-reperfusion (I/R) injury occurs in a variety of clinical settings and generates the release of endogenous noninfectious ligands called damage-associated molecular pattern (DAMP) signal molecules from damaged cells. This study investigates the effect of DAMP molecules released by Kupffer cells (KC) in I/R injury on the expression of liver manganese superoxide dismutase (MnSOD), a key mitochondrial antioxidant enzyme. We show that MnSOD expression levels are increased in rats and remain high for 24 h after 30 min of ischemia. KC were damaged and depleted after I/R, in association with MnSOD upregulation. Causality was established by treatment with gadolinium chloride, known to selectively destroy KC, which also increased MnSOD levels. Recovery from the early damage (6 h) to the liver tissue was evidenced after 24 h. A physiological protective role for MnSOD was also confirmed by the increased susceptibility of MnSOD-knockdown AML-12 hepatocyte cells to I/R-induced cell death. Inhibition of DAMP molecule high-mobility group box-1 activity by injection of neutralizing antibody partially abolished the increase in liver MnSOD after I/R. Direct injection of ATP, to animals or cells, stimulated MnSOD upregulation. Another DAMP molecule, monosodium urate, also induced MnSOD expression in hepatocyte AML 12 and FaO cell cultures. In conclusion, a connection between danger signals and upregulation of the antioxidant defense system is shown here for the first time in the context of I/R liver injury. PMID- 18675900 TI - Dietary oxyresveratrol prevents parkinsonian mimetic 6-hydroxydopamine neurotoxicity. AB - Oxyresveratrol (OXY) is a polyhydroxylated stilbene existing in mulberry. Increasing lines of evidence have shown its neuroprotective effects against Alzheimer disease and stroke. However, little is known about its neuroprotective effect in Parkinson disease (PD). Owing to its antioxidant activity, blood-brain barrier permeativity, and water solubility, we hypothesized that OXY may exert neuroprotective effects against parkinsonian mimetic 6-hydroxydopamine (6-OHDA) neurotoxicity. Neuroblastoma SH-SY5Y cells have long been used as dopaminergic neurons in PD research. We found that both pretreatment and posttreatment with OXY on SH-SY5Y cells significantly reduced the release of lactate dehydrogenase, the activity of caspase-3, and the generation of intracellular reactive oxygen species triggered by 6-OHDA. Compared to resveratrol, OXY exhibited a wider effective dosage range. We proved that OXY could penetrate the cell membrane by HPLC analysis of cell extracts. These results suggest that OXY may act as an intracellular antioxidant to reduce oxidative stress induced by 6-OHDA. Western blot analysis demonstrated that OXY markedly attenuated 6-OHDA-induced phosphorylation of JNK and c-Jun. Furthermore, we proved that OXY increased the basal levels of SIRT1, which may disclose new pathways accounting for the neuroprotective effects of OXY. Taken together, our results suggest OXY, a dietary phenolic compound, as a potential nutritional candidate for protection against neurodegeneration in PD. PMID- 18675901 TI - Neurobehavioral effects of sodium tungstate exposure on rats and their progeny. AB - The use of tungsten as a replacement for lead and depleted uranium in munitions began in the mid 1990's. Recent reports demonstrate tungsten solubilizes in soil and can migrate into drinking water supplies and therefore is a potential health risk to humans. This study evaluated the reproductive and neurobehavioral effects of sodium tungstate in Sprague-Dawley rats following 70 days of daily pre- and postnatal exposure. Adult male and female rats were orally dosed with diH(2)O vehicle, 5 or 125 mg/kg/day of sodium tungstate through mating, gestation, and weaning (PND 0-20). Daily administration of sodium tungstate produced no overt evidence of toxicity and had no apparent effect on mating success or offspring physical development. Distress vocalizations were elevated in the highest dose group. There was no treatment related effect on righting reflex latencies, however, the males had significantly shorter latencies than the females. Locomotor activity was affected in both the low and high dose groups of F0 females. Those in the low dose group showed increased distance traveled, more time in ambulatory movements, and less time in stereotypic behavior than controls or high dose animals. The high dose group had more time in stereotypical movements than controls, and less time resting than controls and the lowest exposure group. Maternal retrieval was not affected by sodium tungstate exposure and there were no apparent effects of treatment on F1 acoustic startle response or water maze navigation. Overall, the results of this study suggest pre- and postnatal oral exposure to sodium tungstate may produce subtle neurobehavioral effects in offspring related to motor activity and emotionality. These findings warrant further investigation to characterize the neurotoxicity of sodium tungstate on dams and their developing pups. PMID- 18675902 TI - ATP depletion alters the mode of cell death induced by benzyl isothiocyanate. AB - Pro-inflammatory death is presumably an undesirable event in cancer prevention process, thus biochemical comprehension and molecular definition of this process could have important clinical implications. In the present study, we examined the cytophysiological conversion of cell death mode by benzyl isothiocyanate (BITC) in human cervical cancer HeLa cells. The detailed studies using flow cytometric and morphological analyses demonstrated that the cells treated with appropriate concentration (25 microM) of BITC showed apoptotic feature, such as chromatin condensation, DNA fragmentation, and preserved plasma membrane integrity, whereas these features were disappeared by treatment with higher concentration (100 microM). The treatment with 2-deoxyglucose, an inhibitor of ATP synthesis, drastically increased in the ratio of necrotic dead cells, while it influences little that of apoptotic cells. Moreover, an analysis using the mitochondrial DNA deficient HeLa cells demonstrated that the rho degrees cells were more susceptible to the BITC-induced necrosis-like cell death compared to the wild type (rho(+)) cells, whereas the ROS production was significantly inhibited in the rho degrees cells. It is likely that the BITC-induced ROS is derived from mitochondrial respiratory chain and ruled out the contribution to the mechanism of cell death mode switching. In addition, the BITC treatment resulted in a more rapid depletion of ATP in the rho degrees cells than in the rho(+) cells. Furthermore, a caspase inhibitor, Z-VAD-fmk counteracted not only apoptosis, but also necrosis-like cell death induced by BITC, suggesting that increment in this cell death pattern might be due to the interruption of events downstream of a caspase-dependent pathway. The obtained data suggested that the decline in the intracellular ATP level plays an important role in tuning the mode of cell death by BITC. PMID- 18675903 TI - Effect of N-betainate and N-piperazine derivatives of chitosan on the paracellular transport of mannitol in Caco-2 cells. AB - The effects of novel quaternary chitosan derivatives on the paracellular transport of mannitol and cell viability were studied in the Caco-2 cell model. The N-betainate derivative with the degree of substitution of 0.05 was very effective at 1.0% (w/v) concentration. The activity decreased as the degree of substitution increased. The cytotoxicity of N-betainates was rather low. The N piperazines were at least equally effective as the N-betainates with a similar degree of substitution (>0.15). Most of the N-piperazines did not exert toxic effects on the cell monolayers. Overall, the inverse proportionality between the degree of substitution and activity suggests that an intact chitosan backbone is essential for the bioactivity of chitosan derivatives. The quaternary group does not substitute for the activity of the free amine group. In particular, the N betainate derivatives of chitosan should contain only the minimum number of substituents required for water solubility. PMID- 18675904 TI - Intestinal lymphatic transport of halofantrine in rats assessed using a chylomicron flow blocking approach: the influence of polysorbate 60 and 80. AB - The aim of the study was to examine the effects of polysorbate 60 and 80 on intestinal lymphatic transport of a poorly water-soluble compound, halofantrine, using a chylomicron flow blocking approach in rats. Male Sprague-Dawley rats were pretreated intraperitoneally with 3.0 mg/kg cycloheximide or saline. One hour later, rats were dosed with 6.7 mg/kg halofantrine in 0.4 or 1.0 g/kg polysorbate 60 or 80, 0.33 g/kg soybean oil or 0.33 g/kg soybean oil+1.0 g/kg polysorbate 80 by gavage, and plasma samples were collected. The fraction of halofantrine transported to the lymphatic system was determined as the difference between the bioavailability in saline-pretreated rats and cycloheximide-pretreated rats. No significant differences in halofantrine transport to the systemic blood and in the deduced lymphatic transport were observed between the two polysorbates, at the tested dosages. The lymphatic transport of halofantrine was the same whether dosing with polysorbate 60, polysorbate 80 or soybean oil; accordingly both surfactants can be used as lymphotropic carriers. Furthermore, there was a good correlation between the halofantrine transport to blood and lymphatics in the chylomicron flow blocking model and published results from the mesenteric lymph cannulated model. PMID- 18675905 TI - Pulmonary delivery of scutellarin solution and mucoadhesive particles in rats. AB - The objectives of this study were to investigate the effects of mucoadhesive excipients on systemic bioavailability of an inhaled drug and to evaluate the feasibility of using the pulmonary route for non-invasive systemic delivery of scutellarin, a poorly orally absorbed flavonoid glucuronide. Following intratracheal spray of the scutellarin solution, the bioavailability was found to be approximately 77% in rats, which was >30-fold higher than that via the peroral route. In addition, the pulmonary absorption of scutellarin appeared to avoid the intestinal first-pass metabolism accompanied by peroral administration. Spray dried scutellarin particles with the presence of mucoadhesive excipients were found to affect the corresponding mucociliary transport rate (MTR) as evaluated by a frog palate model. The pharmacokinetic results indicated that the magnitude of AUC(0-480) of intrapulmonary delivered drug particles was not correlated to the fine particle fraction (FPF) but inversely related to the MTR. Incorporating mucoadhesive polymeric mixtures into the scutellarin particles, the MTR decreased by sixfold, and the absolute bioavailability of the drug was found to increase from 70.1% to 97.9% despite a decrease in the FPF. Moreover, in vitro results evaluated using Calu-3 and A549 cell lines showed that scutellarin and spray dried particles with or without the presence of mucoadhesives exhibited no local cell cytotoxic effects in the tested concentration range. In conclusion, the conducting airway is well permeable to scutellarin, and scutellarin may be effectively delivered systemically through inhalation of respirable droplets or particles. PMID- 18675906 TI - Extraction of amino acids by reverse iontophoresis: simulation of therapeutic monitoring in vitro. AB - Reverse iontophoresis across the skin has been investigated as alternative, non invasive method for clinical and therapeutic drug monitoring. This research investigated the reverse iontophoretic extraction of 19 amino acids present at clinically relevant levels in the subdermal compartment of an in vitro diffusion cell. Over a simulated, systemic concentration range of 0-500 microM, the extraction of amino acids was linear. Charged amino acids were extracted towards the electrode of opposite polarity, while zwitterionic species were extracted to both anode and cathode with the latter predominating. The reverse iontophoretic extraction flux was a linear function of amino acid isoelectric point, reflecting the different contributions of electromigration and electroosmosis to electrotransport. Overall, the results confirm the feasibility of monitoring amino acids at clinically relevant levels and provide an incentive for in vivo research to further explore the clinical potential of reverse iontophoresis for the non-invasive monitoring of amino acids. PMID- 18675907 TI - Permeation enhancer dodecyl 6-(dimethylamino)hexanoate increases transdermal and topical delivery of adefovir: influence of pH, ion-pairing and skin species. AB - Adefovir (9-(2-phosphonomethoxyethyl)adenine) is an acyclic nucleoside phosphonate currently used for the treatment of hepatitis B. The aim of this study was to evaluate the effect of permeation enhancer DDAK (6 dimethylaminohexanoic acid dodecyl ester) on the transdermal and topical delivery of adefovir. In porcine skin, DDAK enhanced adefovir flux 42 times with maximum at pH 5.8 suggesting ion pair formation. DDAK increased thermodynamic activity and stratum corneum/vehicle distribution coefficient of adefovir, as well as it directly decreased the skin barrier resistance. Maximal flux was observed already at 2% adefovir+1% DDAK. The results were confirmed in freshly excised human skin where DDAK enhanced adefovir flux 179 times to 8.9 microg/cm(2)/h. This rate of percutaneous absorption would allow for reaching effective plasma concentrations. After the topical application, adefovir concentrated in the stratum corneum with low penetration into the deeper skin layers from either aqueous or isopropyl myristate vehicle without the enhancer. With 1% DDAK, adefovir concentrations in the viable epidermis and dermis were 33-61 times higher. These results offer an attractive alternative to established routes of administration of adefovir and other acyclic nucleoside phosphonates. PMID- 18675908 TI - Amino acids within loops D, E and F of insect nicotinic acetylcholine receptor beta subunits influence neonicotinoid selectivity. AB - Nicotinic acetylcholine (ACh) receptors (nAChRs) are ligand-gated ion channels which mediate fast cholinergic synaptic transmission in insect and vertebrate nervous systems. The nAChR agonist-binding site is present at the interface of adjacent subunits and is formed by loops A-C present in alpha subunits together with loops D-F present in either non-alpha subunits or homomer-forming alpha subunits. To investigate the mechanism of neonicotinoid selectivity, we have examined the effects of altering insect-specific loops D, E and F in hybrid nAChRs containing insect and mammalian subunits (Nlalpha1 from the brown planthopper Nilaparvata lugens and beta2 from rat). Introduction of the insect specific loops D, E and F, singly or together, into rat beta2 subunit resulted in a leftward shift of the imidacloprid dose-response curves for nAChRs Nlalpha1 beta2 chimeras, reflecting decreases in EC(50), compared to wildtype nAChRs Nlalpha1-beta2. By contrast, the influences on ACh potency were minimal or negligible. The effects of loop D could be interpreted by the earlier findings of Shimomura et al. [2006. Role in the selectivity of neonicotinoids of insect specific basic residues in loop D of the nicotinic acetylcholine receptor agonist binding site. Mol. Pharmacol. 70, 1255-1263.], in which T77R and E79V were shown to be responsible for neonicotinoid selectivity. In the present study, S131Y(R) and D133N in loop E and T191W and P192K in loop F were found to contribute to the neonicotinoid selectivity of insect-specific loops E and F. These results indicated the insect-specific loops D, E and F each play important roles in neonicotinoids selectivity. This study contributes to our understanding of the molecular mechanism underlying selectivity of neonicotinoids against insects over vertebrates. PMID- 18675909 TI - Ecdysteroid signaling in ecdysteroid-resistant cell lines from the polyphagous noctuid pest Spodoptera exigua. AB - Although dibenzoylhydrazine-type non-steroidal ecdysone agonists such as methoxyfenozide (RH-2485) have an excellent performance record, the emergence of resistance could severely compromise the efficacy of these compounds in integrated pest management programs. To investigate possible mechanisms of resistance, cell lines derived from the polyphagous noctuid pest Spodoptera exigua (Se4 cells) were selected for continuous growth in the presence of high concentrations of 20-hydroxyecdysone (20E) or methoxyfenozide. Here we describe an analysis of ecdysteroid receptor signaling in the ecdysteroid-resistant Se4 cell lines. In contrast to other ecdysteroid-resistant cell lines described in literature, our data support the existence of a normal functioning ecdysteroid receptor complex in the resistant Se4 cell lines: (1) using a recombinant BmNPV baculovirus as a transduction tool, activation of an ecdysone-responsive luciferase cassette was demonstrated; (2) the early gene HR3 is constitutively expressed in the resistant cell lines that are grown in the presence of 20E or methoxyfenozide. Quantitative RT-PCR experiments indicated that expression levels of SeEcR mRNA were comparable among sensitive and resistant cell lines. Sequencing of PCR fragments also revealed the presence of SeEcR mRNA with a wild type ligand-binding domain in resistant cells. Finally, a possible role for the gene FTZ-F1, whose expression correlates with the absence of circulating ecdysteroids during insect development, in the resistance mechanism was investigated. However, it was observed that FTZ-F1, in contrast to what is observed during insect development, is constitutively expressed in Se4 cells and that its expression is not regulated by the addition of ecdysteroid. It is proposed that the resistance mechanism in Se4 cells resides at the coupling between the conserved hierarchical cascade of early and early-late gene expression and the differentiation program in the Se4 cell line. The use of insect cell lines for the investigation of resistance against dibenzoylhydrazine ecdysone agonists and their relevance for uncovering resistance mechanisms in insects during pest control programs is discussed. PMID- 18675910 TI - A novel clade of cysteinyl leukotriene scavengers in soft ticks. AB - Inflammation is an important vertebrate defense mechanism against ecto-parasites for which ticks have evolved numerous mechanisms of modulation. AM-33 and TSGP4, related lipocalins from the soft ticks Argas monolakensis and Ornithodoros savignyi bind cysteinyl leukotrienes with high affinity as measured by isothermal titration calorimetry. This was confirmed in a smooth muscle bioassay that measured contraction of guinea pig ileum induced by leukotriene C4 where both proteins inhibited contraction effectively. Conservation of this function in two diverse soft tick genera suggests that scavenging of cysteinyl leukotrienes evolved in the ancestral soft tick lineage. In addition soft ticks also evolved mechanisms that target other mediators of inflammation that include scavenging of histamine, serotonin, leukotriene B4, thromboxane A2, ATP and inhibition of the complement cascade. Inhibitors of blood-coagulation and platelet aggregation were also present in the ancestral soft tick lineage. Because histamine and cysteinyl leukotrienes are mainly produced by mast cells and basophils, and these cells are important in the mediation of tick rejection reactions, these findings indicate an ancient antagonistic relationship between ticks and the immune system. As such, modulation of the hemostatic system as well as inflammation was important adaptive responses in the evolution of a blood-feeding lifestyle in soft ticks. PMID- 18675911 TI - Characterization of the multicopper oxidase gene family in Anopheles gambiae. AB - The multicopper oxidase (MCO) family of enzymes includes laccases, which oxidize a broad range of substrates including diphenols, and several oxidases with specific substrates such as iron, copper or ascorbic acid. We have identified five putative MCO genes in the genome of Anopheles gambiae and have cloned cDNAs encompassing the full coding region for each gene. MCO1 mRNA was detected in all developmental stages and in all of the larval and adult tissues tested. We observed an increase in MCO1 transcript abundance in the midguts and Malphighian tubules of adult females following a blood meal and in adult abdominal carcasses in response to an immune challenge. Two alternatively spliced isoforms of MCO2 mRNA were identified. The A isoform of MCO2 was previously detected in larval and pupal cuticle where it probably catalyzes sclerotization reactions (He, N., Botelho, J.M.C., McNall, R.J., Belozerov, V., Dunn, W.A., Mize, T., Orlando, R., Willis, J.H., 2007. Proteomic analysis of cast cuticles from Anopheles gambiae by tandem mass spectrometry. Insect Biochem. Mol. Biol. 37, 135-146). The B isoform was transcriptionally upregulated in ovaries in response to a blood meal. MCO3 mRNA was detected in the adult midgut, Malpighian tubules, and male reproductive tissues; like MCO1, it was upregulated in response to an immune challenge or a blood meal. MCO4 and MCO5 were observed primarily in eggs and in the abdominal carcass of larvae. A phylogenetic analysis of insect MCO genes identified putative orthologs of MCO1 and MCO2 in all of the insect genomes tested, whereas MCO3, MCO4 and MCO5 were found only in the two mosquito species analyzed. MCO2 orthologs have especially high sequence similarity, suggesting that they are under strong purifying selection; the A isoforms are more conserved than the B isoforms. The mosquito specific group shares a common ancestor with MCO2. This initial study of mosquito MCOs suggests that MCO2 may be required for egg development or eggshell tanning in addition to cuticle tanning, while MCO1 and MCO3 may be involved in metal metabolism or immunity. PMID- 18675912 TI - Differential translational regulation of IRE-containing mRNAs in Drosophila melanogaster by endogenous IRP and a constitutive human IRP1 mutant. AB - Insects, like vertebrates, express iron regulatory proteins (IRPs) that may regulate proteins in cellular iron storage and energy metabolism. Two mRNAs, an unspliced form of ferritin H mRNA and succinate dehydrogenase subunit b (SDHb) mRNA, are known to comprise an iron responsive element (IRE) in their 5' untranslated region making them susceptible to translational repression by IRPs at low iron levels. We have investigated the effect of wild-type human IRP1 (hIRP1) and the constitutively active mutant hIRP1-S437 in transgenic Drosophila melanogaster. Endogenous Drosophila IRE-binding activity was readily detected in gel retardation assays. However, translational repression assessed by polysome gradients was only visible for unspliced IRE-containing ferritin H mRNA, but not for SDHb mRNA. Upon expression of exogenous hIRP1-S437 both mRNAs were strongly repressed. This correlated with a diminished survival rate of adult flies with hIRP1 and complete lethality with hIRP1-S437. We conclude that constitutive IRP1 expression is deleterious to fly survival, probably due to the essential function of SDHb or proteins encoded by yet unidentified target mRNAs. PMID- 18675913 TI - Quantitative determination of catecholic degradation products from insect sclerotized cuticles. AB - Acid hydrolysates of cuticle from various insect species were quantitatively analyzed for five catecholic amino acid adducts. Four of the adducts are ketocatechols; in three of them the amino acid moiety, either lysine, glycine or beta-alanine, is connected via its amino group to the alpha-carbon atom of 3,4 dihydroxyacetophenone, in the fourth a tyrosine residue is connected to the same position via its phenolic group. The fifth adduct contains histidine linked via its imidazole-ring to the beta-position of the dopamine sidechain. The three ketocatecholic adducts containing alpha-amino acids were obtained in significant yields from adult cuticles of the locust Schistocerca gregaria, the cockroaches Blaberus craniifer and Periplaneta americana, and the beetles Pachynoda sinuata and Tenebrio molitor, but only in trace amounts from larval and pupal cuticles of T. molitor, pupal cuticles of the moths Manduca sexta and Hyalophora cecropia, and puparia of the blowfly Calliphora vicina. The beta-alanine-containing ketocatechol was not obtained from cuticle of locusts and T. molitor larvae and pupae, but it was present in the hydrolysates of the other cuticles. The beta histidine-dopamine adduct was obtained from all the cuticles, the highest yield was obtained from adult P. sinuata and the lowest yield was from adult S. gregaria. The beta-histidine-dopamine adduct is derived from the product formed by reaction of p-quinone methides of N-acetyldopamine (NADA) or N-beta alanyldopamine (NBAD) with histidine residues in the cuticular proteins. The ketocatecholic adducts are assumed to be degradation products of crosslinks formed when oxidized dehydro-NADA reacts with the cuticular proteins. The insect species investigated appear to use both pathways for sclerotization, but to widely differing extents; the dehydro-NADA pathway dominates in cuticles which are exposed to strong deforming forces, such as those of adult locusts and cockroaches, and the p-quinone methide pathway dominates in cuticle of lepidopteran pupae and blowfly puparia, which are not exposed to strong mechanical forces but have to be effectively protected against microbial and fungal attacks. PMID- 18675914 TI - Wild-type LRRK2 but not its mutant attenuates stress-induced cell death via ERK pathway. AB - Leucine-rich repeat kinase 2 (LRRK2) is a recently identified gene that, when mutated at specific locations, results in the onset of parkinsonian symptoms with clinical features indistinguishable from idiopathic Parkinson's disease. Based on structural and domain analysis, LRRK2 is predicted to function as a stress responsive protein scaffold mediating the regulation of mitogen activating protein kinase (MAPK) pathways. Consistent with this notion, our results supported the notion that expression of wild-type LRRK2 but not Y1699C or G2019S mutants enhanced the tolerance of HEK293 and SH-SY5Y cells towards H(2)O(2) induced oxidative stress. This increase in stress tolerance was dependent on the presence of the kinase domain of the LRRK2 gene and manifested through the activation of the ERK pathway. Collectively, our results indicated that cells expressing LRRK2 mutants suffer a loss of protection normally derived from wild type LRRK2, making them more vulnerable to oxidative stress. PMID- 18675915 TI - Does the hydrolysis of 2-arachidonoylglycerol regulate its cellular uptake? AB - Very little is known about the processes regulating the cellular uptake of the endogenous cannabinoid 2-arachidonoylglycerol (2-AG). In the present study, we investigated whether inhibition of 2-AG hydrolysis reduced its uptake, i.e. whether this compound behaves in a manner analogous to the related endocannabinoid anandamide. The selective fatty acid amide hydrolase inhibitor URB597 (3'-(aminocarbamoyl)[1,1'-biphenyl]-3-yl)-cyclohexylcarbamate) completely blocked the hydrolysis of anandamide and reduced its uptake by about half in RBL2H3 basophilic leukaemia cells. In contrast, in these cells, in PC3 and R3327AT-1 prostate cancer cells and in Neuro-2a neuroblastoma cells, the compound had more modest effects upon the hydrolysis of 2-AG and did not affect its cellular uptake at all, indicating that in these cells fatty acid amide hydrolase does not regulate the uptake of 2-AG. The serine hydrolase inhibitor methylarachidonoyl fluoronophosphonate behaved like URB597 with respect to anandamide uptake by RBL2H3 and Neuro-2a cells, and inhibited the hydrolysis of 2 AG with IC50 values of 0.014, 0.052, 0.41 and approximately 1 microM for RBL2H3, PC3, AT-1 and Neuro-2a cells, respectively. MAFP (1 microM) did not significantly reduce the uptake of 2-AG by RBL2H3, PC3 and AT-1 cells but did reduce the uptake of this endocannabinoid by Neuro-2a cells. Arachidonoyl trifluoromethyl ketone and URB602 ([1,1'-biphenyl]-3-yl-carbamic acid, cyclohexyl ester) reduced the uptake of 2-AG by both RBL2H3 and Neuro-2a cells, but at the high concentrations needed, the compound also blocked the retention of these ligands by wells. It is concluded that unlike the situation for anandamide, hydrolysis of 2-AG does not regulate its cellular uptake in RBL2H3, AT-1 and PC3 cells, but may gate the uptake in Neuro-2a cells. PMID- 18675917 TI - Oscillatory brain activity before and after an internal context change--evidence for a reset of encoding processes. AB - Prior behavioral work has shown that changing context during encoding can influence long-term memory performance. The present study examined the neural correlates of such context effects by analyzing oscillatory brain activity during the encoding of words, both before and after a context change. Participants studied two lists of items and, after the presentation of List 1, were either cued to change their internal context through a simple imagination task, or not. Replicating the behavioral work, the change in context led to forgetting of the first list (List 1) and to memory enhancement of the second (List 2). Measuring EEGs during encoding of the two lists, the context change was found to affect oscillatory brain activity. Whereas an increase of theta and alpha power from List-1 to List-2 encoding was found when the context was left unchanged, a slight theta and alpha power decrease was found when the context was changed. In addition, median split analysis revealed that alpha power during List-2 encoding was related to the enhancement effect of the context change. The results suggest that a change in internal context can lead to a reset of encoding processes, thus pointing to a crucial role of encoding processes in context-dependent memory. PMID- 18675916 TI - Functional dissection of SYG-1 and SYG-2, cell adhesion molecules required for selective synaptogenesis in C. elegans. AB - Cell adhesion molecules of the Immunoglobulin superfamily (IgCAMs) play diverse functions during neural development. Previously, we have identified SYG-1/Neph1 and SYG-2/Nephrin, IgCAMs necessary for synaptic specificity in Caenorhabditis elegans. Here, we conduct an in vivo structure-function analysis of SYG-1 and SYG 2 to identify domains of SYG-1 and SYG-2 necessary for heterophilic binding as well as synaptic specificity. We find the first Ig domain of SYG-1 and the first 5 Ig domains of SYG-2 are necessary and sufficient for their binding in vivo, as well as for synapse formation. We also find the SYG-2 cytoplasmic domain is required for SYG-2 subcellular trafficking, while the intracellular region of SYG 1 is required for synaptic function at earlier developmental stages, but is dispensable for later stages. This study defines the domain requirements for SYG 1/SYG-2 heterophilic binding and suggests that unknown SYG-1 extracellular interactors may play a role in SYG-1-mediated synaptic specificity. PMID- 18675919 TI - The surprising evolutionary history of South American deer. AB - To clarify the systematic relationships and evolutionary history of South American deer, we conducted a comprehensive phylogenetic analysis using representative species of all of the genera of Neotropical deer. Our results revealed high levels of molecular and cytogenetic divergence between groups of morphologically similar species of brockets (Mazama), and suggest a polyphyletic origin. At least eight ancestral forms of deer invaded South America during the late Pliocene (2.5-3 MYA), and members of the red brockets had an independent early explosive diversification soon after their ancestor arrived there, giving rise to a number of morphologically cryptic species. PMID- 18675918 TI - Validation of a fully automated 3D hippocampal segmentation method using subjects with Alzheimer's disease mild cognitive impairment, and elderly controls. AB - We introduce a new method for brain MRI segmentation, called the auto context model (ACM), to segment the hippocampus automatically in 3D T1-weighted structural brain MRI scans of subjects from the Alzheimer's Disease Neuroimaging Initiative (ADNI). In a training phase, our algorithm used 21 hand-labeled segmentations to learn a classification rule for hippocampal versus non hippocampal regions using a modified AdaBoost method, based on approximately 18,000 features (image intensity, position, image curvatures, image gradients, tissue classification maps of gray/white matter and CSF, and mean, standard deviation, and Haar filters of size 1x1x1 to 7x7x7). We linearly registered all brains to a standard template to devise a basic shape prior to capture the global shape of the hippocampus, defined as the pointwise summation of all the training masks. We also included curvature, gradient, mean, standard deviation, and Haar filters of the shape prior and the tissue classified images as features. During each iteration of ACM - our extension of AdaBoost - the Bayesian posterior distribution of the labeling was fed back in as an input, along with its neighborhood features as new features for AdaBoost to use. In validation studies, we compared our results with hand-labeled segmentations by two experts. Using a leave-one-out approach and standard overlap and distance error metrics, our automated segmentations agreed well with human raters; any differences were comparable to differences between trained human raters. Our error metrics compare favorably with those previously reported for other automated hippocampal segmentations, suggesting the utility of the approach for large-scale studies. PMID- 18675920 TI - Radiation of southern African daisies: biogeographic inferences for subtribe Arctotidinae (Asteraceae, Arctotideae). AB - The majority of the approximately 80-90 species in subtribe Arctotidinae occur in southern Africa with the centre of diversity in the winter-rainfall region. Three species are restricted to afromontane eastern Africa and three species are endemic to Australia. To investigate biogeographic and phylogenetic relationships within Arctotidinae, sequence data from four cpDNA regions (psbA-trnH, trnT-trnL and trnL-trnF spacers and trnL intron) and the ITS nrDNA region for 59 Arctotidinae species were analyzed with parsimony and Bayesian-inference approaches. Eight well-supported major lineages were resolved. The earliest diverging extant lineages are afromontane or inhabit mesic habitats, whereas almost all sampled taxa from the winter-rainfall and semi-arid areas have diverged more recently. Molecular dating estimated that the major clades diverged during the Miocene and Pliocene, which is coincident with the trend of increasing rainfall seasonality, aridification and vegetation changes in southwestern Africa. Trans-oceanic dispersal to Australia was estimated to have occurred during the Pliocene. PMID- 18675921 TI - IGF2-driven PI3 kinase and TGFbeta signaling pathways in chondrogenesis. AB - Insulin-like growth factor-2 (IGF2) is essential for fetal development as well as maintenance of adult organs such as brain and liver. Although genetic polymorphisms of IGF2 are linked to cytoskeletal variations little is known about the mechanisms of IGF2 action in proliferation and differentiation of chondrocytes for skeletal growth. A genome-wide mRNA expression analysis using C28/I2 chondrocyte cells studied potential signaling pathways underlying the responses to IGF2. Microarray data predicted involvement of the phosphatidylinositol 3-kinase (PI3K) and transforming growth factor beta (TGFbeta) signaling pathways. Protein analyses revealed IGF2 administration activated phosphorylation of Akt and GSK3beta in the PI3K pathway. LY294002 (selective inhibitor of PI3K) blocked Akt phosphorylation and abolished IGF2 driven elevation of the mRNA levels of the proteoglycans, Aggrecan and Versican. LY294002 did not suppress upregulation of TGFbeta mRNA induced by IGF2, so IGF2 activates PI3K and TGFbeta pathways. IGF2-driven transcriptional activation of proteoglycan genes such as Aggrecan and Versican is mediated by the PI3K pathway. PMID- 18675922 TI - Identification of potential genes/proteins regulated by Tiam1 in colorectal cancer by microarray analysis and proteome analysis. AB - Tiam1 (T-cell lymphoma invasion and metastasis-inducing protein 1), a guanine nucleotide exchange factor that activates Rac, is a colorectal cancer metastasis related gene. In this study, we aimed to better understand the mechanism underlying Tiam1-mediated metastasis. We applied gene microarray and proteome analysis and compared expression of genes and proteins in a stable Tiam1 silencing colorectal cancer cell line and in a control cell line. Our analysis identified three genes, high-mobility group box1 (HMGB1), annexin IV (ANXA4) and phosphoglycerate mutase 1 (PGAM1) that were associated with Tiam1. Analysis of these proteins, which may be directly or indirectly regulated by Tiam1, may provide insight into the role and mechanism of Tiam1 in colorectal cancer metastasis. PMID- 18675923 TI - Puerarin protects PC12 cells against beta-amyloid-induced cell injury. AB - beta-Amyloid protein (Abeta), a major protein component of brain senile plaques in Alzheimer's disease, is known to be directly responsible for the production of reactive oxygen species (ROS) and induction of apoptosis. In this study, the protective effect of puerarin, an isoflavone purified from the radix of the Chinese herb Pueraria lobata, on Abeta-induced rat pheochromocytoma (PC12) cultures was investigated. Although exposure of PC12 cells to 50 microM Abeta25 35 caused significant viability loss and apoptotic rate increase, pretreatment of the cells with puerarin for 24h reduced the viability loss and apoptotic rate. Puerarin (1 microM) significantly inhibited Abeta25-35-induced apoptosis of PC12 cells. Preincubation of the cell with puerarin also restored the ROS and mitochondrial membrane potential levels that had been altered as a result of Abeta25-35 treatment. Puerarin was also found to increase the Bcl-2/Bax ratio and reduce caspase-3 activation. These results suggest that puerarin could attenuate Abeta25-35-induced PC12 cell injure and apoptosis and could also promote the survival of PC12 cells. Therefore, puerarin may act as an intracellular ROS scavenger, and its antioxidant properties may protect against Abeta25-35-induced cell injury. PMID- 18675924 TI - Interactions between the budding yeast IQGAP homologue Iqg1p and its targets revealed by a split-EGFP bimolecular fluorescence complementation assay. AB - A split-EGFP based bimolecular fluorescence complementation (BiFC) assay has been used to detect interactions between the Saccharomyces cerevisiae cytoskeletal scaffolding protein Iqg1p and three targets: myosin essential light chain (Mlc1p), calmodulin (Cmd1p) and the small GTPase Cdc42p. The format of the BiFC assay used ensures that the proteins are expressed at wild type levels thereby avoiding artefacts due to overexpression. This is the first direct in vivo detection of these interactions; in each case, the complex is localised to discrete regions of the yeast cytoplasm. The labelling with EGFP fragments results in changes in growth kinetics, cell size and budding frequency. This is partly due to the reassembled EGFP locking the complexes into essentially permanent interactions. The consequences of this for Iqg1p interactions and BiFC assays in general are discussed. PMID- 18675925 TI - Influence of age and sex on the spontaneous DNA damage detected by micronucleus test and comet assay in mice peripheral blood cells. AB - We have investigated the normal variations in basal DNA damage detected by Comet assay in leukocytes and micronucleated erythrocytes (MNE) using the Micronucleus test (MN) in peripheral blood cells from 45 female and male mice from different age groups (newborns, 3.5, 12, and 104 weeks) to clarify age and sex-related changes. Comparison of basal DNA damage detected by Comet assay showed significantly increased values in 104 weeks old mice in relation to the other ages (P < or = 0.01), and newborn mice showed higher values in MNE frequency when compared to all the other groups (P < or = 0.01). A positive correlation was observed between Damage Frequency (r =0.382, P = 0.010) and Damage Index (r = 0.640, P < 0.001) and age. Age was also correlated with the ratio of polychromatic erythrocytes/normachromatic erythrocytes (PCE/NCE) (r = -0.473, P = 0.001), and the MNE frequency was positively correlated with the ratio of PCE/NCE (r = 0.454, P = 0.002). These results suggest an age-related slow down of DNA repair efficiency of DNA damage and/or DNA damage accumulation. Furthermore, data on the spontaneous MNE frequency indicate that the reticuloendothelial system matures with age, and there is a close relationship between erythropoiesis and micronucleus induction in erythrocytes. The influence of sex in the parameters analyzed was less clear. In conclusion, age seems to influence in basal DNA damage and should be considered in genotoxicity studies using mice. Finally, comparisons between assays must be made with care when different cells are compared (e.g. leukocytes and erythrocytes), as found with the Comet assay and MN test. PMID- 18675926 TI - Long-term exposure to environmental enrichment since youth prevents recognition memory decline and increases synaptic plasticity markers in aging. AB - Aging-associated brain changes include functional alterations that are usually related with memory decline. Epidemiological reports show that a physically and intellectually active life provides a protective effect on this decline and delays the onset of several neurodegenerative diseases. The cellular mechanisms behind the behavioral-based therapies, such as environmental enrichment (EE) exposure, as a method for alleviating age-related memory impairments, are still unknown. Although some reports have shown the benefits of EE exposure in cognitive outcomes in old mice and in animals with experimental neurodegenerative conditions, the effects of lifelong animal exposure to EE have not been explored in detail. In the present work we tested in a rat model the effects of intermittent lifelong exposure since youth to EE on behavioral performance, object recognition memory and anxiety level, as well as on some morphological and biochemical markers of brain plasticity such as hippocampal neurogenesis, synaptophysin content and synaptic morphology. We found that environmental factors have a positive impact on short-memory preservation, as well as on the maintenance of synapses and in the increase in number of new generated neurons within the hippocampus during aging. PMID- 18675927 TI - Habituation of the orienting reflex and the development of Preliminary Process Theory. AB - The orienting reflex (OR), elicited by an innocuous stimulus, can be regarded as a model of the organism's interaction with its environment, and has been described as the unit of attentional processing. A major determinant of the OR is the novelty of the eliciting stimulus, generally operationalized in terms of its reduction with stimulus repetition, the effects of which are commonly described in habituation terms. This paper provides an overview of a research programme, spanning more than 30 years, investigating psychophysiological aspects of the OR in humans. The major complication in this research is that the numerous physiological measures used as dependent variables in the OR context fail to jointly covary with stimulus parameters. This has led to the development of the Preliminary Process Theory (PPT) of the OR to accommodate the complexity of the observed stimulus-response patterns. PPT is largely grounded in autonomic measures, and current work is attempting to integrate electroencephalographic measures, particularly components in the event-related brain potentials reflecting aspects of stimulus processing. The emphasis in the current presentation is on the use of the defining criteria of the habituation phenomenon, and Groves and Thompson's Dual-process Theory, in the development of PPT. PMID- 18675928 TI - Habituation and migraine. AB - The most reproducible and ubiquitous interictal abnormality of the migraineurs' brain is lack of habituation in neuronal information processing. The underlying mechanisms are uncertain. Increased neuronal excitability, decreased inhibition or decreased pre-activation levels have all been proposed as possible culprits. The present review summarizes the available data on habituation in migraine patients obtained with different methodological approaches. We will discuss the change of habituation level over the migraine cycle and we will show that these data congruently indicate that the sensory cortices of migraineurs react excessively to repetitive, but not to a small series of stimuli. Although the precise cellular and molecular mechanisms of this hyper-responsivity must still be determined, we will describe experimental data suggesting that a dysrhythmic thalamo-cortical activity due to inadequate monoaminergic control might be a plausible explanation. PMID- 18675929 TI - NO scavenging by 3-hydroxyanthranilic acid and 3-hydroxykynurenine: N-nitrosation leads via oxadiazoles to o-quinone diazides. AB - The tryptophan metabolites kynurenine, 3-hydroxykynurenine, anthranilic, 3 hydroxyanthranilic and 3-methoxyanthranilic acids were compared with regard to diazotation by .NO or NO+, using three different donors, nitrite at pH 5, PAPA NONOate at pH 7.4 and NO+SbF(6)- at pH 2.0. With all three sources of NO species, 3-hydroxykynurenine and 3-hydroxyanthranilic acid were readily nitrosated, thereby forming an intensely yellow compound. Nitrosation of the non-hydroxylated analogs did not lead to colored products within the period of observation. Competition experiments, using PAPA-NONOate as NO donor, showed that 3 hydroxyanthranilic acid is a more potent NO scavenger than N-acetylcysteine. Nitrosation of 3-hydroxykynurenine and 3-hydroxyanthranilic acid leads, presumably via a nitrosamine intermediate, to a diazonium ion, which forms an oxadiazole tautomerizing to a yellow o-quinone diazide. While the diazonium derived quinone diazide is apparently the sole product detected directly after incubation of 3-hydroxyanthranilic acid, additional substances are formed from 3 hydroxykynurenine. Contrary to rapid carbenium ion formation from diazonium ions of non-hydroxylated anilines, nitrogen is practically not released from oxadiazoles/quinone diazides at moderate temperatures. Since carbenium ions are known to cause adduct formation with other biomolecules, and since non hydroxylated anilines and their aminophenol analogs differ in their reactions following diazotation, these findings should be of relevance for the relative toxicity of anilines. PMID- 18675930 TI - Thiol activation of a model O2-aryl diazeniumdiolate prodrug in phospholipid vesicle media. AB - Thiolysis of the model diazeniumdiolate prodrug, O2-(2,4-dinitrophenyl) 1-(N,N diethylamino)diazen-1-ium-1,2-diolate (DNP-DEA/NO), by glutathione (GSH), cysteine (CYSH) and 1-heptanethiol (heptylmercaptan, HM) has been examined in anionic (DOPG), neutral (DPPC, DOPE) and cationic (DOTAP) vesicle media and in glycine buffered aqueous solutions. DOTAP vesicles accelerate the bimolecular reaction with glutathione, cysteine and 1-heptanethiol by factors of 81, 8.2 and 4630, respectively, while reaction is inhibited 5- to 10-fold in the presence of neutral and anionic vesicles. The intrinsic nucleophilicity of the thiols has been compared through the second-order rate constants, 22.9, 5.24 and 43.1M(-1)s( 1), for nucleophilic attack on 1 by GS(-), CYS(-) and M(-), respectively, obtained in buffered aqueous media. Analysis of the catalysis by DOTAP vesicles, using pseudophase ion-exchange formalism, suggests that the rate increase is due to reactant concentration in the bilayer and interfacial region coupled with enhanced dissociation of the thiol at the vesicle surface. Some contribution from enhanced nucleophilic reactivity at the vesicle interface may also contribute to the greater catalysis by HM. Inhibition of the thiolysis reaction by phospholipid liposomes is attributed to repulsion of the thiolate anions by the negatively charged acyl phosphate of the lipid head group. DOPG=1,2-dioleoyl-sn-glycero-3 [phospho-rac-(1-glycerol)], DPPC=1,2-dipalmitoyl-sn-glycero-3-phosphocholine, DOPE=1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, DOTAP=1,2-dioleoyl-3 trimethylammonium-propane. PMID- 18675931 TI - Is ascorbate able to reduce disulfide bridges? A cautionary note. AB - The biotin switch assay has recently been proposed as the eligible method to identify different S-nitrosated proteins in biological matrices. However, notwithstanding its wide application, a thorough validation of this method is still lacking. In particular, it has been suggested that ascorbate concentrations higher than 1mM (as proposed in the original method) are needed since ascorbate reaction with S-nitrosothiols is slow. But the selectivity of ascorbate as a cleaving agent of the S-N bond under these conditions has not been well characterized. Our data indicate that ascorbate is able to reduce disulfide bridges of DTNB, cystine, cystinylglycine, glutathione disulfide, protein mixed disulfides and biotin-HPDP with pH and concentration dependent rates. Additionally, we tested the effect of indirect sunlight on ascorbate-mediated cleavage of both disulfides and S-nitrosothiols. PMID- 18675932 TI - Nitric oxide synthase isoforms and NF-kappaB activity in normal and osteoarthritic human chondrocytes: regulation by inducible nitric oxide. AB - To elucidate the role of endogenous inducible nitric oxide (NO) on the regulation of NF-kappaB activity in human chondrocytes, we evaluated (i) the pattern of expression of the neuronal (nNOS) and inducible (iNOS) NO synthase isoforms and the basal NF-kappaB activity in normal and osteoarthritic (OA) human chondrocytes, (ii) the role of cytokines and growth factors in modulating the protein levels of the two NOS isoforms, and (iii) the effect of inhibiting endogenous inducible NO production on the ability of interleukin-1beta (IL-1) to induce NF-kappaB activation. nNOS was more frequently expressed in normal than in OA chondrocytes, whereas the opposite was found for iNOS. IL-1 induced the degradation of both enzymes, but iNOS disappeared more rapidly. Although IkappaB alpha was present in all the normal samples and in the majority of the OA samples, NF-kappaB-DNA binding activity in OA chondrocytes was increased approximately twofold relatively to normal cells. Addition of a NOS inhibitor, after induction of iNOS expression, induced IkappaB-alpha degradation and potenciated the effect of IL-1, indicating that endogenous inducible NO inhibits NF-kappaB activation. Taken together, these findings favor an inhibitory role of high NO levels on the regulation of NF-kappaB activation in chondrocytes, indicating that NF-kappaB activity is regulated, at least in part, by the balanced production of NO resulting from a dynamic process that, at any given moment, determines the availability of the constitutive and inducible NOS isoforms. Moreover, the down-regulatory role of NO on NF-kappaB activation warrants caution as to the possible utilization of NO inhibitors in the therapy of OA. PMID- 18675933 TI - How it all began: microbes and infection 120years later. PMID- 18675935 TI - Internal reproductive anatomy of the praying mantid Ciulfina klassi (Mantodea: Liturgusidae). AB - Using light and scanning electron microscopy, the internal male and female reproductive anatomy of the praying mantid Ciulfina klassi is identified and described. This is the first detailed study to investigate the internal reproductive morphology of any Mantodea. The female structures identified were (1) paired ovaries with primitive panoistic type ovarioles, (2) a single blind ended spermatheca with secretory gland cells and surrounding layer of striated muscle, and (3) female accessory glands associated with the production of the ootheca (the egg casing). The male structures identified were (1) paired multi tubular testes, in which different stages of spermatogenesis were observed, (2) tubular vasa deferentia, (3) seminal vesicles, (4) male accessory glands and (5) a single muscular ejaculatory duct. Knowledge of basic reproductive morphology can be used to infer function and so provide key information for future research into reproductive behavior and physiology in the Mantodea. PMID- 18675934 TI - Tissue-specific oxidative imbalance and mitochondrial dysfunction during Trypanosoma cruzi infection in mice. AB - In this study, we examined the tissue specificity of inflammatory and oxidative responses and mitochondrial dysfunction in mice infected by Trypanosoma cruzi. In acute mice, parasite burden and associated inflammatory infiltrate was detected in all tissues (skeletal muscle>heart>stomach>colon). The extent of oxidative damage and mitochondrial decay was in the order of heart>stomach>skeletal muscle>colon. In chronic mice, a low level of parasite burden and inflammation continued in all tissues; however, oxidant overload and mitochondrial inefficiency mainly persisted in the heart tissue (also detectable in stomach). Further, we noted an unvaryingly high degree of oxidative stress, compromised antioxidant status, and decreased mitochondrial respiratory complex activities in peripheral blood of infected mice. A pair-wise log analysis showed a strong positive correlation in the heart-versus-blood (but not other tissues) levels of oxidative stress markers (malonyldialdehyde, glutathione disulfide), antioxidants (superoxide dismutase, MnSOD, catalase), and mitochondrial inhibition of respiratory complexes (CI/CIII) in infected mice. T. cruzi-induced acute inflammatory and oxidative responses are widespread in different muscle tissues. Antioxidant/oxidant status and mitochondrial function are consistently attenuated in the heart, and reflected in the peripheral-blood of T. cruzi-infected mice. Our results provide an impetus to investigate the peripheral-blood oxidative responses in relation to clinical severity of heart disease in chagasic human patients. PMID- 18675936 TI - Sperm ultrastructure of the European hornet Vespa crabro (Linnaeus, 1758) (Hymenoptera: Vespidae). AB - This study represents the first sperm description of a Vespinae species (Vespa crabro). The acrosome consists of an acrosomal vesicle and a perforatorium. The nucleus has compact chromatin and shows lenticular structures on the nuclear envelope. These structures, which have never been observed in a hymenopteran sperm, could be clusters of nuclear pores. The centriolar adjunct has an asymmetric pattern and shows a structured periphery. The centriole consists of 9 accessory microtubules and 9 doublet microtubules devoid of arms and spokes. The axoneme has a 9+9+2 microtubule pattern and the accessory microtubules have 16 protofilaments. The mitochondrial derivatives differ in length and diameter. The larger one is adjacent to the nuclear base, while the smaller one begins below the centriolar adjunct. They possess three distinct areas and a large paracrystalline region, which occurs only in the large one. The large mitochondrial derivative ends first, followed by the small one. The axoneme gradually disorganizes: first the central microtubules disappear, then the doublets, which show opened B-tubules, and finally the accessory microtubules. The sperm morphology of V. crabro is very similar to that of the polistine wasp, Agelaia vicina. This can indicate that, in Vespidae, sperm morphology is maintained without important variations among subfamilies and/or that this similarity indicates close phylogenetic relationship between these two subfamilies. Although Vespidae phylogenetically related to Formicidae, these data suggest that the former more closely related to Apoidea than to Formicidae. PMID- 18675937 TI - Neuroimmunology of the gut: physiology, pathology, and pharmacology. AB - It has been increasingly appreciated that an intimate interaction between cells of the nervous and immune systems takes place in the gut, and may have a role in diverse inflammatory disorders. Thus, for instance, activation of the enteric nervous system may reduce intestinal epithelial permeability, via several mediators including S-nitrosoglutathione and vasoactive intestinal peptide (VIP). Moreover, ablation of glial cells instigated enterocolitis in murine models. These neuronal effects are particularly intriguing given our current understanding of the immunopathogenesis of Crohn's disease, in which intestinal barrier defect is suspected to at least partly drive the immune hyper-reactivity and ensuing inflammation. Parasympathetic nicotinic signaling, primarily via nicotinic acetylcholine receptor alpha7 (alpha7 nACHr), also exerts immunomodulatory effects, possibly underlaying the detrimental effects of smoking on Crohn's disease, and its beneficial impact on ulcerative colitis. These, and others, neuro-immune interactions may pave the way to the design of novel therapeutic agents for the treatment of chronic inflammatory bowel disorders. PMID- 18675938 TI - Residual cognitive effects of uncomplicated idiopathic and cryptogenic epilepsy. AB - We assessed residual cognitive deficits in young people with idiopathic and cryptogenic epilepsy. In the setting of an ongoing prospective study, we invited participants initially diagnosed and enrolled in the cohort 8-9 years earlier to undergo standardized neuropsychological assessment. Sibling controls were invited when available. We analyzed 143 pairs in which cases had idiopathic or cryptogenic epilepsy and both case and control had normal intelligence. Compared with that for siblings, the Full Scale IQ for cases was 3.3 points lower (P=0.01) mainly due to slower processing speed, which was 5.6 points lower (P=0.0004). Word reading (P=0.04) and spelling (P=0.01), but not other scores, were also lower in cases. Remission status and drug use did not influence findings. In young people of normal intelligence with idiopathic or cryptogenic childhood onset epilepsy, substantial residual effects of epilepsy appear to be confined largely to slower processing speed. PMID- 18675939 TI - Detecting recombination and diversifying selection in human alpha-papillomavirus. AB - Intragenic recombination and selection analyses were performed in DNA sequences of human alpha-papillomavirus. Recombination was estimated and the corresponding breakpoints obtained by re-analyzing data grouped by phylogenetic and epidemiological criteria, using different alignment methods. Diversifying or positive selection has been scarcely studied in these viruses probably due to the high divergence rates. We have applied maximum likelihood, empirical Bayesian and maximum parsimony methods to detect the presence of positive selection. Within the HPV 16 type, significant positive selection was detected at the time of the separation of the African 1 and African 2 branches from the other populations. At the inter-type level, positive selection can be traced in some codons of the gene L2 of the high and low risk groups. These results indicate that positive selection could have been important in the evolution of HPV both at inter- and intra-type levels. PMID- 18675940 TI - Immunomodulatory pretreatment with Kalanchoe pinnata extract and its quercitrin flavonoid effectively protects mice against fatal anaphylactic shock. AB - Previously, we reported the immunosuppressive action of the aqueous extract of Kalanchoe pinnata (Kp) in mice. In the present study, we report on the protective effect of Kp in fatal anaphylactic shock, likewise a Th2-driven immunopathology, and the identification of its active component. Mice daily treated with oral Kp during hypersensitization with ovalbumin were all protected against death when challenged with the allergen, as compared with the 100% mortality in the untreated group. A single intraperitoneal dose 3 h prior to challenge was partially effective. Oral protection was accompanied by a reduced production of OVA-specific IgE antibodies, reduced eosinophilia, and impaired production of the IL-5, IL-10 and TNF-alpha cytokines. In vitro, Kp prevented antigen-induced mast cell degranulation and histamine release. Oral treatment with the quercitrin flavonoid isolated from Kp prevented fatal anaphylaxis in 75% of the animals. These findings indicate that oral treatment with Kp effectively downmodulates pro anaphylactic inducing immune responses. Protection achieved with quercitrin, although not maximal, suggests that this flavonoid is a critical component of Kp extract against this extreme allergic reaction. PMID- 18675941 TI - Comparison of nonhomologous end joining and homologous recombination in human cells. AB - The two major pathways for repair of DNA double-strand breaks (DSBs) are homologous recombination (HR) and nonhomologous end joining (NHEJ). HR leads to accurate repair, while NHEJ is intrinsically mutagenic. To understand human somatic mutation it is essential to know the relationship between these pathways in human cells. Here we provide a comparison of the kinetics and relative contributions of HR and NHEJ in normal human cells. We used chromosomally integrated fluorescent reporter substrates for real-time in vivo monitoring of the NHEJ and HR. By examining multiple integrated clones we show that the efficiency of NHEJ and HR is strongly influenced by chromosomal location. Furthermore, we show that NHEJ of compatible ends (NHEJ-C) and NHEJ of incompatible ends (NHEJ-I) are fast processes, which can be completed in approximately 30 min, while HR is much slower and takes 7h or longer to complete. In actively cycling cells NHEJ-C is twice as efficient as NHEJ-I, and NHEJ-I is three times more efficient than HR. Our results suggest that NHEJ is a faster and more efficient DSB repair pathway than HR. PMID- 18675942 TI - PHOX2B mutations and ventilatory control. AB - The transcription factor PHOX2B is essential for the development of the autonomic nervous system. In humans, polyalanine expansion mutations in PHOX2B cause Congenital Central Hypoventilation Syndrome (CCHS), a rare life-threatening disorder characterized by hypoventilation during sleep and impaired chemosensitivity. CCHS is combined with comparatively less severe impairments of autonomic functions including thermoregulation, cardiac rhythm, and digestive motility. Respiratory phenotype analyses of mice carrying an invalidated Phox2b allele (Phox2b+/- mutant mice) or the Phox2b mutation (+7 alanine expansion) found in patients with CCHS (Phox2b(27Ala/+) mice) have shed light on the role for PHOX2B in breathing control and on the pathophysiological mechanisms underlying CCHS. Newborn mice that lacked one Phox2b allele (Phox2b+/-) had sleep apneas and depressed sensitivity to hypercapnia. However, these impairments resolved rapidly, whereas the CCHS phenotype is irreversible. Heterozygous Phox2b(27Ala/+) pups exhibited a lack of responsiveness to hypercapnia and unstable breathing; they died within the first few postnatal hours. The generation of mouse models of CCHS provides tools for evaluating treatments aimed at alleviating both the respiratory symptoms and all other autonomic symptoms of CCHS. PMID- 18675943 TI - Oxidative metabolism of dopamine: a colour reaction from human midbrain analysed by mass spectrometry. AB - In order to identify the protein responsible for a dopamine peroxidizing activity, previously described in human normal and parkinsonian substantia nigra by our group, we developed non-denaturing polyacrylamide gel electrophoresis conditions, mimicking the characteristic colour in vitro reaction, resulting from cyclic oxidation of dopamine (DA). After separating protein mixtures from human normal midbrain homogenates on two sets of identical native gels, one gel set was subjected to specific activity staining by using DA and hydrogen peroxide. An activity red/orange band appeared in midbrain tissue lanes, similarly to the lane where commercial horseradish peroxidase (HRP) was present as control of peroxidative activity. The second set of gels, stained with Coomassie Blue, showed other, not enzymatically active protein bands. Mass spectrometry analysis of the bands containing the activity and the corresponding Coomassie Blue bands revealed the presence of proteins that may play a role in neurodegenerative disease, highlighting a possible functional link among dopamine/dopaminochrome redox cycle and protein metabolism. PMID- 18675944 TI - Substrate-induced conformational changes of the truncated catalytic domain of Geobacillus stearothermophilus lysyl-tRNA synthetase as examined by fluorescence. AB - The substrate-induced conformational change of the truncated C-terminal catalytic domain (CAT) of Geobacillus stearothermophilus lysyl-tRNA synthetase was examined by measuring tryptophan fluorescence of the truncated CAT domain in the presence or absence of the truncated N-terminal tRNA anticodon-binding domain (TAB). The fluorescence spectrum of CAT was not changed by the addition of l-lysine or ATP, whereas the intensity increased by adding a lysyl-adenylate analogue, suggesting that the CAT fluorescence increases when lysyl-adenylate is formed in the active site of CAT in l-lysine activation. In the presence of TAB, the addition of l lysine to CAT decreased the fluorescence, and the subsequent addition of ATP recovered partially the decreased intensity, as is similar to the case of the intact enzyme. The static parameters of the CAT-TAB complex were similar to those of the intact enzyme, suggesting that a somewhat impaired structure of CAT is repaired on the formation of the complex with TAB. The mutational analysis of the fluorescence showed that Trp314 but not Trp332 is responsible for the observed fluorescence changes. The role of the TAB domain in the intact enzyme is considered to enhance the binding efficiency of lysyl-adenylate to the CAT domain. PMID- 18675945 TI - The extra C-terminal tail is involved in the conformation, stability changes and the N/C-domain interactions of the calmodulin-like protein from pearl oyster Pinctada fucata. AB - Pearl oyster Pinctada fucata calmodulin-like protein (PfCaLP), containing an extra tail (D150-K161) at the C-terminal, is a novel protein involved in the regulation of oyster calcium metabolism. The purpose of this study is to gain insight into the conformational characteristics of the N/C-domain of PfCaLP, especially the detailed contribution of the extra tail to the Ca(2+)/Mg(2+) induced conformational changes, the stability of the intact PfCaLP molecule and its C-domain, as well as to the interdomain communications in PfCaLP. Our results demonstrate that a strong interaction exists between the hydrophilic tail and the C-domain of PfCaLP. The extra tail, through affecting the C-domain conformational changes, further influences the migration rate, conformational changes, N/C domain interactions and exposure of the hydrophobic patches of the intact PfCaLP molecule. Furthermore, the tail could actively regulate the stability of PfCaLP and its C-domain. Our studies are helpful to explain our previous finding that the tail plays important roles in PfCaLP-target interaction in the oyster calcium metabolism. PMID- 18675946 TI - AKT3 as a candidate gene for corpus callosum anomalies in patients with 1q44 deletions. PMID- 18675947 TI - A 12Mb deletion at 7q33-q35 associated with autism spectrum disorders and primary amenorrhea. AB - An interstitial deletion of about 12Mb at 7q33-q36 was found in an adult female affected by autism and primary amenorrhea. Two genes, CNTNAP2 and NOBOX, both contained within the deletion region, have been recently associated with autism susceptibility and premature ovarian failure, respectively. Our findings reinforce the hypothesis that haploinsufficiency of both these genes is sufficient for autism development and occurrence of primary amenorrhea, confirming a previous case in which CNTNAP2 had been disrupted by a chromosome inversion and possibly enlarging the phenotype of ovarian function disturbances already demonstrated for NOBOX mutations. PMID- 18675949 TI - The effects of intention and practice on a non-motor duration estimation task. AB - It has already been shown that the intention to reproduce a motor task in the future improves its learning. Here, we tested whether intention had a similar effect on learning to perform a non-motor duration estimation task, and if intention interacted with the amount of practice. Participants learned to estimate and compare two different durations, with or without intending to reproduce this learning in the future, and with two different amounts of practice. The results showed that intention and practice enhanced task learning independently. Moreover, only practice enhanced learning in over-estimation strategy. These finding are discussed with respect to a possible mechanism by which intention and practice might improve the learning of a duration task. PMID- 18675950 TI - [Right ventricular infarction caused by isolated right ventricular branch occlusion: a case report]. AB - It is rare to observe right ventricular infarction caused by isolated right ventricular branch occlusion. Isolated right ventricular infarction accounts for less than three percent of all cases of infarction. Generally, it is associated with occlusion of a non dominant right coronary artery or of a right ventricular branch. ECG can be misleading with ST segment elevation in anterior leads. We describe a patient admitted for chest pain with ST segment elevation in leads V1 to V3 associated with ST segment elevation in leads V3R and V4R. Coronary angiography demonstrated isolated total occlusion of the right ventricular branch. Thus, right precordial leads need to be done in every patient presenting with ST segment elevation in precordial leads V1 to V3 and not only in inferior myocardial infarction. PMID- 18675951 TI - Evaluation of musculoskeletal health outcomes in the context of job rotation and multifunctional jobs. AB - Higher physical demands at work are associated with health outcomes such as discomfort, disorders and sick leave. Variations in work exposure patterns, introduced by multifunctional jobs and ergonomic interventions, bring confounders into the complex relation between occupational risks and musculoskeletal disorders. This study compared whole-body rating of perceived exertion (RPE), discomfort, ergonomic workplace analysis (EWA) and sick leave due to musculoskeletal disorders, among workers exposed to diversified work. The results showed that EWA performed by the observer differed from workers' ratings. There were no differences between groups of workers taking or not taking sick leave regarding RPE and discomfort at their current workstations. Workers significantly discriminated between progressive workload levels, and RPE scores for specific tasks were nonlinear during shifts. These differences might be associated with exposure variability. Thus, in the context of diversified work, the RPE scale seems more appropriate for evaluating acute effects of work variability. PMID- 18675948 TI - Chromatin loops in gene regulation. AB - The control of gene expression involves regulatory elements that can be very far from the genes they control. Several recent technological advances have allowed the direct detection of chromatin loops that juxtapose distant genomic sites in the nucleus. Here we review recent studies from various model organisms that have provided new insights into the functions of chromatin loops and the mechanisms that form them. We discuss the widespread impact of chromatin loops on gene activation, repression, genomic imprinting and the function of enhancers and insulators. PMID- 18675952 TI - Influence of tooth removal on mandibular bone response to mastication. AB - The quantification of biomechanical response of mandibular bone to mastication is an integral component for a key in understanding the biological consequence of masticatory functions. Understanding the response of mandibular bone to external loading may also well explain the mechanisms of bone turnover. In this study, three finite element (FE) models simulating the lower second premolar, first and second molars along with their supporting structures were developed to determine stress/strain levels and distribution under different occlusal loading. The changes in stress/strain values and profiles have been investigated in three scenarios: pre-extraction of the lower first molar, post-extraction and after full healing of the extracted socket. The mastication induced equivalent strains within the supporting mandibular bone at each of these three scenarios were quantified and compared against the Frost's mechanostat theory. The results of stress/strain profiles show considerably lower magnitudes in the post-extracted and healed scenarios compared with the pre-extraction case. Following the Frost's MES hypothesis, the initial equivalent strains are related to local bone remodelling. It is found that in the extracted case the bone near the tooth socket undergoes resorption from lingual respect whilst filling the cavity, whereas in the healed case bone turnover reaches equilibrium. The results provide important data for clinical assessment of constructing dentures or other restorative devices. PMID- 18675953 TI - Words may not be enough! No increased emotional Stroop effect in obsessive compulsive disorder. AB - Conflicting evidence has been obtained whether or not patients diagnosed with obsessive-compulsive disorder (OCD) share an attentional bias towards disorder related stimuli. Some of these inconsistencies can be accounted for by suboptimal stimuli selection. In consideration of the heterogeneity of OCD, we investigated Stroop interference effects for two classes of OCD items (i.e., washing and checking) in 23 OCD patients and 23 healthy controls. In order to cover prevalent OCD concerns, item compilation was based on experts' appraisals. Patients neither displayed greater immediate as well as delayed Stroop interference nor any bias for OCD and subtype-congruent stimuli. On the contrary, for washing-related items, OCD patients, and here especially washers, displayed facilitation relative to healthy controls. Although the present study at first sight refutes the notion of an attentional bias in OCD in contrast to other anxiety disorders, several potential moderators need to be considered before this account is ultimately dismissed. In particular, an attentional bias may only be elicited using visual material that is more attention-grabbing than verbal stimuli. Finally, blockwise instead of random item administration and greater consideration of individual relevance may be crucial prerequisites for the effect to emerge. PMID- 18675954 TI - Anxiety sensitivity and perceived control over anxiety-related events: evaluating the singular and interactive effects in the prediction of anxious and fearful responding to bodily sensations. AB - The present investigation examined the singular and interactive effects of anxiety sensitivity and perceived control over anxiety-related events in the prediction of panic symptoms using a CO(2)-enriched air biological challenge. Two hundred and twenty-nine adult participants (M(age)=21.02, SD=7.55, 124 females) were recruited from the greater Burlington, Vermont community. Results indicated that pre-challenge anxiety sensitivity, but not perceived control over anxiety related events, significantly predicted post-challenge panic attack symptoms, anxiety focused on bodily sensations, and, interest in returning for another challenge (behavioral avoidance). In regard to physiological findings, anxiety sensitivity was significantly related to skin conductance level whereas perceived control over anxiety-related events was related to respiration rate. Neither anxiety sensitivity nor perceived control over anxiety-related events was related to heart rate. There also were no interactive effects between anxiety sensitivity and perceived control over anxiety-related events for any of the studied dependent variables. Results are discussed in relation to multi-risk factor models of cognitive vulnerability for panic psychopathology. PMID- 18675956 TI - Anergic cells generated by blocking CD28 and CD40 costimulatory pathways in vitro ameliorate collagen induced arthritis. AB - Collagen type II induced arthritis is an experimental model for studying the pathological mechanisms of therapeutic agents for human rheumatoid arthritis. In this study, we have investigated the effects of anergic cells on the development and disease progression of CIA. Anergic cells inhibited the proliferative response to collagen II primed lymphocytes, whereas the supernatant from anergic cell culture had no suppressive effect. Administration of anergic cells reduced the clinical score of CIA and ameliorated the development of CIA. The messenger RNA levels of IL-2 and IFN-gamma were significantly decreased, whereas mRNA levels of IL-4, IL-10 and TGF-beta showed no significant differences. A decrease in the collagen-specific Th1 associated IgG(2a) response was observed, whereas IgG(1) was unaffected. This effective treatment is associated with a reduction in IFN-gamma production and through cell-cell contact. Our results suggest that anergic cells may be beneficial for the treatment of rheumatoid arthritis. PMID- 18675955 TI - Acute hippocampal brain-derived neurotrophic factor restores motivational and forced swim performance after corticosterone. AB - BACKGROUND: Alterations in cellular survival and plasticity are implicated in the neurobiology of depression, based primarily on the characterization of antidepressant efficacy in naive rodents rather than on models that capture the debilitating and protracted feelings of anhedonia and loss of motivation that are core features of depression. METHODS: In adult male mice, we evaluated persistent effects of oral corticosterone (CORT) exposure on anhedonic-like behavior, immobility in the forced swim test (FST), motivational performance in the progressive ratio task, and later endogenous CORT secretion. After verifying long term decreases in hippocampal brain-derived neurotrophic factor (BDNF) and cAMP Response Element Binding protein phosphorylation (pCREB), the ability of direct hippocampal BDNF microinfusion after CORT exposure to reverse deficits was investigated. RESULTS: Prior CORT exposure decreased sucrose consumption, appetitive responding, and FST mobility without long-term effects on water/quinine discrimination and endogenous CORT secretion. Critically, BDNF replacement mimicked chronic antidepressant treatment (ADT) by reversing CORT induced reductions in instrumental performance and FST mobility. CONCLUSIONS: Together these findings link persistent alterations in hippocampal BDNF expression and CREB transcriptional activity with a persistent depressive-like state-as opposed to ADT efficacy. These results identify hippocampal BDNF as an essential molecular substrate that bidirectionally regulates appetitive instrumental behavior. Additionally, we suggest this CORT model might provide a powerful tool for future investigation into the neurobiology of complex stress associated depressive symptoms that persist long after stress exposure itself. PMID- 18675957 TI - Electrophysiological evidence for the time-course of verifying text ideas. AB - We examined how verb factivity influences the ability of readers to detect and resolve the mismatch of receiving false referents in relation to true referents in discourse contexts. Factive verbs (e.g., know), but not nonfactive verbs (believe), entail the truth of their complements. Recent research by Singer [Singer, M. (2006). Verification of text ideas during reading. Journal of Memory and Language, 54, 574-591] suggests that there are pragmatic costs associated with knowing something that is clearly false and only believing something that is clearly true. However, because Singer measured reading times for full sentences, it could not be determined whether these costs were initiated upon the appearance of the critical target word (i.e., the word that validated or invalidated previous text ideas) or at a later point in the sentences. In the present research we recorded event-related brain potentials while people read the same passages for comprehension and analyzed potentials evoked to the critical target words. Our results demonstrate that the brain distinguishes between true and false target words by at least 200ms after their onset, and that the pragmatic costs identified by Singer lead to interactions between verb factivity and truth in both early (P2) and later occurring brain components (late phase of N400 and late frontal positivity). In general, the results suggest readers had greater difficulty integrating false nouns than true nouns following factive than nonfactive verbs, and that detection of this mismatch also occurred earlier following factive verbs. Our results provide insight into the time-course of the processes that underlie the verification of text ideas, and extend neurocognitive research on anaphoric resolution. PMID- 18675958 TI - Incorporating PCA and fuzzy-ART techniques into achieve organism classification based on codon usage consideration. AB - To recognize the DNA sequence and mine the hidden information to achieve the classification of organisms are viewed as a difficult work to biologists. As we know, the amino acids are the basic elements to construct DNA. Hence, if the codon usage of amino acids can be analyzed well, the useful information about classification of organisms may be obtained. However, if we choose too many amino acids to perform the clustering analysis, the high dimensions also lead the clustering analysis to be a complicated structure. Hence, in this study, we will incorporate the principle component analysis and fuzzy-ART clustering techniques into constructing an integrated approach. The useful information about organisms classification based on the codon usage can be mined by using the proposed approach. Finally, we also employ a case including 18 bacteria to demonstrate the rationality and feasibility of our proposed approach. PMID- 18675959 TI - Gonadotropin-releasing hormone agonists for prevention of chemotherapy-induced ovarian damage: prospective randomized study. AB - OBJECTIVE: To determine whether GnRHa administration before and during combination chemotherapy for breast cancer could preserve posttreatment ovarian function in young women or not. DESIGN: Prospective randomized controlled study. SETTING: Department of Obstetrics and Gynecology, Mansura University Hospital, Mansura, Egypt. PATIENT(S): Eighty patients with unilateral adenocarcinoma of the breast and with no metastasis who had undergone modified radical mastectomy or breast-conserving surgery plus full axillary lymph node dissection were included in the study. Patients were assigned randomly to receive combined GnRHa and chemotherapy or chemotherapy alone. One woman in each group dropped out. MAIN OUTCOME MEASURE(S): Return of spontaneous menstruation and ovulation. Hormonal changes (FSH, LH, E(2), P) during and after the course of treatment. RESULT(S): In the study group, 89.6% resumed menses and 69.2% resumed spontaneous ovulation within 3-8 months of termination of the GnRHa/chemotherapy cotreatment; 11.4% experienced hypergonadotrophic amenorrhoea and ovarian failure 8 months after treatment. In the control group (chemotherapy without GnRHa), 33.3% resumed menses and 25.6% resumed normal ovarian activity. The median FSH and LH concentrations, 6 months after completion of the GnRHa/chemotherapy cotreatment group, were significantly less than the control group. During the GnRHa/chemotherapy cotreatment the concentrations of FSH, LH, and P decreased to almost prepubertal levels. However, within 1-3 months after the last GnRHa injection, an increase in LH and FSH concentrations was detected, followed several weeks later in by an increase in P concentrations to within normal levels. CONCLUSION(S): GnRHa administration before and during combination chemotherapy for breast cancer may preserve posttreatment ovarian function in women <40 years. Long-term studies are required. PMID- 18675960 TI - Infertility among women working in horticulture. A follow-up study in the Danish Occupational Hospitalization Register. AB - The possible association between employment in horticulture with potential exposure to pesticides and female infertility was examined by identification of women with hospital contact due to infertility and working in horticulture through the Danish Occupational Hospitalization Register. This follow-up study gave a standardized incidence ratio of 1.06 (95% confidence interval: 0.84-1.32) for treatment of infertility in women working in horticulture compared with the standard population and did not confirm that women working in the horticultural industry are at increased risk for infertility. PMID- 18675961 TI - Comparison of luteal estradiol patch and gonadotropin-releasing hormone antagonist suppression protocol before gonadotropin stimulation versus microdose gonadotropin-releasing hormone agonist protocol for patients with a history of poor in vitro fertilization outcomes. AB - OBJECTIVE: To compare IVF outcomes in poor-responder patients undergoing stimulation after luteal phase E(2) patch/GnRH antagonist (LPG) protocol versus microdose GnRH agonist protocol. DESIGN: Retrospective analysis. SETTING: University-based IVF center. PATIENT(S): Forty-five women undergoing ovarian stimulation for IVF using the LPG protocol were compared with 76 women stimulated with the microdose GnRH agonist protocol from May 2005 to April 2006. MAIN OUTCOME MEASURE(S): Cancellation rate, number of oocytes retrieved, and clinical pregnancy rates. RESULT(S): The mean number of oocytes (9.1 +/- 4.1 vs. 8.9 +/- 4.3) and mature oocytes (6.7 +/- 3.5 vs. 6.8 +/- 3.1) retrieved were similar, as were the fertilization rates (70.0% +/- 24.2% vs. 69.9% +/- 21.5%) and the number of embryos transferred (2.5 +/- 1.1 vs. 2.7 +/- 1.3). The cancellation rate was not significantly different between the groups (13/45, 28.9% vs. 23/76, 30.3%). Likewise, there were no significant differences among the implantation rate (15.0% vs. 12.5%), clinical pregnancy rate (43.3% vs. 45.1%), and ongoing pregnancy rate per transfer (33.3% vs. 26.0%) between both groups. CONCLUSION(S): This study demonstrates that the use of an E(2) patch and a GnRH antagonist during the preceding luteal phase in patients with a history of failed cycles can provide similar IVF outcomes when compared with the microdose GnRH agonist protocol. PMID- 18675962 TI - The value of repeat testicular sperm retrieval in azoospermic men. AB - To determine the predictive value of a previous testicular biopsy to the chance of sperm retrieval in the next testicular sperm extraction (TESE) procedure, we retrospectively analyzed the outcome of past sperm collection procedures and histopathology diagnoses of patients with nonobstructive azoospermia. Repeated TESE ensured a high recovery rate (96%) when the first recovery procedure had been successful and when hypospermatogenesis was diagnosed (77%); when no spermatozoa were found on the first attempt, a repeat TESE procedure was successful in one-third of the patients. PMID- 18675963 TI - Transurethral partial cystectomy and laparoscopic reconstruction for the management of bladder endometriosis. AB - OBJECTIVE: To report the successful management of bladder endometriosis with laparoscopic and transurethral partial cystectomy. DESIGN: Case report. SETTING: Tertiary-care university hospital. PATIENT(S): A 36-year-old woman with bladder endometriosis. INTERVENTION(S): Combined laparoscopic and transurethral excision of endometriotic lesions and bladder repair. MAIN OUTCOME MEASURE(S): Symptoms remission. RESULT(S): A hypoestrogenic agent with gonadotropin-releasing hormone (GnRH) agonist was administered for 6 months after the surgery. The patient found to be in good health with normal voiding and full continence during 14 months of regular follow-up evaluations. CONCLUSION(S): Combined laparoscopy and transurethral resectoscopy can be an alternative treatment to traditional laparotomy in women with bladder endometriosis, especially in those who have simultaneous pelvic endometriosis. PMID- 18675964 TI - Long-term ovarian function and fertility after heterotopic autotransplantation of cryobanked human ovarian tissue: 8-year experience in cancer patients. AB - OBJECTIVE: To assess the long-term ovarian function and fertility after heterotopic autotransplantation of frozen-thawed ovarian tissue in cancer patients. DESIGN: Prospective clinical case series. SETTING: Academic medical center PATIENT(S): Four young cancer patients who completed cancer treatment. INTERVENTION(S): Cryopreserved ovarian tissue (2000-2002) was thawed and transplanted to the heterotopic site (between the rectus muscle and fascia) between 2002 and 2005. MAIN OUTCOME MEASURE(S): [1] Serial blood tests (FSH, LH, estradiol, progesterone, testosterone) and ultrasound examinations. [2] Oocyte retrieval and in vitro fertilization. RESULT(S): The hormonal profiles were consistent with the postmenopausal level before transplantation. The return of the ovarian function was evidenced by hormonal profiles between 12 and 20 weeks after transplantation. Three patients underwent a second transplantation, as restored ovarian function lasted only 3 to 5 months. After the second transplantation, long-term ovarian function (lasting for 15-41 months) was established in all three patients. Six oocytes (one GV, four MI, one MII) were retrieved from the grafts. Three MI oocytes were developed to full maturity in vitro. Four MII oocytes were fertilized and developed to the cleavage stage embryos (up to six-cell). CONCLUSION(S): Autotransplantation of frozen-thawed ovarian tissue to a heterotopic site restored long-term ovarian function (for >40 months), and showed a potential to restore fertility in cancer patients. PMID- 18675965 TI - Meiotic spindle recovery is faster in vitrification of human oocytes compared to slow freezing. AB - OBJECTIVE: To investigate spindle behavior during and after slow freezing at room temperature (RT) and vitrification at different temperatures. DESIGN: Randomized, comparative study. SETTING: University hospital. PATIENT(S): Patients undergoing IVF treatment volunteered for the study and donated part of their supernumerary oocytes. INTERVENTION(S): Metaphase II oocytes were divided into group A: slow freezing RT /thawing RT; group B: vitrification RT/warming RT; group C: vitrification RT/warming 37 degrees C; and group D: vitrification 37 degrees C/warming 37 degrees C. Spindle presence was evaluated at each step of the four procedures and in culture. MAIN OUTCOME MEASURE(S): Cumulative spindle recovery rate comparing warming phase of the three vitrification groups and culture phase among the four groups. RESULT(S): During warming, the three vitrification groups showed a significantly fast spindle recovery rate compared to the thawing of the slow freezing group. A progressively significant fast cumulative recovery rate was observed in the three vitrification groups by increasing the number of phases at physiological temperature (hazard rate = 2.68; 95% confidence interval 1.71 4.02). CONCLUSION(S): The present study demonstrates that spindle recovery is faster in vitrification than in slow freezing. These data support a possible protective effect of vitrification/warming at 37 degrees C on the meiotic spindle structure and, therefore, on the subsequent clinical outcome of the procedure, although comparative clinical studies are needed. PMID- 18675966 TI - Effect of ormeloxifene, a selective estrogen receptor modulator, on biomarkers of endometrial receptivity and pinopode development and its relation to fertility and infertility in Indian subjects. AB - OBJECTIVE: To compare expression of endometrial estrogen (ER) and progesterone (PR) receptors, beta(3)-integrin subunit, leukemia inhibitory factor (LIF), interleukin-6 (IL-6), and pinopodes in Indian women using ormeloxifene as a contraceptive with fertile and infertile subjects during the window of implantation. DESIGN: Controlled, prospective, clinical study. SETTING: Hospital based reproductive health unit and research laboratories. PATIENT(S): Thirteen fertile women, 6 using ormeloxifene (30 mg/week), 29 with primary and 10 with secondary infertility. INTERVENTION(S): Transvaginal ultrasonography, midluteal endometrial biopsies, and blood samples. MAIN OUTCOME MEASURE(S): Histologic dating, endometrial thickness, immunohistochemical localization of ER, PR, beta(3)-integrin subunit, LIF, and IL-6 and scanning electron microscopy. RESULT(S): Ormeloxifene significantly reduced endometrial thickness, pinopode density, and caused histologic delay with increased epithelial ER and PR and unaltered epithelial beta(3)-integrin subunit expression. Appearance of fully developed pinopodes, down-regulation of epithelial PR, and increased epithelial beta(3)-integrin subunit expression was observed in in-phase endometrium from fertile and infertile women. LIF and IL-6 expression and serum estradiol and progesterone levels remained unaltered between groups. CONCLUSION(S): Ormeloxifene-induced effects might produce asynchrony between endometrial and embryo development resulting in implantation failure. Based on unaltered luteal phase serum progesterone concentration, ormeloxifene did not appear to prevent ovulation in any participant. PMID- 18675967 TI - Proportional change of CD4+CD25+ regulatory T cells after lymphocyte therapy in unexplained recurrent spontaneous abortion patients. AB - OBJECTIVE: To investigate the proportional changes of CD4(+)CD25(+) regulatory T cells in peripheral blood after lymphocyte therapy in unexplained recurrent spontaneous abortion (URSA) patients. DESIGN: Prospective cohort study. SETTING: University Hospital. PATIENT(S): Twenty-five URSA patients. INTERVENTION(S): Measurements of CD4(+)CD25(+) regulatory T cells in peripheral blood before and after paternal or third-party lymphocyte immunization. MAIN OUTCOME MEASURE(S): The proportion of CD4(+)CD25(bright) regulatory T cells and the percentage of CD25(bright) cells in the CD4(+) T-cell population. RESULT(S): The proportion of CD4(+)CD25(bright) T cells in peripheral blood from URSA patients was increased significantly after paternal or third-party lymphocyte immunization therapy, whereas the percentage of CD4(+)CD25(dim) cells were decreased significantly. The percentage of CD4(+)CD25(bright) cells in the CD4(+) T-cell population was significantly increased, and the proportion of CD4(+)CD25(bright) T cells was significantly higher in successfully pregnant women than in those with pregnancy loss after lymphocyte therapy. CONCLUSION(S): Allogeneic lymphocyte therapy can enhance the percentage of CD4(+)CD25(bright) regulatory T cells in peripheral blood, therefore CD4(+)CD25(+) regulatory T cells may serve as a novel biomarker for monitoring allogeneic lymphocyte therapy in URSA patients. PMID- 18675968 TI - A case report of successful pregnancy in a patient with pure 46,XY gonadal dysgenesis. AB - OBJECTIVE: To report a successful pregnancy in a patient with pure 46,XY gonadal dysgenesis. DESIGN: Case report. SETTING: Academic reproductive endocrinology and infertility unit. PATIENT(S): A patient with pure 46,XY gonadal dysgenesis and a desire to become pregnant. INTERVENTION(S): Laparoscopic gonadectomy, in vitro fertilization using donor oocytes, transfer of cryopreserved blastocysts, and cesarean delivery. MAIN OUTCOME MEASURE(S): Successful pregnancy and live birth. RESULT(S): Successful pregnancy and delivery of a healthy infant following in vitro fertilization using donor oocytes and embryo transfer. CONCLUSION(S): With the use of donor oocytes, patients with pure 46,XY gonadal dysgenesis can anticipate successful pregnancy. PMID- 18675969 TI - Rapidly progressing, late-onset obstructive azoospermia linked to herniorrhaphy with mesh. AB - OBJECTIVE: To report a case of a man presenting with rapidly progressing inguinal vasal obstruction 5 years after bilateral herniorrhaphy with polypropylene mesh. DESIGN: Case report with a review of the scientific literature. SETTING: Male infertility clinic, Kobe University Hospital (Kobe, Japan). PATIENT(S): A 30-year old patient who had undergone bilateral inguinal herniorrhaphy using polypropylene mesh 5 years previously. INTERVENTION(S): The patient underwent bilateral vasography and left testicular sperm extraction (TESE). MAIN OUTCOME MEASURE(S): Long-term effects of the adult inguinal herniorrhaphy with polypropylene mesh. RESULT(S): Rapid progression of vas deferens obstruction occurred within several months. We performed left TESE for intracytoplasmic sperm injection (ICSI) and retrieved many motile sperm. CONCLUSION(S): Before azoospermia, men who undergo inguinal herniorrhaphy using polypropylene mesh need to rapidly cryopreserve their sperm for future fertility; TESE-ICSI is also a selectable treatment. PMID- 18675970 TI - Ovarian follicular flushing among low-responding patients undergoing assisted reproductive technology. AB - A randomized comparison trial was performed to evaluate whether follicular reaspiration with use of a double-lumen retrieval needle improves oocyte recovery when compared with direct follicular aspiration among low-responding patients undergoing ART. There were no differences observed in the number of oocytes retrieved (single lumen: 6.5 +/- 2.2 oocytes, double lumen: 7.2 +/- 2.3 oocytes) whereas follicular reaspiration with the double-lumen retrieval needle resulted in a twofold increase in procedure time. PMID- 18675971 TI - High failure rates of medical termination of pregnancy after introduction to a large teaching hospital. AB - The success rates of medical termination of pregnancy in two time periods (2000 2001 and 2002-2003) were compared to assess the effectiveness of medical abortion introduction to a large academic tertiary medical center. The success rates were markedly reduced over time (87.0% vs. 79.3%) probably owing to the difficulty in defining clear sonographic criteria for treatment failure and the complexity of a follow-up program implemented at a large teaching hospital by a broad staff with widely varying experience and knowledge of the new procedure. PMID- 18675972 TI - GnRH agonist versus GnRH antagonist in controlled ovarian hyperstimulation: their role in patients with an unfavorable prognosis a priori. AB - In an attempt to examine the influence of the type of GnRH analogue used during controlled ovarian hyperstimulation on IVF outcome in patients with an unfavorable outcome a priori, we studied 728 consecutive cycles in patients with repeated IVF failure. In patients with repeated failure, the GnRH agonist group showed significantly higher clinical pregnancy rate compared with the GnRH antagonist group (20.8% vs 14.5%). PMID- 18675973 TI - Laparoscopic metroplasty in bicornuate and didelphic uteri. AB - OBJECTIVE: To determine the feasibility of laparoscopic metroplasty in the treatment of bicornuate and didelphic uteri. DESIGN: Case report. SETTING: University and private hospitals. PATIENT(S): Four women with a diagnosis of double uterine cavity (two bicornuate and two didelphic uteri) with a history of two recurrent spontaneous abortions at less than 5 months of pregnancy. INTERVENTION(S): Laparoscopic metroplasty with diagnostic hysteroscopy was performed for the unification of the uterus. Second-look laparoscopy and hysteroscopy was performed about 3 months later. MAIN OUTCOME MEASURE(S): Evaluation of the uterine compliance to high intrauterine pressure and presence of adhesions in the pelvic and uterine cavities. RESULT(S): In all four patients, laparoscopic metroplasty resulted in a unified uterus with a good cavity and tolerance to high intrauterine pressure. Minimal pelvic adhesions were noted in the two patients at the second surgery. CONCLUSION(S): This new technique of laparoscopic metroplasty is an acceptable alternative for abdominal metroplasty, with minimal adhesion formation. PMID- 18675974 TI - Ultrashort flare GnRH agonist combined with flexible multidose GnRH antagonist for patients with repeated IVF failures and poor embryo quality. AB - To evaluate the appropriate controlled ovarian hyperstimulation (COH) protocol in patients with repeated IVF failures and poor embryo quality we compared the stimulation characteristics of ten cycles which included ultrashort flare GnRH agonist combined with flexible multidose GnRH antagonist with the patients' earlier failed IVF attempts. The use of ultrashort GnRH agonist/GnRH antagonist COH protocol resulted in a significantly higher number and proportion of top quality embryos, with a consequent improvement in clinical pregnancy rate (50%). PMID- 18675975 TI - Successful term pregnancy after treatment of a cesarean scar ectopic gestation by endoscopic technique and conservative therapy. AB - OBJECTIVE: To describe a case of cesarean scar pregnancy treated successfully by endoscopic technique and medical therapy. DESIGN: Case report. SETTING: State general hospital in a major city. PATIENT(S): A 32-year-old woman with two cesarean sections and a recent curettage presented with ectopic cesarean scar pregnancy. INTERVENTION(S): Transvaginal ultrasound and hysteroscopy were used for diagnosis, and conservative therapy with methotrexate and hysteroscopy to treat the patient. MAIN OUTCOME MEASURE(S): Modified hysteroscopic technique and outcome. RESULT(S): Sonography and hysteroscopy revealed the presence of a gestational sac in the lower segment of the uterus in a patient with a cesarean section scar. She was successfully treated with systemic methotrexate and aspiration of the sac and local methotrexate injection under endoscopic control. CONCLUSION(S): Endoscopic intervention combined with medical treatment can result in a good therapeutic outcome with preservation of fertility in early ectopic cesarean scar pregnancy. PMID- 18675976 TI - Body mass index, waist to hip ratio and waist/height in adult Polish women in relation to their education, place of residence, smoking and alcohol consumption. AB - Obesity is a complex, multifactorial disorder that develops from genotype and environmental interactions. The aim of this study is to describe the variability of body mass index (BMI), waist to hip ratio (WHR) and waist to height (W/Ht) in adult Polish women, and to determine relationships between these variables and factors such as education, place of residence, smoking and alcohol drinking. The tested group consisted of 10,254 women aged 25-95 years, who voluntarily filled in questionnaires and participated in anthropometric measurements (body height and mass, waist and hip circumferences). The BMI, WHR and W/Ht values were calculated based on these measurements. The participants were differentiated in terms of education, residence and lifestyle (smoking, alcohol drinking). Chi squared test, product-moment correlations, ANOVA, multiple correspondence analysis (MCA) and logistic regression with backward elimination were used to evaluate associations between social and lifestyle factors and BMI, WHR and W/Ht. The results confirm (1) the relationship between low social status and the risk of overweight and obesity as observed in developed countries; (2) higher susceptibility to environmental factors such as education, place of residence, smoking and alcohol drinking in younger (premenopausal) women; (3) the usefulness of simple and practical anthropometric indicators such as WHR and W/Ht for the identification of the higher risk of future metabolic diseases in obese people and those with a normal body mass. PMID- 18675977 TI - Raymond Dart as a pioneering primatologist. AB - Raymond Dart is best known today for his groundbreaking research in palaeoanthropology. It is often forgotten, however, that Dart was a scientist of many interests, who made significant contributions to various disciplines. One of these is the study of living non-human primates. Dart became aware of the importance of primate studies and their relevance for research in other disciplines early in his career. In the late 1920s Dart established a colony of captive baboons in the Anatomy Department, University of the Witwatersrand, Johannesburg. On these animals, members of his Department, most notably Joseph Gillman and Christine Gilbert, carried out a number of significant behavioural and endocrinological researches. In 1930, as a member of an Italian Scientific Expedition, Dart was involved in hunting a mountain gorilla (for research purposes). He was also active in primate field studies. In 1957 he and Phillip Tobias founded a Witwatersrand University Uganda Gorilla Research Unit for the study of the Virunga mountain gorillas. The unit produced pioneering studies, conducted by Jill Donisthorpe, on the behaviour of these primates in their natural habitat. At the same time Dart was actively engaged in conservation of the mountain gorillas. He also studied South African chacma baboons in the wild. In the field of primate studies Raymond Dart figures prominently as a pioneering catalyst as well as researcher and conservationist. PMID- 18675978 TI - Brachial plexus reconstruction based on the new definition of level of injury. AB - SUMMARY: Although brachial plexus reconstruction remains a challenge to microsurgeons, especially when attempting to reconstruct cases with total root avulsion, much improvement in results has been recently achieved by a better understanding of various new methods of reconstruction, such as nerve transfer, functioning free muscle transplantation and prolonged postoperative rehabilitation. To better understand these improved results, we classified our patients into four levels of injury: (1) preganglionic root; (2) postganglionic spinal nerve; (3) pre- and retro-clavicular; (4) infraclavicular brachial plexus injury. Nerve transfer, functioning free muscle transplantation or local muscle transfer are the only possible solutions for the level 1 injury. Nerve transfers include extraplexus, intraplexus, close-target and end-to-side neurotisation. Neurolysis, nerve repair, nerve grafts, C-loop vascularised ulnar nerve grafts, nerve transfer and functioning free muscle transplantation are options for levels 2, 3 and 4 injuries. Tendon transfer, functioning or functional muscle transfer, arthrodesis or orthotics can be used for late palliative reconstruction. Taken together, these options can make brachial plexus surgery a worthy pursuit and make a useless limb useful. PMID- 18675979 TI - Survival following resuscitative thoracotomy for combined left ventricle and left atrium ruptures secondary to blunt trauma. AB - Improvements in pre-hospital care and the development of integrated Trauma Systems have streamlined access for the severely injured to sophisticated, specialist Trauma Centre reception and resuscitation. We describe the initial care of a survivor of combined ruptures of the left ventricle and left atrium secondary to blunt injury. This case emphasises the contribution of such a Trauma System in achieving a favourable outcome for a severely injured trauma patient with injuries previously considered non-survivable. PMID- 18675980 TI - Novel genetic variants linked to coronary artery disease by genome-wide association are not associated with carotid artery intima-media thickness or intermediate risk phenotypes. AB - BACKGROUND: It is uncertain whether the novel single nucleotide polymorphisms (SNPs) that have recently been associated with coronary artery disease (CAD) in genome-wide studies also influence carotid atheroma and stroke risk. The mechanisms of their association with CAD are unknown; relationships to other cardiovascular phenotypes may give mechanistic clues. Carotid artery intima-media thickness (CIMT) is a subclinical marker of atherosclerosis associated with stroke. We investigated association of reported CAD risk variants with CIMT, and with other intermediate phenotypes that may implicate causative pathways. METHODS: We studied 1425 members of 248 British Caucasian families ascertained through a hypertensive proband. We genotyped CAD risk SNPs on chromosomes 9 (rs1333049, rs7044859, rs496892, rs7865618), 6 (rs6922269) and 2 (rs2943634) using TaqMan. Merlin software was used for family-based association testing. RESULTS: No significant association was found between genotype at any SNP and CIMT in 846 individuals with acceptable measurements. Nor were SNPs significantly associated with blood pressure, obesity, cholesterol, CRP, interleukin-6, TNF alpha, or leptin. CONCLUSIONS: These novel CAD variants are not associated with CIMT and do not appear to mediate the risk of atherothrombosis through known risk factors. PMID- 18675981 TI - A variable-stiffness shoe lowers the knee adduction moment in subjects with symptoms of medial compartment knee osteoarthritis. AB - The purpose of this study was to evaluate the effectiveness of variable-stiffness shoes in lowering the peak external knee adduction moment during walking in subjects with symptomatic medial compartment knee osteoarthritis. The influence on other lower extremity joints was also investigated. The following hypotheses were tested: (1) variable-stiffness shoes will lower the knee adduction moment in the symptomatic knee compared to control shoes; (2) reductions in knee adduction moment will be greater at faster speeds; (3) subjects with higher initial knee adduction moments in control shoes will have greater reductions in knee adduction moment with the intervention shoes; and (4) variable-stiffness shoes will cause secondary changes in the hip and ankle frontal plane moments. Seventy-nine individuals were tested at self-selected slow, normal, and fast speeds with a constant-stiffness control shoe and a variable-stiffness intervention shoe. Peak moments for each condition were assessed using a motion capture system and force plate. The intervention shoes reduced the peak knee adduction moment compared to control at all walking speeds, and reductions increased with increasing walking speed. The magnitude of the knee adduction moment prior to intervention explained only 11.9% of the variance in the absolute change in maximum knee adduction moment. Secondary changes in frontal plane moments showed primarily reductions in other lower extremity joints. This study showed that the variable-stiffness shoe reduced the knee adduction moment in subjects with medial compartment knee osteoarthritis without the discomfort of a fixed wedge or overloading other joints, and thus can potentially slow the progression of knee osteoarthritis. PMID- 18675982 TI - Thermo-responsive monolithic materials. AB - One of the recent major improvements of HPLC was the introduction of monolithic silica columns, which have the advantage of faster separation and lower back pressure as compared to common silica beads. Here, we present an interesting alternative to such reversed-phase monolithic columns by a convenient coupling route of a thermo-responsive polymer to hydrophilic silica monoliths. Poly(N isopropylacrylamide) (PNIPAM) was polymerized in solution via a reversible addition fragmentation chain transfer (RAFT) polymerization technique and coupled then in situ onto an amino-modified silica monolithic column. These columns were compared with RP-18 monolithic columns in the separation of steroids under isocratic condition in aqueous mobile phase. Separation is optimized just by changing the temperature, instead of changing the mobile phase composition. PMID- 18675983 TI - Development of packed-column suppressor system for capillary ion chromatography and its application to environmental waters. AB - A novel suppressor unit for capillary ion chromatography was designed to reduce the background conductivity and at the same time to increase the analyte signal. Regeneration of the suppressor was carried out on-line by passing an appropriate acidic solution through the column to displace the accumulated eluent cations. By using two 6-port microswitching valves and two packed capillary column suppressors, the background conductivity of sodium carbonate-bicarbonate mobile phase was maintained at low conductivity for continuous chromatographic runs, and the detection limits at low ppb levels were achieved. The relative standard deviations (RSDs) for the retention time, peak area and peak height of six common inorganic anions (0.05 mM each of F(-), Cl(-), NO(2)(-), Br(-), NO(3)(-) and SO(4)(2-)) were between 0.5-0.9, 1.1-4.6 and 0.7-4.9%, respectively. The present system was successfully applied to the determination of inorganic anions in river water and tap water samples. PMID- 18675984 TI - Kinetic plot equations for evaluating the real performance of the combined use of high temperature and ultra-high pressure in liquid chromatography. Application to commercial instruments and 2.1 and 1 mm I.D. columns. AB - The use of ultra-high pressure liquid chromatography (UHPLC) with pressures up to 1000 bar and columns packed with sub-2-microm particles combined with high temperature mobile phase conditions (up to 90 degrees C) is assessed according to the current available instrumentation via constrained kinetic plot equations. It is shown that the gain in separation speed, theoretically expected from high temperature UHPLC (HT-UHPLC), is significantly reduced when taking into account the existing instrumental constraints (extra-column band broadening, flow-rate and column length limitations). This study also shows that significant improvements could be expected on the current commercial instruments by increasing the flow-rate limit and/or using packing columns with particle size in the range 2.5-3.5 microm instead of the current sub-2 microm. These particles should obviously withstand very high pressure. PMID- 18675985 TI - Dynamic adhesion behavior of micrometer-scale particles flowing over patchy surfaces with nanoscale electrostatic heterogeneity. AB - The dynamic adhesion behavior of micrometer-scale silica particles is investigated numerically for a low Reynolds number shear flow over a planar collecting wall with randomly distributed electrostatic heterogeneity at the 10 nanometer scale. The hydrodynamic forces and torques on a particle are coupled to spatially varying colloidal interactions between the particle and wall. Contact and frictional forces are included in the force and torque balances to capture particle skipping, rolling, and arrest. These dynamic adhesion signatures are consistent with experimental results and are reminiscent of motion signatures observed in cell adhesion under flowing conditions, although for the synthetic system the particle-wall interactions are controlled by colloidal forces rather than physical bonds between cells and a functionalized surface. As the fraction of the surface (Theta) covered by the cationic patches is increased from zero, particle behavior sequentially transitions from no contact with the surface to skipping, rolling, and arrest, with the threshold patch density for adhesion (Theta(crit)) always greater than zero and in quantitative agreement with experimental results. The ionic strength of the flowing solution determines the extent of the electrostatic interactions and can be used to tune selectively the dynamic adhesion behavior by modulating two competing effects. The extent of electrostatic interactions in the plane of the wall, or electrostatic zone of influence, governs the importance of spatial fluctuations in the cationic patch density and thus determines if flowing particles contact the wall. The distance these interactions extend into solution normal to the wall determines the strength of the particle-wall attraction, which governs the transition from skipping and rolling to arrest. The influence of Theta, particle size, Debye length, and shear rate is quantified through the construction of adhesion regime diagrams, which delineate the regions in parameter space that give rise to different dynamic adhesion signatures and illustrate selective adhesion based on particle size or curvature. The results of this study are suggestive of novel ways to control particle-wall interactions using randomly distributed surface heterogeneity. PMID- 18675986 TI - Solvent-tuned multiple self-assembly of a new sugar-appended gelator. AB - 4-(4'-Butoxyphenyl)phenyl-beta-O-D-glucoside was synthesized as a low-molecular mass gelator. It was able to immobilize a variety of aqueous and organic solvents in large amounts through the formation of three-dimensional self-assembled fibrillar networks. The morphologies of the aggregates depended on the nature of solvent where they were formed. Planar ribbons were observed in water, while helical ribbons were dominant in toluene and sheets in CHCl(3). The xerogel from water exhibited a lamellar structure with a d-spacing of 2.45 nm as demonstrated by 1D-WAXD, indicating a bilayer structure interdigitated with butoxy tails, whereas the xerogel from CHCl(3) or toluene yielded a lamellar structure with a d spacing of 3.05 nm, implying a bilayer structure interdigitated with glycosyl heads. Increasing the content of 1,4-dioxane in water caused a gradual transformation from planar to twisted ribbons and then tubes. PMID- 18675987 TI - Characterization of physical structure of silica nanoparticles encapsulated in polymeric structure of polyamide films. AB - Polyamide nanocomposite films were prepared from nanometer sized silica particles and trimesoyl chloride-m-phenylene diamine based polyamides. The type of silica nanoparticles used is commercial LUDOX HS-40 and the particle size characterized by the radius of gyration (R(g)) is about 66 A. The immediately prepared films were easily broken into particles to form colloidal-like dilute suspension of the silica-polyamide composite particles in D(2)O-H(2)O solutions for SANS measurements, that in this dilute system SANS data the complication of scattering data from the interacting particles is minimized. At about 60% D(2)O of the sample solution, the silica is contrasted out, therefore the SANS profiles are predominantly from the organic polyamide scattering. The SANS profile of the sample solutions measured at 90% D(2)O clearly indicates scattering from both silica and polymer. The scattering heterogeneities for two-phase system were evident from the validity of the Debye-Bueche expression in case of the nanocomposite with high silica loading. At limited silica loading of the nanocomposite, interaction between the silica and polymer chains forming core shell morphology was observed. The transport properties of the membranes made from the nanocomposite films were measured on a batch type test kit with an aqueous solution of 500 ppm dioxane concentration at pressures ranging from 50 to 200 psig, and correlated to their composite structure. PMID- 18675988 TI - Synthesis of ordered mesoporous silica membrane on inorganic hollow fiber. AB - This paper reports on a new method for the preparation of mesoporous silica membranes on alumina hollow fibers. A surfactant-silica sol is filled in the lumen of an alpha-alumina hollow fiber. The filtration technique combined with an evaporation-induced self-assembly (EISA) process results in the formation of a continuous ordered mesoporous silica layer on the outer side of alpha-alumina hollow fibers. X-ray diffraction (XRD), transmission electron microscopy (TEM), and nitrogen isothermal adsorption measurements reveal that these membranes possess hexagonal (P6mm) mesostructures with pore diameters of 4.48 nm and BET surfaces of 492.3 m(2) g(-1). Scanning electron microscopy (SEM) studies show that the layers are defect free and energy-dispersive spectroscopy (EDS) mapping images further confirm the formation of continuous mesoporous silica layer on the outer side of alpha-alumina hollow fibers. Nitrogen and hydrogen permeance tests show that the membranes are defect free. PMID- 18675989 TI - Influence of long-term aqueous exposure on surface properties of plasma-sprayed oxides Cr2O3 and Cr2O3-25 wt% TiO2. AB - The influence of water exposure on the surface properties of plasma-sprayed Cr(2)O(3) and Cr(2)O(3)-25 wt% TiO(2) was studied. It was shown that both plasma sprayed materials contained Cr(VI) hydrous oxide phases, which dissolved rapidly at the beginning of water exposure. The dissolution continued slowly during the whole water exposure time. The Cr(VI) dissolution was accompanied by a rapid increase in surface IEP value. Both Cr(2)O(3) and Cr(2)O(3)-25 wt% TiO(2) showed similar dissolution, zeta potential, and surface oxidation states. Thus the addition of TiO(2) did not influence the surface properties of the plasma-sprayed Cr(2)O(3). PMID- 18675990 TI - Lymphoepithelioma-like carcinoma of the urinary bladder in a cow associated with bovine papillomavirus type-2. AB - Lymphoepithelioma-like carcinoma (LELCA) of the urinary bladder is reported in a 7-year-old cow that had grazed pasture rich in bracken fern and had suffered from severe intermittent haematuria from 3 to 4 years of age. On necropsy examination there were multiple haemorrhagic foci scattered over the mucosal surface of the urinary bladder. Microscopically there were nests, cords and sheets of neoplastic cells infiltrating the lamina propria and muscularis propria. These had a syncytial appearance with ill-defined cytoplasmic borders, large nuclei and prominent nucleoli. There was a prominent associated inflammatory infiltrate comprising lymphocytes and plasma cells with sparse histiocytes and granulocytes. Immunohistochemically, LELCA cells expressed cytokeratin but not vimentin. The LELCA was focally admixed with a concomitant papillary high-grade carcinoma that also infiltrated the lamina propria. A diffuse carcinoma in situ was also present. Bovine papillomavirus type-2 (BPV-2) DNA was amplified from frozen neoplastic tissue and from selected areas of formalin-fixed, paraffin wax embedded tissue obtained by laser capture microdissection. Microbiological culture of a urine sample resulted in isolation of Weeksella virosa, Rhizobium radiobacter and Staphylococcus warneri. Flow cytometric analysis performed on blood mononuclear cells revealed down-regulation of a panel of markers including CD3, CD4, CD8alpha, CD45, MHC class I and MHC class II (HLA-DRalpha, HLA-DQ, HLA DP). This report extends the spectrum of neoplastic urothelial lesions described in cattle and provides further evidence that some features of these tumours are similar to human counterparts. PMID- 18675991 TI - Detection and characterization of chondroid metaplasia in canine atrioventricular valves. AB - The atrioventricular valves of 25 dogs of different breeds and age were examined grossly and microscopically following histochemical staining and immunohistochemical labelling for collagen types I, III and VI, and for fibronectin and laminin. Foci of cartilage were identified in the tricuspid septal leaflet within the fibrosa (n=21) or spongiosa (n=3). These were further characterized as either fibrocartilage, predominantly composed of collagens I and VI, or hyaline cartilage consisting of laminin and collagens III and VI. Eighteen of the dogs were of large breed and seven of small breed. Retrospective echocardiographic findings were available from five cases and in three of these a hyperechogenic structure was identified corresponding to the cartilage focus (0.1, 1.12 and 5.63 mm(2) in size). The clinical significance and mechanism of formation of these cartilaginous foci remain undetermined, although factors such as breed, size and concurrent chronic valvular disease may be significant. PMID- 18675992 TI - Serine/threonine kinase Pim-2 promotes liver tumorigenesis induction through mediating survival and preventing apoptosis of liver cell. AB - BACKGROUND: It has been proven that serine/threonine kinase pim-2 mediates cell survival and prevents apoptosis in hematopoietic system tumors and lymphomas, but its role in solid organ tumor induction is still unclear. In this study, we investigated its effects and underlying mechanisms in tumorigenesis of hepatocellular carcinoma. METHODS: We first examined the pim-2 gene expression and its protein levels in human hepatocellular carcinoma, paired noncancerous liver, and normal liver tissues. Then, we cultured human liver cancer cells and immortalized liver cells to examine the effects of pim-2 gene on the cell viability, growth, and apoptosis in different culture conditions. For further investigation of the molecular events in the pim-2 signal pathway, we also explored pim-2 kinase activity on phosphorylation of the two downstream signal mediators: 4E-BP1 and Bad. RESULTS: Pim-2 gene and protein were notably expressed in human liver cancer tissues and HepG2 cells. The ectopic pim-2 overexpressing L02 cells were able to survive in interleukin-3 (IL-3)-deprived circumstance but not in glucose-free medium. Compared with HepG2 cells, pim-2 knock-down HepG2 cells lost survival ability in IL-3 starvation medium. In pim-2-expressing cells, both the total protein expressions of 4E-BP1 and Bad were kept stable; however, their phosphorylated patterns were notably increased. CONCLUSIONS: Our results indicate that pim-2 acts as a pro-survival kinase to inhibit apoptosis and keep liver cell survival in IL-3-deprived medium. Pim-2 might participate in the tumorigenesis of hepatocellular carcinoma induction through its downstream molecules 4E-BP1 and Bad. PMID- 18675993 TI - The role of P38 MAPK and PKC in BLP induced TNF-alpha release, apoptosis, and NFkappaB activation in THP-1 monocyte cells. AB - BACKGROUND: P38 mitogen activated protein kinase (p38 MAPK) is a critical mediator of the inflammatory response, which makes it a suitable candidate as a novel therapeutic strategy for inflammatory conditions. In this study, we set out to examine the precise role of both protein kinase C (PKC) and P38 MAPK signaling kinases in bacterial lipoprotein (BLP) induced nuclear factor-kappa B (NFkappaB) activation and tumor necrosis factor-alpha (TNFalpha) release in THP-1 monocytic cell line. MATERIALS AND METHODS: THP-1 cells were incubated with BLP(0-1000 ng/mL), phorbol myristate acetate (PMA; 0-100 microg/mL) or a combination of both for 6 and 24 h, with or without pretreatment with SB202190, a specific inhibitor of p38 MAPK and bisindolylmaleimide I, a specific inhibitor of PKC (0-200 microm). Cell supernatants were analyzed for TNF-alpha release and apoptosis. NFkappaB activity was analyzed by electromobility supershift assay. RESULTS: BLP induced TNF-alpha release was significantly reduced by pretreatment with SB202190 at all concentrations (428.7 +/- 5.9 versus 51 +/- 0.8 rhog/mL, P < 0.05). Pretreatment with bis I significantly inhibited TNF-alpha release at higher concentrations (200 microM) (429.7 +/- 5.9 versus 194.9 +/- 42.68 rhog/mL, P < 0.05) but this was much less effective than SB202190. PMA induced TNF-alpha release was not inhibited at 6 h by either SB202190 or bis I, but was significantly so at 24 h (148.5 +/- 9.8 versus 24 +/- 1.7 and 25.1 +/- 4.4 rhog/mL, P < 0.05). BLP or lipopolysaccharide (LPS) did not result in apoptosis in THP-1 cells (P > 0.05) with PMA inducing apoptosis in a time- and dose dependent manner. In combination with BLP (1000 ng/mL) but not LPS (1000 ng/mL), low dose PMA resulted in a significant increase in apoptosis, 6% +/- 0.5% (Control) versus 9.2% +/- 0.3% (P < 0.05) and 7% +/- 2.2% (Control) versus 7.7% +/- 0.3% (P > 0.05), respectively. This synergistic effect was inhibited by bisindolylmaleimide 100 nm, 8.9% +/- 0.9% (Control) versus 9.8% +/- 0.2% (P > 0.05). PMA and BLP induced rapid nuclear translocation of NFkappaB, which was inhibited by pretreatment with both SB-202190 and bis I, and SB202190 but not bis I, respectively. CONCLUSIONS: P38 is a critical mediator of BLP induced TNF-alpha release and NFkappaB activation, whereas PKC is only partially responsible for its response. P38 and PKC are both critical mediators of PMA induced TNF-alpha release and NFkappaB activation. PMID- 18675994 TI - Establishment of an acute superior mesenteric artery injury model for damage control surgery. AB - BACKGROUND: Managements of superior mesenteric artery (SMA) injuries are difficult and often result in a disappointing outcome. Damage control surgery (DCS) has been approved to be an effective and reliable strategy for severe trauma victims. We aimed to build up a severe trauma-shock-hypothermia model of SMA injuries for DCS study and determine the optimal time to institute DCS. METHODS: Pigs were anesthetized and instrumented with arterial and a thermodilution cardiac output catheter. SMA flow was interrupted while animals were hemorrhaged to 45% estimated blood volume. Pigs were maintained shock and intestine ischemia for three durations: intestine ischemia for 30 min (I-30; n = 6), 60 min (I-60; n = 6), and 90 min (I-90; n = 6). Cold lactated Ringer's (10 mL/kg) was infused to induce hypothermia. SMA was then declamped and kept in reperfusion for 6 h. Hemodynamic data and serum samples were collected during shock and resuscitation. Distal ileum was collected at the end of ischemia and reperfusion. RESULTS: All animals presented with disastrous conditions at the end of ischemia: low temperature, severe acidosis, decreased blood pressure, depressed cardiac output, and oxygen delivery. I-90 animals suffered the lowest temperature, the most severe acidosis, lowest blood pressure, and depressed cardiac output and oxygen delivery. Coagulopathy developed in I-90, whereas normal prothrombin time and thrombin time were detected in I-30 and I-60. Aspartate aminotransferase, lactate dehydrogenase, creatine kinase, and alkaline phosphatase were equally within groups (P > 0.05). All (6/6) of I-30, 83.3% (5/6) of I-60, and 16.7% (1/6) of I-90 pigs survived (P < 0.01). Base excess in I-90 was much lower than that in I-30 and I-60 animals. CONCLUSIONS: We first built up an acute SMA injury animal model for DCS investigations and determined that the optimal institution time of DCS was before 60 min after SMA injury in the trauma shock-hypothermia swine model. PMID- 18675995 TI - Functional mapping in PD and PSP for sustained phonation and phoneme tasks. AB - AIM: To elucidate the central basis of articulatory speech disorders in Parkinsonian syndromes using functional magnetic resonance imaging (fMRI). METHODS: Twenty-two patients with Parkinson's disease (PD) and 18 with progressive supranuclear palsy (PSP) were clinically evaluated for speech dysfunction. Functional magnetic resonance imaging (fMRI) was carried out in these patients using sustained phonation and phoneme tasks. Individual and group analysis using SPM2 was done for eight patients with PD, 7 with PSP and 6 healthy controls. SETTING: Tertiary Medical Teaching Institute. RESULTS: For sustained phonation paradigm, superior temporal gyrus area was activated in PD patients, and occipital cortex in PSP subjects in comparison to controls. For phoneme paradigm, the patients with PD recruit lingual gyrus obviating the need for more efforts for the task. Also wider areas as well as more clusters were activated in PD patients compared to controls. Lingual gyrus was found to be strongly activated in PSP patients. Reduced activation of the primary areas with recruitment of remote areas was another prominent finding in PSP. Due to excessive motion (>1.5 mm, >1 degrees ) in all the MSA patients, they could not be considered for analysis. CONCLUSION: The failure of the executive fronto striatal network would lead to increased activation of other areas in PD, but in PSP, there is a widespread cortical dysfunction. PMID- 18675996 TI - [123I]-FP-CIT and [99mTc]-HMPAO single photon emission computed tomography in a new sporadic case of rapid-onset dystonia-parkinsonism. AB - Rapid-onset dystonia-parkinsonism (RDP) is a rare, autosomal-dominantly inherited syndrome characterized by abrupt onset, over hours to days, of dystonic and parkinsonian symptoms. To date, RDP has been described in a small number of families, and in only four sporadic cases. METHODS: We here report a new sporadic case of RDP who has a novel de novo mutation in the ATP1A3 gene. Striatal dopamine transporters have been assessed quantitatively using [123I]-FP-CIT SPECT. A volume of interest (VOI) was drawn within the occipital cortex to obtain non-specific activity and specific to non-specific binding ratios (BR) were calculated. A single template of predefined VOI 3D-drawn on right and left caudate nucleus and putamen was applied to the spatially normalized BR images. BR values were compared to those obtained from an age-matched control group and from a group of patients suffering from Parkinson's disease (Hoehn and Yahr score 2 or 3). A [99mTc]-HMPAO cerebral blood flow study was also performed. RESULTS: In the control group, BR values (mean+/-Standard Deviation) were 3.5+/-0.4 for the left striatum and 3.3+/-0.3 for the right one. RDP patient's values were 3 and 2.7, respectively. In the Parkinson group, values were 1.6+/-0.3 and 1.7+/-0.4, respectively. [99mTc]-HMPAO scan showed homogeneous cortical and sub-cortical perfusion. CONCLUSION: Quantification of striatal [123I]-FP-CIT uptake in a new sporadic case of RDP with a novel mutation in the ATP1A3 gene showed values just within the range of normality. [99mTc]-HMPAO scan was normal. PMID- 18675997 TI - Flow cytometric assay for quantitative and qualitative evaluation of adhesive interactions of tumor cells with endothelial cells. AB - The purpose of the study was to develop a flow cytometric assay for quantitative determination of adhesive interactions of human endothelial cells (ECs) with tumor cells. EC lines established from human lymph node, appendix, lung, skin and intestine microvessels, labeled with PKH26-GL fluorescent dye, were grown to confluency in 24-well TC plates. Human colon adenocarcinoma cell suspension was overlaid onto labeled ECs, and allowed to adhere for 20 min at 4 degrees C under static conditions. Non-adhering cells were collected first, and adhering tumor cells together with ECs were detached from the culture plate. Collected cell fractions were evaluated by flow cytometry. Results were re-calculated as a ratio (R) of adhering colon carcinoma cells per one EC. We demonstrated that immortalized human microvascular ECs preserved their organ specificity. Colon carcinoma cells adhere preferentially to ECs of intestine origin. The immunofluorescent staining of adhering and non-adhering cancer cell subpopulations has revealed an augmented level of Lewis(x) antigen on adhering cancer cells. The organ specificity of endothelial cell interactions with colon carcinoma cells demonstrated in static conditions was verified and confirmed with flow adhesion assay. The method elaborated is suitable for quantifying of tumor cells adhering to ECs, with simultaneous evaluation of cell surface phenotypic markers of both partner cells participating in adhesive interactions. Validated by comparison to dynamic shear stress adhesion assay in blood flow reconstituted conditions this assay greatly facilitates evaluation of tumor cell-endothelial cell mutual interactions taking place during metastatic process. PMID- 18675998 TI - [Acute disseminated encephalomyelitis associated with hepatitis A virus infection]. AB - INTRODUCTION: Hepatitis virus A (HVA) infection is usually a benign infection, but it can lead to severe manifestations and neurological symptoms. CASE REPORT: We report the case of a 44-year-old man who was admitted for pyramidal tetraparesis, loss of proprioceptive sensitivity and cranial nerve involvement. He had developed concomitally jaundice and fatigue. Brain MRI and cerebrospinal fluid examination were normal. Blood tests revealed elevated serum transaminase and anti-hepatitis A virus (IgM and IgG) levels. Acute disseminated encephalomyelitis (ADEM) was diagnosed and the patient was treated with high dose intravenous then oral corticosteroid therapy. The clinical condition continued to deteriorate and the patient died at eight months. DISCUSSION: ADEM is exceptionally associated with HVA infection or after vaccination for hepatitis A. Other neurological complications, including either peripheral or central nervous system, are reported. The clinical presentation and the outcome of our patient are atypical. PMID- 18675999 TI - Ventricular septal defects at the University of Nigeria Teaching Hospital, Enugu: a review of echocardiogram records. AB - This research aimed to evaluate the prevalence of ventricular septal defect (VSD) and to determine the age and gender distribution of this defect and its common associated congenital cardiac anomalies among patients referred for echocardiographic examination at the echocardiography laboratory of the University of Nigeria Teaching Hospital, Enugu, Nigeria. Echocardiogram reports from the 10-year period February 1991-September 2001 were analysed. There were 2486 echocardiogram scans with an additional 334 repeat scans; 593 subjects had congenital anomalies, of which 165 (28%; 98 males and 67 females) had VSDs. The highest prevalence of VSDs (118; 70%) was in children age 0-9 years although VSDs were found in adults aged up to 50 years and above. Persistent ductus arteriosus and atrial septal defect were the commonest associated congenital anomalies. The study has shown that VSD is a relatively common congenital heart disease and that many patients present with VSD in adult life. There is a need for thorough scrutiny to be made for other cardiac anomalies in all patients with VSD, as well as for corrective surgery to be made affordable to those affected by this defect and other associated anomalies. PMID- 18676000 TI - The capsid-coding region hairpin element (cHP) is a critical determinant of dengue virus and West Nile virus RNA synthesis. AB - Dengue virus (DENV) and West Nile virus (WNV) are members of the Flavivirus genus of positive-strand RNA viruses. RNA sequences and structures, primarily in the untranslated regions, have been shown to modulate flaviviral gene expression and genome replication. Previously, we demonstrated that a structure in the DENV coding region (cHP) enhances translation start codon selection and is required for viral replication. Here we further characterize the role of the cHP in the DENV life cycle. We demonstrate that the cHP is required for efficient viral RNA synthesis in a sequence-independent manner. Viruses with a disrupted cHP are rescued by a spontaneous compensatory mutation that restabilizes the structure. Furthermore, the cHP, which is predicted to be conserved among arthropod-borne flaviviruses, is required for WNV replication. We propose that the cHP is a multifunctional determinant of flavivirus replication, functioning in both translation and RNA synthesis. PMID- 18676001 TI - Soil particle-size dependent partitioning behavior of pesticides within water soil-cationic surfactant systems. AB - Cationic surfactants have been proposed for enhanced sorption zones to contain hydrophobic organic compound (HOC) contamination. Benzalkonium chloride (BC), a cationic surfactant, was selected to study the particle-size dependent sorption behavior of the surfactant and its role in the immobilization of two hydrophobic pesticides (atrazine and diuron) within soil-water-surfactant systems for this application. Five different soils were considered in this study. Our results showed significant particle-size dependent behavior for surfactant sorption and pesticide immobilization in the presence of the sorbed cationic surfactant. The cation exchange capacity (CEC) of the bulk soils and their size fractions (clay, silt, and sand fractions) determined BC sorption capacity. In the absence of BC the sand fractions were the least effective sorbent for the pesticides compared with silts and clays. However, at relatively low BC mass sorbed (<10,000mg/kg) to any of the soil fractions, well below sorption saturation, the sand fractions became more effective sorbents for either pesticide than the clay and silt fractions. The pesticide partitioning coefficient onto sorbed BC (K(ss)) was not constant. Particle CEC, availability of CEC sites for sorption of the cationic surfactant, and the amount of the BC sorbed determined the phase of K(ss). The maximum K(ss) occurred before BC saturation sorption capacity was reached and at different % CEC occupancy for the different size fractions. For the clay fractions, the maximum K(ss) occurred at lower % CEC occupancy ( approximately 30 40%) than for the silt and sand fractions. The maximal K(ss) for the sand fractions occurred at the highest % CEC occupancy among all fractions ( approximately 50-60%). These findings suggest that for an in situ surfactant enhanced sorption zone it may be better to operate well below the saturation sorption of the cationic surfactant. This would enhance sorption of the HOCs onto the immobile fractions (silt and sand fractions) rather than the potentially mobile clay fractions. PMID- 18676002 TI - Toxicity and metal speciation characterisation of waste water from an abandoned gold mine in tropical northern Australia. AB - The decommissioned Mount Todd gold mine, located in the wet-dry tropics of northern Australia, consists of a large waste water inventory and an acid rock drainage problem, which has the potential to impact upon freshwater ecosystems of the Edith River catchment. The toxicity of retention pond 1 (RP1) water was determined using six local freshwater species (duckweed, alga, cladoceran, snail, hydra and a fish). RP1 water was very toxic to all species, with the percentage dilution of RP1 water inhibiting 10% of organism response (IC10), or lethal to 5% of individuals (LC5), ranging from 0.007 to 0.088%. The percentage dilution of RP1 water inhibiting 50% of organism response (IC50), or lethal to 50% of individuals (LC50), ranged from 0.051% to 0.58%. Based on chemical analyses and geochemical speciation modelling of the test waters, Cu, Zn and Al were the most likely toxic components at acidic dilutions (i.e. > or =1%), while Cu and Zn were the most likely toxic components at 0.1% RP1 water, where pH was 6.5. Species sensitivity distributions (SSDs) were used to predict dilutions of RP1 water that would protect or unacceptably affect the downstream aquatic ecosystems. A dilution ratio of 1 part RP1 water to 20000 parts Edith River water (0.005% RP1 water) was calculated to be required for the protection of at least 95% of species. This information can be used in conjunction with field chemical and biological data to better predict the ecological risks of RP1 waste water downstream of the Mount Todd mine. PMID- 18676003 TI - Assessment of the functionality of a pilot-scale reactor and its potential for electrochemical degradation of calmagite, a sulfonated azo-dye. AB - Electrochemical degradation (ECD) is a promising technology for in situ remediation of diversely contaminated environmental matrices by application of a low level electric potential gradient. This investigation, prompted by successful bench-scale ECD of trichloroethylene, involved development, parametric characterization and evaluation of a pilot-scale electrochemical reactor for degradation of calmagite, a sulfonated azo-dye used as a model contaminant. The reactor has two chambers filled with granulated graphite for electrodes. The system has electrical potential, current, conductivity, pH, temperature, water level and flow sensors for automated monitoring. The reactor supports outdoor and fail-safe venting, argon purging, temperature regulation and auto-shutdown for safety. Treatment involves recirculating the contaminated solution through the electrode beds at small flow velocities mimicking low fluid-flux in groundwater and submarine sediments. The first phase of the investigation involved testing of the reactor components, its parametric probes and the automated data acquisition system for performance as designed. The results showed hydraulic stability, consistent pH behavior, marginal temperature rise (<5 degrees C) and overall safe and predictable performance under diverse conditions. Near complete removal of calmagite was seen at 3-10V of applied voltage in 8-10h. The effects of voltage and strength of electrolyte on degradation kinetics have been presented. Further, it was observed from the absorption spectra that as calmagite degrades over time, new peaks appear. These peaks were associated with degradation products identified using electrospray ionization mass spectrometry. A reaction mechanism for ECD of calmagite has also been proposed. PMID- 18676004 TI - Atmospheric pollution for trace elements in the remote high-altitude atmosphere in central Asia as recorded in snow from Mt. Qomolangma (Everest) of the Himalayas. AB - A series of 42 snow samples covering over a one-year period from the fall of 2004 to the summer of 2005 were collected from a 2.1-m snow pit at a high-altitude site on the northeastern slope of Mt. Everest. These samples were analyzed for Al, V, Cr, Mn, Co, Ni, Cu, Zn, As, Rb, Sr, Cd, Sb, Pb, and Bi in order to characterize the relative contributions from anthropogenic and natural sources to the fallout of these elements in central Himalayas. Our data were also considered in the context of monsoon versus non-monsoon seasons. The mean concentrations of the majority of the elements were determined to be at the pg g(-1) level with a strong variation in concentration with snow depth. While the mean concentrations of most of the elements were significantly higher during the non-monsoon season than during the monsoon season, considerable variability in the trace element inputs to the snow was observed during both periods. Cu, Zn, As, Cd, Sb, and Bi displayed high crustal enrichment factors (EFc) in most samples, while Cr, Ni, Rb, and Pb show high EFc values in some of the samples. Our data indicate that anthropogenic inputs are potentially important for these elements in the remote high-altitude atmosphere in the central Himalayas. The relationship between the EFc of each element and the Al concentration indicates that a dominant input of anthropogenic trace elements occurs during both the monsoon and non-monsoon seasons, when crustal contribution is relatively minor. Finally, a comparison of the trace element fallout fluxes calculated in our samples with those recently obtained at Mont Blanc, Greenland, and Antarctica provides direct evidence for a geographical gradient of the atmospheric pollution with trace elements on a global scale. PMID- 18676005 TI - Description of a novel mutation leading to MYH9-related disease. PMID- 18676006 TI - Immunolocalization of insulin-like growth factors and their receptors in the diabetic mouse oviduct and uterine tissues during the preimplantation period. AB - The aim of the present study was to analyze the immunolocalization of insulin like growth factor (IGF)-1 and IGF-2 and their receptors in the oviduct and uterus of control and diabetic mice. Sexually mature female ICR mice aged 6-8 weeks were rendered diabetic by streptozotocin (200 mg/kg, administered intraperitoneally). Oviductal and uterine tissues were obtained from the superovulated control and diabetic mice at 48, 72 and 96 h post-human chorionic gonadotropin (hCG) treatment. Localization of IGF-1, IGF-2, IGF-1R and IGF-2R was determined by immunohistochemistry and a semi-quantitative scoring of immunolabelling was performed using a standardized 5-point system. The immunohistochemical scorings for both IGF-1 and IGF-1R were significantly decreased in the oviducts of diabetic mice at 96 h post-hCG treatment. The scores for IGF-2 were significantly increased in the oviducts of diabetic mice at 48 and 72 h post-hCG treatment, and for IGF-2R at 72 h post-hCG treatment. However, there was no significant difference in the scores of IGFs and their receptors in the uterus of control and diabetic mice. In conclusion, the oviductal immunolabelling for IGFs and their receptors was significantly altered by maternal diabetes, which may be of importance in the pathogenesis of preimplantation diabetic embryopathy. PMID- 18676007 TI - Adaptive features of skeletal muscles of mole rats (Spalax ehrenbergi) to intensive activity under subterranean hypoxic conditions. AB - Mole rats of the Spalax ehrenbergi superspecies are blind subterranean rodents that live under fluctuating oxygen supply, reduced to a measured 6% O(2), and mostly probably lower, during the rainy season. Fiber typing of muscles of the neck (trapezius) and leg (gastrocnemius, quadriceps) using standard histochemical techniques (succinic dehydrogenase, myosin ATPase) showed that the muscle fibers of mole rats in natural settings, as well as after extended captivity, were predominantly type IIa. The same muscles in laboratory rats showed the full range of fiber types. In contrast, the hearts of the mole rats and the laboratory rats were very similar. Our results indicate that skeletal muscle in the mole rats appears to have evolved in response to specific environmental demands to permit intensive endurance burrowing activities under conditions of severe or chronic hypoxia. PMID- 18676008 TI - Distribution of the interstitial Cajal-like cells in the gallbladder and extrahepatic biliary duct of the guinea-pig. AB - It has been suggested that interstitial Cajal-like cells (ICLC) may be involved in the spontaneous rhythmic electrical activities of the extrahepatic bile duct system. The present study investigated the distribution and characteristics of ICLC, which are immunopositive for CD117/ Kit receptor tyrosine kinase, using immunohistochemistry employing a monoclonal antibody raised against CD117/Kit on whole-mount preparations. The Kit-positive ICLC were examined using confocal laser scanning microscopy or fluorescence microscopy. ICLC, immunoreactive for Kit, were pleiomorphic and/or spindle-shaped cells with a few bipolar processes and distributed in the smooth muscle layers of the gallbladder and bile duct system. They were scattered in the hepatic duct, cystic duct and gallbladder as well as in the upper part of the common bile duct. The ICLC gradually increased in number and formed a completed cellular network in the lower part of the common bile duct and ampulla. The numbers of ICLC in the ampulla were similar to that of the duodenum and significantly much greater in number than in the gallbladder and bile ducts. The density of the ICLC in the common bile duct was significantly higher than that of other bile ducts. Our results suggested that the ICLC might contribute to the regulation of the spontaneous rhythmic contraction and development of motility disorders of the bile duct system. PMID- 18676009 TI - Immunolocalization of 8-OHdG and OGG1 in pancreatic islets of streptozotocin induced diabetic rats. AB - This study examined whether oxidative DNA damage and its repair system contribute to the occurrence of diabetes in an experimental rat model. The changed morphological findings of the 8-hydroxydeoxyguanosine (8-OHdG) and 8-oxoG-DNA glycosylase (OGG1) were examined in the pancreatic islets in streptozotocin induced diabetic rats (60 mg/kg, i.p.). The patterns of immunolocalization were mainly observed in the periphery of the normal pancreatic islet: 8-OHdG in the nucleus and OGG1 in the cytoplasm. The altered immunolocalization of 8-OHdG and OGG1 were greatest in the first hours after streptozotocin injection, and then declined in parallel with the morphological observations of pancreatic beta cell destruction. These results suggested that increased oxidative DNA damage might play a role as the inducer of diabetes and that OGG1 may not successfully mediate DNA repair in streptozotocin-induced diabetic rat pancreas. PMID- 18676010 TI - Boar semen variability and its effects on IVF efficiency. AB - In vitro fertilization (IVF) in pigs is still considered sub-optimal, due to the occurrence of polyspermy, as well as the inter- and intra-boar variability in sperm characteristics. Numerous studies have investigated the relationship between fresh and frozen-thawed semen parameters, such as motility, morphology and viability with in vitro fertility in order to develop methods of selecting boars for use in IVF. These studies have clearly shown that sperm parameters have limited value in predicting IVF efficiency. On the other hand, it has been demonstrated that the requirements of boar sperm during co-incubation with the oocytes (sperm:oocyte ratio, substances added to the fertilization medium and co incubation time) vary among boars. Preliminary assays required for individual males will be discussed with the objective of reaching maximum efficiency of in vitro fertilization. PMID- 18676011 TI - Iodine-deficiency disorders. AB - 2 billion individuals worldwide have insufficient iodine intake, with those in south Asia and sub-Saharan Africa particularly affected. Iodine deficiency has many adverse effects on growth and development. These effects are due to inadequate production of thyroid hormone and are termed iodine-deficiency disorders. Iodine deficiency is the most common cause of preventable mental impairment worldwide. Assessment methods include urinary iodine concentration, goitre, newborn thyroid-stimulating hormone, and blood thyroglobulin. In nearly all countries, the best strategy to control iodine deficiency is iodisation of salt, which is one of the most cost-effective ways to contribute to economic and social development. When iodisation of salt is not possible, iodine supplements can be given to susceptible groups. Introduction of iodised salt to regions of chronic iodine-deficiency disorders might transiently increase the proportion of thyroid disorders, but overall the small risks of iodine excess are far outweighed by the substantial risks of iodine deficiency. International efforts to control iodine-deficiency disorders are slowing, and reaching the third of the worldwide population that remains deficient poses major challenges. PMID- 18676012 TI - Molecular epidemiology of Staphylococcus aureus from implant orthopaedic infections: ribotypes, agr polymorphism, leukocidal toxins and antibiotic resistance. AB - Staphylococcus aureus is a leading pathogen of implant-related infections. In the field of biomaterials a variety of alternative approaches are currently proposed for prophylaxis and treatment of implant infections, but little is known on the role of the different pathogenetic mechanisms and spreading strategies that lead selected S. aureus clones to prevail and become epidemic. This study aimed at identifying and characterizing the major clones in a collection of 200 S. aureus isolates from implant orthopaedic infections. Strain typing by automated ribotyping identified 98 distinct ribogroups. Ribogroups corresponded to specific accessory gene regulatory (agr) polymorphisms and possessed peculiar arrangements of toxins. The agr type II allele was more represented in epidemic clones, while agr type I in sporadic clones. A clear trend was observed, where epidemic clones resisted antibiotics more than sporadic ones. Conversely, the gene for lukD/lukE leukotoxin, found in 68% of the isolates, was unrelated to the level of clonal spreading. Surprisingly, the isolates of the most prevalent ribogroup were susceptible to almost all antibiotics and never possessed the lukD/lukE gene, thus suggesting the role of factors other than antibiotic resistance and the here investigated toxins in driving the major epidemic clone to the larger success. PMID- 18676013 TI - Inflammatory reaction and implantation: the new entries PTX3 and D6. AB - Successful embryonic implantation implies anchoring the conceptus in the maternal uterine wall, establishing a vascular supply to enable optimal growth and development of the conceptus, and promoting tolerance of fetal alloantigens encoded by paternal genes. To achieve these goals, complex molecular dialogues take place among the maternal endometrium, the conceptus, and the placenta. Several factors are involved in the fetal-maternal interaction, including hormones, growth factors, cytokines, chemokines, adhesion molecules, extracellular matrix components, and matrix-degrading enzymes. This complex cross talk results in the induction of a local inflammatory response and a state of systemic inflammation, as revealed by leukocytosis, endothelium activation, increased activity of innate immune cells, and increased levels of inflammatory cytokines and chemokines. The enriched cytokine milieu associated to implantation is likely to control trophoblast migration and differentiation, leukocyte influx and activation, complement activation, as well as angiogenic and angiostatic processes in the implantation site. Finally, these mediators play a key role in tuning the immune responses to protect the fetus from infections as well as from maternal rejection. Here, the role of pro-inflammatory networks activated in implantation will be discussed. In particular, emphasis will be put on two new players involved in regulating inflammation at the maternal-fetal interface: the long pentraxin PTX3 and the decoy receptor for inflammatory chemokines D6. PMID- 18676014 TI - Calorimetric study as a potential test for choosing treatment of B-cell chronic lymphocytic leukemia. AB - Differential scanning calorimetry (DSC) and complementary techniques were utilized to evaluate the sensitivity of B-cell chronic lymphocytic leukemia (B CLL) cell samples in vitro exposed to cladribine or fludarabine in combination with mafosfamide. Mafosfamide, the active in vitro form of cyclophosphamide with both purine analogs produced the cytotoxic effect on mononuclear cell probes, however, to a different degree. Our results indicated that higher sensitivity of examined leukemic cell samples to the used drug combinations was usually accompanied by a marked decrease or even a complete loss of thermal transition at 95+/-3 degrees C in DSC scans of nuclear preparations as well as by more significant reduction of cell viability, higher extent of DNA damage estimated by the comet assay and by dropping/disappearance of anti-apoptotic protein Mcl-1 in comparison with untreated cells. We have also observed that the reduction of transition at 95+/-3 degrees C in thermal scans of nuclear preparations isolated from blood of B-CLL randomized patients who showed response to cladribine or fludarabine combined with cyclophosphamide, i.e., CC and FC, respectively, corresponded with the decrease or disappearance of anti-apoptotic proteins Bcl-2 and/or Mcl 1. In conclusion, these in vitro and in vivo studies revealed that quick DSC technique, usually supplemented by other methods, is a potent tool to distinguish efficacy of B-CLL treatment and could be helpful in choosing the most effective manner of treatment for this type of leukemia. PMID- 18676015 TI - Analysis of prognostic factors in patients with refractory anemia with excess of blasts (RAEB) reclassified according to WHO proposal. AB - The WHO classification subdivides the FAB RAEB category into RAEB-1 (bone marrow (BM) blasts <10%, peripheral blasts <5%) and RAEB-2 (bone marrow blasts >10% and peripheral blasts >5%). We reclassified according to WHO criteria 228 RAEB patients and analysed them in terms of haematological, karyotypic and prognostic features. We used the database of 680 MDS patients referred to our Institution from 1990 to 2000. Clinical features at presentation, such as sex, age, leukocyte count, polymorphonuclear cell count (PMN), platelet count, haemoglobin level, presence of one or more lineage dysplasia were tested in univariate and multivariate analysis in the two groups of RAEB-1 and RAEB-2 reclassified patients. In multivariate analysis we identified prognostic significant factors in the two patient groups, which consisted of age >70 years and platelet count <100 x 10(9)l(-1) for RAEB-1 category, while for RAEB-2 group parameters negatively influencing survival and risk of progression were haemoglobin <10g/dl, platelet count <100 x 10(9)l(-1), bone marrow blastosis >15% and complex karyotype. We also found differences in cytogenetic data (more balanced translocations and complex karyotypes in RAEB-2 group, p=0.02), and in survival (23.3 months in RAEB-1 vs. 16.1 months in RAEB-2 group, p=0.001). WHO classification provides valuable prognostic information for RAEB patient population, and can identify those subjects with more unfavourable prognosis who should be offered alternative therapeutic strategies. PMID- 18676016 TI - In vivo maintenance of synergistic cytarabine:daunorubicin ratios greatly enhances therapeutic efficacy. AB - We demonstrate here that cytarabine and daunorubicin, a standard drug combination used in the treatment of leukaemia, exhibits drug ratio-dependent synergistic antitumor activity in vitro and in vivo. A cytarabine:daunorubicin molar ratio of 5:1 displayed the greatest degree of synergy and minimum antagonism in a panel of 15 tumor cell lines in vitro. Co-encapsulating cytarabine and daunorubicin inside liposomes maintained the synergistic drug ratio in plasma for 24h post-injection. Liposome-encapsulated cytarabine:daunorubicin combinations exhibited drug ratio dependent in vivo efficacy with the 5:1 molar drug ratio (designated CPX-351) having the greatest therapeutic index, despite using sub-MTD daunorubicin doses. CPX-351 exhibited superior therapeutic activity compared to free-drug cocktails, with high proportions of long-term survivors, consistent with in vivo synergy. The therapeutic advantage of CPX-351 was associated with prolonged maintenance of synergistic drug ratios in bone marrow. These results indicate that in vitro informatics on cytarabine:daunorubicin cytotoxicity can be translated in vivo to optimize the efficacy of anticancer drug combinations by controlling the exposure of drug ratios with drug delivery vehicles. PMID- 18676017 TI - Concurrent B-cell chronic lymphocytic leukemia and multiple myeloma treated successfully with lenalidomide. AB - The occurrence of multiple myeloma and chronic lymphocytic leukemia in the same patient is very uncommon. The immunomodulatory agent lenalidomide has been shown to have high response rates in multiple myeloma and appears to be quite active in advanced CLL. We report two patients with concurrent CLL and MM who were both treated successfully with lenalidomide. PMID- 18676018 TI - Investigation of an NQO1 polymorphism as a possible risk and prognostic factor for chronic lymphocytic leukemia. AB - NAD(P)H:quinoneoxidoreductase 1 (NQO1) inhibits some cancers and increases p53 and apoptosis in cells. Due to an inactivating polymorphism, 10% of humans have no NQO1 activity. A case:control study suggested that chronic lymphocytic leukemia (CLL) patients may have an increased incidence of the NQO1 null genotype compared with controls. NQO1 genotype did not correlate with various CLL prognostic factors, but we observed a trend toward lower drug response in patients with the NQO1 null genotype. Inhibiting NQO1 activity decreased p53 levels and drug induced apoptosis in CLL cells. These results raise the possibility that the NQO1 polymorphism may be a risk factor for CLL and a predictor of response to chemotherapy. PMID- 18676019 TI - Genetic aberrations including chromosome 1 abnormalities and clinical features of plasma cell leukemia. AB - Plasma cell leukemia (PCL) is a rare form of plasma cell malignancy, few large series reported underlying genetic abnormalities. We systematically evaluated the genomic aberrations in 41 PCL patients by combining fluorescence in situ hybridization with cytoplasmic light chain immunofluorescence and correlated with their clinical outcome. The genomic aberrations in the 15 primary PCL (pPCL) and 26 secondary PCL (sPCL) were compared with 220 newly diagnosed multiple myeloma (MM) patients. There was no significant difference in the prevalence of genetic abnormalities in pPCL and sPCL but del(13q), t(4;14), 1q21 amplification and del(1p21) were more common in PCL than MM. Patients with pPCL had higher creatinine and beta(2)-microglobulin levels and tended to have a longer overall survival than patients with sPCL. In univariant analysis, PCL patients with t(4;14) (p=0.006) and del(1p21) (p=0.003) had shorter overall survivals. In multivariant analysis adjusting for all tested genetic factors as well as clinico biologically relevant factors including C-reactive protein, calcium and beta(2) microglobulin, t(4;14) remained a significant predictor for adverse overall survival in PCL (p=0.008). PMID- 18676020 TI - Polybrominated diphenyl ethers (PBDEs) in free-range domestic fowl from an e waste recycling site in South China: levels, profile and human dietary exposure. AB - To evaluate the status of polybrominated diphenyl ethers (PBDEs) contamination in poultry and sequentially human exposure through consumption of poultry in an e waste recycling site in South China, two kinds of free-range domestic birds, chicken and duck, were collected and their muscle and liver tissues were analyzed for 16 PBDE congeners. Chicken shows higher PBDE concentrations (summation of 16 PBDE congeners) in both muscle and liver tissues, ranged from 5.7 to 4381 and from 1.5 to 7897 ng/g (lipid weight, the same hereinafter), respectively, compared to duck, ranged from 2.4 to 51 and from 1.9 to 134 ng/g. Different living habitat and feeding habits between the two species might be responsible for this observation. No sex-related differences in PBDE concentrations were found for the two species, while the PBDE concentrations in muscle were higher than those in liver for chicken. The PBDE concentrations in muscle of chicken in the present study were higher than the levels of PBDEs in chicken from other studies reported by far. BDE209 and nona-BDEs were the major congeners in poultry. Comparison of PBDE profiles between birds and environmental matrix implied that the biodebromination of BDE209 might occur in poultry. The intake of PBDEs through consumption of poultry ranges from 7.8 ng/day to 3582 ng/day with a medial 68 ng/day, which is comparable to the calculated values through consumption of all foodstuffs in other studies. The present study suggested that the total dietary PBDEs intake for local residents might be considerably enhanced due to the e-waste recycling activity. PMID- 18676021 TI - Molecular recognition of botulinum neurotoxin B heavy chain by human antibodies from cervical dystonia patients that develop immunoresistance to toxin treatment. AB - We determined the entire profile of the continuous antigenic regions recognized by blocking antibodies (Abs) in sera from 30BoNT/B-treated cervical dystonia (CD) patients who developed unresponsiveness to treatment. The sera protected mice against a lethal dose of BoNT/B. We analyzed Ab binding to a panel of 60 synthetic 19-residue peptides (peptide C31 was 24 residues) that overlapped consecutively by 5 residues and encompassed the entire BoNT/B heavy (H) chain (residues 442-1291). Most Abs recognized a limited set of peptides but the pattern and Ab levels bound varied with the patient, consistent with genetic control of immune responses and with responses to each epitope being separately controlled. Abs were bound by peptides (in decreasing order): C1 (residues 848 866), C10 (974-992), C16 (1058-1076), C14 (1030-1048), N15 (638-656), N21/N22 (722-740/736-754), N24/N25 (764-782/778-796) and N29 (834-852). Peptides N3/N4 (470-488/484-502), N27 (806-824), C2 (862-880), C4 (890-908), C6/C7 (918-936/932 950), C17 (1072-1090), C24 (1170-1188), C29 (1240-1258) and C31 (1268-1291) exhibited low Ab binding. The remaining peptides bound little or no Abs. Of the 30 antisera, 28 (93.3%) had Abs that bound to peptides C1, C10, C14 or C16, and 27 (90.0%) bound to peptide N22. No peptide was recognized by all the antisera, but peptide combinations N24+C1, N22+N24+C1, N24+C1+C10, C10+C14+C16, N22+N24+C1+C10, C1+C10+C14+C16 or N22+N24+C1+C10+C14 bound blocking Abs in 30 (100%) antisera. BoNT/B-treated CD patients had higher Ab levels and bound to more epitopes (at least 11) than did BoNT/A-treated patients (5 regions). The regions recognized by anti-BoNT/B Abs occupied surface areas that displayed no correlation to surface electrostatic potential, hydrophilicity, hydrophobicity, or temperature factor. These regions afford candidates for epitope-specific manipulation of anti-toxin immune responses. PMID- 18676022 TI - Isoflavone-rich extracts from wooly glycine Glycine tomentella inhibits LPS induced TNF-alpha expression in a macrophage cell line of Atlantic salmon (Salmo salar L.). AB - The immunomodulatory effects of an isoflavone-rich extract from the root of wooly glycine Glycine tomentella (GTE) were studied in a macrophage-like cell line from Atlantic salmon (TO cells). The TO cell line was stimulated with defined concentrations of lipopolysaccharide (LPS) from Escherichia coli (serotype O127:B8) for defined time periods to induce expression of pro-inflammatory enzymes and cytokines. Cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were measured by real-time PCR methods and combined with analyses of eicosanoid production in cell extracts and evaluation of molecules of the TNF-alpha cell signaling pathway. The results showed that TNF-alpha was strongly induced by LPS, while GTE (25miicrog/ml) inhibited 67% of the TNF-alpha response when added to the cells together with LPS. Incubation of LPS in combination of GTE in TO cells caused increased intracellular prostaglandin E2 (PGE2), and reduced activation of p38 MAP kinase compared to LPS alone. GTE seemed to arrest NADPH oxidation, the coenzyme for carbonyl reductase and the prostaglandin-E2 9-reductase converting PGE2 to PGF2. We suggest that the mechanism of increased intracellular PGE2 levels following GTE treatment is caused by reduced breakdown of PGE2. GTE did not inhibit the other pro-inflammatory responses in LPS stimulated cells studied herein. IL-1beta and COX-2 showed moderately increased levels of expression likely caused by the increased PGE2. PMID- 18676024 TI - Psychotherapeutic intervention and suicide risk reduction in bipolar disorder: a review of the evidence. AB - BACKGROUND: 25-50% of bipolar patients attempt suicide at least once in their lifetime and completed suicide in this population is about 1% annually, about 60 times the rate of the general population. Psychotherapy may be an effective adjunctive option in preventing suicide in bipolar patients. It has been suggested that interpersonal, cognitive and behavioural techniques may be effective in controlling mood shifts, increasing compliance with pharmacotherapy, and maintaining morale in the face of therapeutic adversity and incomplete response. The aim of our study was to systematically review the literature concerning the efficacy of psychosocial interventions in reducing the risk for attempting or committing suicide. METHODS: We searched MEDLINE with the combination of the key words 'psychotherapy' or 'psychoeducation' or 'cognitive therapy' or 'behavio(u)ral therapy', 'cognitive-behavio(u)ral' or 'family therapy' or 'social rhythm' or 'rhythm' with 'suicide' and 'bipolar', limited to English language papers published between 1990 and January 2008. Papers were selected based on the criterium that they provided definite data on the role of psychotherapy in suicide prevention, and specifically in bipolar disorder. RESULTS: Our search returned 481 references, of which 17 were selected based on the above criteria. The selected papers were classified according to the area of suicide prevention they were dealing with as 1. Psychosocial and demographic factors, 2. Psychological profile and 3. Efficacy of psychotherapies. DISCUSSION: Our paper summarizes specific features and correlates of suicide in bipolar patients and possible targets of psychosocial intervention in the prevention of suicide in bipolar patients. Although studies researching the effect of psychosocial interventions on suicidal behaviour are virtually non-existent, hard data concerning the effectiveness of psychosocial interventions in bipolar disorder are emerging, but still suffer from methodological drawbacks. PMID- 18676023 TI - Treatment of cells with n-alpha-tosyl-L-phenylalanine-chloromethyl ketone induces the proteolytic loss of STAT6 transcription factor. AB - The implication of the STAT6 transcription factor in several human diseases makes the regulation of its activity a topic of great biological interest. The activation of this transcription factor is tightly regulated by kinases, phosphatases, and proteases. The initial aim of this study was to investigate the utility of protease inhibitors in controlling STAT6 activation. Among all inhibitors analyzed, n-alpha-tosyl-L-phenylalanine-chloromethyl ketone (TPCK) was found to inhibit the IL-4-induced STAT6 activation. Unexpectedly, this inhibition was accompanied by a loss of STAT6 protein. Thus, TPCK promoted the loss of STAT6 by a mechanism sensitive to the serine-protease inhibitor 4-(2-aminoetyl) benzenesulfonyl fluoride. However, the effects of TPCK seemed not to be mediated by its protease inhibitory activity since multiple protease inhibitors tested had no effect on STAT6 expression. The results found suggest that the effect of TPCK was mediated by its alkylating activity. Thus, cysteine reactive and thiol antioxidant compounds prevented the loss of STAT6 induced by TPCK. The reactivity of thiol groups on STAT6 was moreover demonstrated with biotinylated sulfhydryl reactive compounds. Analysis of other signaling molecules indicated that STAT5, but not other STATs, Shc, or c-Rel, was also affected by TPCK, suggesting a common downregulatory mechanism for STAT6 and STAT5. These results reveal a novel mechanism of action of TPCK in inducing a selective loss of STAT proteins. These findings may have implications for diseases in which STAT proteins are involved. PMID- 18676025 TI - Allergen-specific IgE in Icelandic horses with insect bite hypersensitivity and healthy controls, assessed by FcepsilonR1alpha-based serology. AB - Insect bite hypersensitivity (IBH) and atopy can both be causes of pruritus in horses and are associated with allergen-specific IgE to biting insects and environmental allergens respectively. Information with respect to differences in IgE levels in diseased and healthy animals is crucial in enabling an understanding of the clinical relevance of results of allergen-specific IgE tests. The aim of this study was (i) to evaluate and compare levels of allergen specific IgE, using an ELISA method, in Icelandic horses, with and without IBH, from Iceland and Sweden respectively; (ii) to investigate patterns of allergen specific IgE to insects, pollens, moulds and mites in those groups of horses; and (iii) to investigate the clinical significance of employing two different cut-off levels for the ELISA. The study compromised a total number of 99 horses from Iceland and Sweden, with and without IBH, divided in 5 groups. Sera from the horses were analysed blindly with the use of Allercept , a non-competitive, solid phase ELISA-test, designed to detect the presence of allergen-specific IgE in sera using the recombinant alpha chain of the high-affinity IgE receptor (FcepsilonR1alpha). The distribution of the ELISA values was shown for each insect, mould, mite and pollen allergen, in the different groups using 10th, 50th and 90th percentiles. The use of two cut-off levels, 150 EA and 300 EA, did not eliminate the false positives. Horses with IBH had a higher number of positive reactions, counting all the 29 allergens, than healthy controls and this was borderline significant (P=0.053). In this study it was shown that serological testing with an ELISA that uses the high-affinity IgE receptor (FcepsilonR1alpha) is presently not suitable as a tool for establishing a diagnosis of IBH or equine atopy. The importance of establishing a correct cut-off level for the ELISA for the different allergens is emphasised. PMID- 18676026 TI - Characterization of anti-channel catfish MHC class IIbeta monoclonal antibodies. AB - This study characterizes four monoclonal antibodies (mAb) developed against the major histocompatibility complex (MHC) class II beta chain of the channel catfish, Ictalurus punctatus. Immunoprecipitations using catfish clonal B cells revealed that each of these mAbs immunoselected proteins of approximately 32 and 36 kD, which are of the appropriate sizes for MHC class II alpha and beta chains, respectively. Cell distribution studies using a fluorescence-activated cell sorter (FACS) combined with RT-PCR analyses demonstrated that MHC class II beta is expressed at a high density on catfish clonal macrophage, B and T cell lines, on alloantigen stimulated leukocytes, and on lipopolysaccharide-induced B-cell blasts. Collectively, these results demonstrate the potential importance of these antibodies as reagents in future studies dealing with the functional role of MHC class II molecules in immune recognition of self from non-self. PMID- 18676027 TI - Gene expression in the skin of Bos taurus and Bos indicus cattle infested with the cattle tick, Rhipicephalus (Boophilus) microplus. AB - The cattle tick Rhipicephalus microplus (formerly Boophilus microplus) is responsible for severe production losses to the cattle industry worldwide. It has long been known that different breeds of cattle can resist tick infestation to varying degrees; however, the mechanisms by which resistant cattle prevent heavy infestation are largely unknown. The aim of this study was to determine whether gene expression varied significantly between skin sampling sites (neck, chest and tail region), and whether changes in gene expression could be detected in samples taken at tick attachment sites (tick attached to skin sample) compared with samples taken from non-attachment sites (no tick attachment). We present here the results of an experiment examining the expression of a panel of forty-four genes in skin sections taken from Bos indicus (Brahman) cattle of known high resistance, and Bos taurus (Holstein-Friesian) cattle of known low resistance to the cattle tick. The forty-four genes chosen for this study included genes known to be involved in several immune processes, some structural genes, and some genes previously suggested to be of importance in tick resistance by other researchers. The expression of fifteen gene transcripts increased significantly in Holstein Friesian skin samples at tick attachment sites. The higher expression of many genes involved in innate inflammatory processes in the Holstein-Friesian animals at tick attachment sites suggests this breed is exhibiting a non-directed pathological response to infestation. Of the forty-four genes analysed, no transcripts were detected in higher abundance at tick attachment sites in the Brahman cattle compared with similar samples from the Holstein-Friesian group, nor difference between attachment site and non-attachment site samples within the Brahman group. The results presented here suggest that the means by which these two cattle breeds respond to tick infestation differ and warrant further investigation. PMID- 18676028 TI - Exploring the pathogenesis of IIH: an inflammatory perspective. AB - Idiopathic intracranial hypertension (IIH) is a common blinding condition amongst the young obese female population (20 per 100,000) characterised by elevated intracranial pressure (ICP). The aetiology of IIH is not known. In this review we explore the literature investigating the pathogenesis of IIH and suggest additional hypotheses. Chronic inflammation is emerging as an aetiological factor in the pathogenesis of obesity and we propose that this may be a feature of IIH. Obesity is also related to dysregulation of cortisol production by the pre receptor enzyme, 11beta-hydroxysteroid dehydrogenase, and we speculate that this may have a role in the pathogenesis of obesity and raised ICP seen in IIH. PMID- 18676029 TI - Inflammatory pseudotumor in the middle ear cavity. AB - Inflammatory pseudotumor includes a diverse group of lesions characterized by inflammatory cell infiltration and variable fibrotic responses. It is extremely rare in the middle ear alone. A 7-year-old girl presented right hearing impairment. Because an otitis media with effusion was first suspected, a myringotomy was performed, but it found a mass that was different from a congenital cholesteatoma. Canal wall-down tympanomastoidectomy removed the mass successfully. The pathologic study of the specimen confirmed an inflammatory pseudotumor. We report an extremely rare case of the inflammatory pseudotumor in the middle ear with a review of the poor literature about this subject. PMID- 18676030 TI - Early myelination patterns in the central auditory pathway of the higher brain: MRI evaluation study. AB - OBJECTIVE: The purpose of this study was to use magnetic resonance imaging (MRI) to investigate the early myelination patterns of the central auditory pathway and then compare the data with past histological research. We observe the MRI signal intensity of the central auditory pathway and clarify the time course difference between MRI and previous histological research studies. METHODS: A total of 192 infants ranging in age from -4 to 224 corrected postnatal weeks were included in the study. Images were obtained using a 1.5 T MR unit. We chose three sites (medial geniculate body, auditory radiation, and splenium of the corpus callosum) of the central auditory pathway for analysis. Three cross sections were obtained perpendicular to the long axis of the brain and used to analyze the signal changes of the T1- and T2-weighted MRI by employing a region-of-interest (ROI) methodology that was corrected for postnatal age. RESULTS: At 10 corrected postnatal weeks, the medial geniculate body showed myelinated intensity changes on T2-weighted images. Auditory radiation showed myelinated intensity changes at 19 corrected postnatal weeks on the T1-weighted images and at 24 corrected postnatal weeks on the T2-weighted images. The splenium of the corpus callosum showed myelinated intensity changes at 16 corrected postnatal weeks on T1 weighted images and at 24 corrected postnatal weeks on T2-weighted images. CONCLUSIONS: As compared to the histological literature, the MRI documented signal intensity changes caused by myelination occurred approximately 3 weeks later for the medial geniculate body, 7-24 weeks later for the auditory radiation and 7-15 weeks later for the splenium of the corpus callosum. Since myelination is a process that occurs gradually, substantial changes of the myelin sheath makeup, a loss of water and the addition of lipids are more required in order to be detectable by MRI than myelin staining of histological study. PMID- 18676031 TI - Signaling at G-protein-coupled serotonin receptors: recent advances and future research directions. AB - The broadly distributed monoaminergic neurotransmitter serotonin (5 hydroxytryptamine, 5-HT) exerts its actions via 14 classes of receptor. With the exception of 5-HT3 receptors, which gate a cation-permeable ion channel, all 5-HT receptors are coupled to G proteins. The core features of transduction via 5-HT receptors are well established, but much still remains to be learned, in particular, with regard to native populations in the brain. In this article, we survey the current knowledge of cellular signaling at G-protein-coupled 5-HT receptors and focus on several novel (and surprising) insights that have emerged over the past few years. We also highlight several promising directions for future research that should improve the understanding of serotonin signaling and ultimately permit its therapeutic exploitation in the control of central nervous system disorders. In view of the diversity of transduction mechanisms engaged by 5-HT, much of this discussion is relevant to other classes of G-protein-coupled receptors. PMID- 18676032 TI - Death receptor Fas (CD95) signaling in the central nervous system: tuning neuroplasticity? AB - For over a decade, neuroscientific research has focused on processes of apoptosis and its contribution to the pathophysiology of neurological diseases. In the central nervous system, the degree of intrinsic mitochondrial-mediated apoptotic signaling expresses a cell's individual metabolic stress, whereas activation of the extrinsic death receptor-induced cascade is regarded as a sign of imbalanced cellular networks. Under physiological conditions, most neurons possess death receptors without being sensitive to receptor-mediated apoptosis. This paradox raises two questions: what is the evolutionary advantage of expressing potentially harmful proteins? How is their signaling controlled? This review summarizes the functional relevance of FasL-Fas signaling--a quintessential death ligand/receptor system--in different neurological disease models ranging from traumatic, inflammatory and ischemic to neurodegenerative processes. Furthermore, it outlines alternative non-apoptotic Fas signaling, shedding new light on its neuroplastic capacity. Finally, receptor-proximal regulatory proteins are introduced and identified as potential protagonists of disease-modifying neurological therapies. PMID- 18676033 TI - fMRI and its interpretations: an illustration on directional selectivity in area V5/MT. AB - fMRI is a tool to study brain function noninvasively that can reliably identify sites of neural involvement for a given task. However, to what extent can fMRI signals be related to measures obtained in electrophysiology? Can the blood oxygen-level-dependent signal be interpreted as spatially pooled spiking activity? Here we combine knowledge from neurovascular coupling, functional imaging and neurophysiology to discuss whether fMRI has succeeded in demonstrating one of the most established functional properties in the visual brain, namely directional selectivity in the motion-processing region V5/MT+. We also discuss differences of fMRI and electrophysiology in their sensitivity to distinct physiological processes. We conclude that fMRI constitutes a complement, not a poor-resolution substitute, to invasive techniques, and that it deserves interpretations that acknowledge its stand as a separate signal. PMID- 18676034 TI - Expression of glutathione S-transferase (GST) genes in the marine copepod Tigriopus japonicus exposed to trace metals. AB - The intertidal copepod, Tigriopus japonicus has been recognized as a potential model species for marine pollution toxicity testing. Toxicity ranges of several biocides, endocrine-disrupting chemicals (EDCs), and trace metals are known in T. japonicus. A large number of expressed sequence tags (ESTs) and genomic DNA are also sequenced from T. japonicus. In this study, expression of ten glutathione S transferase (GST) genes was studied in the copepods exposed to trace metals. Expression of these genes was also studied against exposure to hydrogen peroxide (H(2)O(2)) used as a positive control with prooxidant activity. Of all genes, expression of GST-Sigma (GSTS) was highly upregulated in H(2)O(2) as well as trace metal-exposed copepods. In the time-course study, expression of GSTS mRNA was more consistent compared to other GSTs such as GST-Omega, GST-Delta1, GST Theta3 or microsomal GST1 (mGST1). GSTS is predominantly reported from the insects. Coupled with the previous study of the in vitro antioxidant role of T. japonicus GSTS, these findings imply an antioxidant role for GSTS and highlight its importance as a biomarker of exposure to trace metals in T. japonicus. However, further validation and field trials would be necessary to propose GSTS gene expression as biomarker of exposure to trace metals, as for some trace metals such as silver the response was not consistent in concentration and time series exposure experiments. PMID- 18676035 TI - Investigating a potential mechanism of Cd resistance in Chironomus riparius larvae using kinetic analysis of calcium and cadmium uptake. AB - The uptake kinetics of waterborne Ca and Cd, both independently and in combination, were examined in C. riparius larvae, which are extremely Cd tolerant. Larvae exposed to Ca (100-2500 micromol L(-1)), exhibited classic Michaelis-Menten saturation kinetics for Ca influx, measured using (45)Ca as a radio-tracer. The maximum rate of Ca influx (J(max)(Ca)) was 0.39 micromol g( 1)h(-1), and the Ca concentration where the carrier reached half saturation (K(M)(Ca)) was 289 micromol L(-1). Cd influx was measured using (109)Cd as a radio-tracer in larvae exposed to Cd (0-1400 micromol L(-1)) while the Ca concentration was set to the K(M)(Ca). This revealed a J(max)(Cd) (2.26 micromol g(-1)h(-1)) which was nearly 6-fold higher that of Ca. This unusually high capacity for Cd uptake is in accordance with the huge tissue Cd burdens that chironomid larvae are able to accumulate during high level exposures. The apparent K(M)(Cd) (1133 micromol Cd L(-1)), when recalculated to account for the background Ca level, was still high (567 micromol Cd L(-1)), suggesting that this organism has a low affinity for Cd relative to most aquatic animals, indeed lower or comparable to its affinity for Ca. In consequence, even well above environmentally relevant Cd exposures, C. riparius does not accumulate Cd at the expense of Ca, thereby avoiding internal hypocalcaemia, in contrast to most other organisms which are much more sensitive to Cd. However, Ca influx was significantly reduced when 1200 micromol Cd L(-1) was added to Ca exposures (96 2410 micromol L(-1)). Michaelis-Menten analysis revealed a similar J(max)(Ca) in Cd-exposed and control larvae (i.e. exposed only to Ca), but that the apparent K(M)(Ca) was many-fold higher in larvae which were simultaneously exposed to Ca and Cd. Conversely, increasing Ca concentrations (96-2410 micromol L(-1)) progressively inhibited Cd uptake from a Cd exposure concentration (1200 micromol L(-1)), providing additional support for a common transport system. These results suggest that the interaction of Cd and Ca in C. riparius is one of simple competitive interaction, and that the unusual Cd transport kinetics (low affinity, high capacity) relative to fairly standard Ca transport kinetics help explain the unusual tolerance that this organism has to acute Cd exposure. PMID- 18676036 TI - Suspicion of elder abuse in South Eastern Spain: the extent and risk factors. AB - Elder abuse is recognized internationally as a growing problem. Recent years have seen an increase in the number of authors recommending that the MDs systematically question old people concerning possible abuse. The aim of our study was to ascertain the extent of suspicion of elder abuse and the different types of abuse. We design a cross-sectional survey including 460 patients > or = 65 years at different health centers (South East Spain). A face-to-face interview and a physical examination was carried out. Extent of suspected abuse was 44.6%. Female sex, > or = 75 years, widowhood, living alone or with children, accommodation in house of relatives and income < or = 300 euros/month were the associated sociodemographic variables. The risk factors associated are recent worsening of health, living with a mentally ill person, excessive consumption of alcohol or illegal drugs, arguing frequently with relatives or the dependence on someone to carry out a daily activity. The signs in the physical examination associated are dehydration/malnutrition, pressure ulcers and poor body and/or mouth hygiene. There is a high extent of suspicion of elder abuse and the keyword in this respect is prevention. PMID- 18676037 TI - Inducibility of atrial fibrillation during electrophysiologic evaluation is associated with increased dispersion of atrial refractoriness. AB - The impact of atrial dispersion of refractoriness (Disp_A) in the inducibility and maintenance of atrial fibrillation (AF) has not been fully resolved. AIM: To study the Disp_A and the vulnerability (A_Vuln) for the induction of self-limited (<60 s) and sustained episodes of AF. METHODS AND RESULTS: Forty-seven patients with paroxysmal AF (PAF): 29 patients without structural heart disease and 18 with hypertensive heart disease. Atrial effective refractory period (ERP) was assessed at five sites--right atrial appendage and low lateral right atrium, high interatrial septum, proximal and distal coronary sinus. We compared three groups: group A - AF not inducible (n=13); group B - AF inducible, self-limited (n=18); group C - AF inducible, sustained (n=16). Age, lone AF, hypertension, left atrial and left ventricular (LV) dimensions, LV systolic function, duration of AF history, atrial flutter/tachycardia, previous antiarrhythmics, and Disp_A were analysed with logistic regression to determine association with A_Vuln for AF inducibility. The ERP at different sites showed no differences among the groups. Group A had a lower Disp_A compared to group B (47+/-20 ms vs 82+/-65 ms; p=0.002), and when compared to group C (47+/-20 ms vs 80+/-55 ms; p=0.008). There was no significant difference in Disp_A between groups B and C. By means of multivariate regression analysis, the only predictor of A_Vuln was Disp_A (p=0.04). CONCLUSION: In patients with PAF, increased Disp_A represents an electrophysiological marker of A_Vuln. Inducibility of both self-limited and sustained episodes of AF is associated with similar values of Disp_A. These findings suggest that the maintenance of AF is influenced by additional factors. PMID- 18676039 TI - Coronary artery aneurysm related to percutaneous coronary intervention. AB - We report a case of coronary artery aneurysm possibly related to percutaneous coronary intervention. The various clinical features, epidemiology and management of the condition are also discussed in detail. PMID- 18676038 TI - Favourable effects of exercise-based Cardiac Rehabilitation after acute myocardial infarction on left atrial remodeling. AB - BACKGROUND: Left atrial enlargement is an important predictor of cardiovascular outcomes in patients after acute myocardial infarction. While the favourable effect of exercise exercise-based Cardiac Rehabilitation (CR) on postinfarction LV remodeling has been well documented, those on LA remodeling have yet to be defined. This study investigated the effects of CR on LA remodeling in postinfarction patients with moderate left ventricular (LV) dysfunction. METHODS: Sixty postinfarction patients were randomised randomized into two groups, each composed of 30 patients: group T (LV ejection fraction (EF) 43.7+/-4.2%, mean+/ SD) entered a 6-month CR program, whereas group C (EF 44.7+/-4.4%, P=ns) did not. Doppler echocardiography and cardiopulmonary exercise test were performed upon enrolment and at 6-month. RESULTS: At 6-month, trained patients showed a significant (P<0.001) improvement in peak oxygen consumption (DeltaVO(2peak)=+5.2+/-2.1 ml/kg/min) and a reduction in LA (DeltaLAV(MAX)=-1.9+/ 3.7 ml/m(2)) and in LV volumes (DeltaLVEDV=-3.6+/-4.4 ml/m(2)). At 6-month, untrained patients showed LAV(MAX) (+3.6+/-4.4 ml/m(2), P<0.001) and LV dilation (+4.2+/-5.1 ml/m(2), P<0.001; group T vs. C, P<0.001); whereas no significant changes in VO(2peak) were observed. Multiple linear regression analysis showed that age (beta=0.442, P<0.001), inclusion in the training group (beta=-0.599, P<0.001), E/A ratio (beta=-0.210, P=0.038), LVEDV (beta=0.376, P<0.001), and LVEF (beta=-0.279, P=0.007) are significant predictors of LA remodeling. CONCLUSIONS: Six-month exercise-based CR in postinfarction patients with mild to moderate LV dysfunction induced a favourable LA remodeling. PMID- 18676040 TI - Endothelium dysfunction classification: why is it still an open discussion? AB - In the nineties, the present author developed a didactic endothelium dysfunction classification based on personal experience while waiting for a consensus about the need of such an accomplishment. As time went by and no publications regarding this subject were released, he published a text entitled "An open discussion about endothelial dysfunction: is it timely to propose a classification? Eight years later and the question about endothelium dysfunction classification remains "an open discussion". Nevertheless, we still keep using our proposed classification although always questioning its suitability and wondering reasons for why the scientific communities avoid discussing this very subject. Based on these thoughts we attempted to hypothesize the causes of why this proposed discussion is not an inciting issue: (I) First hypothesis: An endothelium dysfunction classification is not important; (II) Second hypothesis: Such classification is still a "premature reductionism"; (III) Third hypothesis: Endothelium dysfunction is a multifaceted disorder and involves uncountable variations becoming impossible to propose an adequate classification; (IV) Fourth hypothesis: Vasoplegic syndrome and massive nitric oxide release accounts for the functional classification, thus, it seems that cardiovascular journals feel apprehensive about this since their point of view is closely associated with impaired endothelium nitric oxide release as a cause to increased risk of spasm and thrombosis and; (V) Fifth hypothesis: The classification is so fulfilling that it discards the need of discussion or the pioneer attitude to propose such classification has incited some degree of jealousy. Obviously, the fifth hypothesis is pretentiously and overwhelmed, but we again attempt to reopen this discussion because it is time to update the presented classification. In this manner, we are sure that this issue demands special attention of the scientific community. Therefore, the aim of this text is exclusively to "reopen" the discussion about the need of an effective endothelium dysfunction classification. PMID- 18676041 TI - An anomalous coronary artery: bypass grafting based on CT angiography. AB - This report illustrates a patient in whom coronary artery bypass grafting (C.A.B.G) was based on multi-row detector computerized tomography (MDCT) rather than coronary angiography. She had symptomatic ischaemic heart disease and an anomalous origin of her right coronary artery. Angiography of this vessel was not possible but MDCT contrast cardiac angiography defined the diseased right coronary artery, allowing an appropriate interventional therapeutic decision for C.A.B.G. to be made, with a successful clinical outcome. PMID- 18676042 TI - Relationship between pattern of Tako-Tsubo cardiomyopathy and age: from midventricular to apical ballooning syndrome. AB - Several patterns of Tako-Tsubo cardiomyopathy (TTC) have been recently published. However, its physiopathology is still unclear. We report the case of a 63-year old woman who had a previous history of transient midventricular ballooning and was admitted 11 years after the first TTC, for a recurrence of typical pattern of TTC. Our report suggests that age could affect the left ventricular pattern of TTC and should be taken into account for a correct diagnosis of TTC. PMID- 18676043 TI - Sudden death in type 1 diabetes: the mystery of the 'dead in bed' syndrome. AB - Sudden cardiac death is an unpredictable and devastating event, particularly in the young. A significant proportion of sudden deaths in the young are unexplained no cause is identified either during life or at post-mortem. This is seen in a subgroup of young patients with type 1 diabetes who have dead in bed syndrome, where these victims are in good health, retire to bed, only to be found dead the following morning in a bed which is undisturbed, suggesting no terminal struggle or seizure. The underlying cause of dead in bed syndrome remains unknown, but is likely to be due to a terminal malignant arrhythmia. A plausible hypothesis is that it may be secondary to QT interval prolongation (followed by a degenerate ventricular tachycardia), caused by a number of factors including acute hypoglycaemia, on a background of cardiac autonomic neuropathy, and possible genetic influences. It is envisaged that understanding the causes and triggers of dead in bed syndrome will allow appropriate therapeutic interventions to be initiated in high-risk patients with type 1 diabetes, with the ultimate goal to prevent sudden death. PMID- 18676044 TI - Athlete's heart or hypertrophic cardiomyopathy: usefulness of N-terminal pro brain natriuretic peptide. AB - BACKGROUND: Distinguishing physiological hypertrophy from hypertrophic cardiomyopathy in athletes remains difficult, but vital in view of the risk of sudden death under effort. NT-proBNP was previously reported to be usually normal in healthy athletes, but often elevated in case of hypertrophic cardiomyopathy. OBJECTIVES: To assess the interest of NT-proBNP in differential diagnosis of left ventricular hypertrophy in athletes. METHODS: NT-proBNP levels were measured at rest and after effort in trained athletes referred for suspectedly abnormal (>or=13 mm) left ventricular hypertrophy. RESULTS: 17 patients were included, 10 of whom were diagnosed with hypertrophic cardiomyopathy (group I) while the other 7 presented typical signs of athlete's heart (group II). NT-proBNP levels did not significantly differ between groups, whether at rest or after effort. NT-proBNP levels were, however, significantly elevated in 3 subjects in group I, while being consistently normal in group II. CONCLUSIONS: In active athletes presenting with ambiguous left ventricular hypertrophy, abnormal NT-proBNP levels indicate hypertrophic cardiomyopathy, whereas normal values are inconclusive. PMID- 18676045 TI - Metabolic syndrome and left ventricular dysfunction: new player of the game. AB - The metabolic syndrome (MS) is associated with impaired global left ventricular function. These preclinical cardiac abnormalities could be ascribed to the interplay of the metabolic components characterizing the MS. The MS can be clinically manifested in a variety of ways. A sizable number of metabolic changes thus occur in people with clinical evidence of the syndrome.We believe that the identification of these changes should provide a broader picture of the metabolic status of an affected individual. PMID- 18676046 TI - Association between polymorphism of the AGTR1 and cardiovascular events in a Japanese general sample (The Shigaraki Study). AB - We examined whether the A1166C polymorphism of the angiotensin II type I receptor gene (AGTR1) affects cardiovascular event occurrence in a Japanese prospective cohort study. The 2212 participants who gave informed consent for genetic analysis were enrolled in this study (the Shigaraki Study). The average observation period was 1954 days. Cardiovascular events occurred in 37 individuals (1.7%). The independent factors which specified cardiovascular events were age (hazard ratio (HR)=1.13; 95% confidence interval (CI): 1.10-1.16; p<0.0001) and sex (HR=2.18; 95%CI: 1.23-3.85; p=0.007). However, the A1166C polymorphism of AGTR1 was not a predictive factor for cardiovascular events (HR=1.11; 95%CI: 0.61-2.02; p=0.731). PMID- 18676047 TI - Evolution of a regulatory framework for pharmaceuticals derived from genetically modified plants. AB - The use of genetically modified (GM) plants to synthesize proteins that are subsequently processed, regulated and sold as pharmaceuticals challenges two very different established regulatory frameworks, one concerning GM plants and the other covering the development of biotechnology-derived drugs. Within these regulatory systems, specific regulations and guidelines for plant-made pharmaceuticals (PMPs)--also referred to as plant-derived pharmaceuticals (PDPs)- are still evolving. The products nearing commercial viability will ultimately help to road test and fine-tune these regulations, and might help to reduce regulatory uncertainties. In this review, we summarize the current state of regulations in different countries, discuss recent changes and highlight the need for further regulatory development in this burgeoning, new industry. We also make the case for the harmonization of international regulations. PMID- 18676048 TI - A case of hypoglycemic brittle diabetes with peripheral edema successfully managed by conversion from insulin lispro to insulin aspart. PMID- 18676049 TI - Breast milk as the "water that supports and preserves life"--socio-cultural constructions of breastfeeding and their implications for the prevention of mother to child transmission of HIV in sub-Saharan Africa. AB - OBJECTIVES: Complementary breastfeeding represents an important source of risk of HIV infection for infants born to HIV positive mothers. The World Health Organisation recommends that infants born to HIV positive mothers receive either replacement feeding or exclusive breastfeeding (EBF) followed by early weaning. Beyond the clinical and epidemiological debate, it remains unclear how acceptable and feasible the two options are for rural populations in sub-Saharan Africa. This qualitative study aims to fill this gap in knowledge by exploring both the socio-cultural construction and the practice of breastfeeding in the Nouna Health District, rural Burkina Faso. METHODS: Information was collected through 32 individual interviews and 3 focus group discussions with women of all ages, and 6 interviews with local guerisseurs. RESULTS: The findings highlight that breastfeeding is perceived as central to motherhood, but that women practice complementary, rather than exclusive, breastfeeding. The findings also indicate that women recognise both the nutritional value of breast milk and its potential to act as a source of disease transmission. CONCLUSIONS: The findings suggest that given the socio-cultural importance attributed to breastfeeding and the prevailing poverty, it may be more acceptable and more feasible to promote EBF followed by early weaning than replacement feeding. A set of operational strategies are proposed to favour the prevention of mother to child transmission of HIV in the respect of the local socio-cultural setting. PMID- 18676050 TI - Evolution of complex structures: minicollagens shape the cnidarian nematocyst. AB - The generation of biological complexity by the acquisition of novel modular units is an emerging concept in evolutionary dynamics. Here, we review the coordinate evolution of cnidarian nematocysts, secretory organelles used for capture of prey, and of minicollagens, proteins constituting the nematocyst capsule. Within the Cnidaria there is an increase in nematocyst complexity from Anthozoa to Medusozoa and a parallel increase in the number and complexity of minicollagen proteins. This complexity is primarily manifest in a diversification of N- and C terminal cysteine-rich domains (CRDs) involved in minicollagen polymerization. We hypothesize that novel CRD motifs alter minicollagen networks, leading to novel capsule structures and nematocyst types. PMID- 18676051 TI - Robust projective filtering of time-warped ECG beats. AB - In this paper a new version, more immune to noise, of the nonlinear projective filtering is presented. The method employs an algorithm of robust principal component analysis to signal subspaces construction and as a result it achieves high performance in real electromyographic noise environment. Two aspects of the method's action are investigated: its ability to suppress noise and its influence on the precision of the QT interval measurement. Then the method influence on evaluation of the beat-to-beat variability of the repolarization duration is presented. A comparison to the previous versions of the nonlinear projective filtering and to the classical linear one is carried out. PMID- 18676052 TI - Estimating the lifetime utilization rate of radiotherapy in cancer patients: the Multicohort Current Utilization Table (MCUT) method. AB - Extensive research has been carried out to establish the appropriate proportion of cancer patients requiring radiotherapy at some point during their illness. However, it is difficult to compare the actual rates against the appropriate rate, because calculating the actual rates requires life-long follow up of cancer patients. We have developed a method, referred to as the Multicohort Current Utilization Table (MCUT) method, to estimate the predicted lifetime utilization rates based on current medical practice. We implemented the method in SAS as a macro, and validated it by comparing the predicted and the actual utilization rates of radiotherapy in lung, breast, and prostate cancer cases diagnosed in Ontario, Canada. The MCUT method could be used to predict lifetime utilization rate of any medical services. PMID- 18676053 TI - An artificial ant colonies approach to medical image segmentation. AB - The success of image analysis depends heavily upon accurate image segmentation algorithms. This paper presents a novel segmentation algorithm based on artificial ant colonies (AC). Recent studies show that the self-organization of ants is similar to neurons in the human brain in many respects. Therefore, it has been used successfully for understanding biological systems. It is also widely used in many applications in robotics, computer graphics, etc. Considering the features of artificial ant colonies, we present an extended model for image segmentation. In our model, each ant can memorize a reference object, which will be refreshed when it finds a new target. A fuzzy connectedness measure is adopted to evaluate the similarity between target and the reference object. The behavior of an ant is affected by the neighbors and the cooperation between ants is performed by exchanging information through pheromone updating. Experimental results show that the new algorithm can preserve the detail of the object and is also insensitive to noise. PMID- 18676054 TI - Improved attenuation correction via appropriate selection of respiratory correlated PET data. AB - PURPOSE: We propose a respiratory-correlated PET data processing method (called "BH-CT-based") based on breath-hold CT acquisition to reduce the smearing effect and improve the attenuation correction. The resulting images are compared with the ungated PET images acquired using a standard, free-breathing clinical protocol. METHODS: The BH-CT-based method consisted of a list-mode acquisition with simultaneous respiratory signal recording. An additional breath-hold CT acquisition was also performed in order to define a tissue position from which PET events can be selected. A phantom study featured a 0.5-ml sphere (filled with 18F-fluorodeoxyglucose ((18)F-FDG) solution) pushed onto a rubber balloon (filled with (18)F-FDG solution and iodinated contrast agent). The feasibility of the BH CT-based method was also assessed in two patients. RESULTS: In the phantom study, the contrast-to-noise ratios (CNRs) were -1.6 for the Ungated volume and 5.1 for the BH-CT-based volume. For patients, CNRs were higher for BH-CT-based volumes than those for Ungated volumes (17.3 vs. 6.3 and 7.3 vs. 3.8, for patients 1 and 2, respectively). Bias-variance measurements were performed and yielded bias reduction of 40% with BH-CT-based. CONCLUSION: The application of a BH-CT-based method decreases motion bias in PET images. This method resolves issues related to both PET-to-CT misregistration and erroneous attenuation correction and increases lesion detectability. PMID- 18676055 TI - Significance of the urinary 8-OHdG level as an oxidative stress marker in lung cancer patients. AB - OBJECTIVE: In the present study, we investigated the relationship between the urinary excretion rate of the oxidized nucleoside 8-hydroxydeoxyguanosine (8 OHdG) and clinical factors in lung cancer patients. METHODS: The present study included 100 patients, who underwent a lung surgery. The patients included 62 men and 38 women with a mean age of 65.5 years ranging from 35 to 82. The diagnosis included 81 primary lung cancers, 9 metastatic lung cancers and 10 benign lung diseases. Urine samples collected for 24h were analyzed for the content of 8-OHdG using an ELISA assay. RESULTS: The urinary excretion rate of 8-OHdG in smokers was significantly higher than that in never-smokers. Specifically, the 8-OHdG excretion rate of current smokers was higher than that of patients who had quit smoking for longer than 1 month. Excluding current smokers, the urinary excretion rate of 8-OHdG did not relate to age or gender, but to the malignant potential of the disease. The urinary 8-OHdG level increased in the order of metastatic lung cancer, primary lung cancer and benign disease. In lung cancer patients, furthermore, the mean urinary 8-OHdG level of patients with stages II-IV disease was significantly lower than that of patients with stage I disease. CONCLUSIONS: Smoking significantly increased the urinary excretion rate of 8-OHdG, suggesting that smoking causes an increased rate of oxidative DNA modifications. On the other hand, the capacity to repair oxidative DNA modifications might be impaired to some extent in cancer patients. PMID- 18676056 TI - Induction therapy of loco-regional non-small-cell lung cancer with reliable response and low toxicity (low dose radiotherapy sensitizes tumor to subsequent chemotherapy?). AB - INTRODUCTION: For the induction therapy of non-small-cell lung cancer, we need to look for a regimen which produces a reliable high response rate with a low treatment related morbidity and mortality. METHODS: Patients in clinical stages IB, IIA and B, IIIA and B received a course of therapy with 20Gy of radiation in 2 weeks. This was followed by two courses of chemotherapy consisting of paclitaxel 180mg/m(2), cisplatin 45mg/m(2), and ifosfamide 1000mg/m(2). Two to 3 weeks after chemotherapy, the patients were re-evaluated and, if suitable, underwent surgical therapy. RESULTS: A total of 35 patients were entered into the study. The overall response rate was 82.86% (95% confidence interval, 66.35 94.5%). Complete response (CR) was 20% (95% confidence interval, 8.44-36.94%). Twenty-five patients had surgical resection. Subsequently 18 patients received completion radiotherapy of additional 45Gy. The median follow up is 30 months. In 12 patients with stages IB, IIA and B, the median survival was 61 months, and 5 year survival was 55%. In 23 patients with stages IIIA and B, the median survival was 26 months, and 5-year survival was 9.5%. There was 1 patient with Grade 4 and 13 patients with Grade 3 leukopenia, and half of them received granulocyte colony stimulating factor. By the completion radiotherapy, 6 out of 18 patients had less than Grade 2 esophagitis. Five patients had Grade 2 radiation pneumonitis and one Grade 5 (one mortality). There was no postoperative death. The survival results were comparable to those reported recently by others, however the regimen produced a high response rate with low treatment related morbidity/mortality. CONCLUSION: It is a suitable regimen for induction therapy to include earlier stage resectable non-small-cell lung cancers. PMID- 18676057 TI - Radiotherapy for small-cell lung cancer-Where are we heading? AB - Radiotherapy has an established role in the management of limited-disease small cell lung cancer. However, essential questions related to the optimisation of thoracic radiotherapy remain unanswered including (i) optimal total dose, (ii) fractionation, (iii) timing and sequencing of radiation, (iv) volume of irradiation, and (v) concurrent chemotherapy combinations. The role of thoracic radiotherapy for extensive-disease small-cell lung cancer is more poorly understood but evidence suggests radiotherapy may have an important role in this setting. This review highlights the need for well-designed multi-national trials aimed at the optimisation and standardisation of radiotherapy for SCLC. PMID- 18676058 TI - Converging hydrostatic and hydromechanic concepts of preferential flow definitions. AB - The boundary between preferential flow and Richards-type flow is a priori set at a volumetric soil water content theta* at which soil water diffusivity D (theta*) = eta (= 10(-6) m(2) s(-1)), where eta is the kinematic viscosity. First we estimated with a hydrostatic approach from soil water retention curves the boundary, theta(K), between the structural pore domain, in which preferential flow occurs, and the matrix pore domain, in which Richards-type flow occurs. We then compared theta(K) with theta* that was derived from the respective soil hydrological property functions of same soil sample. Second, from in situ investigations we determined 96 values of theta(G) as the terminal soil water contents that established themselves when the corresponding water-content waves of preferential flow have practically ceased. We compared the frequency distribution of theta(G) with the one of theta* that was calculated from the respective soil hydrological property functions of 32 soil samples that were determined with pressure plate apparatuses in the laboratory. There is support of the notion that theta(K) approximately = theta(G) approximately = theta*, thus indicating the potential of theta* to explain more generally what constitutes preferential flow. However, the support is assessed as working hypothesis on which to base further research rather than a procedure to a clear-cut identification of preferential flow and associated flow paths. PMID- 18676059 TI - Investigation of surfactant-enhanced mass removal and flux reduction in 3D correlated permeability fields using magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) was used to visualize the NAPL source zone architecture before and after surfactant-enhanced NAPL dissolution in three dimensional (3D) heterogeneously packed flowcells characterized by different longitudinal correlation lengths: 2.1 cm (aquifer 1) and 1.1 cm (aquifer 2). Surfactant flowpaths were determined by imaging the breakthrough of a paramagnetic tracer (MnCl(2)) analyzed by the method of moments. In both experimental aquifers, preferential flow occurred in high permeability materials with low NAPL saturations, and NAPL was preferentially removed from the top of the aquifers with low saturation. Alternate flushing with water and two surfactant pulses (5-6 pore volumes each) resulted in approximately 63% of NAPL mass removal from both aquifers. However, overall reduction in mass flux (Mass Flux 1) exiting the flowcell was lower in aquifer 2 (68%) than in aquifer 1 (81%), and local effluent concentrations were found to increase by as high as 120 times at local sampling ports from aquifer 2 after surfactant flushing. 3D MRI images of NAPL revealed that NAPL migrated downward and created additional NAPL source zones in previously uncontaminated areas at the bottom of the aquifers. The additional NAPL source zones were created in the direction transverse to flow in aquifer 2, which explains the higher mass flux relative to aquifer 1. Analysis using a total trapping number indicates that mobilization of NAPL trapped in the two coarsest sand fractions is possible when saturation is below 0.5 and 0.4, respectively. Results from this study highlight the potential impacts of porous media heterogeneity and NAPL source zone architecture on advanced in-situ flushing technologies. PMID- 18676060 TI - Lymphomonocyte alpha-synuclein levels in aging and in Parkinson disease. AB - In this study we employed an ELISA assay to measure alpha-synuclein protein in lymphomonocytes from 78 PD patients and 78 controls. We correlated protein levels with demographic and clinical characteristics and with the chymotryptic and tryptic activities of the 20S proteasome. Alpha-synuclein levels were not significantly different between patients and controls. In control subjects, alpha synuclein protein levels increased significantly with age and were significantly higher in men compared to women. Proteasome activity was not significantly different between cases and controls. In control group, the 20S chymotryptic activity tended to decrease significantly with increasing age, though it was not correlated to alpha-synuclein levels. The 20S tryptic activity was not significantly correlated to age, but was inversely correlated to alpha-synuclein levels. Our findings suggest that alpha-synuclein levels in lymphomonocytes are affected by age, gender, and by the 20S proteasome activity in control subjects, but they are not useful as a diagnostic biomarker for PD. PMID- 18676061 TI - Regulation of M1-receptor mRNA stability by smilagenin and its significance in improving memory of aged rats. AB - The purpose of this work is to study the effect of smilagenin on the mRNA stability of muscarinic receptor subtype 1 (M(1); m1 mRNA) in aged rat brains and its significance in improving memory. The Y-maze avoidance task showed that oral administration of smilagenin significantly improved spatial memory performance in aged rats. Mechanistic studies showed that smilagenin was neither a ligand of the M receptors nor a cholinesterase inhibitor, while radioligand binding assays revealed that smilagenin significantly increased the M(1)-receptor density. The increase of M(1)-receptor density correlated with memory improvement. Real-time polymerase chain reaction (RT-PCR) revealed that the m1 mRNA in m1 gene transfected CHO cells increased significantly, and the average half-life of m1 mRNA was approximately doubled by smilagenin treatment. These results suggest that smilagenin improves memory of aged rats at least partially by increasing the stability of m1 mRNA. However since the ChAT activity in the cortex of aged rats was also elevated by smilagenin, it cannot be excluded that the increase of intrinsic acetylcholine excretion also plays a role in the memory-improvement effect of smilagenin. PMID- 18676062 TI - Synthesis of some new 1,2,4-triazoles, their Mannich and Schiff bases and evaluation of their antimicrobial activities. AB - 4-Phenyl-5-pyridin-4-yl-4H-1,2,4-triazole-3-thiol (3) was obtained in basic media via the formation of 2-isonicotinoyl-N-phenylhydrazinecarbothioamide (2), and converted to some alkylated derivatives (4a,b) and Mannich base derivatives (5a c). 2-[(4-Phenyl-5-pyridin-4-yl-4H-1,2,4-triazol-3-yl)thio]acetohydrazide (7) that was obtained by using compound 3 as precursor in two steps was converted to thiosemicarbazide derivative (8), Schiff base derivatives (9) and 5-{[(4-phenyl-5 pyridin-4-yl-4H-1,2,4-triazol-3-yl)thio]methyl}-1,3,4-oxadiazole-2-thiol (10). Moreover, 5-{[(4-phenyl-5-pyridin-4-yl-4H-1,2,4-triazol-3-yl)thio]methyl}-3-{[(2 morpholin-4-ylethyl)amino]methyl}-1,3,4-oxadiazole-2(3H)-thione (11) was synthesized via reaction of compound 10 with 2-(4-morpholino)ethylamine. The treatment of compound 8 with NaOH gave 4-(4-methylphenyl)-5-{[(4-phenyl-5 pyridine-4-yl-4H-1,2,4-triazol-3-yl)thio]methyl}-4H-1,2,4-triazole-3-thiol (12), while the acidic treatment of compound 8 afforded 5-{[(4-phenyl-5-pyridin-4-yl-4H 1,2,4-triazol-3-yl)thio]methyl}-2(4-methylphenyl)-amino-1,3,4-thiadiazole (14). N Methyl derivative of compound 14 and a Mannich base derivative of compound 12 were synthesized from the reactions of these precursors with methyl iodide and methyl piperazine, respectively. All newly synthesized compounds were screened for their antimicrobial activities. The antimicrobial activity study revealed that all the compounds screened showed good or moderate activity except compounds 3, 5c, 7, 9c, 9e, 9g, 9h, 11, and 13. PMID- 18676063 TI - Synthetic, structural and biochemical studies of polynuclear platinum(II) complexes with heterocyclic ligands. AB - "Non-classical" di- and trinuclear Pt(II) complexes with polydentate nitrogen ligands; ionic [(PtCl(2))(2)(tptz)(2)(mu-PtClNCPh)]Cl (1) [tptz =2,4,6-tris(2 pyridyl)-1,3,5-triazine], [(PtCl(2))(2)(bptz)(2)(mu-Pt)]Cl(2) (2) [bptz = 3,6 bis(2-pyridyl)-1,2,4,5-tetrazine] and neutral [(PtCl(2))(2)(tptz)(2)(mu PtCl(2))](H(2)O)(4) (3), [(PtCl(2))(2)(mu-tppz)](CHCl(3)) (4) [tppz = 2,3,5,6 tetra(2-pyridyl)pyrazine] complexes, have been prepared and structurally characterized. The neutral tptz and tppz complexes present three and two separate PtCl(2) moieties, respectively, in a cis position, presumably acting in a bifunctional mode towards DNA; the cationic tptz and bptz complexes contain monofunctional and bifunctional bridging Pt(II) moieties, respectively, (other Pt(II) moieties in the complexes are bifunctional). All complexes were tested for their biological activity. Both tptz complexes, neutral and ionic, show a potent cytotoxic activity and reduced cell viability in a concentration-dependent manner that was evaluated in a panel of different cancer cell lines: human HT29 colon rectal carcinoma, HepG2 hepatoma, MDA-MB-231 breast cancer and MG63 osteosarcoma cells; their activity was higher than cisplatin, IC50 values have been calculated for the active compounds and flow cytometric analysis for the tptz complexes performed. Therefore, these new platinum drugs warrant further investigation into their antitumor activity against different types of tumors. PMID- 18676064 TI - [Mesenteric artery involvement in giant cell arteritis: two cases and literature review]. AB - INTRODUCTION: Mesenteric involvement has been rarely reported in giant cell arteritis (GCA). Our literature search using the Medline database (1966-2008), reviewing all articles in English and French languages identified only 31 cases of mesenteric ischemia related to GCA. We report two additional cases. CASE REPORTS: We report two men with GCA. At diagnosis of GCA-associated mesenteric involvement, patients exhibited: fever/fatigue (n=2), abdominal complaints (n=1), clinical signs of GCA (n=1). In both patients, abdominal CT-scan showed circumferential halo around the superior mesenteric artery. At systematic follow up, CT-scan revealed improvement of mesenteric damage in both patients. CONCLUSION: Our study indicates that CT-scan is a useful technique in diagnosis of GCA-associated mesenteric involvement. Furthermore, we suggest that CT-scan may also be helpful in both the monitoring and the medical management of GCA related mesenteric artery involvement. PMID- 18676065 TI - [Cutaneous lupus induced by etanercept in rheumatoid arthritis]. AB - Therapy with anti-TNFalpha in rheumatoid arthritis may induce autoimmune disorders. Induction of autoantibodies is frequently observed, but lupus-like syndrome is rare and few cases only have been reported. We report a 41-year-old female, treated with etanercept for a rheumatoid arthritis, who developed a cutaneous lupus induced without any other organ involvement, associated with high ANA and DNA antibody titres. The skin biopsy and the histological analysis with immunofluorescence confirmed the diagnosis. The anti-TNFalpha treatment was stopped. Corticosteroids were increased and hydroxychloroquine administered because skin lesions persist after three months. PMID- 18676066 TI - Multifrequency bioelectrical impedance analysis and bioimpedance spectroscopy for monitoring fluid and body cell mass changes after gastric bypass surgery. AB - BACKGROUND & AIMS: There is a growing need for clinically applicable body composition assessment tools for extremely obese individuals. The objective of this research was to evaluate several bioimpedance techniques for monitoring changes in fluid, particularly intracellular water (reflecting body cell mass) after bariatric surgery. DESIGN: Fifteen extremely obese women (body mass index: 48.9+/-7.0 kg/m(2); age: 48+/-9 years) were assessed before (baseline; T1), and approximately 6 weeks after gastric bypass surgery (T2) by several multifrequency bioelectrical impedance analysis approaches (MFBIA; QuadScan 4000), a bioimpedance spectroscopy device (BIS; Hydra 4200), and multiple dilution. RESULTS: BIS provided intracellular water estimates that were comparable to criterion, based on mean comparisons, at both time points (T1: criterion: 24.2+/ 3.1L, BIS: 24.0+/-3.7 L; T2: criterion: 20.6+/-3.7 L, BIS: 19.7+/-3.2L). MFBIA (with Deurenberg equations) provided comparable measures to criterion only at T2 (criterion: 20.3+/-3.7L, MFBIA: 20.6+/-2.7 L). Both MFBIA (with QuadScan proprietary equations) and BIS produced estimates of intracellular water change that were comparable to dilution. There was substantial variability in individual volume measures. CONCLUSIONS: Although MFBIA and BIS hold promise as convenient techniques for assessing fluid changes, individual variability in measurements makes them impractical for assessment of extremely obese patients in the clinical setting. PMID- 18676067 TI - Will global warming affect soil-to-plant transfer of radionuclides? AB - Recent assessments of global climate/environmental change are reaching a consensus that global climate change is occurring but there is significant uncertainty over the likely magnitude of this change and its impacts. There is little doubt that all aspects of the natural environment will be impacted to some degree. Soil-to-plant transfer of radionuclides has long been a significant topic in radioecology, both for the protection of humans and the environment from the effects of ionising radiation. Even after five decades of research considerable uncertainty exists as to the interplay of key environmental processes in controlling soil-plant transfer. As many of these processes are, to a lesser or greater extent, climate-dependent, it can be argued that climate/environmental change will impact soil-to-plant transfer of radionuclides and subsequent transfers in specific environments. This discussion attempts to highlight the possible role of climatic and climate-dependent variables in soil-to-plant transfer processes within the overall predictions of climate/environmental change. The work is speculative, and intended to stimulate debate on a theme that radioecology has either ignored or avoided in recent years. PMID- 18676068 TI - Isotope tracing of submarine groundwater discharge offshore Ubatuba, Brazil: results of the IAEA-UNESCO SGD project. AB - Results of groundwater and seawater analyses for radioactive (3H, 222Rn, 223Ra, 224Ra, 226Ra, and 228Ra) and stable (D and 18O) isotopes are presented together with in situ spatial mapping and time series 222Rn measurements in seawater, direct seepage measurements using manual and automated seepage meters, pore water investigations using different tracers and piezometric techniques, and geoelectric surveys probing the coast. This study represents first time that such a new complex arsenal of radioactive and non-radioactive tracer techniques and geophysical methods have been used for simultaneous submarine groundwater discharge (SGD) investigations. Large fluctuations of SGD fluxes were observed at sites situated only a few meters apart (from 0 cm d(-1) to 360 cm d(-1); the unit represents cm3/cm2/day), as well as during a few hours (from 0 cm d(-1) to 110 cm d(-1)), strongly depending on the tidal fluctuations. The average SGD flux estimated from continuous 222Rn measurements is 17+/-10 cm d(-1). Integrated coastal SGD flux estimated for the Ubatuba coast using radium isotopes is about 7x10(3) m3 d(-1) per km of the coast. The isotopic composition (deltaD and delta18O) of submarine waters was characterised by significant variability and heavy isotope enrichment, indicating that the contribution of groundwater in submarine waters varied from a small percentage to 20%. However, this contribution with increasing offshore distance became negligible. Automated seepage meters and time series measurements of 222Rn activity concentration showed a negative correlation between the SGD rates and tidal stage. This is likely caused by sea level changes as tidal effects induce variations of hydraulic gradients. The geoelectric probing and piezometric measurements contributed to better understanding of the spatial distribution of different water masses present along the coast. The radium isotope data showed scattered distributions with offshore distance, which imply that seawater in a complex coast with many small bays and islands was influenced by local currents and groundwater/seawater mixing. This has also been confirmed by a relatively short residence time of 1-2 weeks for water within 25 km offshore, as obtained by short lived radium isotopes. The irregular distribution of SGD seen at Ubatuba is a characteristic of fractured rock aquifers, fed by coastal groundwater and recirculated seawater with small admixtures of groundwater, which is of potential environmental concern and has implications on the management of freshwater resources in the region. PMID- 18676069 TI - Retrospective review of donor site complications after harvest of cancellous bone from the anteriomedial tibia. AB - Donor site morbidity is important in deciding the site for harvest of cancellous bone for alveolar bone grafts. Many studies have supported the view that tibia is safe with few complications in the short term. We know of no studies on the long term complications to the donor site after tibial bone grafting in children with alveolar clefts. The casenotes of 40 children who had had tibial bone grafts for alveolar clefts were studied retrospectively, and parents or patients were contacted by telephone and a standardised questionnaire was used to assess any long term complications at the donor site. We found none. We found no evidence of long-term complications at the donor site in children who had had proximal tibial bone grafting for secondary repair of alveolar clefts. We conclude that the proximal tibia is a safe site from which to obtain cancellous bone graft for alveolar clefts in children. This study is preliminary, and highlights the need for a randomised trial. PMID- 18676070 TI - Current status of replacement of the temporomandibular joint in the United Kingdom. AB - Total replacement of the temporomandibular joint (TMJ) has been done in the UK since 1987. The three currently available systems are the Christensen system, the TMJ Concepts system and the Lorenz (BMF) system. Data from surgeons who replace TMJ were collated up to May 2007. There were nine units (eight NHS, one private) offering replacement. The TMJ Concepts system is the most popular of the three systems. Units are treating between five and 12 patients each year with an estimated total annual workload of 60-65 patients. The current total costs range from pound 15 000 to pound 19 000 for bilateral replacement. The most worrying complication is infection, which may affect up to 2.6% of patients. PMID- 18676071 TI - The power of language: a secondary analysis of a qualitative study exploring English midwives' support of mother's baby-feeding practice. AB - OBJECTIVE: to explore the use of language by midwives reporting their experiences of baby-feeding practice. DESIGN: A qualitative approach incorporating a secondary analysis of data previously collected in a study based on grounded theory principles. Data were collected using in-depth interviews. The secondary analysis involved a content analysis method. SETTING: two maternity services in the north of England. PARTICIPANTS: 30 midwives who worked across all clinical areas. FINDINGS: these midwives' explanations of how they supported women with baby feeding suggest that they used language to direct women towards decisions that the midwives thought best, without prior exploration of the woman's understanding and beliefs of baby feeding. Women were often described as 'girls' and 'ladies' indicating the power differentials in their relationship. The midwives were aware that, at times, the language they used was contradictory to woman-centred maternity care. KEY CONCLUSIONS: language may be used to control childbearing women and direct them to decisions that the midwife wants, rather than enabling the woman to make her own decisions. The terms used by midwives to relate to women, such as 'girls' and 'ladies', may be a strategy used to improve a midwife's perception of her status in maternity care. IMPLICATIONS FOR PRACTICE: language may be used to undermine women. It is important that the language used when interacting with women is considered carefully in order to facilitate an unbiased perspective and to promote partnership. The word 'women', rather than 'girls' or 'ladies', should be used when referring to users of the maternity services. PMID- 18676072 TI - Residential runoff as a source of pyrethroid pesticides to urban creeks. AB - Pyrethroid pesticides occur in urban creek sediments at concentrations acutely toxic to sensitive aquatic life. To better understand the source of these residues, runoff from residential neighborhoods around Sacramento, California was monitored over the course of a year. Pyrethroids were present in every sample. Bifenthrin, found at up to 73 ng/L in the water and 1211 ng/g on suspended sediment, was the pyrethroid of greatest toxicological concern, with cypermethrin and cyfluthrin of secondary concern. The bifenthrin could have originated either from use by consumers or professional pest controllers, though the seasonal pattern of discharge from the drain was more consistent with professional use as the dominant source. Stormwater runoff was more important than dry season irrigation runoff in transporting pyrethroids to urban creeks. A single intense storm was capable of discharging as much bifenthrin to an urban creek in 3h as that discharged over 6 months of irrigation runoff. PMID- 18676073 TI - Comparison of annual dry and wet deposition fluxes of selected pesticides in Strasbourg, France. AB - This work summarizes the results of a study of atmospheric wet and dry deposition fluxes of Deisopropyl-atrazine (DEA), Desethyl-atrazine (DET), Atrazine, Terbuthylazine, Alachlor, Metolachlor, Diflufenican, Fenoxaprop-p-ethyl, Iprodione, Isoproturon and Cymoxanil pesticides conducted in Strasbourg, France, from August 2000 through August 2001. The primary objective of this work was to calculate the total atmospheric pesticide deposition fluxes induced by atmospheric particles. To do this, a modified one-dimensional cloud water deposition model was used. All precipitation and deposition samples were collected at an urban forested park environment setting away from any direct point pesticide sources. The obtained deposition fluxes induced by atmospheric particles over a forested area showed that the dry deposition flux strongly contributes to the total deposition flux. The dry particle deposition fluxes are shown to contribute from 4% (DET) to 60% (cymoxanil) to the total deposition flux (wet+dry). PMID- 18676074 TI - Kidney disease in childhood-onset diabetes. PMID- 18676075 TI - Progression of kidney disease in moderately hypercholesterolemic, hypertensive patients randomized to pravastatin versus usual care: a report from the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT). AB - BACKGROUND: Dyslipidemia is common in patients with chronic kidney disease. The role of statin therapy in the progression of kidney disease is unclear. STUDY DESIGN: Prospective randomized clinical trial, post hoc analyses. SETTING & PARTICIPANTS: 10,060 participants in the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (lipid-lowering component) stratified by baseline estimated glomerular filtration rate (eGFR): less than 60, 60 to 89, and 90 or greater mL/min/1.73 m(2). Mean follow-up was 4.8 years. INTERVENTION: Randomized; pravastatin, 40 mg/d, or usual care. OUTCOMES & MEASUREMENTS: Total, high-density lipoprotein, and low-density lipoprotein cholesterol; end-stage renal disease (ESRD), eGFR. RESULTS: Through year 6, total cholesterol levels decreased in the pravastatin (-20.7%) and usual-care groups (-11.2%). No significant differences were seen between groups for rates of ESRD (1.36 v 1.45/100 patient-years; P = 0.9), composite end points of ESRD and 50% or 25% decrease in eGFR, or rate of change in eGFR. Findings were consistent across eGFR strata. In patients with eGFR of 90 mL/min/1.73 m(2) or greater, the pravastatin arm tended to have a higher eGFR. LIMITATIONS: Proteinuria data unavailable, post hoc analyses, unconfirmed validity of the Modification of Diet in Renal Disease Study equation in normal eGFR range, statin drop-in rate in usual-care group with small cholesterol differential between groups. CONCLUSIONS: In hypertensive patients with moderate dyslipidemia and decreased eGFR, pravastatin was not superior to usual care in preventing clinical renal outcomes. This was consistent across the strata of baseline eGFR. However, benefit from statin therapy may depend on the degree of the cholesterol level decrease achieved. PMID- 18676076 TI - Prevalence of estimated GFR reporting among US clinical laboratories. AB - BACKGROUND: Routine laboratory reporting of estimated glomerular filtration rate (eGFR) may help clinicians detect kidney disease. The current national prevalence of eGFR reporting in clinical laboratories is unknown; thus, the extent of the situation of laboratories not routinely reporting eGFR with serum creatinine results is not quantified. DESIGN: Observational analysis. SETTING: National Kidney Disease Education Program survey of clinical laboratories conducted in 2006 to 2007 by mail, web, and telephone follow-up. PARTICIPANTS: A national random sample, 6,350 clinical laboratories, drawn from the Federal Clinical Laboratory Improvement Amendments database and stratified by 6 major laboratory types/groupings. PREDICTORS: Laboratory reports serum creatinine results. OUTCOMES: Reporting eGFR values with serum creatinine results. MEASUREMENTS: Percentage of laboratories reporting eGFR along with reporting serum creatinine values, reporting protocol, eGFR formula used, and style of reporting cutoff values. RESULTS: Of laboratories reporting serum creatinine values, 38.4% report eGFR (physician offices, 25.8%; hospitals, 43.6%; independents, 38.9%; community clinics, 47.2%; health fair/insurance/public health, 45.5%; and others, 43.2%). Physician office laboratories have a reporting prevalence lower than other laboratory types (P < 0.001). Of laboratories reporting eGFR, 66.7% do so routinely with all adult serum creatinine determinations; 71.6% use the 4 variable Modification of Diet in Renal Disease Study equation; and 45.3% use the ">60 mL/min/1.73 m(2)" reporting convention. Independent laboratories are least likely to routinely report eGFR (50.6%; P < 0.05) and most likely to report only when specifically requested (45.4%; P < 0.05). High-volume laboratories across all strata are more likely to report eGFR (P < 0.001). LIMITATIONS: Self reporting by laboratories, federal database did not have names of laboratory directors/managers (intended respondents), assumed accuracy of federal database for sample purposes. CONCLUSIONS: Routine eGFR reporting with serum creatinine values is not yet universal, and laboratories vary in their reporting practices. PMID- 18676077 TI - West Nile virus encephalitis in a renal transplant recipient: the role of intravenous immunoglobulin. AB - West Nile virus is a common viral infection in endemic areas. Although the disease has a benign course in immunocompetent individuals, it tends to run a more malignant course in immunocompromised patients such as solid-organ transplant recipients. In this study, a renal transplant recipient presented with fever, impaired speech, obtundation, and features suggestive of meningitis on cerebrospinal fluid examination. Although initial serological study results were negative, the patient was treated promptly with intravenous immunoglobulin (IVIG) based on a strong clinical suspicion of West Nile virus encephalitis. Therapy with IVIG was associated with complete recovery of neurological features, and the patient was discharged on day 7 after resolution of neurological complications. The favorable outcome of this patient may be related to early treatment with IVIG. PMID- 18676079 TI - Social support, material circumstance and health behaviour: influences on health in First Nation and Inuit communities of Canada. AB - An expansive literature describes the links between social support and health. Though the bulk of this evidence emphasizes the health-enhancing effect of social support, certain aspects can have negative consequences for health (e.g., social obligations). In the Canadian context, the geographically small and socially interconnected nature of First Nation and Inuit communities provides a unique example through which to explore this relationship. Despite reportedly high levels of social support, many First Nation and Inuit communities endure broad social problems, thereby leading us to question the assumption that social support is primarily health protective. We draw from narrative analysis of interviews with 26 First Nation and Inuit Community Health Representatives to critically examine the health and social support relationship, and the social structures through which social support influences health. Findings indicate that there are health-enhancing and health-damaging properties of the health-social support relationship, and that the negative dimensions can significantly outweigh the positive ones. Social support operates at different structural levels, beginning with the individual and extending toward family and community. These social structures are important as they reinforce an individual's sense of belonging, however, these high-density networks can also exert conformity pressures and social obligations that promote health-damaging behaviours such as domestic violence and smoking. The poor material circumstances that characterize so many First Nation and Inuit communities add another layer of complexity as limited resources can trap individuals within the confines of their immediate social contexts. Research and policy interventions must pay close attention to the social context within which social support, health behaviours and material circumstances interact to influence health outcomes among First Nation and Inuit communities. PMID- 18676078 TI - Adolescent alcohol involvement and suicide attempts: toward the development of a conceptual framework. AB - The present article provides a conceptual framework of the relation between alcohol involvement (A) and suicide attempts (S). This framework can be broadly construed to reflect two dimensions: directionality (direction of causality; A- >S, S-->A, or a spurious relation) and temporality (distinguishing between proximal and distal effects of both behaviors). We review and evaluate the evidence on the association between A and S among adolescents using this conceptual framework as a guide. The extant data suggest that this relation is complex and not fully understood. Further, it seems unlikely that a single approach will be found to determine direction of causality, and the specification and validation of hypothesized mechanisms will involve a variety of different types of evidence. Suggestions for additional research using informative designs are discussed. PMID- 18676080 TI - The clomiphene citrate challenge test (CCCT) in women with elevated basal FSH: biological significance and predictive value. AB - OBJECTIVE: The rationale of the clomiphene citrate challenge test (CCCT) is that day 10 serum FSH is influenced by the quality of the recruited oocytes. Biological evidence supporting this assumption is, however, lacking. The aim of this study is to investigate the relationship between results from the CCCT and the quantity and the quality of the recruited oocytes. STUDY DESIGN: Patients selected for in vitro fertilization (IVF) and who were found to have elevated basal FSH (n=114) underwent an IVF cycle using follicles developing during CCCT. Subsequently, a subgroup of patients (n=89) underwent a second cycle receiving high doses of gonadotropins. The main outcome considered was the transfer of viable embryos. RESULTS: During the CCCT cycle, the area under the receiver operating characteristics (ROC) curves for day 3 and day 10 serum FSH to predict embryo-transfer was 0.48 (95% CI, 0.37-0.60) and 0.74 (95% CI, 0.63-0.82), respectively. In the subsequent cycle, the area under the ROC curves for the two variables was 0.58 (0.43-0.72) and 0.58 (0.43-0.72), respectively. CONCLUSIONS: CCCT effectively mirrors the quantity and the quality of the recruited oocytes but its predictive value is low. PMID- 18676081 TI - Probing conformational changes in orphan nuclear receptor: the NGFI-B intermediate is a partially unfolded dimer. AB - Human nerve growth factor-induced B (NGFI-B) is a member of the NR4A subfamily of orphan nuclear receptors (NRs). Lacking identified ligands, orphan NRs show particular co-regulator proteins binding properties, different from other NRs, and they might have a non-classical quaternary organization. A body of evidence suggests that NRs recognition of and binding to ligands, DNA, homo- and heterodimerization partners and co-regulator proteins involve significant conformational changes of the NR ligand-binding domains (LBDs). To shed light on largely unknown biophysical properties of NGFI-B, here we studied structural organization and unfolding properties of NGFI-B ligand (like)-binding domain induced by chemical perturbation. Our results show that NGFI-B LBD undergoes a two-state guanidine hydrochloride (GndHCl) induced denaturation, as judged by changes in the alpha-helical content of the protein monitored by circular dichroism spectroscopy (CD). In contrast, changes in the tertiary structure of NGFI-B LBD, reported by intrinsic fluorescence, reveal a clear intermediate state. Additionally, SAXS results demonstrate that the intermediate observed by intrinsic fluorescence is a partially folded homodimeric structure, which further unfolds without dissociation at higher GndHCl concentrations. This partially unfolded dimeric assembly of NGFI-B LBD might resemble an intermediate that this domain access momentarily in the native state upon interactions with functional partners. PMID- 18676082 TI - Reversible brainstem hypertensive encephalopathy (RBHE): Clinicoradiologic dissociation. AB - We report two cases of reversible brainstem hypertensive encephalopathy (RBHE) with unusual magnetic resonance (MR) findings. Patient 1, an 85-year-old man without a history of hypertension, developed acute severe hypertension and mild consciousness disturbance as the only symptoms. Patient 2, a 46-year-old man with an untreated hypertension, presented with extremely high blood pressure and general fatigue, vertigo, and mild dysarthria as the initial manifestations. In these patients, fluid-attenuated inversion recovery (FLAIR) and T2-weighted MR images revealed diffuse hyperintensities in the brainstem. Diffusion-weighted imaging (DWI) findings were normal, and apparent diffusion coefficient (ADC) values were increased in the brainstem. The supratentorial regions were largely spared, and mildly diffuse hyperintensities were noted in the white matter. There were no accompanying changes in the occipital lobe and cerebellum. The lesions completely resolved after stabilization of blood pressure. The normal DWI findings and high ADC values were consistent with vasogenic edema due to severe hypertension. The characteristics of RBHE are a very high blood pressure, mild clinical and neurologic symptoms, rapidly improved MR findings after initial treatment with the control of hypertension, and a marked clinicoradiologic dissociation. PMID- 18676083 TI - Dicamba-induced genotoxicity in Chinese hamster ovary (CHO) cells is prevented by vitamin E. AB - In the present study the cytogenetic and genotoxic effect of benzoic herbicide dicamba and its Argentinean commercial formulation banvel (57.71% dicamba) was evaluated and whether this effect is mediated through oxidative damage or not. The protective role of vitamin E was also studied. Sister chromatid exchange (SCE) frequency, cell-cycle progression, and cell viability analyses in CHO cells were used as in vitro end-points. Treatments with the test compounds were performed either during 24h (Protocol A) or 12h (Protocol B) before harvesting. Protocol A showed that vitamin E decreased pesticide SCE induction, corrected the cell-cycle delay and partially protected cell-death only in 500 microg/ml dicamba treated cultures. A similar trend was found in banvel-treated cultures. Protocol B revealed similar protective role of vitamin E only for dicamba-induced geno- and cytotoxicity. Based on these observations it could be suggested that dicamba injures DNA by delivering reactive oxygen species rather than by another type of mechanism/s. Although banvel mimics the effect observed by dicamba, its formulation contains other xenobiotic/s agents able to induce cellular and DNA damage by a different mechanism/s. Further investigations are needed to acquire a comprehensive knowledge of the possible mechanism/s through dicamba and banvel exert their toxic effects. PMID- 18676084 TI - Removal of paraquat from synthetic wastewater using phillipsite-faujasite tuff from Jordan. AB - A phillipsite-faujasite tuff (faujasite) from Jordan has been activated and characterized to evaluate its efficiency in removing paraquat from synthetic wastewater and to specify optimum conditions with maximum efficiency of the faujasite tuff. Noticed variation in CEC is indicated and explained by selectivity of the faujasite to K(+) and Na(+). Removal of paraquat by using faujasite directly gave average removal efficiency equal to 59%. Thermal activation of the samples enhances their performance. The higher is the temperature of activation the higher the efficiency. Treatment of paraquat effluent using faujasite is a good alternative. Na- or K-loading of the original sample give better results. Loading with Ca and Mg however is not recommended because they always exhibit lower efficiency, which might be related to their lower CEC value. Removal of paraquat from wastewater using charcoal and the non activated faujasite increased efficiency to about 82%. Thermal activated faujasite at 200 degrees C and 300 degrees C increased the uptake up to about 93% and 99.5%, respectively. Therefore thermal activation of faujasite is necessary to improve its uptake performance. PMID- 18676085 TI - Coupling between neuronal nitric oxide synthase and glutamate receptor 6-mediated c-Jun N-terminal kinase signaling pathway via S-nitrosylation contributes to ischemia neuronal death. AB - S-nitrosylation, as a post-translational protein modification, recently has been paid more and more attention in stroke research. S-nitrosylation regulates protein function by the mechanisms of covalent attachment that control the addition or the removal of nitric oxide (NO) from a cysteine thiol. The derivation of NO is established by the demonstration that, in cerebral neurons, NO mainly generates from neuronal nitric oxide synthase (nNOS) during the early stages of reperfusion. In the past researches, we demonstrate that global ischemia-reperfusion facilitates the activation of glutamate receptor 6 (GluR6) mediated c-Jun N-terminal kinase (JNK) signaling pathway. The objective of this study is primarily to determine, during the early stages of reperfusion in rat four-vessel occlusion (4-VO) ischemic model, whether nNOS-derived NO affects the GluR6-mediated JNK signaling route via S-nitrosylation which is performed mainly by the biotin switch assay. Here, we show that administration of 7-nitroindazole, an inhibitor of nNOS, or ketamine, an antagonist of N-methyl-d-aspartate receptor (NMDAR), diminishes the increased S-nitrosylation of GluR6 induced by cerebral ischemia-reperfusion. In contrast, 2-amion-5,6-dihydro-6-methyl-4H-1,3-thiazine, an inhibitor of inducible NO synthase does not affect S-nitrosylation of GluR6. Moreover, treatment with sodium nitroprusside (SNP), an exogenous NO donor, increases the S-nitrosylation and phosphorylation of nNOS, leading to the attenuation of the increased S-nitrosylation of GluR6 and the assembling of GluR6* postsynaptic density protein 95 (PSD95)* mixed lineage kinase 3 (MLK3) signaling module induced by cerebral ischemia-reperfusion. The results also show that GluR6 downstream MLK3* mitogen activated protein kinase kinase 4/7* JNK signaling module and nuclear or non-nuclear apoptosis pathways are involved in the above signaling route. However, dithiothreitol (DTT) antagonizes the neuroprotection of SNP. Treatment with DTT alone, as a negative control, prevents S-nitrosylation of proteins, which indicates the existence of endogenously produced S-nitrosylation. These data suggest that GluR6 is S-nitrosylated by endogenous NO in cerebral ischemia-reperfusion, which is possibly correlated with NMDAR* PSD95* nNOS signaling module, and further activates GluR6* PSD95* MLK3 signaling module and JNK signaling pathway. In contrast, exogenous NO donor antagonizes the above action of endogenous NO generated from nNOS. Thus, our results provide the coupling of nNOS with GluR6 by S-nitrosylation during the early stages of ischemia-reperfusion, which can be a new approach for stroke therapy. PMID- 18676086 TI - Behavioral effects of the CRF1 receptor antagonist R121919 in rats selectively bred for high and low activity in the swim test. AB - This study assessed effects of a CRF(1) receptor antagonist, R121919, on the behavior of rats that have been selectively bred to exhibit very high or very low activity in a swim test. Following treatment with R121919 (10 mg/kg, s.c.) or vehicle, several types of behavior were examined including: (1) spontaneous ambulatory activity in a novel environment, (2) swim-test activity, (3), and responses in an elevated plus maze. The most pronounced effects were observed in the swim test. Although R121919 had little effect on the swim-test behavior of normal, non-selected rats, Swim High-active rats (SwHi), characterized by being very active and exhibiting pronounced struggling behavior in the swim test, showed increased activity (more struggling) after R121919; in contrast, Swim Low active (SwLo) rats, characterized by being very inactive and exhibiting pronounced floating behavior in the swim test, showed decreased activity (more floating) after R121919. This effect was observed in both male and female rats. No differences between strains or the effects of R121919 were observed for spontaneous ambulation or in the elevated plus maze test. PMID- 18676088 TI - Inflammatory malignant fibrous histiocytoma of kidney: a case report. AB - Among the renal sarcomas, inflammatory malignant fibrous histiocytoma (MFH) is an extremely rare presentation. A 45-year-old woman presented with acute retention urine following an episode of gross hematuria. Computerized tomography showed a solid mass at the lower pole of the left kidney. The patient underwent left nephrectomy. Histologically and immunohistochemically, the tumor was diagnosed as an inflammatory subtype of MFH. Histological appearances of inflammatory MFH vary widely and frequently overlap with benign reactive conditions such as Xanthogranulomatous pyelonephritis (XGPN) and malignant lesions, e.g. malignant lymphoma and, less frequently, a sarcomatoid variant of renal cell carcinoma. It is important, though difficult, to differentiate inflammatory MFH from these lesions. Careful morphological examination and immunohistochemical findings of the lesion are of great value, in particular in excluding it from its mimics. We discuss the pathological features and challenges involved in differentiating inflammatory MFH from its masquerader. PMID- 18676087 TI - Progesterone reduces wakefulness in sleep EEG and has no effect on cognition in healthy postmenopausal women. AB - Sleep is frequently impaired in postmenopausal women. Progesterone prompted benzodiazepine-like effects on sleep EEG in young normal male subjects. Aim of this study was to test if treatment with progesterone improves sleep after menopause. A randomised double blind crossover design study with 2 treatment intervals of 21 days duration separated by a 2 weeks washout was performed. An oral dose of 300 mg micronized progesterone was given each for 21 days. At the beginning and the end of the two intervals a sleep EEG was recorded and cognitive performance was assessed in 10 healthy postmenopausal women (age: 54-70 years). Progesterone treatment led to a decrease of intermittent time spent awake. During the first third of the night rapid eye movement (REM) sleep increased. The spectral analysis of the EEG resulted in no significant differences of the power spectra. Progesterone did not affect cognitive performance. In summary progesterone demonstrated a distinct sleep promoting effect by reduction of time of wake without impairing cognitive functions during daytime. As possible mechanisms of progesterone a GABA-agonistic effect and the regulation of gene expression via the progesterone receptor are discussed. Progesterone might be useful in the treatment of sleep disturbances of postmenopausal women. PMID- 18676089 TI - Surveillance of craniopharyngioma cyst growth in children treated with proton radiotherapy. AB - PURPOSE: Craniopharyngiomas are benign, slow-growing tumors that frequently contain a cystic component. Even with gross total resection, the cyst can reform and cause symptoms. Fluctuations in cyst volume during radiotherapy (RT) can affect treatment planning and delivery. The aim of this study was to report our experience with cyst enlargement during conformal proton RT for children with craniopharyngioma and to make recommendations regarding mid-treatment surveillance. METHODS AND MATERIALS: Between January 2001 and August 2007, 24 children (aged 7). Half of this population had already intended to stop, they declared to smoke mostly because of stress. The global cessation rate after a median follow-up of 32 weeks is 27%. The cessation rate lasting six months for 83 patients followed regularly was 23%. The good prognostic factors of cessation, after multivariate analysis, were (when taking out from the model the regular follow-up), to be confidant on the possibility to stop (OR=0.87 [0.78-0.97]). On the contrary, a high score (>7) for Fagerstrom test (OR=1.9 [1.1-3.4), the use of smoking to fight against stress (OR=1.08 [1.0-1.2) and a smoking environment at home (OR=4.5 [1.1-18.9]) were prognostic of a failure in quitting smoking. CONCLUSION: These results show that the cessation rate, which is still too low as compared with the literature, could be increased by a better follow-up of smokers associated with an information campaign on the existing possibilities to quit smoking in Tunisia. PMID- 18676109 TI - [Diabetes always hits the heart]. PMID- 18676110 TI - ["Antibiotics only when necessary" campaign in the Alpes-Maritimes district: no negative impact on invasive infections in children in the community 1998-2003]. AB - OBJECTIVES: A multidisciplinary group (for the study and prevention of infections in children, GEPIE) conducted a local public health campaign to improve antibiotic prescribing in pediatric outpatient care in the Alpes-Maritimes district. This campaign included peer-conducted academic-detailing (educational outreach) visits in 2000 and 2003. Practitioners often report during both focus groups and the GEPIE visits that they prescribe antibiotics because of a fear of subsequent complications. This study therefore sought to explore the trends in invasive diseases related to bacterial respiratory pathogens. METHODS: A retrospective survey examined the incidence in the district of invasive infections due to Streptococcus pneumoniae, Haemophilus influenzae, Streptococcus pyogenes, and Neisseria meningitidis from 1998 through 2003 among children aged 1 month to 15 years. All local laboratories were asked for all reports of blood cultures (as well as spinal, synovial, pericardiac, peritoneal, and pleural fluid cultures) positive for these bacteria in the study population over the study period. Resistance and serotype data were recorded. RESULTS: Over the 6-year study period, there were 113 cases of invasive infections: 64% due to S. pneumoniae, 26% to N. meningitidis, 6% to H. influenzae, and 4% to S. pyogenes. There was no statistically significant difference in the annual average incidence rate before and during the campaign. The overall incidence rate was 11.2 cases per 100,000; meningitis accounted for 4.2, and invasive pneumococcal diseases 7.2 (42.7 per 100,000 children younger than 2 years). Pneumococcal resistance rates to penicillin and erythromycin did not change significantly. DISCUSSION: Two local cross-sectional studies at daycare centers in 1999 and 2002 showed a 9% reduction in the proportion of children who received antibiotics. The stability of invasive infection in children and of bacterial resistance during the campaign is reassuring. The campaign's risk/benefit ratio appears positive. CONCLUSION: These results encourage continuing promotion of rational antibiotic prescribing and ought to allay practitioners' fears about the consequences of prescribing antibiotics less often. PMID- 18676111 TI - Magnetic resonance imaging in the diagnosis and management of hip pain after total hip arthroplasty. AB - Evaluation of pain following total hip arthroplasty (THA) can be challenging in the absence of radiographic pathology. This study aimed to examine the diagnostic utility of magnetic resonance imaging (MRI) in the evaluation of enigmatic hip pain following THA. We reviewed a series of patients who were evaluated with MRI after presenting with enigmatic hip pain following THA. MRI was able to demonstrate pathology in the periprosthetic tissues in all hips with minimal artifact. Patients underwent a range of conservative and operative interventions depending on the underlying pathology. If used discriminately in situations where pathology cannot be detected by conventional methods, MRI is a highly effective modality that can aid in the diagnosis of a wide range of disorders thereby allowing the clinician to determine the most appropriate intervention. PMID- 18676112 TI - Comparison of clinical outcome in primary total hip arthroplasty by conventional anterolateral transgluteal or 2-incision approach. AB - Studies comparing the minimally invasive surgery with 2-incision technique (MIS 2) technique with the conventional transgluteal technique are lacking in the literatures. We compared the clinical outcomes in a series of 166 hips, including 83 MIS-2 total hip arthroplsty (THA) (group I) and another 83 matched-pair series (group II) using conventional transgluteal approach. Demographic data, hospital course, radiographic data, and functional outcome (Western Ontario and McMaster University Osteoarthritis Index and Harris hip score [HHS]) were investigated. The 2 groups significantly differed in 3 and 6-month HHS with a higher HHS in MIS 2 group. But the MIS-2 group had longer operation time, increased blood loss, and more complications. The current study indicates that the benefit of MIS-2 technique was only short-term with quicker functional recovery and shorter duration use of nonsteroid antiinflammatory drugs postoperatively. PMID- 18676113 TI - Hip resurfacing in an 88-year-old patient? Highlighting selection criteria for hip resurfacings in patients older than 65 years. AB - The success of hip resurfacings in the older population is still to be determined, although it has been an attractive option in the much younger patients with arthrosis of the hip. We present a 95-year-old active, independent patient who underwent a Birmingham hip resurfacing at 88 years of age. More than 7 years after surgery, the original components are still in situ, and the patient is pain-free with a very active lifestyle at the age of 95 years. Hip resurfacings are not without risks, and we highlight the evolution of strict selection criteria for patients older than 65 years who may be suitable candidates for hip resurfacing arthroplasty. PMID- 18676114 TI - Comparison of the model-based and marker-based roentgen stereophotogrammetry methods in a typical clinical setting. AB - Roentgen stereophotogrammetric analysis (RSA) is an established method for the precise measurement of implant migration. Model-based RSA (MBRSA) alleviates the need to attach tantalum markers to the prosthesis, which has prevented wider application of RSA. The goal of this study was to investigate the equivalence of both methods for the clinical measurement of implant migration. Tibial component migration was measured in 24 patients using both methods from the same set of radiographs. The maximum agreement interval, mean (+/-2 standard deviations), of the difference between both methods was modest, at 0.002 mm (0.144 mm) and -0.078 degrees (0.782 degrees ). The results suggest that MBRSA can be used interchangeably with the marker-based method and that the advantages of MBRSA do not come at the cost of a loss in accuracy. PMID- 18676115 TI - Vascular injuries after total joint arthroplasty. AB - Vascular injuries, although highly feared, can occur after total joint arthroplasty, often resulting in legal suits. This study evaluates the circumstances related to vascular injuries after joint arthroplasty. Using prospectively collected data on 13,517 patients undergoing total joint arthroplasty at our institution, 16 (0.1%) vascular injuries were identified. Eleven injuries occurred after total knee arthroplasty (TKA) and 5 after total hip arthroplasty (THA). Indirect injury was the most common mechanism in TKA. In contrast, direct injury was most prevalent in THA. One patient died of complications related to vascular injury. Of 16 patients, 8 (50%) had launched a legal suit against the operating surgeon. There appears to be no further specific measure that can be taken to absolutely avoid this complication. Patient awareness regarding this real problem may play a role in defraying the high likelihood of legal suits associated with this complication. PMID- 18676116 TI - Assessing persistent cancer pain: a comparison of current pain ratings and pain recalled from the past week. AB - Recent guidelines developed by the U.S. Food and Drug Administration for the use of patient-reported outcomes discuss the rating of pain and other symptoms at their current level of severity versus rating these symptoms using a recall period, such as the past 24 hours or past week. To explore whether the overall experience of cancer patients is better represented by ratings of current pain or pain recalled from the past week, we conducted a secondary analysis of Eastern Cooperative Oncology Group data from 1147 patients with cancer who had reported having persistent pain during the past week. Patients used the Brief Pain Inventory (BPI) to rate their current pain along with their pain at its worst, least, and average during the past week. T-tests were used to compare ratings of current pain and pain recalled from the past week. Linear regressions described the extent to which the various pain ratings contributed to overall pain interference, also derived from the BPI. Overall, patients rated their current pain as less severe than their worst or average pain recalled from the past week. Worst pain recalled from the past week contributed most to ratings of pain interference. These findings indicate that ratings of recalled worst pain, rather than ratings of current pain, might better reflect the overall experience of pain and its impact on function in cancer patients with persistent pain. Our results provide information that might guide the choice of recall period for cancer clinical trials with pain as a self-reported outcome. PMID- 18676117 TI - Systematic review of the effectiveness of botulinum toxin or radiotherapy for sialorrhea in patients with amyotrophic lateral sclerosis. AB - Fifty percent of patients with amyotrophic lateral sclerosis (ALS) experience problems handling serous saliva and 20% fail to achieve adequate control of sialorrhea with anticholinergic medications, or experience intolerable adverse effects from these drugs. Both botulinum and radiotherapy have been suggested in the literature as treatments for intractable sialorrhea. In this review, we assess the evidence for the effectiveness and toxicity of botulinum toxin and radiotherapy for sialorrhea in patients with ALS. Relevant studies were retrieved from Medline, Embase and Cochrane Databases. Handsearching of Neurology, Journal of Pain and Symptom Management, and Palliative Medicine and of reference lists, was carried out. Five studies (28 patients) were included in the analysis of botulinum. Of the four studies using an intraglandular method of injection, no adverse effects occurred. Two of these had positive findings of the effect of botulinum in salivary secretion rate and quality of life. In contrast, significant adverse effects were experienced by two patients in a study of retrograde injections into the salivary ducts. Two studies were included in the analysis of radiotherapy (27 patients). Both demonstrated a positive effect of radiotherapy on salivary secretion rate. Some patients experienced mild acute side effects. Because of the small numbers of studies, small sample sizes, and poor quality of reporting, it is not possible to draw firm conclusions. There is some evidence indicating that both botulinum and radiotherapy are well tolerated, effective treatments for persistent sialorrhea in patients with ALS and that the duration of action is up to three months with botulinum and six months with radiotherapy. PMID- 18676118 TI - Development and preliminary validation of the NePIQoL: a quality-of-life measure for neuropathic pain. AB - Neuropathic pain is frequently associated with negative effects on quality of life (QoL), affecting physical, social, and psychological functioning. Of many existing scales used to measure QoL, none have been validated in a neuropathic pain patient population. This study reports on the development and preliminary psychometric evaluation of the Neuropathic Pain Impact on Quality-of-Life questionnaire (NePIQoL), a measure to assess QoL in neuropathic pain. In Phase I, focus groups with 27 patients and a panel of experts identified QoL issues for inclusion in the measure. Initial items (152) and response categories were pretested using cognitive interviewing (18 patients). Following this, the number of items was reduced to 91. In Phase II, the 91-item version of the NePIQoL was administered to a further 112 patients, poorly performing items were identified, and internal consistency was examined. In Phase III, the revised NePIQoL was administered to a further 110 patients on two occasions to examine validity and test-retest reliability. Qualitative and quantitative pretesting led to extensive revision, resulting in a final measure of 42 items. Finally, Phase IV tested the concurrent validity and responsiveness of the NePIQoL. The authors conclude that the NePIQoL is an acceptable, patient-derived, neuropathic pain-specific measure with evidence of reliability, validity, and temporal stability. PMID- 18676119 TI - Coping strategies in the presence of one's own impending death from cancer. AB - An incurable cancer is a threat to life itself. This study focused on how native born Swedes, who define themselves as nonreligious, actually reflect and act when they try to create helpful strategies in the presence of their own impending deaths and how the strategies serve their purposes. Twenty patients were interviewed in depth. The patients were enrolled in an advanced hospital-based home care team. The interviews were taped, transcribed and analyzed with a qualitative, hermeneutic interpretative method. The informants' efforts to develop useful strategies to restrain death could be symbolized as a cognitive and emotional pendulum, swinging between the extremes of life and death. During the swings of the pendulum, the informants used every means available: their own resources, other people, animals, nature, a transcendent power, hope, imagination and magical thinking. They strove to find factors that fitted their conceptual system and supported their inner balance and structure, all to keep death at a discreet distance and preserve their links to life. These links were togetherness, involvement, hope and continuance, and they served as a shield against hurtful feelings connected to their impending death. The new knowledge about how strategies in the presence of one's own impending death can develop and be used is perhaps the most novel and clinically relevant contribution of this study. PMID- 18676120 TI - Clinical utility of the MDASI-BT in patients with brain metastases. AB - Symptom occurrence has been shown to predict treatment course and survival in cancer patients. The M. D. Anderson Symptom Inventory-Brain Tumor Module (MDASI BT) was recently validated as a tool for primary brain tumor patient self-report of symptoms. This study evaluated the reliability and validity of the MDASI-BT in patients with brain metastases. Data collection included demographic and clinical factors, and the MDASI-BT (0-10 scale). Construct validity was assessed using confirmatory factor analysis, and known-group validity was evaluated by detecting group differences due to disease severity and treatment approach. For reliability, Cronbach's alpha values were computed for each subscale. A sample of 124 patients participated, of which 53.2% were women. Participants were primarily white (79.8%) and married (78.2%), and a variety of solid tumor malignancies were represented. Factor analysis revealed six underlying constructs, including affective symptoms, cognitive dysfunction, focal neurologic deficits, constitutional and gastrointestinal symptoms, and interference with life. The solution with these factors explained 68.4% of the variance. Mean symptom scores were 1.2 and 2.6, and mean interference scores were 1.8 and 4.3 for patients with good and poor Karnofsky scores, respectively (P<0.001). These subscales were also sensitive to opioid analgesic use, with group differences of 1.5 and 2.2 (P<0.001). Cronbach's alpha was 0.9 for each of the two subscales. Fatigue, sleep disturbance, drowsiness, distress, and dry mouth were the most severe symptoms. The MDASI-BT demonstrated validity and reliability in brain metastases patients and can be used to identify and monitor symptom occurrence in relation to treatment course and survival. PMID- 18676122 TI - Oil-in-water lecithin-based microemulsions as a potential delivery system for amphotericin B. AB - In this work the structural features of microemulsions (MEs) containing the pharmaceutical biocompatible Soya phosphatidylcholine/Tween 20 (1:1) as surfactant (S), Captex 200 as oil phase (O), and phosphate buffer 10mM, pH 7.2 as aqueous phase (W) were studied. Systems obtained with different proportions of the components were described by pseudo-ternary phase diagrams in order to characterize the microemulsions studied here. MEs were prepared with and without the polyene antifungal drug amphotericin B (AmB). The maximum AmB incorporation into the ME system was dependent on both the oil phase and surfactant proportions with 6.80 and 5.7 mg/mL in high contents, respectively. The incorporation of AmB into the ME systems significantly increased the profile of the droplet size of the ME for all ranges of surfactant proportions used in the formulations. The microstructures of the system were characterized by dynamic light scattering (DLS) and rheological behavior. The DLS results showed that the size of the oil droplets increases 4.6-fold when AmB is incorporated into the ME system. In all cases the increase in the proportion of the oil phase of the ME leads to a slight increase in the diameter of the oil droplets of the system. Furthermore, for both the AmB-loaded and AmB-unloaded MEs, the size of the oil droplets decrease significantly with the increase of the S proportion in the formulations, demonstrating the efficiency of the surfactant in stabilizing the ME. Depending on the ME composition, an anti-thixotropic behavior was found. The maximum increases of the consistency index caused by the increase of the oil phase of the ME were of 17- and 25-times for the drug-loaded and drug-unloaded MEs, respectively. However, the observed effect for the drug-loaded ME was about 4.6 times higher than that for the drug-unloaded one, demonstrating the strong effect of the drug on the rheological characteristics of the ME system. Therefore, it is possible to conclude that the investigated ME can be used as a very promising vehicle for AmB. PMID- 18676121 TI - Post-traumatic stress disorder, social anxiety disorder, and depression in survivors of the Kosovo War: experiential avoidance as a contributor to distress and quality of life. AB - Few studies have been conducted on psychological disorders other than post traumatic stress disorder (PTSD) in war survivors. The aim of this study was to examine PTSD, social anxiety disorder (SAD), and major depressive disorder (MDD) and their associations with distress and quality of life in 174 Albanian civilian survivors of the Kosovo War. This included testing of conceptual models suggesting that experiential avoidance might influence associations between anxiety and mood disorders with psychological functioning. Each of the three psychiatric disorders was associated with greater experiential avoidance and psychological distress, and lower quality of life. Being a refugee was associated with a higher likelihood of having SAD and MDD. We found evidence for experiential avoidance as a partial mediator of the respective effects of SAD and PTSD on quality of life; experiential avoidance did not mediate the effects of disorders on global distress. We also found support for a moderation model showing that only war survivors without SAD and low experiential avoidance reported elevated quality of life; people with either SAD or excessive reliance on experiential avoidance reported compromised, low quality of life. This is the third independent study, each using a different methodology, to find empirical support for this moderation model [Kashdan, T. B., & Breen, W. E. (2008). Social anxiety and positive emotions: a prospective examination of a self-regulatory model with tendencies to suppress or express emotions as a moderating variable. Behavior Therapy, 39, 1-12; Kashdan, T. B., & Steger, M. F. (2006). Expanding the topography of social anxiety: an experience sampling assessment of positive emotions and events, and emotion suppression. Psychological Science, 17, 120 128]. Overall, we provided initial evidence for the importance of addressing PTSD, SAD, MDD, and experiential avoidance in primarily civilian war survivors. PMID- 18676123 TI - Taurine provides antioxidant defense against NaF-induced cytotoxicity in murine hepatocytes. AB - Fluoride is an environmental and industrial pollutant that affects various organs in humans and animals. The present study was conducted to investigate the protective role of taurine (2-aminoethane sulphonic acid) against fluoride induced cytotoxicity in murine hepatocytes. Sodium fluoride (NaF) was used as the source of fluoride for this particular study. Dose-dependent studies suggest that incubation of hepatocytes with NaF (100mM) for 1h significantly decreased the cell viability as well as intracellular antioxidant power. Increased activities of alanine transaminase (ALT) and alkaline phosphatase (ALP) due to the same dose of toxin exposure confirmed membrane damage. Toxin-induced increased level of intracellular reactive oxygen species (ROS) was confirmed by intracellular ROS production assay using a fluorescent probe 2',7'-dichlorofluorescein diacetate (DCF-DA). In addition, the activities of the antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST) were also decreased by toxin treatment at the previous dose. The same treatment also reduced the level of glutathione (GSH) and total thiols, elevated the level of oxidized glutathione (GSSG) and increased the level of lipid peroxidation end products, protein carbonyl content and extent of DNA fragmentation. Incubation of hepatocytes with taurine, both prior to and in combination with NaF, altered all the NaF-induced parameters. A known antioxidant, vitamin C was taken to compare the cytoprotective activity of taurine against fluoride poisoning. Combining all, the results suggest that taurine protects mouse hepatocytes against fluoride induced cytotoxity. PMID- 18676124 TI - [A rare case of polyorchidism: review of the literature]. AB - Polyorchidism is a rare congenital anomaly. We report 1 case in a Malagasy young man revealed by left scrotal mass. The diagnostic was suspected by clinical examination and confirmed by testicular sonography. Management is conservative, based on clinical and sonography surveillance of supernumerary testicle. PMID- 18676125 TI - [Oral corticosteroids are efficient in recurrent intussusception associated with intestinal lymphoid hyperplasia]. AB - We report a case of recurrent intussusception in a 6-year-old boy associated with Intestinal Lymphoid Hyperplasia (ILH). A single course of steroids resulted in resolution of both symptoms and hyperplasia. We conclude that steroids treatment must be considered in recurrent intussusception associated with ILH and no other lead point identified (lymphoma or intestinal malformation) instead of a more radical surgical approach by a ileum resection. PMID- 18676126 TI - [Juvenile dermatomyositis: 4 case studies]. AB - Juvenile dermatomyositis is a rare disease. The estimated incidence is 2 to 3 cases per million population. Before the use of corticosteroids, prognosis was poor for most patients. Today, despite a better prognosis, the side effects of long-term treatment still burden the outcome of the disease. The diagnosis of this entity is difficult and often delayed due to its rarity. It has been shown that the delay in diagnosis and care is a major factor for poor prognosis. Cutaneous findings are often inaugural, allowing early diagnosis. This study's aim was to share our 10-year experience with juvenile dermatomyositis. PMID- 18676127 TI - [Management of autoimmune thrombocytopenic purpura: a prospective study of 147 children from the regional network RHemaP]. AB - AIM: Assessment of the impact of guidelines from a regional pediatric network to standardize the management of childhood immune thrombocytopenic purpura (ITP). MATERIALS AND METHODS: Consensus guidelines were drawn up in centers of the pediatric network for hematological diseases, RHemaP, and a cohort of children referred for ITP in these centers was set up. A 1-year follow-up was recorded for each patient over a 43-month period. RESULTS: We report data from a cohort of 147 children. At diagnosis, we recorded severe thrombocytopenia (median=8G/l) and 141 children had hemorrhagic symptoms (96%). Only 23 children had a bone marrow aspiration (BMA) at diagnosis (16.3%), which meant a high level of implementation of the RHemaP recommendations (96%) since indications of BMA were limited to rare indications. For 135 children (91.8%), treatment fulfilled the RHemaP guidelines that were mainly based on the platelet count: 121 received intraveinous immunoglobulin (IVIG) and 14 were not treated. Among those who received IVIG, 110 were good responders (91%) at the 96-h evaluation (platelet count greater than 20G/l), nine (7.4%) were poor responders, and 1 died of intracranial hemorrhage. At 6 months, chronic ITP was observed in 40 children (32.8%). Chronic ITP was associated with a higher platelet count at diagnosis and an older age (p<10(-3) and p=10(-3), respectively). CONCLUSION: The practices recorded over a 43-month period in our cohort fulfilled the RhemaP guidelines and we conclude that we managed to standardize regional practices for children with ITP. We observed conventional epidemiological characteristics in this cohort. Older children and higher platelet count at diagnosis were significantly associated with higher frequency of chronic ITP. PMID- 18676128 TI - [A cause of intracerebral calcifications that should be known: pseudohypoparathyroidism Ib]. AB - In children, pseudohypoparathyroidism (PHP) is a rare but classical cause of basal ganglia calcifications. It is caused by resistance to parathormone (PTH). Hypocalcemia, which may be symptomatic, is its main feature. We report the case of a 13-year-old boy, affected by type Ib PHP revealed by hypocalcemia and seizures, with basal ganglia calcifications on the CT scan. We describe the characteristics of the 2 main types of PHP and emphasize the search for this disease when basal ganglia calcifications are discovered, even fortuitously, on a cerebral CT scan. PMID- 18676129 TI - Risk stratification for failure in patients with advanced cervical cancer after concurrent chemoradiotherapy: another way to optimise treatment results. AB - AIMS: To identify risk factors for disease-free survival (DFS) and para-aortic lymph node (PALN) metastasis in advanced cervical cancer patients after concurrent chemoradiotherapy (CCRT) using risk stratification. MATERIALS AND METHODS: In total, 148 patients with stage IB2-IVA cervical cancer without PALN metastasis treated with a full course of CCRT were included for analysis. Radiotherapy consisted of external beam irradiation followed by four courses of high-dose rate intracavitary brachytherapy using 6.0 Gy to point A. Chemotherapy consisted of weekly cisplatin at a dose of 40mg/m(2) for a planned six cycles. Cox's proportional hazards model was used for risk stratification for DFS and PALN relapse-free survival. RESULTS: Patients were divided into low- and high risk groups. The low-risk group was composed of patients with stage IB-IIB disease without enlarged pelvic nodes, whereas the high-risk group was comprised of patients with stage IB2-IIB tumours with enlarged nodes or those with stage III-IVA disease. The 4-year DFS for the low- and high-risk groups was 83 and 52%, respectively (P=0.0001, relative risk 4.51, 95% confidence interval 1.3-10.7), whereas the 4-year PALN metastasis-free survival for the low- and high-risk groups was 92 and 61%, respectively (P=0.0003, relative risk 4.93, 95% confidence interval 1.2-12.5). CONCLUSION: The risk of failure in advanced cervical cancer patients treated in the CCRT era can be predicted. For patients with high risk of PALN relapse, this study can provide patient selection criteria when considering prophylactic PALN irradiation. PMID- 18676130 TI - Review of hypofractionated small volume radiotherapy for early-stage non-small cell lung cancer. AB - A review of the technical aspects of high-dose hypofractionated radiotherapy for localised non-small cell lung cancer was carried out to allow correlation with outcome measures and with a consensus view of the technique. A Pubmed search carried out between January 2001 and April 2007 identified 15 studies for inclusion. The clinical and technical aspects of treatment were extracted and their effect on survival, progression-free survival and toxicity were assessed using the summary statistic of weighted means. A comparison was made with the RTOG 0236 consensus study protocol. The range of variables in the studies precluded correlation of outcome with tumour parameters, dose fractionation and technical aspects such as immobilisation, techniques dealing with breathing motion, beam number and arrangement and organ at risk dose constraints. Robust data to justify a consensus view were not found, which suggests that further studies are required. They should focus on developing the treatment technique of stereotactic body radiation therapy for early-stage non-small cell lung cancer and correlating it with outcome to provide a rational basis for future randomised trials, comparing the technique with conformal radiotherapy and surgery, and the introduction of the technique into routine clinical practice. PMID- 18676131 TI - Lower fasting blood glucose, glucose variability and nocturnal hypoglycaemia with glargine vs NPH basal insulin in subjects with Type 1 diabetes. AB - BACKGROUND AND AIMS: To compare switching from NPH insulin (NPH) to insulin glargine (glargine) with continuing NPH for changes in fasting blood glucose (FBG) in patients with Type 1 diabetes on basal-bolus therapy with insulin lispro as bolus insulin. Secondary objectives included self-monitoring blood glucose, mean daily blood glucose (MDBG) and mean amplitude glucose excursion (MAGE) values alongside changes in HbA(1c) and safety profiles. METHODS AND RESULTS: This was a 30-week, parallel, open-label, multicentre study. Seven-point profiles were used to calculate MDBG and MAGE. Hypoglycaemia and adverse events were recorded by participants. FBG improved significantly with both glargine (baseline endpoint change: -28.0 mg/dL; 95% CI: -37.3, -18.7 mg/dL; p<0.001) and NPH (-9.8 mg/dL; 95% CI: -19.1, -0.5 mg/dL; p=0.0374). The improvement was significantly greater with glargine than NPH (mean difference: -18.2 mg/dL; 95% CI: -31.3, -5.2 mg/dL; p=0.0064). MDBG (-10.1 mg/dL; 95% CI: -18.1, -2.1 mg/dL; p=0.0126) and MAGE (-20.0 mg/dL; 95% CI: -34.5, -5.9 mg/dL; p=0.0056) decreased significantly with glargine, but not NPH although endpoint values were no different with the two insulins. Baseline to endpoint change in HbA(1c) was similar (-0.56 vs 0.56%) with no differences at endpoint. Overall hypoglycaemia was no different, but glargine reduced nocturnal hypoglycaemia ("serious episodes" with BG < 42 mg/dl, p=0.006) whereas NPH did not (p=0.123), although endpoint values were no different. CONCLUSION: Switching from NPH to glargine is well tolerated and results into lower FBG, and lower glucose variability while reducing nocturnal hypoglycaemia. These data provide a rationale for more aggressive titration to target with glargine in Type 1 diabetes. PMID- 18676132 TI - Down regulation of CD11b and CD18 expression in children with hypercholesterolemia: a preliminary report. AB - BACKGROUND AND AIM: Cell adhesion molecules play an important role in the development of atherosclerosis mediating the attachment of monocytes to the endothelium. The aim of our study was to assess the cell surface expression of CD11b/CD18 integrin on the phagocytes of children affected by hypercholesterolemia. METHODS AND RESULTS: Twenty-six children with hypercholesterolemia (15 males, mean age 8.3, range 2-18) with a family history of early cardiovascular disease, as well as 26 children with normocholesterolemia matched for gender and age (15 males, mean age 8.3) were studied. Cell surface expression of CD11b/CD18 on peripheral blood mononuclear cells (PBMC) were analyzed by flow cytometry. The geometric mean percentages of CD11b and CD18 expression were significantly lower in the hypercholesterolemic group [52 (95% confidence intervals, 40-68) and 88 (84-93)] than in the control group [87 (83 91), P<0.0001 and 93 (89-96), P<0.05], respectively. After correction for age, gender, and pubertal status, CD11b cell surface expression on PBMC was inversely and independently correlated with total cholesterol concentrations (r=-0.395; P<0.01) and LDL (r=-0.307; P<0.05), as well as with triglycerides (r=-0.406; P<0.01). CONCLUSIONS: In children with hypercholesterolemia, cell surface expression of CD11b and CD18 on PBMC was significantly decreased. Follow-up studies are necessary to determine the clinical implications of these findings in the context of the natural course and progression of atherosclerosis in high risk children. PMID- 18676133 TI - Hyperuricaemia is associated with increased C-reactive protein concentrations in a large cohort of unselected outpatients. PMID- 18676134 TI - Dietary fructose, fruits, fruit juices and glucose tolerance status in Japanese Brazilians. AB - BACKGROUND AND AIMS: Evidence suggests that fructose and sweetened beverages may be a risk factor for obesity and type 2 diabetes, but the role of sweetened fruit juices in glucose disturbances has been minimally explored. The aim of this study was to examine the association of total fructose, fresh fruit and sweetened fruit juice intake with glucose tolerance homeostasis in Japanese-Brazilians. METHODS AND RESULTS: A total of 475 men and 579 women aged >or=30 years were evaluated in a cross-sectional population-based survey with a standardized protocol including a 2-h oral glucose tolerance test (WHO criteria). Habitual food consumption was obtained using a validated food frequency questionnaire for Japanese-Brazilians. After adjustments for potential confounding variables, the odds ratio (OR; 95%CI) for impaired glucose tolerance was 2.1 (1.0-4.5; P for trend=0.05) for the highest as compared to the lowest tertile intake of total fructose and 2.3 (1.1 5.1; P for trend=0.05) for the highest as compared to the lowest tertile intake of sweetened fruit juices. CONCLUSION: Our results showed that high intakes of dietary fructose and sweetened fruit juices, but not whole fresh fruits, were associated with impaired glucose tolerance among genetically susceptible individuals. PMID- 18676135 TI - The effect of sibutramine therapy on occurrence of depression symptoms among obese patients. PMID- 18676136 TI - Educational impact of pulmonary rehabilitation: Lung Information Needs Questionnaire. AB - INTRODUCTION: The Lung Information Needs Questionnaire (LINQ) assesses, from the patient's perspective, their need for education. This questionnaire yields a total score and scores in six domains: disease knowledge, medicine, self management, smoking, exercise and diet. The aim of this study was to assess the sensitivity of the LINQ to change before and after pulmonary rehabilitation (PR). METHOD: PR programmes across the UK recruited 158 patients (male=94; 59%). The participants completed the LINQ and other measures as used by the individual sites pre- and post-PR, including the Shuttle Walking Test, Chronic Respiratory Disease Questionnaire, the Hospital Anxiety and Depression Scale. RESULTS: Data were analysed on 115 patients who completed data collection pre- and post-PR. The LINQ total scores, and subscales scores across all sites improved significantly with large effect sizes, except for the smoking domain as information needs about smoking were well met prior to PR. There were similar patterns of information needs at baseline and after PR in all sites. DISCUSSION: This study shows that the LINQ is a practical tool for detecting areas where patients need education and is sensitive to change after PR. The quality of the education component of PR can be assessed using the LINQ, which could be considered as a routinely collected outcome measure in PR. The LINQ may also be a useful tool for general practitioners to assess their patients' educational needs. PMID- 18676137 TI - Effects of red wine polyphenolic compounds on paraoxonase-1 and lectin-like oxidized low-density lipoprotein receptor-1 in hyperhomocysteinemic mice. AB - Hyperhomocysteinemia, or abnormally high plasma homocysteine (Hcy) concentration, has often been associated with vascular thrombosis and the development of premature atherosclerosis. Many studies have shown that moderate wine consumption has potential beneficial effects related to the prevention of atherosclerosis, in part attributed to the biological properties of polyphenolic components, mainly flavonoids. The aim of the present study is to determine the effects of a red wine polyphenolic extract (PE) administration on hyperhomocysteinemia due to cystathionine beta-synthase (CBS) deficiency and on the associated biochemical markers of hepatic and endothelial dysfunctions in mice. Red wine PE was added for 4 weeks to the drinking water of heterozygous CBS-deficient mice fed a high methionine diet, a murine model of hyperhomocysteinemia. Red wine PE supplementation at low dose significantly reduced plasma Hcy levels and restored the hepatic and plasma-decreased paraoxonase-1 activity induced by chronic hyperhomocysteinemia. Moreover, aortic expression of proinflammatory cytokines and adhesion molecules and levels of soluble lectin-like oxidized low-density lipoprotein receptor-1 were reduced in hyperhomocysteinemic mice fed the red wine PE supplementation. These findings suggest that red wine PE administration in low quantities has beneficial effects on biochemical markers of endothelial dysfunction due to hyperhomocysteinemia. PMID- 18676138 TI - Quantitative evaluation of waste prevention on the level of small and medium sized enterprises (SMEs). AB - Waste prevention is a principle means of achieving the goals of waste management and a key element for developing sustainable economies. Small and medium sized enterprises (SMEs) contribute substantially to environmental degradation, often not even being aware of their environmental effects. Therefore, several initiatives have been launched in Austria aimed at supporting waste prevention measures on the level of SMEs. To promote the most efficient projects, they have to be evaluated with respect to their contribution to the goals of waste management. It is the aim of this paper to develop a methodology for evaluating waste prevention measures in SMEs based on their goal orientation. At first, conceptual problems of defining and delineating waste prevention activities are briefly discussed. Then an approach to evaluate waste prevention activities with respect to their environmental performance is presented and benchmarks which allow for an efficient use of the available funds are developed. Finally the evaluation method is applied to a number of former projects and the calculated results are analysed with respect to shortcomings and limitations of the model. It is found that the developed methodology can provide a tool for a more objective and comprehensible evaluation of waste prevention measures. PMID- 18676139 TI - "Macromolecules to PDMS transfer" as a general route for PDMS biochips. AB - "Macromolecules to PDMS transfer" technique relying on the direct entrapment of macromolecules spots during PDMS polymerisation is proposed as an alternative for the easy and simple PDMS surface modification. In the present work, the development of three different applications based on this procedure is presented as proof of the method potentialities. First, C-reactive protein (CRP) sandwich immunoassay using immobilised monoclonal anti-CRP antibodies was developed for sepsis diagnosis. The preserved integrity of the immobilised monoclonal immunoglobulin permitted the sensitive detection of free CRP in human sera (LOD=12.5 microg/L, detection ranging over two decades). Then, rheumatoid arthritis diagnosis through the rheumatoid factor (RF) detection based on rabbit immunoglobulins immobilisation allowed the detection of specific antibodies in human sera samples down to low RF levels (detection range 5.3-485 IU/mL). Finally, the "Macromolecules to PDMS transfer" procedure was used to easily and rapidly produce fibronectin-based cell culture arrays. The successful attachment of HeLa and BALB/3T3 cells was demonstrated with optical microscopy and specific staining of actin and vinculin. PMID- 18676140 TI - Symptom and quality of life results of an international randomised phase III study of adjuvant vaccination with Bec2/BCG in responding patients with limited disease small-cell lung cancer. AB - AIMS: This study reports the symptom and HRQOL results in which standard treatment was compared to standard therapy plus Bec2, an anti-idiotypic antibody that mimics GD3, a ganglioside antigen. METHODS: Five hundred and fifteen LD SCLC patients were randomised to receive five vaccinations of Bec2 (2.5mg)/BCG vaccine arm (VA) or an observational arm (OA) administered over a 10-week period. Survival was the primary end-point; HRQOL was a secondary end-point, assessed using the EORTC QLQ-C30/LC 13. RESULTS: There was no improvement in survival or progression-free survival in the vaccination arm. At baseline patients in both arms demonstrated significantly impaired scores on the global QOL scale, when compared to a normative population. However, HRQOL and symptom scores between the two treatment arms were not statistically different at any time point. CONCLUSION: We found no benefits to patient HRQOL by additional vaccination with Bec2/BCG to LD SCLC for patients who have been undergoing standard therapy. PMID- 18676141 TI - A statistical approach to optimization of fermentative production of poly(gamma glutamic acid) from Bacillus licheniformis NCIM 2324. AB - This paper reports on the optimization of poly(gamma-glutamic acid) (PGA) production by Bacillus licheniformis NCIM 2324 using a statistical approach. One factor-at-a-time method was used to investigate the effect of carbon sources, nitrogen sources and pH on PGA production. Plackett-Burman design was adopted to select the most important nutrients influencing the yield of PGA. After identifying effective nutrients, response surface methodology was used to develop a mathematical model to identify the optimum concentrations of the key nutrients for higher PGA production, and confirm its validity experimentally. PGA production increased significantly from 5.27 to 26.12 g/l when the strain was cultivated in the optimal medium developed by using statistical approach, as compared to basal medium. PMID- 18676142 TI - The chemical and hydroxyl radical scavenging activity changes of ginsenoside-Rb1 by heat processing. AB - The chemical and hydroxyl radical (*OH) scavenging activity changes of ginsenoside Rb(1) (Rb(1)) by heat processing were investigated in this study. Rb(1) was changed into 20(S)-Rg(3), 20(R)-Rg(3), Rk(1), and Rg(5) by heat processing through glucosyl elimination and epimerization of carbon-20 by SN1 reaction. The glucosyl moiety, separated from Rb(1), made Maillard reaction product (MRPs) with glycine. The generations of 20(S)-Rg(3) and MRPs were related to the increased OH scavenging activity of Rb(1) by heat processing. PMID- 18676143 TI - Potent and selective pyrazole-based inhibitors of B-Raf kinase. AB - Herein we describe a novel pyrazole-based class of ATP competitive B-Raf inhibitors. These inhibitors exhibit both excellent cellular potency and striking B-Raf selectivity. A subset of these inhibitors has demonstrated the ability to inhibit downstream ERK phosphorylation in LOX tumors from mouse xenograft studies. PMID- 18676144 TI - Discovery of imidazole vinyl pyrimidines as a novel class of kinase inhibitors which inhibit Tie-2 and are orally bioavailable. AB - Tie-2 is a receptor tyrosine kinase which is involved in angiogenesis and thereby growth of human tumours. The discovery and SAR of a novel class of imidazole vinyl-pyrimidine kinase inhibitors, which inhibit Tie-2 in vitro is reported. Their synthesis was carried out by condensation of imidazole aldehydes with methyl pyrimidines. These compounds are lead-like, with low molecular weight, good physical properties and oral bioavailability. PMID- 18676145 TI - Gait and kinematics of the trunk and lower extremities in stair ascent using quadricane in healthy subjects. AB - Different cane placement methods require different gait patterns and ranges of motion either at the trunk or lower extremities. The aim of this study was to examine the effect of cane placement on body biomechanics in stair ascent (SA) in 16 healthy adults (9 women, 7 men) aged 27.2+/-3.2 years old. The height and weight of the women and men were 160.8+/-5.4 cm and 54.1+/-8.1 kg, and 170.8+/ 3.9 cm and 69.6+/-5.6 kg, respectively. Three-dimensional motion data were collected in non-reciprocally SA associated with following methods: (1) dominant foot stepped up first, then the opposite foot without a cane (NC); (2) forward placement of a quadricane followed by the ipsilateral foot, then contralateral foot (FCI); (3) forward cane placement followed by the contralateral foot, then ipsilateral foot (FCC); (4) ipsilateral foot stepping up first, followed by the contralateral foot and the cane (LCI); (5) contralateral foot stepping up, followed by the ipsilateral foot and cane (LCC). LCI and LCC were considered as lateral cane placement. Temporal gait parameters, kinematics of the trunk and lower extremities were calculated. The results indicate that the cane placement had significant effect on the kinematics of the trunk and lower extremities. The main differences between forward and lateral cane placement were flexion and side flexion of the trunk, and flexion of the leading and opposite hip and knee. Hip rotation and abduction, and ankle plantar and dorsiflexion were different between contralateral and ipsilateral cane placements. PMID- 18676146 TI - Determination of gait patterns in children with spastic diplegic cerebral palsy using principal components. AB - This study developed an objective graphical classification method of spastic diplegic cerebral palsy (CP) gait patterns based on principal component analysis (PCA). Gait analyses of 20 healthy and 20 spastic diplegic CP children were examined to define gait characteristics. PCA was used to reduce the dimensionality of 27 parameters (26 selected kinematics variables and age of the children) for the 40 subjects in order to identify the dominant variability in the data. Fuzzy C-mean cluster analysis was performed plotting the first three principal components, which accounted for 61% of the total variability. Results indicated that only the healthy children formed a distinct cluster; however it was possible to recognise gait patterns in overlapping clusters in children with spastic diplegia. This study demonstrates that it is possible to quantitatively classify gait types in CP using PCA. Graphical classification of gait types could assist in clinical evaluation of the children and serve as a validation of clinical reports as well as aid treatment planning. PMID- 18676147 TI - Foot contact event detection using kinematic data in cerebral palsy children and normal adults gait. AB - Initial contact (IC) and toe off (TO) times are essential measurements in the analysis of temporal gait parameters, especially in cerebral palsy (CP) gait analysis. A new gait event detection algorithm, called the high pass algorithm (HPA) has been developed and is discussed in this paper. Kinematics of markers on the heel and metatarsal are used. Their forward components are high pass filtered, to amplify the contact discontinuities, thus the local extrema of the processed signal correspond to IC and TO. The accuracy and precision of HPA are compared with the gold standard of foot contact event detection, that is, force plate measurements. Furthermore HPA is compared with two other kinematics methods. This study has been conducted on 20 CP children and on eight normal adults. For normal subjects all the methods performed equally well. True errors in HPA (mean+/-standard deviation) were found to be 1+/-23 ms for IC and 2+/-25 ms for TO in CP children. These results were significantly (p<0.05) more accurate and precise than those obtained using the other algorithms. Moreover, in the case of pathological gaits, the other methods are not suitable for IC detection when IC is flatfoot or forefoot. In conclusion, the HPA is a simple and robust algorithm, which performs equally well for adults and actually performs better when applied to the gait of CP children. It is therefore recommended as the method of choice. PMID- 18676148 TI - Synthesis and structure-activity relationship studies of theophylline analogs on population responses in the rat hippocampus in vitro. AB - We synthesized several theophylline analogs and tested the hypothesis that these compounds may be nootropic or cognitive enhancers by examining their effects on evoked population spikes recorded extracellularly in the CA1 region of the rat hippocampus. Whereas the length of the carbon chain on N7 had no effect, different size of the terminal lactam ring strongly influenced neuroactivity. Our results suggest that hexahydroazepin-2-one analogs have potential for further development as cognitive enhancers. PMID- 18676149 TI - Nakijiquinones G-I, new sesquiterpenoid quinones from marine sponge. AB - Three new sesquiterpenoid quinones, nakijiquinones G-I (1-3), containing a different amino group derived from amino acids have been isolated from Okinawan marine sponges of the family Spongiidae, and the structures and relative stereochemistry of 1-3 were elucidated on the basis of the spectral data. Nakijiquinones G-I (1-3) showed modest cytotoxicity and inhibitory activity against HER2 kinase, while nakijiquinone H (2) exhibited antimicrobial activity. PMID- 18676150 TI - Synthesis and antimicrobial activity of dermaseptin S1 analogues. AB - Dermaseptins are peptides found in the skin secretions of Phyllomedusinae frogs. These peptides exert lytic action on some microorganisms without substantial haemolysis. In an attempt to understand the antimicrobial activity of these peptides we designed several dermaseptin S1 (ALWKTMLKKLGTMALHAGKAALGAAADTISQGTQ) (DS1) analogues. All peptides were tested on the growth of prokaryotic (Gram positive and Gram-negative bacteria) and eukaryotic (the yeast Candida albicans and the protozoon Leishmania major) organisms. Our data showed a dose-dependent killing effect by most DS1 derivatives. Maximal antibacterial activity was displayed by a 16-mer peptide that was more active than native DS1. PMID- 18676151 TI - Syntheses and biological evaluation of topoisomerase I-targeting agents related to 11-[2-(N,N-dimethylamino)ethyl]-2,3-dimethoxy-8,9-methylenedioxy-11H isoquino[4,3-c]cinnolin-12-one (ARC-31). AB - Several 11-ethyl-2,3-dimethoxy-8,9-methylenedioxy-11H-isoquino[4,3-c]cinnolin-12 ones with varied functionality on the ethyl substituent have exhibited potent topoisomerase I (TOP1) targeting activity and antitumor activity. The influence of various polar substituents at the 2-position of the 11-ethyl substituent, including N-methylamine, N-isopropylamine, hydroxyl, and hydroxylamino groups, on TOP1-targeting activity and cytotoxicity was assessed. The N-methylamine and N isopropylamine derivatives were also evaluated as antitumor agents in athymic nude mice with MDA-MB-435 human tumor xenografts. Both compounds were active as antitumor agents upon either parenteral or oral administration. PMID- 18676152 TI - Heme activates artemisinin more efficiently than hemin, inorganic iron, or hemoglobin. AB - Artemisinin derivatives appear to mediate their anti-malarial through an initial redox-mediated reaction. Heme, inorganic iron, and hemoglobin have all been implicated as the key molecules that activate artemisinins. The reactions of artemisinin with different redox forms of heme, ferrous iron, and deoxygenated and oxygenated hemoglobin were analyzed under similar in vitro conditions. Heme reacted with artemisinin much more efficiently than the other iron-containing molecules, supporting the role of redox active heme as the primary activator of artemisinin. PMID- 18676153 TI - Monoquaternary pyridinium salts with modified side chain-synthesis and evaluation on model of tabun- and paraoxon-inhibited acetylcholinesterase. AB - Acetylcholinesterase reactivators are crucial antidotes for the treatment of organophosphate intoxication. Eighteen monoquaternary reactivators of acetylcholinesterase with modified side chain were developed in an effort to extend the properties of pralidoxime. The known reactivators (pralidoxime, HI-6, obidoxime, trimedoxime, methoxime) and the prepared compounds were tested in vitro on a model of tabun- and paraoxon-inhibited AChE. Monoquaternary reactivators were not able to exceed the best known compounds for tabun poisoning, but some of them did show reactivation better or comparable with pralidoxime for paraoxon poisoning. However, extensive differences were found by a SAR study for various side chains on the non-oxime part of the reactivator molecule. PMID- 18676154 TI - Cell survival and chromosomal aberrations in CHO-K1 cells irradiated by carbon ions. AB - Chinese hamster ovary CHO-K1 cells were exposed to high LET (12)C-beam (LET: 830 keV/microm) in the dose range of 0-6 Gy and to (60)Co irradiation and the RBE value was obtained. Effects of (12)C-beam exposure on cell survival and chromosomal aberrations were calculated. The chromosomal aberration data were fitted with linear equation. The distribution of aberration in cells was examined with a standard u-test and used to evaluate the data according to Poisson probabilities. The variance to the mean ratio sigma(2)/Y and the dispersion index (u) were determined. Overdispersion was significant (p<0.05) when the value of u exceeded 1.96. PMID- 18676155 TI - WCS120 protein family and frost tolerance during cold acclimation, deacclimation and reacclimation of winter wheat. AB - We studied how long-term cold acclimation of winter wheat (variety Mironovskaya 808), interrupted by deacclimation and then followed by reacclimation, affected the levels of cold-induced WCS120 proteins, dry-weight content, and frost tolerance in leaves. Two experiments were performed: (1) plants undergoing long term cold acclimation (up to 112days) were quickly deacclimated (for 5days), and then reacclimated again to cold; (2) plants vernalized for varying periods of time in an early stage of their development were, after a longer deacclimation of about 14days, exposed for the same time period to cold. Five members of the WCS120 protein family were detected and quantified by image analysis in protein gel blots (in the first experiment); as well as in two-dimensional electrophoresis gels (in the second experiment). In both experiments, partially vernalized plants, after reacclimation, re-established their frost tolerance to levels similar to plants having had the same duration of cold treatment, but without deacclimation. On the other hand, these partially and fully vernalized plants reaccumulated WCS120 proteins to lower levels than plants that were not deacclimated. Further, using a mathematical model (the peak four-parameter Weibull equation), the same type of response curve was observed during plant cold treatment not only for the level of frost tolerance, but also for dry-weight content and accumulation of WCS120 proteins, with the maximum values reached at about the same time as vernalization saturation. PMID- 18676156 TI - The biological properties of cetuximab. AB - Cetuximab is a recombinant chimeric human murine immunoglobulin G1 antibody that binds to the extra-cellular domain of epidermal growth factor receptor with a higher affinity than either endogenous ligand. This binding inhibits receptor phosphorylation and activation and it leads to receptor internalization and degradation. Several studies have shown that cetuximab is able to inhibit growth of epidermal growth factor receptor (EGFR)-expressing tumour cells in vitro. Moreover, treatment with cetuximab results in a marked inhibition of tumour growth in nude mice bearing xenografts of human cancer cell lines. These results are linked to cetuximab biological effects as inhibition of cell cycle, tumour progression, neo-angiogenesis, invasion and metastatization, as well as increase and activation of pro-apoptotic molecules. Additionally, cetuximab potentiates, in combination, the effects of chemotherapy and radiation therapy in eradicating well-established tumours in nude mice and it may even reverse the resistance to some cytotoxic agents in these xenografts. Moreover, numerous clinical trials demonstrated cetuximab efficacy in different tumour types. It has been approved by Food and Drugs Administration in the treatment of metastatic colorectal cancer as single agent or in combination with chemotherapy, in locally and regionally advanced head and neck squamous cell carcinoma in combination with radiation, and as monotherapy for recurrent and metastatic head and neck squamous cell carcinoma after failing platinum-based chemotherapy. This paper will overview all the experimental and pre-clinical data on the biological properties of cetuximab. PMID- 18676157 TI - Role of lipid peroxidation and oxidative stress in the association between thyroid diseases and breast cancer. AB - Evidence is accumulating for a role of the thyroid in the natural history of breast cancer, although no plausible mechanism has been advanced to explain this association. We believe that the thyroid disease-breast cancer relationship provides a unique opportunity to find out the causes of breast cancer. Both diseases are female predominant, with specifically identified biological pathways and genetic and environmental determinants, and seeing them in concert provides an opportunity to identify the most relevant mechanistic pathways. In this communication, we advance a plausible mechanism to explain the thyroid disease breast cancer relationship. We specifically propose that the reduction in risk associated with hyperthyroidism or increased levels of thyroid hormones, or iodine, may derive from the pro-oxidant properties of these compounds, i.e., from its ability to generate oxidative stress-induced apoptosis. Conversely, the increased risk from hypothyroidism may derive from its ability to inhibit this stress-mediated apoptotic process. PMID- 18676158 TI - Calibration of replacement international standards of diphtheria and tetanus toxoids for use in flocculation test. AB - The 1st International Reference Reagents (IRR) of Diphtheria and Tetanus Toxoids for Flocculation Test (DIFT and TEFT) were established by the WHO in 1988. These reagents are essential for the standardization of assays used to calculate Lf units of toxoids. Candidate replacement materials were provided by several European vaccine manufacturers and were formulated and freeze-dried at NIBSC. This paper provides a summary of the results of an international collaborative study including 18 laboratories from 16 countries, which examined the candidate replacement materials in a variety of methods. Materials 02/176 and 04/150 were proposed and adopted by the Expert Committee on Biological Standardization of WHO in October 2007 as 2nd WHO International Standards of Diphtheria and Tetanus Toxoid for use in Flocculation Test. The replacement standards were assigned the value of 1100 and 690Lf/ampoule, respectively, based on results of flocculation tests carried out using provided reagents. Material coded 02/176 fully complied with the WHO specifications for stability, residual moisture content, precision of fill and sterility. Stability of material coded 04/150 was slightly lower than expected but predictions were based only on 2-year data and were to be further monitored, post-adoption. PMID- 18676159 TI - Evaluation of antibody levels during simultaneous aflatoxicosis and vaccination against infectious laryngotracheitis in pullets. AB - Chickens fed 200 ppb aflatoxin from 10 days of age were evaluated for their immune response to a modified live infectious laryngotracheitis vaccine. Vaccination was administered at age 4 and 12 weeks. Antibody titers to the vaccine were reduced in chickens given dietary aflatoxin. After 7 weeks, aflatoxin feeding was continued for one month in a treated group and was withdrawn in another. Serology indicated significant differences between the two treated groups relative to whether aflatoxin was fed or not. Significant reduction in body weights, antibody titers and elevated SGOT and SGPT levels were found in chickens treated with aflatoxin. The impact of aflatoxin on reduced body weight, decreased SGOT and SGPT levels and lower antibody titers was shown to be significant in the treated group fed on a ration of aflatoxin until throughout the experiment. PMID- 18676160 TI - Phenobarbital withdrawal seizures may occur over several weeks before remitting: human data and hypothetical mechanism. AB - This case is the first report of a patient who had phenobarbital (PB) withdrawal seizures after having been seizure-free for 3 years following temporal lobe surgery. The patient had been taking PB for 14 years when a gradual taper of PB was started. When PB was at 60 mg/d, a titration of lamotrigine (LTG) was started. However, typical complex seizures occurred when the patient was on PB 60 mg/d, along with LTG 25mg/d. PB was increased back to 90 mg/d and levetiracetam (LEV) was titrated. Seizures appeared when the patient was on PB 30 mg/d and LEV 750 mg BID and continued for 3 weeks after PB was stopped and the patient was on LEV 1,000 mg BID. For the following 6 months, her aura frequency remained elevated in comparison to her baseline aura of two auras per month for the previous year before the start of the PB taper. She was followed for 24 months after her last PB withdrawal seizure. During the last 8 months, her aura frequency returned to her baseline. As suggested by animal studies, the PB withdrawal seizures and increase in aura frequency in this patient may be explained by changes in her levels of GABA(A) receptor subunits. PMID- 18676161 TI - Correlation between arthroscopic and histopathological grading systems of articular cartilage lesions in knee osteoarthritis. AB - PURPOSE: Arthroscopic and particularly histopathological assessments have been used to evaluate alterations of knee cartilage in osteoarthritis (OA). The aim of this study was to examine the correlation between an arthroscopic method to grade the severity of chondropathies and the histological/histochemical grading system (HHGS) applied to the corresponding articular cartilage areas in knee OA. METHODS: The articular cartilage surface was examined by chondroscopy using the Beguin and Locker severity criteria, analysing the lesions in 72 chondroscopic areas. Afterwards, samples were obtained by dividing the cartilage surface of the medial tibiofemoral compartment of three OA knee joints into equal squares and they were evaluated histologically using the HHGS. The correlation between both grading methods was assessed using the weighted Kappa coefficient (K(w)). RESULTS: The results obtained with both scores showed good agreement (K(w): mean+/-standard deviation, 0.619+/-0.071). While the average HHGS scores of the chondral samples showed a better agreement with arthroscopic grades 0, I and II, the arthroscopic evaluation has a tendency to overestimate chondral lesions for histological grades III and IV. The intra- and inter-observer reliability of the HHGS evaluation of chondral lesions was excellent (Intraclass Correlation Coefficient: 0.909 and 0.941, respectively). CONCLUSION: In this study, we found a good quantitative correlation between established arthroscopic severity and histopathological scoring systems, particularly in less advanced lesions. Our results suggest that the arthroscopic method is a valuable tool in clinical research to score chondropathies in the medial femorotibial compartment of the OA knee, although some limitations should not be overlooked. PMID- 18676162 TI - The anemia of ageing is not associated with increased plasma hepcidin levels. AB - It has been proposed that the anemia of ageing may be caused, at least in part, by elevated hepcidin levels, representing a response to increased IL-6 levels. Using a recently developed immunoassay, we have measured the plasma hepcidin levels of eight patients with the anemia of ageing and sex- and age-matched controls, and found that the levels of hepcidin were not increased in patients with the anemia of ageing. In contrast, patients with the anemia of inflammation have higher hepcidin levels than sex- and age-matched controls. In the overall group there was a strong correlation between serum ferritin levels and hepcidin levels, as has been found previously. PMID- 18676163 TI - Congenital hypofibrinogenemia: characterization of two missense mutations affecting fibrinogen assembly and secretion. AB - Congenital hypofibrinogenemia is a rare bleeding disorder characterized by abnormally low levels of fibrinogen in plasma, generally due to heterozygous mutations in one of the three fibrinogen genes (FGA, FGB, and FGG, coding for Aalpha, Bbeta, and gamma chain, respectively). Hypofibrinogenemic patients are usually asymptomatic, whereas individuals bearing similar mutations in the homozygous or compound heterozygous state develop a severe bleeding disorder: afibrinogenemia. The mutational spectrum of these quantitative fibrinogen disorders includes large deletions, point mutations causing premature termination codons, and missense mutations affecting fibrinogen assembly or secretion, distributed throughout the 50-kb fibrinogen gene cluster. In this study, we report the mutational screening of two unrelated hypofibrinogenemic patients leading to the identification of two missense mutations, one hitherto unknown (alphaCys45Phe), and one previously described (gammaAsn345Ser). The involvement of alphaCys45Phe and gammaAsn345Ser in the pathogenesis of hypofibrinogenemia was investigated by in-vitro expression experiments. Both mutations were demonstrated to cause a severe impairment of intracellular fibrinogen processing, either by affecting half-molecule dimerization (alphaCys45Phe) or by hampering hexamer secretion (gammaAsn345Ser). PMID- 18676164 TI - Insight into the structural requirements of proton pump inhibitors based on CoMFA and CoMSIA studies. AB - In the present study, a series of 179 quinoline and quinazoline heterocyclic analogues exhibiting inhibitory activity against Gastric (H+/K+)-ATPase were investigated using the comparative molecular field analysis (CoMFA) and comparative molecular similarity indices (CoMSIA) methods. Both the models exhibited good correlation between the calculated 3D-QSAR fields and the observed biological activity for the respective training set compounds. The most optimal CoMFA and CoMSIA models yielded significant leave-one-out cross-validation coefficient, q(2) of 0.777, 0.744 and conventional cross-validation coefficient, r(2) of 0.927, 0.914 respectively. The predictive ability of generated models was tested on a set of 52 compounds having broad range of activity. CoMFA and CoMSIA yielded predicted activities for test set compounds with r(pred)(2) of 0.893 and 0.917 respectively. These validation tests not only revealed the robustness of the models but also demonstrated that for our models r(pred)(2) based on the mean activity of test set compounds can accurately estimate external predictivity. The factors affecting activity were analyzed carefully according to standard coefficient contour maps of steric, electrostatic, hydrophobic, acceptor and donor fields derived from the CoMFA and CoMSIA. These contour plots identified several key features which explain the wide range of activities. The results obtained from models offer important structural insight into designing novel peptic-ulcer inhibitors prior to their synthesis. PMID- 18676165 TI - Short-chain acyl-CoA dehydrogenase (SCAD) deficiency: an examination of the medical and neurodevelopmental characteristics of 14 cases identified through newborn screening or clinical symptoms. AB - The medical and neurodevelopmental characteristics of 14 children with short chain acyl-CoA dehydrogenase deficiency (SCADD) are described. Eight were detected as neonates by newborn screening. Three children diagnosed on the basis of clinical symptoms had normal newborn screening results while three were born in states that did not screen for SCADD. Treatment included frequent feedings and a low fat diet. All children identified by newborn screening demonstrated medical and neuropsychological development within the normative range on follow-up, although one child had a relative weakness in the motor area and another child exhibited mild speech delay. Of the three clinically identified children with newborn screening results below the cut-off value, two were healthy and performed within the normal range on cognitive and motor tests at follow-up. Four clinically identified children with SCADD experienced persistent symptoms and/or developmental delay. However, in each of these cases, there were supplementary or alternative explanations for medical and neuropsychological deficits. Results indicated no genotype-phenotype correlations. These findings suggest that SCADD might be benign and the clinical symptoms ascribed to SCADD reflective of ascertainment bias or that early identification and treatment prevented complications that may have occurred due to interaction between genetic susceptibility and other genetic factors or environmental stressors. PMID- 18676166 TI - Liver hepatoblastoma and multiple OXPHOS deficiency in the follow-up of a patient with methylmalonic aciduria. AB - A boy who was diagnosed with methylmalonic aciduria (MMA) at the age of 10 days developed persistent hepatomegaly and raised transaminases from the age of 4 years. He was subsequently diagnosed with Leigh syndrome and required a kidney transplantation for end-stage renal failure. A massive hepatoblastoma led to his death by the age of 11 years. Methylmalonyl-CoA mutase activity was undetectable on both cultured skin fibroblasts and kidney biopsy and multiple respiratory chain deficiency was demonstrated in the kidney. Mitochondrial dysfunction and/or post-transplant immunosuppressive therapy should be considered as a possible cause of liver cancer in this patient. PMID- 18676167 TI - The molecular basis of pyruvate carboxylase deficiency: mosaicism correlates with prolonged survival. AB - Pyruvate carboxylase (PC) deficiency (OMIM, 266150) is a rare autosomal recessive disease. The revised PC gene structure described in this report consists of 20 coding exons and four non-coding exons at the 5'-untranslated region (5'-UTR). The gene codes for three transcripts due to alternative splicing: variant 1 (NM_000920.3), variant 2 (NM_022172.2) and variant 3 (BC011617.2). PC deficiency is manifested by three clinical phenotypes-an infantile form (Type A), a neonatal form (Type B), and a benign form (Type C). We report the molecular basis for eight cases (one Type A, five Type B and two Type C) of PC deficiency. Eight novel complex mutations were identified representing different combinations of missense mutations, deletions, a splice site substitution and a nonsense mutation. The classical phenotypes (A, B and C) correlated poorly with clinical outcomes. Mosaicism was found in five cases (one Type A, three Type B and one Type C) and four of these cases had prolonged survival. Death in the fifth case resulted from unrelated medical complications. The discrepancy between the current findings and the existing classification system should be addressed to accommodate these new observations. PMID- 18676168 TI - Infection of the subcutis of the nose in a cat caused by Mucor species: successful treatment using posaconazole. AB - A 14-year-old neutered female domestic shorthair cat was presented for investigation of a non-painful subcutaneous swelling of the nasal dorsum at the site of a scratch injury. Cytological evaluation demonstrated a granulomatous reaction and many variably shaped organisms consistent with yeasts/fungi. Subsequent biopsy and culture yielded a pure growth of a Mucor species. The cat was treated with the second-generation triazole antifungal agent posaconazole for 5 months. Complete resolution was seen with no recurrence 12 months after discontinuing treatment. PMID- 18676169 TI - [What is the future of the children after therapeutic irradiation of their mother during their pregnancy?]. PMID- 18676170 TI - [Luxatio erecta of the shoulder. Report of eight cases]. AB - The luxatio erecta is a rare form of shoulder dislocation frequently described in young patients. The purpose of our study is to review the mechanism of this affection as well as its clinical, radiographic and therapeutic features. We also report eight cases of luxatio erecta observed between 1995 and 2006. We report six males and two females with an average age of 40 years. The left shoulder was dislocated in seven cases and a direct mechanism was reported by two patients. All of our cases presented with the typical attitude described for this dislocation; an upper limb in fixed abduction with the hand raised and inability to bring the elbow back to the body. No neurovascular injury was reported in this study. Radiographic examination showed inferior dislocation of the humeral head and the axis of the shaft above the horizontal line in all of our cases. The treatment consisted of closed reduction under general anaesthesia followed by a Dujarier's bandage for three weeks. Luxatio erecta is the commonest type of inferior dislocation. Its mechanism is a fall on the upper limb in great abduction or antepulsion. The clinical diagnosis is easy, confirmed by an anteroposterior radiograph. The functional long-term prognosis is excellent. PMID- 18676171 TI - Use of the Higuchi's fractal dimension for the analysis of MEG recordings from Alzheimer's disease patients. AB - Alzheimer's disease (AD) is an irreversible brain disorder of unknown aetiology that gradually destroys brain cells and represents the most prevalent form of dementia in western countries. The main aim of this study was to analyse the magnetoencephalogram (MEG) background activity from 20 AD patients and 21 elderly control subjects using Higuchi's fractal dimension (HFD). This non-linear measure can be used to estimate the dimensional complexity of biomedical time series. Before the analysis with HFD, the stationarity and the non-linear structure of the signals were proved. Our results showed that MEG signals from AD patients had lower HFD values than control subjects' recordings. We found significant differences between both groups at 71 of the 148 MEG channels (p<0.01; Student's t-test with Bonferroni's correction). Additionally, five brain regions (anterior, central, left lateral, posterior and right lateral) were analysed by means of receiver operating characteristic curves, using a leave-one-out cross-validation procedure. The highest accuracy (87.8%) was achieved when the mean HFD over all channels was analysed. To sum up, our results suggest that spontaneous MEG rhythms are less complex in AD patients than in healthy control subjects, hence indicating an abnormal type of dynamics in AD. PMID- 18676172 TI - Body fluid volumes measurements by impedance: A review of bioimpedance spectroscopy (BIS) and bioimpedance analysis (BIA) methods. AB - This paper reviews various bioimpedance methods permitting to measure non invasively, extracellular, intracellular and total body water (TBW) and compares BIA methods based on empirical equations of the wrist-ankle resistance or impedance at 50 kHz, height and weight with BIS methods which rely on an electrical model of tissues and resistances measured at zero and infinite frequencies. In order to compare these methods, impedance measurements were made with a multifrequency Xitron 4200 impedance meter on 57 healthy subjects which had undergone simultaneously a Dual X-ray absorptiometry examination (DXA), in order to estimate their TBW from their fat-free-mass. Extracellular (ECW) and TBW volumes were calculated for these subjects using the original BIS method and modifications of Matthie[Matthie JR. Second generation mixture theory equation for estimating intracellular water using bioimpedance spectroscopy. J Appl Physiol 2005;99:780-1], Jaffrin et al. [Jaffrin MY, Fenech M, Moreno MV, Kieffer R. Total body water measurement by a modification of the bioimpedance spectroscopy method. Med Bio Eng Comput 2006;44:873-82], Moissl et al. [Moissl UM, Wabel P, Chamney PW, Bosaeus I, Levin NW, et al. Body fluid volume determination via body composition spectroscopy in health and disease. Physiol Meas 2006;27:921-33] and their TBW resistivities were compared and discussed. ECW volumes were calculated by BIA methods of Sergi et al. [Sergi G, Bussolotto M, Perini P, Calliari I, et al. Accuracy of bioelectrical bioimpedance analysis for the assessment of extracellular space in healthy subjects and in fluid retention states. Ann Nutr Metab 1994;38(3):158-65] and Hannan et al. [Hannan WJ, Cowen SJ, Fearon KC, Plester CE, Falconer JS, Richardson RA. Evaluation of multi-frequency bio-impedance analysis for the assessment of extracellular and total body water in surgical patients. Clin Sci 1994;86:479-85] and TBW volumes by BIA methods of Kushner and Schoeller [Kushner RF, Schoeller DA. Estimation of total body water by bioelectrical impedance analysis. Am J Clin Nutr 1986;44(3):417-24], Lukaski et al. [Lukaski HC, Bolonchuk WW. Estimation of body fluid volumes using tetrapolar bioelectrical impedance measurements. Aviat Space Environ Med 1988;59:1163-9], Hannan et al. [Hannan WJ, Cowen SJ, Fearon KC, Plester CE, Falconer JS, Richardson RA. Evaluation of multi-frequency bio-impedance analysis for the assessment of extracellular and total body water in surgical patients. Clinical Science 1994;86:479-85], Deurenberg et al. [Deurenberg P, van der Koy K, Leenen R, Westrate JA, Seidell JC. Sex and age specific prediction formulas for estimating body composition from bioelectric impedance: a cross validation study. Int J Obesity 1991;15:17-25] These volumes were compared against those given by BIS method and, in the case of TBW, with those by DXA. For ECW, a good agreement was found between various BIS methods and that of Sergi while Hannan's values were higher. Both Matthie's and Moissl's methods gave mean TBW resistivities and volumes lower than those of Jaffrin's and DXA methods. Kushner et al. method gave values of TBW not significantly different from those of Jaffrin et al. and DXA, as Hannan's method in men, but Lukaski and Deurenberg methods led to an underestimation. PMID- 18676173 TI - Primary epithelial submandibular salivary gland tumours--review of management in a district general hospital setting. AB - Primary epithelial submandibular gland (SMG) tumours are uncommon, accounting for 8-12% of all salivary gland neoplasms, and most studies come from large specialised centres. There is little published about the relative frequency and outcome of SMG tumours treated in general hospitals. A retrospective study from two district general hospitals over a 10 year period (1996-2005). Seventeen benign (68%) and eight malignant (32%) tumours were included. A painless mass was the most common presentation. Pain and paraesthesia were not pathognomonic of malignancy. Pleomorphic adenoma accounted for all 17 benign tumours and adenoid cystic carcinoma was the commonest malignant tumour. Fine needle aspiration cytology (FNAC) accurately identified 78% of the benign tumours but none of the malignant tumours. Pre-operative imaging was also unable to distinguish malignant from benign tumours. Incomplete excision was reported in 20% of cases and was more common for malignant tumours. It may be difficult to distinguish benign from malignant SMG tumours on clinical examination and pre-operative investigations. Any suspected submandibular tumour should be considered for early treatment even when FNAC is suggestive of a benign tumour. PMID- 18676174 TI - Ab initio determination of molecular geometries and vibrational frequencies of CX3 COOH (X=H, F, Cl, Br). AB - Using Gaussian 03 Revision C.02 version of the quantum chemical program ab initio and DFT computations have been carried out at the rhf/6-31+g*, b3lyp/6-31+g*, b3lyp/6-31++g** and b3lyp/6-311++g** levels to compute optimized geometries, harmonic vibrational frequencies along with intensities in IR and Raman spectra and atomic charges for the acetic (ethanoic) acid and its 1,1,1-tri-halo (fluoro, chloro and bromo) derivatives. The optimized molecular structures for all the four molecules are found to possess Cs point group symmetry. The symmetric stretching mode is found to have lowest magnitude of the three CX3 stretching modes for all the four molecules, whereas the symmetric deformation mode is found to have the lowest magnitude for EA and TFEA and the highest magnitude for TCEA and TBEA. The parallel rocking mode of the CX3 group is found to have lower magnitude than the perpendicular rocking mode for EA and TFEA where reverse is found for TCEA and TBEA. The modes of the COOH group are substituent sensitive except the OH stretching mode. Moreover, the maximum effect is found for the TFEA molecule. The CF3 group is found to have the characteristic frequencies as 235 505, 787, 1150-1190 and 1400 cm(-1) which are due to the modes delta s(CF3), nu s(CF3), nu as(CF3) and nu(C-CF3), respectively. PMID- 18676175 TI - Synthesis, spectral investigation and thermal aspects of coordination polymeric chain assemblies of some transition metal ions with bis-pyrazolones. AB - A few coordination chain polymeric assemblies of the type [M(SB)(H(2)O)(2)](n).xH(2)O or [VO(SB)(H(2)O)](n).H(2)O [where M=Mn(II), Cu(II) and Zn(II), x=1; Co(II) and Ni(II) x=2, H(2)SB=(4Z,4'Z)-4,4'-(2,2'-(4,4' methylenebis(4,1-phenylene)bis(azanediyl))bis(1-hydroxy ethan-2-yl-1 ylidene))bis(3-methyl-1-phenyl-1H-pyrazol-5(4H)-one)] have been investigated. Structural and spectroscopic properties have been studied on the basis of elemental analyses, infrared spectra, (1)H and (13)C NMR spectra, electronic spectra, magnetic measurements and thermo gravimetric analyses. FT-IR, (1)H and (13)C NMR studies reveal that the ligand (H(2)SB) exists in the tautomeric enol form in both the states with intramolecular hydrogen bonding. Magnetic moment and reflectance spectral studies reveal that an octahedral geometry has been assigned to all the prepared coordination polymers. The kinetic parameters such as order of reaction (n) and the energy of activation (E(a)) have been reported using Freeman-Carroll method. The pre-exponential factor (A), the activation entropy (DeltaS(#)), the activation enthalpy (DeltaH(#)) and the free energy of activation (DeltaG(#)) have been calculated. PMID- 18676176 TI - BK virus antibody titers and intensity of infections after renal transplantation. AB - BACKGROUND: The mean urine BK viral load in kidney transplant recipients increases with the intensity of infection as the infection progresses from transient viruria to sustained viremia. OBJECTIVES: This study investigated whether the intensity of infection is associated with the humoral immune response. STUDY DESIGN: We measured BKV-specific IgG antibody titers in stored samples obtained serially over a 1-year period from 70 kidney transplant recipients with BKV infection and 17 control recipients without active BKV infection. RESULTS: The mean pre-transplant BKV antibody level was lower in recipients who developed viremia than the mean level in those who never developed viremia (p=0.004). Mean antibody titers in recipients who never showed evidence of active BKV infection rose slightly after transplant despite immunosuppression. The magnitude of the rise in the mean antibody titers in recipients who developed active BKV infection correlated with the intensity of infection (p<0.001). CONCLUSIONS: The mean antibody level increased in accordance with the intensity of the infection post-transplant. Pre-transplant seropositivity did not protect against sustained viremia and the antibody response was not associated with clearance of the virus. PMID- 18676177 TI - Novel innate immune functions of the whey acidic protein family. AB - Studies on the interaction of HIV with host factors have recently highlighted a potential role in the pathogenesis of AIDS for three distinct members of the whey acidic protein (WAP) family, secretory leukocyte protease inhibitor, Elafin, and ps20. Identified by an evolutionarily conserved canonical four-disulphide structural domain [whey four disulphide core domain (WFDC)], WAP proteins are increasingly being shown to display functions beyond both protease inhibition and anti-infective activity, to which they were originally ascribed. We propose novel mechanisms on why this might be the case based on an analysis of the structure function of its human members. Our analysis suggests that the interaction of HIV with WAP proteins might unravel unknown functions of the ancient WFDC and inform novel immunotherapies for the treatment of HIV and broader virus infections. PMID- 18676178 TI - Natural and TGF-beta-induced Foxp3(+)CD4(+) CD25(+) regulatory T cells are not mirror images of each other. AB - Foxp3(+) CD4(+) CD25(+) regulatory cell (Treg) subsets that maintain immunologic homeostasis have been considered to be a homogeneous population of naturally occurring, thymus-derived CD4(+)CD25(+) cells (nTregs). However, similar Foxp3+ Tregs can be induced from CD25(-) precursors in vivo, and ex vivo with interleukin 2 (IL-2) and transforming growth factor beta (TGF-beta) (iTregs). These two subsets differ in their principal antigen specificities and in the T cell receptor signal strength and co-stimulatory requirements needed for their generation. However, whether iTregs have any unique functions in vivo has been unclear. Although IL-6 can convert nTregs to Th17 cells, iTregs induced by IL-2 and TGF-beta are resistant to this cytokine and thereby might retain suppressive function at inflammatory sites. Thus, nTregs and iTregs may have different roles in the adaptive immune response. PMID- 18676181 TI - "Telethrombolysis": stroke consultation by telemedicine. PMID- 18676180 TI - Efficacy of site-independent telemedicine in the STRokE DOC trial: a randomised, blinded, prospective study. AB - BACKGROUND: To increase the effective use of thrombolytics for acute stroke, the expertise of vascular neurologists must be disseminated more widely. We prospectively assessed whether telemedicine (real-time, two-way audio and video, and digital imaging and communications in medicine [DICOM] interpretation) or telephone was superior for decision making in acute telemedicine consultations. METHODS: From January, 2004, to August, 2007, patients older than 18 years who presented with acute stroke symptoms at one of four remote spoke sites were randomly assigned, through a web-based, permuted blocks system, to telemedicine or telephone consultation to assess their suitability for treatment with thrombolytics, on the basis of standard criteria. The primary outcome measure was whether the decision to give thrombolytic treatment was correct, as determined by central adjudication. Secondary outcomes were the rate of thrombolytic use, 90 day functional outcomes (Barthel index [BI] and modified Rankin scale [mRS]), the incidence of intracerebral haemorrhages, and technical observations. Analysis was by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00283868. FINDINGS: 234 patients were assessed prospectively. 111 patients were randomised to telemedicine, and 111 patients were randomised to telephone consultation; 207 completed the study. Mean National Institutes of Health stroke scale score at presentation was 9.5 (SD 8.1) points (11.4 [8.7] points in the telemedicine group versus 7.7 [7.0] points in the telephone group; p=0.002). One telemedicine consultation was aborted for technical reasons, although it was included in the analyses. Correct treatment decisions were made more often in the telemedicine group than in the telephone group (108 [98%] vs 91 [82%], odds ratio [OR] 10.9, 95% CI 2.7-44.6; p=0.0009). Intravenous thrombolytics were used at an overall rate of 25% (31 [28%] telemedicine vs 25 [23%] telephone, 1.3, 0.7-2.5; p=0.43). 90-day functional outcomes were not different for BI (95-100) (0.6, 0.4 1.1; p=0.13) or for mRS score (0.6, 0.3-1.1; p=0.09). There was no difference in mortality (1.6, 0.8-3.4; p=0.27) or rates of intracerebral haemorrhage after treatment with thrombolytics (2 [7%] telemedicine vs 2 [8%] telephone, 0.8, 0.1 6.3; p=1.0). However, there were more incomplete data in the telephone group than in the telemedicine group (12%vs 3%, 0.2, 0.1-0.3; p=0.0001). INTERPRETATION: The authors of this trial report that stroke telemedicine consultations result in more accurate decision making compared with telephone consultations and can serve as a model for the effectiveness of telemedicine in other medical specialties. The more appropriate decisions, high rates of thrombolysis use, improved data collection, low rate of intracerebral haemorrhage, low technical complications, and favourable time requirements all support the efficacy of telemedicine for making treatment decisions, and might enable more practitioners to use this medium in daily stroke care. PMID- 18676179 TI - Into the wild: simian immunodeficiency virus (SIV) infection in natural hosts. AB - Identifying distinctions between pathogenic HIV and simian immunodeficiency virus (SIV) infections and nonprogressive SIV in natural African primate hosts might provide key insights into HIV pathogenesis. Similar to pathogenic HIV infection in humans, natural SIV infections result in high viral replication and massive acute depletion of mucosal CD4(+) T cells. A key distinction of natural SIV infections is a rapidly developing anti-inflammatory milieu that prevents chronic activation, apoptosis and proliferation of T cells and preserves the function of other immune cell subsets, thus contributing to the integrity of the mucosal barrier and the lack of microbial translocation from the gut to the peritoneum. Immunologic features observed during natural SIV infections suggest approaches for designing new strategies for producing novel second-generation vaccines and therapeutic approaches to inhibit disease progression in HIV-infected humans. PMID- 18676182 TI - Prophylactic and therapeutic suppression of experimental autoimmune encephalomyelitis by a novel bifunctional peptide inhibitor. AB - The objective was to optimize and evaluate the in vivo activities of our novel bifunctional peptide inhibitor (BPI), which alters immune response in autoimmune diseases by modulating the immunological synapse formation. Previously, we have designed PLP-BPI and GAD-BPI by conjugating myelin proteolipid protein (PLP)(139 151) and glutamic acid decarboxylase (GAD)(208-217), respectively, with CD11a(237 246) via a spacer peptide. PLP-BPI and GAD-BPI suppressed the disease progression in experimental autoimmune encephalomyelitis (EAE) and in type-1 diabetes, respectively. In this study, various PLP-BPI derivatives were synthesized and evaluated in the EAE model. Intravenous injections of PLP-BPI derivatives prevented the disease progression more efficiently than did unmodified PLP-BPI. Production of IL-17, a potent proinflammatory cytokine found commonly among MS patients, was significantly low in Ac-PLP-BPI-NH(2)-2-treated mice. Treatment given after the disease onset could dramatically ameliorate the disease. BPI induced anaphylactic responses at a lower incidence than PLP(139-151). In conclusion, PLP-BPI derivatives can effectively suppress the disease severity and morbidity of EAE by post-onset therapeutic treatment as well as prophylactic use. PMID- 18676183 TI - Amyloid-beta detection with saccharide immobilized gold nanoparticle on carbon electrode. AB - The electrochemical sensing of saccharide-protein interactions using a couple of sialic acid derivatives and Alzheimer's amyloid-beta (Abeta) is described. The densely-packed saccharide area for recognition of protein was fabricated onto a carbon electrode by three steps, which were electrochemical deposition of Au nanoparticles on a screen printed strip, self-assembled monolayer (SAM) formation of the acetylenyl group on Au nanoparticles, and the cycloaddition reaction of an azide-terminated sialic acid to the acetylenyl group. The attachment of Abeta peptides to the sialic acid layer was confirmed by electrochemistry and atomic force microscopy imaging. The intrinsic oxidation signal of the captured Abeta(1 40) and (1-42) peptides, containing a single tyrosine (Tyr) residues, was monitored at a peak potential of 0.6 V (vs Ag/AgCl within this sensor) in connection with differential pulse voltammetry. The peak current intensities were concentration dependent. The proposed process provides new routes for analysis of saccharide-protein interactions and electrochemical biosensor development. PMID- 18676184 TI - Mechanism of intestinal lipoprotein overproduction in insulin resistant humans. AB - Triglyceride-rich lipoproteins (TRLs) derived from the intestine are increased in insulin resistant states both in the post-prandial and fasted state and are associated with increased cardiovascular risk. Impaired clearance has long been invoked to explain this accumulation of intestinal TRLs, but more recent studies have highlighted the fact that the production rate of apolipoprotein (apo) B-48 containing particles is also increased in insulin resistance and Type 2 diabetes. Recent data also indicate that the enterocyte, similar to the hepatocyte, overproduces lipoprotein particles in response to an elevation of plasma free fatty acids. It is not yet clear precisely how much this overproduction of intestinal lipoprotein particles contributes to the hypertriglyceridemia of insulin resistant states. We speculate that chronic intestinal overproduction of apoB-48 primes the intestine to rapidly and efficiently facilitate the absorption of ingested fat, enhancing the assembly and secretion of intestinal TRL, and hence contributing to post-prandial lipemia. PMID- 18676185 TI - Three novel mutations in the CFTR gene identified in Galician patients. AB - We report three novel CFTR missense mutations detected in Spanish patients from Galicia (North West of Spain). In the first case, a patient homozygous for a novel S1045Y mutation died due to pulmonary problems. In the other two cases, both heterozygous for novel mutations combined with the F508del mutation, clinical symptoms were different depending on the mutation, detected as M595I and A107V. PMID- 18676186 TI - Stereospecific analysis of sakuranetin by high-performance liquid chromatography: pharmacokinetic and botanical applications. AB - A stereospecific method for analysis of sakuranetin was developed. Separation was accomplished using a Chiralpak AD-RH column with UV (ultraviolet) detection at 288 nm. The stereospecific linear calibration curves ranged from 0.5 to 100 microg/mL. The mean extraction efficiency was >98%. Precision of the assay was <12% (relative standard deviation (R.S.D.)%), and within 10% at the limit of quantitation (0.5 microg/mL). Bias of the assay was lower than 10%, and within 5% at the limit of quantitation. The assay was applied successfully to pharmacokinetic quantification in rats, and the stereospecific quantification in oranges, grapefruit juice, and matico (Piper aduncum L.). PMID- 18676187 TI - Polyester vascular prostheses coated with a cyclodextrin polymer and activated with antibiotics: cytotoxicity and microbiological evaluation. AB - Polyester (PET) vascular grafts are used to replace or bypass damaged arteries. To minimize the risk of infection during and after surgical interventions, a PET vascular prosthesis (Polythese) was functionalized with cyclodextrin polymers (PolyCDs) in order to obtain the controlled release of antibiotics (ABs: ciprofloxacin, vancomcyin and rifampicin). An epithelial cell line (L132) was used to determine the viability of the antibiotics, and human pulmonary microvascular endothelial cells (HPMEC) were used for cell proliferation by cell counting and cell vitality with Alamar Blue fluorescent dye. Staphylococcus aureus, Escherichia coli and Enteroccocus sp. were used to determine the antimicrobial activity of AB-loaded virgin and PolyCD-grafted Polythese by the minimum inhibitory concentration method. The spectrophotometric titration results first showed that a larger amount of ABs was sorbed onto PolyCD-coated Polythese compared to virgin Polythese (26.7 vs. 35.3 mg g(-1), 51.1 vs. 72.4 mg g(-1) and 4.1 vs. 21.0 mg g(-1), respectively, for rifampicin, vancomycin and ciprofloxacin). These results were further confirmed by a microbiological test, which showed AB-loaded PolyCD-coated Polythese displayed better antimicrobial activity. The viability test revealed the toxicity of rifampicin (22 mg l(-1)) and ciprofloxacin (35 mg l(-1)), and the absence of toxicity of vancomycin. These tests allow us to further explain the lower vitality and proliferation of HPMEC on the AB-loaded PolyCD-coated Polythese, which was due not to the functionalization process of prostheses but to the cytotoxicity of certain ABs themselves. Moreover, such a property could be exploited to tackle intracellular bacteria, such as in tuberculosis and other diseases, and will not compromise further in vivo applications of our functionalized vascular prostheses. PMID- 18676188 TI - Silk fibroin film from non-mulberry tropical tasar silkworms: A novel substrate for in vitro fibroblast culture. AB - The silk protein fibroin, isolated from the cocoon of the domesticated mulberry silkworm, Bombyx mori, is used extensively in biomaterial design and in cell and tissue culture. We report here for the first time the potential application of fibroin obtained from the cocoon of non-mulberry tropical silkworm, Antheraea mylitta, as a substrate for in vitro cell culture. The mechanical strength of A. mylitta silk fibers indicates a stronger thread composition. The contact angle of A. mylitta fibroin films suggests that it has lower hydrophilicity and lower solubility in organic solvents compared to B. mori fibroin films. Retention of a secondary structure of fibroin in both A. mylitta and B. mori films is confirmed by Fourier transform infrared analysis. The adherence, growth and proliferation patterns of feline fibroblast cells on A. mylitta fibroin films suggest that this kind of film has a greater ability to support cell growth than B. mori fibroin films and is comparable to that of control. This study demonstrates that, as well as being non-toxic to dermal fibroblast cells, non-mulberry fibroin might be a useful alternative substrate to the more common B. mori fibroin for a variety of biomedical applications. PMID- 18676189 TI - Temperature-responsive glass coverslips with an ultrathin poly(N isopropylacrylamide) layer. AB - A temperature-responsive cross-linked polymer gel was covalently grafted onto glass coverslips by electron beam irradiation. The grafted thickness and amount of polymer as well as the surface wettability increased with the initial monomer concentration. When the monomer concentration was 5 wt.%, the grafted polymer density was 0.84microgcm(-2), and cells adhered and spread on the surface at 37 degrees C, but detached at 20 degrees C. In contrast, when the monomer concentration was 35 wt.%, the polymer density was 1.28microgcm(-2), and the surfaces were cell repellent even at 37 degrees C. These results show a remarkable contrast to those obtained from temperature-responsive polymer-grafted tissue culture polystyrene dishes, since various types of cells showed temperature-dependent cell adhesion/detachment when the grafted density was around 2microgcm(-2) on these surfaces. We discuss the possible molecular mechanisms underlying this discrepancy. PMID- 18676190 TI - Evaluations of osteogenic and osteoconductive properties of a non-woven silica gel fabric made by the electrospinning method. AB - Evaluations of the osteoblast-like cell responses and osteoconductivity of a non woven silica gel fabric were carried out to determine its potential for application as a scaffold material for use in bone tissue engineering. The silica gel solution was prepared by condensation following hydrolysis of tetraethyl orthosilicate under acidic conditions. The solution was spun under a 2kVcm(-1) electric field. The diameters of the as-spun silica gel fibers were in the range of approximately 0.7-6microm. The fabric was then heat-treated at 300 degrees C for 3h. The proliferation of pre-osteoblastic MC3T3-E1 cells evaluated by the MTS assay was lower than on the tissue culture plate (TCP) as many cells leaked through the large voids formed by the randomly placed long, narrow silica gel fibers, which further retarded cell growth. However, the expressions of extracellular signal-regulated kinase and transcriptional factor from the cells were higher when cultured on the non-woven silica gel fabrics than on TCP. The alkaline phosphatase (ALP) activity and differentiation marker expressions assessed by amplication via the reverse transcription-polymerase chain reaction, such as type I collagen, ALP and osteocalcin, were higher for cells cultured on non-woven silica gel fabrics than on TCP. The non-woven silica gel fabric showed good osteoconductivity in the calvarial defect New Zealand white rabbit model. To this end, the non-woven silica gel fabric has good potential as a scaffold material for bone tissue engineering due to its good biological properties. PMID- 18676191 TI - The effect of sterilization processes on the bioadhesive properties and surface chemistry of a plasma-polymerized polyethylene glycol film: XPS characterization and L929 cell proliferation tests. AB - The influence of several sterilization processes (autoclaving, gamma-ray irradiation, ethylene oxide exposure and Ar/H(2) low pressure plasma treatment) on the surface chemistry and the bioadhesive properties of thin films (thickness approximately 20 nm) of plasma-polymerized diethylene glycol dimethyl ether has been studied. X-ray photoelectron spectroscopy (XPS) analysis and cell proliferation tests were used to characterize the surfaces. The XPS results revealed in all cases a change in the surface chemistry of the layer after sterilization, whereas the conservation of non-bioadhesive properties of the coating depends on the type of sterilization process. In particular, the low pressure plasma-based sterilization technique leads to a loss of the non bioadhesive properties of the plasma coating, whereas the coatings are resistant to the other standard decontamination techniques. This property makes them suitable for biomedical applications, provided that an appropriate sterilization process is selected. PMID- 18676192 TI - Bird-bite infection and pyoderma gangrenosum: a rare combination? PMID- 18676193 TI - Merkel cell carcinoma of the face: an analysis of 16 cases in the Japanese. AB - BACKGROUND: There is no agreement regarding a staging system and optimal treatment of Merkel cell carcinoma. Some centres have reported results from larger series of patients, but these do not include Asian or Japanese centres. OBJECTIVE: The purpose of this study was to retrospectively review our experience with the surgical treatment of MCC of the face in the Japanese and to study its management and outcome using the staging system described by Clark et al. METHODS: We report our experiences with 16 cases between 1991 and 2004. Patients and tumour characteristics, treatment variables and outcome were analysed. RESULTS: The follow-up periods ranged from 1 to 180 months. The average was 32.6 months and the median was 17.5 months. The relapse-free survival for all patients was 51% at 2 years. The relapse-free survival was 80% for the patients with Stage I and 33% with Stage II at 2 years. CONCLUSION: This staging system was suggested to reflect prognosis although the number of patients in this series was small. Sentinel lymph node biopsy should be considered to determine the accurate nodal staging, and patients with MCC of the head and neck may be treated according to the revised staging system by Clark et al. PMID- 18676194 TI - Neonatal asymmetric crying face. PMID- 18676195 TI - [Bone imaging in patient with chronic kidney disease: a new tool for managing renal osteodystrophy?]. AB - Bone damage in patients with chronic kidney disease (CKD), in the spectrum of chronic kidney disease-mineral and bone disorders (CKD-MBD), represents a daily challenge for nephrologists. The impact of CKD on bone health may be immediate (biological equilibrium) or delayed (fractures and vascular calcifications). Histomorphometry remains the gold standard to evaluate bone but it is rarely perfomed in clinical practice. A real measurement of bone mineral density (BMD) by dual X-ray absorptiometry (DXA) is currently performed to evaluate bone mass. However, this technique presents some limitations. In 2000, the National Institute of Health (NIH) defined new "quality" criteria for the diagnosis of osteoporosis in addition to a decreased bone mass. Bone strength actually integrates two concepts: bone density and bone quality (i.e., microarchitectural organization, bone turnover, mineralization and microfractures). These quality criteria cannot be evaluated by DXA. New bone imaging techniques have thus been developed, leading to an improvement in bone evaluation. PMID- 18676197 TI - Application of functional CdS nanoparticles in determination of silver ion by resonance light-scattering technique. AB - Based on the strong enhancement effect of silver ion on resonance light scattering intensity of functional CdS nanoparticles, a new direct quantitative determination method for silver ion was established. Under the optimum conditions, the response signal is linearly proportional to the concentration of silver ion. The linear range is 5.0x10(-9)-2.0x10(-6) mol L(-1). The proposed method was applied to determine silver ion in river water samples. PMID- 18676198 TI - Comment on "theoretical acquirement of the red shift of nu(F-H) upon complexation with Ne" by Guoqun Liu and Xianxi Zhang [Spectrochim. Acta Part A 69 (2008) 917 920]. AB - This Comment shows that the agreement between theory and experiment for the vibrational red shift of the FH stretch upon complexation with Ne claimed by the authors of the captioned manuscript is not justified and should be considered as merely fortuitous. PMID- 18676199 TI - Effect of peptidoglycans on erythrocyte survival. AB - Peptidoglycans (PGNs) from bacterial cell walls belong to 'pathogen-associated molecular patterns' (PAMP), which modify the course of an infection with bacterial pathogens. Bacterial infections may lead to anaemia, which at least partially could result from accelerated erythrocyte death. The present study explored the effect of PGNs on eryptosis, a stress-induced suicidal death of erythrocytes, characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the erythrocyte surface. Eryptotic cells are phagocytosed and thus rapidly cleared from circulating blood. Eryptosis is triggered by an increase in the cytosolic Ca(2+) concentration and by formation of ceramide. Erythrocyte Ca(2+) activity was estimated from Fluo3 fluorescence, ceramide formation by fluorescent antibodies, phosphatidylserine exposure from annexin V-binding, and erythrocyte volume from forward scatter in fluorescence activated cell sorting (FACS) analysis. Exposure of erythrocytes to PGNs increased cytosolic Ca(2+) concentration, increased ceramide formation, enhanced the percentage of annexin V-binding erythrocytes, decreased erythrocyte forward scatter, and lowered the intracellular ATP concentration. The effect of peptidoglycans was significantly blunted in the absence of extracellular Ca(2+). The clearance of erythrocytes exposed to PGNs was significantly enhanced in vivo. In conclusion, peptidoglycans induce eryptosis at least partially through an increase in the cytosolic Ca(2+) concentration, an effect presumably contributing to the development of anaemia during bacterial infections. PMID- 18676200 TI - Thoracic hydatid disease; a review of 763 cases. AB - BACKGROUND: Thoracic hydatid disease remains a significant health problem in endemic areas. The aim of this study was to retrospectively evaluate 763 patients diagnosed with thoracic hydatid cysts and treated surgically at Ibn-Alnafis Teaching Hospital between January 1986 and January 2006. METHODS: All patients underwent surgery. The mean age was 26 years. Intact cysts were enucleated in 468 patients after injection with a scolicidal agent and the remaining cavity was left open after securing localised air leaks. Pulmonary resections were performed in 198 patients. RESULTS: Most cysts were removed without lung resection. Resection was reserved for large cysts, severe suppuration and complicated cases with haemoptysis or bronchiectasis. However, lung preservation was always attempted initially. In patients with intact lung cysts, 37% were asymptomatic with incidental diagnosis on routine chest X-ray. Mortality was 1% and morbidity was 12.6%. CONCLUSION: Lung preservation without obliteration of pericystic cavity provides good result in patients with uncomplicated hydatid cysts. Radical resection should be reserved for complicated cases. Medical treatment is indicated postoperatively especially for recurrent or multiple hydatid cysts. PMID- 18676201 TI - Molecular profiling of non-small cell lung cancer: of what value in clinical practice? AB - Future improvements in lung cancer survival are likely to come from delineating its putative oncogenic pathways. The development of microarray technology to perform thousands of simultaneous genetic experiments and the linking of this to clinical information is an imperative for refining our current treatments and developing new ones. This paper reviews the state of this research, describes a typical microarray experiment and the implications for diagnosis and treatment of non-small cell lung cancer. PMID- 18676202 TI - Does combined antegrade-retrograde cardioplegia have any superiority over antegrade cardioplegia? AB - BACKGROUND: In a prospective randomised clinical study we assessed and compared antegrade vs. combined antegrade-retrograde cardioplegia in patients who underwent elective coronary artery by pass grafting. METHODS: Between March 2006 and January 2007, 87 consecutive patients were randomly divided into two groups. Group A (n=45) received antegrade cold (4 degrees C) blood cardioplegia. Besides antegrade cardioplegia, Group B (n=42) received continuous retrograde cardioplegia passively by gravitational force. The need for cardiac support during and after cardiopulmonary bypass, post-operative morbidity, ICU stay, hospital stay and mortality were compared in two groups. RESULTS: There was no significant difference between the two groups in gender, age and pre-operative ejection fraction. Sixteen patients in Group A (35.5%) and eight patients in Group B (19%) needed inotropic support while weaning off cardiopulmonary bypass (p=0.04). Four patients in Group A (8.9%) and two patients in Group B (4.8%) needed intra-aortic balloon pump (p=0.44) in the ICU. We found no statistically important difference between the two groups in post-operative morbidity and mortality. CONCLUSIONS: Retrograde continuous infusion of cardioplegia by gravitational force combined with antegrade cardioplegia, provides satisfactory myocardial protection and eliminates the need for inotropic support compared with antegrade technique alone. PMID- 18676203 TI - Cellular responses to MPT-51, GlcB and ESAT-6 among MDR-TB and active tuberculosis patients in Brazil. AB - Multi-drug resistant pulmonary tuberculosis (MDR-TB) may result from either insufficiency of the host cellular immune response or mycobacterial mechanisms of resistance. Mycobacterium tuberculosis-specific CD8+ and CD4+ T lymphocytes from MDR-TB patients are poorly studied. The aim of this study was to evaluate CD4+IFN gamma+, CD4+IL-10+, CD8(+)IFN-gamma+ and CD8+IL-10+ cell populations by flow cytometry in non-resistant TB and multi-drug resistant tuberculosis (MDR-TB) patients from mid-central Brazil after stimulation with MPT-51, GlcB and ESAT-6 recombinant antigens from M. tuberculosis in comparison to tuberculin skin test negative (TST) healthy individuals. Non-resistant TB patients present specific cellular responses (CD4 and CD8, both IFN-gamma and IL-10) to GlcB, MPT-51 and ESAT-6; while MDR-TB patients present only CD8+IFN-gamma+ responses to ESAT-6 and CD8+IL-10+ responses to GlcB and ESAT-6. The results show that MDR-TB patients present impaired specific CD4 IFN-gamma and IL-10 responses and increased/normal specific CD8 IFN-gamma and IL-10 responses. This suggests an important role for CD8 function in these patients. PMID- 18676204 TI - ROS-deficient monocytes have aberrant gene expression that correlates with inflammatory disorders of chronic granulomatous disease. AB - Chronic granulomatous disease is an immunodeficiency caused by an inability to produce reactive oxygen species. While the mechanism of hyper-sensitivity to infection is well understood in CGD, the basis for debilitating inflammatory disorders that arise in the absence of evident infection has not been fully explained. Herein it is demonstrated that resting and TLR-activated monocytes from individuals with CGD expressed significantly higher levels of inflammatory mediators than control cells; the expression in CGD cells resembled normal cells stimulated with lipopolysaccharide. The lack of acute illness, infection or circulating endotoxin in the blood of the CGD patients at the time of sampling was consistent with infection-free inflammation. The enhanced expression of inflammatory mediators correlated with elevated expression of NF-kappaB and was dependent on ERK1/2 signalling. The results are consistent with the hypothesis that ROS are anti-inflammatory mediators that control gene expression and potentially limit the development of sterile inflammatory disorders. PMID- 18676205 TI - Trends in use of opioids by noncancer pain type 2000-2005 among Arkansas Medicaid and HealthCore enrollees: results from the TROUP study. AB - Use of prescription opioids for noncancer pain has increased significantly in recent years, but it is not known if trends differ among the most common noncancer pain conditions. We examined trends in opioid prescribing for the years 2000 through 2005 for individuals with arthritis/joint pain, back pain, neck pain, and headaches by type and number of pain diagnoses, using data from claims records from 2 health insurers: HealthCore commercially insured members (N = 3,768,223) and Arkansas Medicaid (N = 127,866). Rates of headache, back pain, and neck pain diagnoses increased significantly in Arkansas Medicaid enrollees but more modestly among HealthCore enrollees. Rates of opioid use increased in both groups, with long-term use (>90 days' supply per year) increasing at twice the rate of any use. Rates of opioid use did not differ widely between noncancer pain conditions, but long-term opioid use rates doubled with each additional pain diagnosis. Mean days supply and cumulative yearly dose increased between 2000 and 2005 for all pain types and with increasing number of pain diagnoses, but dose per day supply remained relatively stable. The greatest increases in dose among all the pain conditions were seen in short-acting DEA Schedule II opioids. PERSPECTIVE: This study demonstrates increased use of opioids, particularly long term use, in noncancer pain over a 6-year period among those with multiple pain types. These results appear to reflect a general increase in use of prescription opioids for noncancer pain rather than a condition-specific change in prescribing practices. PMID- 18676206 TI - Anti-glycation activity of gold nanoparticles. AB - Anti-glycation activity of gold nanoparticles (GNPs) has been reported for the first time. Nonenzymatic glycation of alpha-crystallin leads to formation of cataract, or opaque aggregate of proteins. In this article we report prevention of glycation of alpha-crystallin by conjugation with GNPs. Formation of advanced glycosylic end products is prevented even if a strong glycating agent such as fructose is used. In addition, the nanoconjugation can provide some important information on the structural distribution of this dynamic chaperone protein. Because GNPs are biocompatible, their reported anti-glycation activity may have ophthalmological implications. PMID- 18676207 TI - Nanoimmunoliposome delivery of superparamagnetic iron oxide markedly enhances targeting and uptake in human cancer cells in vitro and in vivo. AB - To circumvent the problem of reduction of the supermagnetic properties of superparamagnetic iron oxide (SPIO) nanoparticles after chemical modification to conjugate targeting molecules, we have adapted a tumor-targeting nanoimmunoliposome platform technology (scL) to encapsulate and deliver SPIO (scL SPIO) in vitro and in vivo without chemical modification. Scanning probe microscopy, confocal microscopy, and Prussian blue staining were used to analyze the scL-SPIO and assess intracellular uptake and distribution of SPIO in vitro. In vivo targeting and tumor-specific uptake of scL-SPIO was examined using fluorescent-labeled SPIO. We demonstrated that SPIO encapsulation in the scL complex results in an approximately 11-fold increase in SPIO uptake in human cancer cells in vitro, with distribution to cytoplasm and nucleus. Moreover, the scL nanocomplex specifically and efficiently delivered SPIO into tumor cells after systemic administration, demonstrating the potential of this approach to enhance local tumor concentration and the utility of SPIO for clinical applications. PMID- 18676208 TI - Hollow-fiber-supported liquid phase microextraction with in situ derivatization and gas chromatography-mass spectrometry for determination of chlorophenols in human urine samples. AB - A simple and highly sensitive method that involves hollow-fiber-supported liquid phase microextraction (HF-LPME) with in situ derivatization and gas chromatography-mass spectrometry (GC-MS) was developed for the determination of chlorophenols (CPs) such as 2,4-dichlorophenol (DCP), 2,4,6-trichlorophenol (TrCP), 2,3,4,6-tetrachlorophenol (TeCP) and pentachlorophenol (PCP) in human urine samples. Human urine samples were enzymatically de-conjugated with beta glucuronidase and sulfatase. After de-conjugation, HF-LPME with in situ derivatization was performed. After extraction, 2 microl of extract was carefully withdrawn into a syringe and injected into the GC-MS system. The limits of detection (S/N=3) and quantification (S/N>10) of CPs in the human urine samples are 0.1-0.2 ng ml(-1) and 0.5-1 ng ml(-1), respectively. The calibration curve for CPs is linear with a correlation coefficient of >0.99 in the range of 0.5-500 ng ml(-1) for DCP and TrCP, and of 1-500 ng ml(-1) for TeCP and PCP, respectively. The average recoveries of CPs (n=6) in human urine samples are 81.0 104.0% (R.S.D.: 1.9-6.6%) with correction using added surrogate standards. When the proposed method was applied to human urine samples, CPs were detected at sub ng ml(-1) level. PMID- 18676209 TI - Analysis and purification of alcohol-sensitive chiral compounds using 2,2,2 trifluoroethanol as a modifier in supercritical fluid chromatography. AB - 2,2,2-Trifluoroethanol (TFE) is evaluated as an alternative modifier for the analysis and purification of alcohol-sensitive chiral compounds using supercritical fluid chromatography (SFC). Four chiral compounds, selected for their sensitivity to alcohols, in addition to a variety of standard chiral compounds were analyzed by SFC using TFE with polysaccharide and Pirkle-type chiral stationary phases (CSPs) to produce selectivities (alpha) and resolutions (Rs) as high as 1.4 and 7.2. A preparative isolation of 2-phenylglutaric anhydride was achieved using TFE as the mobile phase modifier to produce clean enantiomers. PMID- 18676210 TI - Convergence of body mass with aging: the longitudinal interrelationship of health, weight, and survival. AB - There has been ongoing debate about the health risks associated with increased body weight among the elderly population. One issue has not been investigated thoroughly is that body weight changes over time, as both the reasons and results of, the development of chronic diseases and functional disabilities. Structural models have the ability to unravel the complicated simultaneous relationship between body weight, disability, and mortality along the aging process. Using longitudinal data from the Medicare Current Beneficiary Survey from 1992 to 2001, we constructed a structural model to estimate the longitudinal dynamic relationship between weight, chronic diseases, functional status, and mortality among the aging population. A simulation of an age cohort from 65 to 100 was conducted to show the changes in weight and health outcomes among the cohorts with different baseline weight based on the parameters estimated by the model. The elderly with normal weight at age 65 experience higher life expectancy and lower disability rates than the same age cohorts in other weight categories. The interesting prediction of our model is that the average body size of an elderly cohort will converge to the normal weight range through a process of survival, senescence, and behavioral adjustment. PMID- 18676211 TI - Bowel preparation before colonoscopy in the era of mass screening for colo-rectal cancer: a practical approach. AB - Colonoscopy constitutes the principal investigation for colo-rectal neoplasms due to its ability to detect and remove most of precancerous lesions; due to the ongoing or planned colon cancer screening programs in many European countries we should expect an enormous increase in colonoscopic demand over the next few years. Diagnostic accuracy and therapeutic safety of colonoscopy strictly depends upon the quality of bowel cleansing which is often perceived as the most unpleasant part of the procedure in individuals undergoing this examination. The ideal preparation for colonoscopy should reliably empty the colon from all faecal material allowing the optimal visualization of the entire colonic mucosa without causing great patient's discomfort nor significant shifts in fluids or electrolytes. Standard PEG solutions and sodium phosphate (NaP) compounds are the most frequently used preparations; both are accepted and relatively well tolerated by the majority of patients undergoing colonoscopy; however, NaP compounds should be avoided in elderly subjects as well as in those with congestive heart failure, renal and hepatic insufficiency or taking diuretics, ACE inhibitors or angiotensin receptor blockers, since they can induce severe electrolyte and/or fluid disturbances. Standard PEG solutions are often taken incompletely due to the low palatability and the high volume of liquids required which induce nausea and vomiting with negative consequences in terms of colon cleansing. Reduced volume and better palatability of PEG solutions, such as those obtained with the newest PEG formulations, as well as improved patient education concerning the importance of bowel cleansing could undoubtedly increase compliance with oral bowel preparations and promote adherence to colo-rectal cancer screening programs. PMID- 18676213 TI - Osteogenic growth peptide incorporated into PLGA scaffolds accelerates healing of segmental long bone defects in rabbits. PMID- 18676212 TI - Regulation of material properties in electrospun scaffolds: Role of cross-linking and fiber tertiary structure. AB - We cross-linked scaffolds of electrospun collagen to varying degrees with glutaraldehyde using an ethanol-based solvent system and subsequently defined how the percentage of cross-linking impacts bulk and microscale material properties and fiber structure. At hydration, electrospun fibers underwent coiling; the extent of coiling was proportional to the percentage of cross-linking introduced into the samples and was largely suppressed as cross-linking approached saturation. These data suggest that electrospun collagen fibers are not deposited in a minimal energy state; fiber coiling may reflect a molecular reorganization. This result has functional/structural implications for protein-based electrospun scaffolds. Changes in fiber topology that develop during post-electrospinning processing may alter monomer organization, mask or unmask receptor binding sites, and/or change the biological properties of these nanomaterials. Hydrated scaffolds were mounted into a custom stretching device installed on a microscope stage and photographed after incremental changes in strain. Changes in fiber alignment were measured using the two-dimensional fast Fourier transform method. Fibers in all scaffolds underwent alignment in response to strain; however, the rate and extent of alignment that could be achieved varied as a function of cross linking. We propose four distinct modes of scaffold response to strain: fiber uncoiling, fiber reorientation, fiber elongation and interfiber sliding. We conclude that bulk material properties and local microscale architecture must be simultaneously considered to optimize the performance of electrospun scaffolds. PMID- 18676214 TI - Functional capacity after pedicled TRAM flap delayed breast reconstruction. PMID- 18676215 TI - Functional reconstruction of the upper and lower lips and commissure with a forearm flap combined with a free gracilis muscle transfer. AB - After resection of an arterio-venous malformation of the upper and lower lips and commissure we performed reconstruction with a forearm flap combined with a free gracilis muscle transfer. First the motor nerve of the gracilis muscle was anastomsed to a buccal nerve branch in the cheek. In a second operation, the red lip was reconstructed with an oral mucosal graft, and the upper lip skin was reconstructed with a local flap. The patient obtained good oral sphincter function for eating, speaking and air inflation. PMID- 18676216 TI - The dead space technique for preparing "surgical concentrations" of adrenaline with local anaesthetic. PMID- 18676217 TI - Re: 'Osmotic tissue expanders in cleft lip and palate surgery: a cautionary tale'. PMID- 18676218 TI - Tigecycline: a critical update. AB - Tigecycline is the first Food and Drug Administration (FDA) approved glycylcycline antibiotic. It has shown remarkable in vitro activity against a wide variety of gram-positive, gram-negative and anaerobic bacteria including many multidrug resistant (MDR) strains. However, it has minimal activity against Pseudomonas aeruginosa and Proteus spp. To date, little resistance to tigecycline has been reported. Clinical trials studying complicated skin and skin-structure infections (cSSSIs) demonstrated that tigecycline has equivalent efficacy and safety compared with the combination of vancomycin and aztreonam. For complicated intra-abdominal infections (cIAIs), tigecycline was found to be as effective as imipenem/cilastatin. Adverse events related to tigecycline therapy, i.e. nausea and vomiting, were tolerable. Currently available data suggest that tigecycline may play an important role in the future as a monotherapy alternative to older broad-spectrum antibiotics, such as advanced generation cephalosporins, carbapenems, fluoroquinolones, piperacillin/tazobactam, and gram-positive directed agents (e.g. daptomycin, linezolid and quinupristin/dalfopristin) for which resistance is being increasingly reported from all parts of the world. PMID- 18676219 TI - Effect of oxazolidinone, RBx 7644 (ranbezolid), on inhibition of staphylococcal adherence to plastic surfaces. AB - Adhesion to biomaterial is assumed to be a crucial step in the pathogenesis of foreign body infection. Slime producing Staphylococcus epidermidis and Staphylococcus aureus have emerged as a preeminent cause of nosocomial bacteremia and infections of prosthetic medical devices. We evaluated the time-dependent anti-adhesive effect of RBx 7644 (ranbezolid), vancomycin, linezolid and quinupristin/ dalfopristin on two isolates each of S. epidermidis and S. aureus. Linezolid and quinupristin/ dalfopristin showed inhibition only at supra inhibitory concentrations (2 and 4X MIC) following 2 and 4 h delayed treatment, whereas RBx 7644 demonstrated significant activity against adhesion of staphylococcal cells that had been treated with 2 to 6 h delay. When vancomycin treatment was delayed by 4 to 6 h, even concentrations above the MIC were unable to prevent adherence. This study indicates that RBx 7644 has anti-adhesion potential and may emerge as an important antibiotic for prevention and treatment of device-related infections caused by staphylococci. PMID- 18676220 TI - The intracellular accumulation of phagocytic and epithelial cells and the inhibitory effect on Chlamydophila (Chlamydia) pneumoniae of telithromycin and comparator antimicrobials. AB - The intracellular accumulation of telithromycin was measured and compared with ciprofloxacin, clarithromycin, minocycline and erythromycin. The activities of telithromycin, clarithromycin and minocycline against Chlamydophila (Chlamydia) pneumoniae were compared in an intracellular killing assay. Maximal telithromycin accumulation (mean intracellular/extracellular concentration ratio) ranged from 6.7 (A549 lung epithelial cells) to 11.8 (THP-1 monocytic cells). This ratio was similar to that of clarithromycin, but less than for minocycline. Minimum inhibitory and minimum lethal telithromycin concentrations against six clinical strains of C. pneumoniae were <0.015-0.03 mg/L and 0.03-0.06 mg/L, respectively, which were lower than those found for minocycline. These results show that telithromycin accumulates rapidly into epithelial cells, similar to previously reported results for phagocytic cells. Intracellular accumulation of telithromycin was lower than that observed for minocycline, but telithromycin demonstrated substantially more potent antichlamydial activity. PMID- 18676221 TI - Rifampicin resistance phenotyping of Brucella melitensis by rpoB gene analysis in clinical isolates. AB - R Rifampicin resistance of Brucella melitensis by rpoB gene analysis has not yet been performed in Turkey, where brucellosis is endemic. In this study, we investigated the efficacy of E-test and single nucleotide polymorphism (SNP) analysis of the B. melitensis rpoB gene, for the detection of mutations conferring rifampicin resistance, by sequencing 21 human B. melitensis strains from the Southeast and Marmara regions of Turkey. On CLSI slow-growing bacteria standards, all isolates were sensitive to rifampicin except for 6 which showed intermediate resistance to rifampicin. MIC(50) and MIC(90)values were 1 microg/ml and 1.5 microg/ml respectively (range 0.50 -1.5 microg/ml). The rifampicin resistant phenotype was investigated at Cd 154 (GTT/TTT), Cd 526 (GAC/TAC, GAC/AAC, GAC/GGC), Cd 536 (CAC/CTC, CAC/TAC), Cd 539 (CGC/AGC), Cd 541 (TCG/TTG) and Cd 574 (CCG/CTG) of the rpoB gene in B. melitensis 16M and B115 strains, and in clinical isolates. No missense mutations were found in any of the B. melitensis isolates, which indicates that all isolates were rifampicin susceptible. In conclusion, SNP analysis was useful as a molecular tool for rifampin resistance testing. Although resistance to rifampicin was not detected in our strains of B. melitensis; the presence of strains with intermediate resistance to rifampicin indicates that susceptibility testing should be performed periodically. PMID- 18676222 TI - Polymorphism in pbp5 gene detected in clinical Enterococcus faecium strains with different ampicillin MICs from a Tunisian hospital. AB - The polymorphism in pbp5 gene was investigated in nine unrelated clinical gentamicin-resistant Enterococcus faecium strains with different minimal inhibitory-concentration values for ampicillin (six ampicillin-resistant and three ampicillin-susceptible). Five alleles were detected when the pbp5 C terminal region was analysed, two of them in the ampicillin-resistant strains showed a new allele characterised by the Thr416Ala and Val462Ala substitutions. Two different alleles were identified when the pbp5 N-terminal region was studied; one of them in the unique strain (E. faecium 83) that presented very low ampicillin MIC (<0.125 microg/ml) and a nucleotidic mutation implicating a stop codon at 451 position. RT-PCR experiments carried out on five E.faecium positive results indication the expression of this gene. Specific mutations in pbp5 gene could be responsible of the high MIC values of some of the E. faecium strains. PMID- 18676223 TI - In vitro and in vivo activity of miltefosine against penicillin-sensitive and resistant Streptococcus pneumoniae strains. AB - The in vitro and in vivo activity of miltefosine against penicillin-sensitive and penicillin-resistant pneumococcal strains was studied. The minimum inhibitory concentrations (MICs) of miltefosine were determined in cation-adjusted Mueller Hinton plus 2% lysed horse blood (CAMHB) and in Todd-Hewitt (TH) broth. The respective MICs were higher using CAMHB (128 and 64 mg/L) than using TH broth (4 and 8 mg/L). The in vivo activity was studied in a murine peritonitis-sepsis model. Miltefosine was orally administered at doses of 15 and 30 mg/kg/day after, at the time of, and before bacterial challenge for 3-5 days. All control and 16 out of the 17 (94.1%) miltefosine-treated animals that were inoculated with the penicillin-sensitive strain died. No survival was observed among control and miltefosine-treated animals inoculated with the penicillin-resistant pneumococcal strain. The in vivo unresponsiveness of miltefosine in this sepsis model could be attributed to some inhibitory effect of blood, inadequate pharmacokinetics and/or the extracellular localization of the pneumococcus. PMID- 18676224 TI - The technical aspects and clinical significance of detecting extended-spectrum beta-lactamase-producing Enterobacteriaceae at a tertiary-care hospital in Kuwait. AB - Extended-spectrum beta-lactamase (ESBL) production by Enterobacteriaceae is an emerging problem. This 3-year prospective study was undertaken to determine the prevalence of such enzymes among the clinically significant isolates of the Enterobacteriaceae family gathered from patients, and to evaluate the different techniques for their detection as well as their clinical significance. Members of the Enterobacteriaceae family isolated from blood, inhibited by the third generation cephalosporins with minimum inhibitory concentrations (MICs) of < or =2 microg/ml and MIC < or =8 microg/ml and isolates from other sources inhibited by MIC < or =8 microg/ml were also investigated for ESBL production by VITEK2 and E test. Their clinical significance in septicemic patients was analyzed. Out of 3,215 isolates, 1018 (31.7%) were ESBL-producers by both VITEK2 and E test. Of these, 428 (42%) were Klebsiella pneumoniae and 376 (37.0%) were Escherichia coli with overall prevalence rates of 13.3% and 11.7%, respectively. There were a total of 184 septicemic patients infected by ESBL-producing Enterobacteriaceae out of which 134 (73%) needed modification of therapy; most (58%) of these patients were initially on third-generation cephalosporin therapy. A total of 58 (31.5%) patients were infected by ESBL-producing blood isolates which were inhibited by cefotaxime/ceftriaxone at MICs =8 microg/ml (within the susceptibility range). Resistance to both aminoglycosides and quinolones were significantly higher among ESBLproducing isolates compared to non-producers (P <0.05). This study highlights a high prevalence of ESBL-producing Enterobacteriaceae in a major tertiary teaching hospital in our country and demonstrates that almost a third of the ESBL-producing Enterobacteriaceae blood isolates would have been released as susceptible by routine susceptibility testing; a finding inimical to optimal therapeutic success. PMID- 18676225 TI - Risk factors for coexistence of fluoroquinolone resistance and ESBL production among Enterobacteriaceae in a Greek university hospital. AB - The purpose of this study was to identify risk factors for fluoroquinolone resistance (QR) among ESBL- producing Enterobacteriaceae causing nosocomial infections. The study was conducted in Laikon General Hospital in Athens, Greece, during the period January 2004 - January 2005. Epidemiological and clinical data were collected from the medical charts of the patients diagnosed with nosocomial infections due to an ESBL-producing Enterobacteriaceae. QR was 60% among the 84 ESBL-producing Enterobacteriaceae isolates. Infection from QR-ESBL bacteria was associated with increased hospital stay (p=0.028); QRESBL bacteria were isolated later during hospitalization than fluoroquinolone susceptible (QS)-ESBL (p=0.089); factors associated with QR were immune-deficiency (p=0.047), previous use of carbapenems (p=0.08) and fluoroquinolones (p=0.067), and admission to the Transplantation Unit (p=0.047). In addition, QR-ESBL bacteria were more likely to be resistant to co-trimoxazole (p<0.001), gentamicin (p=0.054) and tobramycin (p=0.004). Logistic regression analysis indicated that admission to the transplantation unit was an independent risk factor for infection due to a QR ESBL isolate. Results of this study question ciprofloxacin's usefulness as a valid alternative to carbapenems in our hospital for the treatment of infections due to ESBL-producing bacteria. In addition strategies for addressing the QR-ESBL situation should focus on limiting fluoroquinolone and carbapenem consumption and emphasize on barrier precautions in patients with longer hospitalization, immunosuppression, or admission to the transplantation unit. PMID- 18676226 TI - Inhibition of hepatitis B virus replication by Bay 41-4109 and its association with nucleocapsid disassembly. AB - The authors investigate the effects and mechanisms of the anti-hepatitis B virus (HBV) agent Bay 41-4109. HepG2.2.15 cells were used to investigate the antiviral effects of Bay 41-4109 by using real-time polymerase chain reaction (PCR), western blotting, and immunofluorescence. The C terminally truncated core protein was expressed and purified. Changes in hepatitis B capsid formation were assayed by dynamic light scattering and transmission electronic microscopy. Bay 41-4109 was found to be a highly selective and potent inhibitor of hepatitis B virus replication in HepG2.2.15 cells. This compound was equally effective at inhibiting HBV DNA release and the cytoplasmic HBcAg level, with 50% inhibitory concentrations of 32.6 and 132 nM, respectively. HBV DNA and HBcAg were inhibited in a dose-dependent manner, indicating that the anti-HBV mechanisms are associated with and dependent on the rate of HBcAg inhibition. Our results indicate that Bay 41-4109 treatment disassembled the core capsids and separated them into monomers or dimers, the form in which they could be further degraded into peptides. The core protein assembled in a misdirected manner cannot function effectively. Our results suggest that, based on its particular activities, Bay 41 4109 is a promising anti-HBV candidate. PMID- 18676227 TI - Pharmacokinetics of fusidic acid and cefepime in heart tissues: implications for a role in surgical prophylaxis. AB - The pharmacokinetic profiles of fusidic acid and cefepime in heart tissues were assessed in 30 patients undergoing elective valve replacement and cardiopulmonary bypass. Single doses of 1 g of fusidic acid and 2 g of cefepime were administered intravenously to two groups of 15 and 15 patients respectively upon initiation of anesthesia. Samples of serum, heart valves, myocardium, pericardium, mediastinal fat and sternum were collected within <1 hour, 1-2 h and 2-4 h after the end of drug infusion. Drug concentrations were estimated by a microbiological assay. It was found that concentrations of fusidic acid in all specimens were 20-fold higher than the MIC90s of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis, being at such levels throughout all period of sampling. Cefepime concentrations in heart valves collected 1-2 h after drug infusion were higher than the MIC90s of multidrug-resistant Enterobacteriaceae. It is concluded that both fusidic acid and cefepime penetrated heart tissues adequately; however only fusidic acid could also accumulate in the mediastinum. These data suggest that both antibiotics may be a good alternative for prophylaxis in open heart surgery. PMID- 18676228 TI - First and recurrent pediatric urinary tract infections: do they have different antibiotic susceptibilities? AB - Antibiotic susceptibility studies in children rarely differentiate between first and recurrent urinary tract infections (UTI), although the latter, frequently associated with underlying urinary tract anomalies and antibiotic prophylaxis, are more likely to be associated with higher antibiotic resistance of uropathogens as a result. We investigated whether antibiotic resistance was different between first and recurrent UTIs in 250 episodes (145 first and 105 recurrent) in 154 children (2 months to 12 years of age) with culture proven UTI. According to univariate analysis, resistance to cefuroxime and gentamicin was significantly lower in recurrences. This association remained statistically significant in the multivariable analysis, with adjusted odds ratio OR of 0.8 for cefuroxime (p=0.04) and for gentamicin (p=0.003) after adjusting for the role of confounding factors. The risk of resistance to other antibiotics was otherwise similar for first and recurrent UTIs. PMID- 18676230 TI - Emergence of resistance to amphotericin B and triazoles in Candida glabrata vaginal isolates in a case of recurrent vaginitis. AB - Emergence of resistance to triazoles and amphotericin B in Candida glabrata vaginal isolates is documented by Etest. During the 18-month follow-up of a case of vaginitis, 14 consecutive isolates of C. glabrata were examined. The isolates exhibited development of in vitro resistance beginning with itraconazole (>32 microg/ml), followed by fluconazole (>256 microg/ml), amphotericin B (>32 microg/ml), and voriconazole (>32 microg/ml). The DNA sequence analyses and finger printing of the isolates strongly suggest that our patient remained colonized with a single strain. The report underscores the propensity of C. glabrata to acquire resistance during antifungal therapy and the importance of susceptibility testing in the management of infections caused by this species. PMID- 18676229 TI - Clinical features of bacterial meningitis in Italy: a multicenter prospective observational study. AB - We carried out a prospective observational study on clinical features of bacterial meningitis. Between October 2002 and June 2005, 322 adult bacterial meningitis cases in 49 infectious disease wards in Italy (MENTORE study group) were enrolled in the study. 133 cases were due to Streptococcus pneumoniae, 44 to Neisseria meningitidis and 145 to other microorganisms. A high SAPS score and coma on admission, as well as need for mechanical ventilation, were more frequent in the pneumococcal meningitis group. Neurological impairment was present in 151 out of 311 patients, and was more frequent in pneumococcal meningitis. A single antibiotic was employed in only 90 of 315 cases; a combination of ceftriaxone and ampicillin was the most frequently administered treatment. Ceftriaxone was also the single most used drug. Adjunctive treatment with steroids was administered in 210 out of 303 patients for a median duration of 7 days. Median duration of fever was 4 days, and median hospital stay was 16 days; hospitalization was significantly longer in the pneumococcal meningitis group. At discharge, neurological impairment was still present in 59 (21%) of 277 patients. Twenty (6.9%) out of 289 patients died during hospitalization. Distribution of adverse outcome (death and neurological impairment) in patients treated with or without steroids and within different time zones between onset of symptoms and commencement of antibiotics was studied; a trend toward a worse prognosis was seen in patients treated more than 24 hours after onset of the disease.In our study, infectious disease clinicians made extensive use of steroids as adjuvant therapy for bacterial meningitis, even in absence of detailed national and local guidelines. Mortality seemed to be lower in comparison with the literature. PMID- 18676231 TI - A prospective, open-label noncomparative study with piperacillin-tazobactam monotherapy as management of fever in patients with acute leukemia. AB - We evaluated the efficacy of piperacillin-tazobactam monotherapy as empiric therapy of fever in acute leukemia patients in a total of 80 consecutive febrile episodes. The overall success rate was 75% with success without modification in 34% (afebrile at 72 h) and an overall death rate of 10%. No significant differences were seen in correlation between clinical outcome and phases of underlying disease. The success without modification was higher in patients with fever of unknown origin (FUO) than in those with documented infections (47% and 25% respectively). There were no significant differences in correlations between clinical response and degree of neutropenia. Our study suggests that empirical first-line monotherapy with piperacillin-tazobactam may be a reasonable option in patients with acute leukaemia, although in documented infections the response is frequently inadequate. PMID- 18676232 TI - Postoperative chemotherapy in resected gastric cancer: results of a single center experience. AB - Gastric cancer remains a major health problem despite its decline in incidence in Western countries. Although radical surgery represents the primary curative option for gastric cancer patients, most of them relapse and die due to their disease despite an R0 resection. At present the routine use of postoperative adjuvant therapy to reduce disease recurrence is still considered an investigational approach. Out of a total of 275 patients (stage IB through IV M0 AJCC/UICC) who underwent surgery for gastric cancer at our Surgery Unit between 1993 and 2001, 156 were eligible for adjuvant chemotherapy, of whom only 52 accepted to undergo this treatment. This group of patients was retrospectively compared with a control group (1:2) and overall survival was assessed using hazard ratio and Kaplan-Meier estimates. Five-year survival was 40% in the chemotherapy group and 37.8% in the group which underwent surgery alone. Indeed, chemotherapy did not reduce the risk of death (HR 0.87, 95% CI = 0.57-1.34, p=0.54). Serosal involvement and the invasion of more than 6 lymph nodes were the main independent prognostic factors identified by multivariate analysis. The current study did not show a clear advantage of chemotherapy over surgery alone. However, our results can help to define strategies for future clinical trials with the use of new regimens based on more effective and less toxic drugs. PMID- 18676233 TI - FOLFOX2 in the treatment of advanced colorectal cancer: a comparison between elderly and middle aged patients. AB - The effectiveness of chemotherapy in elderly patients is still a matter of debate. We analyzed the toxicity and efficacy of the original FOLFOX2 regimen in middle aged and elderly patients affected by metastatic colorectal cancer. Consecutive patients with metastatic CRC and measurable disease were eligible. Seventy-eight partially pretreated patients were enrolled: 58 patients were defined as middle aged (<70 years) and 20 were elderly patients (>70 years). Elderly patients in comparison to middle-aged patients in a higher percentage were males. No significant differences were found in hematological and non hematological toxicity between the two groups. No significant differences were found in the response rates, time to progression (5.9 vs. 6.0 months respectively), or median overall survival (20.9 and 21.8 months, respectively) between middle aged and elderly patients. The FOLFOX2 regimen provides equivalent feasibility, efficacy, and survival gain in middle-aged and in elderly patients with metastatic CRC. PMID- 18676234 TI - Weekly docetaxel as salvage therapy in patients with gemcitabine-refractory metastatic pancreatic cancer. AB - Gemcitabine-based chemotherapy provides very limited disease control in the treatment of advanced pancreatic cancer. Approximately half of the patients failing upfront treatment present good performance status and are willing to undergo further treatment. Docetaxel activity against pancreatic cancer is reported both in the preclinical and clinical setting. Between November 2004 and November 2005, 10 patients (median age 59; median KPS 80) with metastatic pancreatic adenocarcinoma, progressive disease after gemcitabine-containing chemotherapy, KPS >50, adequate organ function, were treated with weekly docetaxel at 30 mg/m-(2) until progressive disease. Docetaxel dose intensity was 100% of the intended dose. No grade >2 toxicity was observed. No objective response to treatment was obtained. Median progression-free survival was 1.5 months (range 1-3.5 months); median survival was 4.0 months (range 2.0-7.5). Weekly administration of single-agent docetaxel does not seem to have any activity in the treatment of gemcitabine-resistant metastatic pancreatic cancer. PMID- 18676235 TI - Primary ovarian lymphoma: report of two cases and review of the literature. AB - Ovarian lymphoma is a rare entity. Clinicians should be aware of its clinical manifestations and management since surgery alone often is not adequate treatment. Here, we present two cases of ovarian lymphoma and discuss what is known about ovarian lymphoma. PMID- 18676236 TI - Docetaxel therapy for advanced hepatocellular carcinoma developed in healthy liver: report of three cases. AB - Systemic chemotherapy is generally ineffective in patients with advanced hepatocellular carcinoma (HCC). This could be partly explained by the frequent underlying cirrhosis, which induces serious toxicity requiring dose attenuation or drug discontinuation. We present observations of three patients with HCC developed in healthy liver and treated with docetaxel (100 mg/m(2) every 3 weeks in one patient; 30 mg/m(2) weekly, three times every 4 weeks in two patients). An objective partial response with long-term survival was obtained in all cases without severe toxicity. These results suggest that chemotherapy, and especially docetaxel, could be safe and effective in patients with HCC developed in healthy liver, and should be assessed in specific trials. PMID- 18676237 TI - In vitro antifungal activity of sertaconazole nitrate against recent isolates of onychomycosis causative agents. PMID- 18676238 TI - Post-traumatic osteomyelitis due to Aspergillus flavus successfully treated with voriconazole: a case report. PMID- 18676239 TI - [Effect of a compound Danshen preparation in preventing long-term gastric lipid emulsion administration-induced nonalcoholic steatohepatitis in rats]. AB - OBJECTIVE: To investigate the preventive effects of a compound Danshen preparation (DSC) on long-term gastric lipid emulsion administration-induced nonalcoholic steatohepatitis in rats. METHODS: Twenty-seven 3-month-old SD rats were randomized equally into 3 groups and subjected to daily intragastric administration for 20 weeks of distilled water (control), lipid emulsion at 5 ml/kg (model group), and lipid emulsion plus DSC at 5.0 g/kg (DSC treatment group). After blood glucose (BG) determination, the rats were sacrificed for measurement of serum TC, TG, HDL-c, AST, and ALT, and the liver was weighed and pathologically examined. RESULTS: Compared with the control group, the rats in the model group showed significantly increased BG, TC, LDL-c, arteriosclerosis index (AI), AST, ALT, liver weight, and liver index (P<0.01) and decreased HDL-c (P<0.01), while TG remained unchanged. Fatty degeneration, hydropic degeneration and necrosis with inflammatory cell infiltration were observed in the liver of the rats in the model group. Compared with the model group, the rats in DSC groups showed decreased BG, AI (P<0.01), liver weight, liver index, AST, and ALT (P<0.05) and increased HDL-c, with milder pathological changes in the liver. CONCLUSIONS: Long-term gastric perfusion of lipid emulsion causes lipid metabolic disorder and nonalcoholic fatty liver disease in rats characterized by increased TC and decreased HDL-c. DSC can significantly increase HDL-c and provide partial protection of the liver against the damages by the lipid emulsion. PMID- 18676240 TI - [Psychological stress in the earthquake survivors: the psychological aftermath of the Wenchuan earthquake]. AB - OBJECTIVE: To investigate the psychological stress in the survivors of the May 12, 2008 Wenchuan earthquake transferred to Guangzhou for treatment, and explore the association between the psychological stress and the physical injuries related to the earthquake to promote both the physical and psychological recoveries of the survivors. METHODS: Fifty-five earthquake survivors, including 27 with 28 without physical injuries, were transferred in our hospital 1 week after the earthquake. The Posttraumatic Stress Disorder (PTSD) Checklist-Civilian version (PCL-C) and the Self Reporting Questionnaire (SRQ) were used to investigate the psychological stress in these survivors. RESULTS: The earthquake caused serious psychological stress in these survivors, and 43.63% of the survivors had high PCL-C scores (50 or above) and 78.18% had high SRQ scores (over 7), with 41.82% having high scores for both PCL-C and SRQ. The survivors with physical injuries generally had high PCL-C and SRQ scores (49.780-/+14.503 and 12.110-/+4.619, respectively), which, however, were not statistically different from the scores of the survivors without injuries (44.460-/+13.206 and 10.460-/+5.000, respectively, P>0.05). The injured survivors showed high scores for the subscales of persistent avoidance symptoms and of anger and irritability, in which the item scores for "efforts to avoid activities or places that arouse recollections of the earthquake" and "feeling of detachment or estrangement from others" were significantly different between the survivor with physical injuries and those without (P=0.034 and 0.018, respectively). CONCLUSION: The Wenchuan earthquake causes serious psychological stress in the survivors, and the wounded are more inclined to have persistent avoidance symptoms and enhanced irritability. In addition to the care of the physical injuries, psychological counseling and social support may help ameliorate the psychological trauma in the survivors and reduce the incidence of PTSD. PMID- 18676241 TI - [Optimization of tomato genetic transformation, kanamycin-resistant screening and seed selection]. AB - OBJECTIVE: To optimize the process of tomato genetic transformation, screening and seed selection using multiepitope antigenic gene (MAG) and truncated major surface antigen 1 (tSAG1) of Toxoplasma gondii as the target insert genes. METHODS: Tomato high-frequency regeneration system was optimized with different choices of media and explants. The genetic transformation procedure was optimized using different tomato cultivars, explants, culture temperatures, media and acetosyringone (AS) supplements. Three concentrations of kanamycin were utilized for resistant selection of the transgenic candidate roots. The selected lines were trained, transplanted to soil and grown in a greenhouse till maturity. Sterile seeding using kanamycin-incorporated medium was conducted for screening transgenic tomato generations. RESULTS AND CONCLUSION: Cotyledons were better than hypocotyls as the regeneration explants. The regeneration rate of cotyledons reached 98% (59/60) using the optimized regeneration medium ZB3. The culture medium and temperature were the key factors for tomato transgenic shoot induction. The number of transgenic buds increased significantly at the appropriate temperature condition (23-/+1 degrees celsius;), and AS of 100 micromol/L in the medium before inoculation also significantly raised transformation rate. The budding rate of Zhongshu No.5 cotyledons was 35% (28/81) using the medium ZB2 under (23-/+1) degrees celsius;. Kanamycin at 80 mg/L was optimal for transgenic plantlet rooting selection with the rooting rate of 48% (31/65). 117 transgenic lines were obtained. Non-transgenic tomato plant growth, especially the root and elongation, was inhibited obviously with kanamycin at 100 mg/L or above, and the roots became purple and lacked lateral roots. The transgenic tomato seeds could be selected effectively with kanamycin at 150 mg/L. PMID- 18676242 TI - [Neural stem cell transplantation and nerve growth factor promote neurological recovery in rats with ischemic stroke]. AB - OBJECTIVE: To observe the effect of neural stem cell (NSC) transplantation and nerve growth factor (NGF) on neurological function recovery in rats with ischemic stroke induced by transient middle cerebral artery occlusion (MCAO). METHODS: Cultured hippocampal NSCs isolated from neonate rats were labeled with BrdU. Sixty-four rats subjected to transient MCAO were randomized equally into 4 groups, namely group A (MCAO model group), group B (model group with NGF treatment, group C (model group with NSC transplantation), and group D (model group with both NGF and NSC transplantation). The neurological deficits of the rats were evaluated with neurological severity score (NSS) after the treatment, and the brain tissues were examined with immunohistochemical and immunofluorescent staining for BrdU and nestin expression. RESULTS: The NSS of rats in group D 2 and 4 weeks after transplantation was significantly lower than that in the other 3 groups (P<0.05), and the scores in groups B and C were significantly lower than those in group A (P<0.05). The number of BrdU- and nestin-positive cells was significantly greater in group B than in group A (P<0.05), and BrdU-positive cells were markedly more numerous in group D than in group C (P<0.05). CONCLUSION: Both NSC transplantation and NGF treatment can obviously improve the neurological function of rats after MCAO, but their combined use shows stronger effects. NGF can promote autologous NSC activation and proliferation, with also stimulatory effect on the proliferation of transplanted NSCs. PMID- 18676243 TI - [1,2,6-tri-O-galloyl-beta-D-glucopyranose inhibits gp41-mediated HIV envelope fusion with target cell membrane]. AB - OBJECTIVE: To observe the inhibitory effect of 1,2,6-Tri-O-galloyl-beta-D glucopyranose (TGGP) from Balanophora japonica Makino on human immunodeficiency virus (HIV) entry into the host cells and explore the mechanisms. METHODS: TGGP was purified from Balanophora japonica Makino by n-hexane and ethyl acetate extraction and column chromatography. The inhibitory activity of TGGP on HIV gp41 six-helix bundle formation was measured with ELISA, N-PAGE and SE-HPLC, and the inhibitory effect of TGGP on HIV envelope grlycoprotein-induced cell-cell fusion was detected using a non-infectious cell-based assay. RESULTS: TGGP inhibited HIV gp41 six-helix bundle formation, with an IC50 of 1.37-/+0.19 microg/ml as determined by ELISA, and this activity was further confirmed by N-PAGE and SE HPLC. TGGP at 25 microg/ml significantly inhibited syncytium formation between the effector (CHO-WT) and the target (MT-2) cells. CONCLUSION: The HIV transmembrane subunit gp41 mediates the entry of HIV into the target cells. TGGP can inhibit HIV fusion and entry into the target cells by inhibiting the formation of gp41 six-helix bundles, suggesting the potential of TGGP as a microbicide to prevent sexual transmission of HIV. PMID- 18676244 TI - [Effect of adenoviral vectors on the differentiation potential of human bone marrow mesenchymal stem cells]. AB - OBJECTIVE: To investigate the effect of adenoviral vector infection on the differentiation potential of human bone marrow mesenchymal stem cells (hMSCs). METHODS: The third-passage hMSCs were infected with the recombinant adenovirus expressing green fluorescence protein (GFP) for 2, 4, 8 and 16 days. RT-PCR was used to detect the mRNA expression of endodermal marker CYP 51, mesodermal marker SM22alpha, ectodermal marker nestin, pluripotent marker oct-4 and the alternative splicing factor nPTB. Seven days after adenovirus infection, the hMSCs were cultured in the presence of adipogenic agents for 14 days, and the adipose cells differentiated from hMSCs were detected with oil red O staining. RESULTS: Compared with normal hMSCs, the cell infected with the adenovirus for 2, 4, 8 and 16 days showed no obvious down-regulation of CYP51, SM22alpha, nestin, OCT4 or nPTB. The hMSCs 7 days after adenovirus infection were induced to differentiate into adipose cells, with a similar differentiation rate to that of normal hMSCs. CONCLUSION The differentiation potential of hMSCs is not affected by adenovirus infection, suggesting that adenovirus can be used as the gene delivery vector in MSC differentiation studies. PMID- 18676245 TI - [Mechanism of rat sciatic nerve regeneration induced by human hair keratin]. AB - OBJECTIVE: To evaluate the effect of human hair keratin (HHK) in peripheral nerve repair and explore the mechanism of sciatic nerve regeneration. METHODS: Rat models of sciatic nerve damage was established by creating a 10-mm gap in the sciatic nerve, which was bridged with a HHK implant. Histological examinations of the nerve tissues were performed at different time points after the surgery. RESULTS: During the period from 2 days to 2 weeks following HHK implantation, Schwann cells were found to undergo dedifferentiation and proliferate along the HHK implant. Three weeks after HHK implantation, numerous macrophages and megakaryocytes occurred around the HHK, and a large quantity of regenerated Schwann cells aligned in orderly fashion was seen between the fine filaments of partially degraded HHK, where axons and capillaries were also observed. Six weeks later, massive nerve fibers and capillaries developed around the HHK, and at 9 weeks, the HHK implant was substantially degraded and numerous regenerated nerve fibers occurred characterized by obvious epineurium and perineurium. Till 12 weeks after HHK implantation, HHK was almost completely degraded and replaced by the newly regenerated nerve fibers that had grown across the nerve defect. CONCLUSIONS: HHK is an ideal material for nerve injury repair. Apocytosis plays a key role in the differentiation process of highly differentiated Schwann cells into immature Schwann cells following nerve injury. As a protective mechanism, the axons undergo enclosure and dissociation following injuries, and the intact axons give rise to growth cones that extend fibers of growing buds to competitively bind the one or more Schwann cells, but only one such but finally develops into a complete axon. The nerve fiber barrier membrane is derived from the capillary menchymal stem cells and the outmost vascular barrier membrane. The regeneration of the Schwann cells, axons and the nerve membrane is the result of self-organization through a well synchronized and coordinated mechanism. PMID- 18676246 TI - [Detection of serum PICP and PINP and COL1alpha1 DNA sequence analysis in patients with arsenic poisoning in Dazhuyuan Town, Ankang area in southern Shaanxi Province]. AB - OBJECTIVE: To investigate type I collagen metabolism in patients with arsenic poisoning in Dazhuyuan town, Ankang area in southern Shaanxi province known for high-level exposure to arsenic (As), fluoride (F) and selenium (Se). METHODS: This case-control study involved 29 patients with arsenic poisoning from Dazhuyuan, a town in Ankang area of Shaanxi province affected by coal-borne endemic arsenicosis. Thirty individuals without arsenic poisoning from Chang'an county of Xi'an city were included as the control group. The serum concentrations of As and Se were determined using a graphite furnace atomic absorption spectrophotometer, and the levels of F detected with a fluoride ion-selective electrode. Enzyme-linked immunosorbent assay (ELISA) was used for assessing serum contents of PICP and PINP, and the sequence of COL1alpha1 gene was analyzed with ABI sequence analyzer. RESULTS: The contents of PICP and PINP were similar between the two groups, which had also identical COL1alpha1 DNA sequences. After controlling for all the other factors, an inverse correlation was noted between PICP and serum F contents (r=-0.4, P<0.05) and between PINP and hair F content (r=-0.5, P<0.05), but a positive correlation was found between PINP and hair Se content (r=0.5, P<0.05). CONCLUSION: Hair F and Se levels are the major factors affecting PINP synthesis, and increased hair F level is associated with elevated PINP content, whereas hair Se level is inversely correlated to PINP content. But in the patients with arsenic poisoning from Dazhuyuan town, the PICP, PINP contents and COL1alpha1 DNA sequence show no significant alterations, suggesting that hair Se increment abolishes the effect of hair F increment in inducing the decrease in PINP content. PMID- 18676247 TI - [Roles of p38 and ERK signaling pathway in cell apoptosis induced by UVB irradiation]. AB - OBJECTIVE: To explore the involvement of p38 and ERK signal transduction pathways in UVB-induced cell apoptosis. METHODS: HaCat cells were exposed to UVB irradiation for 1, 3, 5, 10, and 15 min, respectively, after which the cell survival was assessed using MTT assay, and the cell apoptosis observed under fluorescent microscope with Hoechst staining. Western blotting was used to examine the possible signal transduction pathway involved in the cell apoptosis following the exposures. RESULTS: For the same incubation time following the exposure, the cell survival rate decreased gradually with the increase of UVB irradiation dose. At a fixed UVB irradiation dose, prolonged cell incubation following the exposure resulted in decreased cell survival rate, which, however, began to increase after the minimum rate was reached. At different UVB doses, cell exposure for 5 min caused the highest cell apoptosis rate, which peaked at 12 h during the post-irradiation incubation. The expressions of p38 and p53 were significantly decreased while p44/42 expression remained unchanged in the exposed cells. CONCLUSION: UVB irradiation can induce growth inhibition and apoptosis of HaCat cells in a dose- and time-dependent manner, and p38 pathway other than ERK pathway is probably involved in UVB-induced cell apoptosis. PMID- 18676248 TI - [Guling tablets promotes the proliferation and mineralization and regulates OPG/RANKL/M-CSF expression of osteoblasts via the p38 MAPK pathway in vitro]. AB - OBJECTIVE: To investigate the effect of Gulingpian (GLP, or Guling tablets) on the proliferation and mineralization of MG-63 cells and the involvement of p38 MAPK pathway in such processes. METHODS: MG-63 cells were cultured in the presence of p38 inhibitor SB203580 and or serum from rats fed with normal saline or GLP. The expression of the OPG, RANKL and M-CSF mRNA, cell proliferation and formation of mineralized bone nodule, the total protein of p38 and phosphorylated p38, were observed or quantitatively measured. RESULTS: Treatment with GLP serum increased the level of p38 phosphorylation but did not affect the total p38 expression. GLP serum significantly promoted MG-63 cell proliferation and differentiation, resulting also in up-regulated OPG and down-regulated RANKL mRNA expressions without obvious alteration in M-CSF expression. These effects were blocked by the specific p38 inhibitor SB203580. CONCLUSION: GLP promotes the proliferation and differentiation of osteoblasts and regulates OPG/RANKL expressions in vitro, which are mediated probably via the p38 MAPK pathway. PMID- 18676249 TI - [Cloning of Rcet3, a novel gene related to family 2 cystatins]. AB - OBJECTIVE: To clone the full-length Rcet3 gene, a novel gene related to family 2 cystatins, from mouse testis or other tissues. METHODS: Rcet3 gene was cloned using digital differential display (DDD) and RT-PCR was performed for cloning the full-length Rcet3 gene from adult mouse testis cDNA library with sequence analysis. RESULTS: Rcet3 cDNA was 610 bp in length, consisting of 4 exons to encode a protein with 140 amino acid residues. The encoded protein contained a potential signal peptide and a cystatin domain, but lacked critical consensus site important for cysteine protease inhibition. These characteristics could be seen in the Cres subgroup related to the family 2 cystatins. Rcet3 was specifically expressed in adult mouse testis, epididymis and the cerebrum, but at higher levels in the testis than in the epididymis and cerebrum. CONCLUSION: Rcet3 may be a new member of Cres subgroup of family 2 cystatins. PMID- 18676250 TI - [Protective effects of estrogen on mitochondria in human umbilical vascular endothelial cells]. AB - OBJECTIVE: To investigate the protective effects of estrogen on the mitochondria in human umbilical vascular endothelial cells (HUVECs). METHODS: HUVECs were exposed to H2O2 at 250 micromol/L for 4 h with or without pretreatment with 17 estradiol (E2) and ICI182780. Complex IV activity of the cells was measured with chromometry, and 2, 7-dichlorofluorescein diacetate (DCFH-DA) was used to determine intracellular reactive oxygen species (ROS). Intracellular adenosine triphosphate (ATP) level was quantified with a luciferin- and luciferase-based assay. RESULTS: Compared to the blank control group, H2O2 caused a decrease in complex IV activity, intracellular ATP level, and the cell viability, but elevated intracellular ROS. E2 pretreatment of cells significantly attenuated these effects of H2O2 exposure. ICI182780 administered prior to E2 pretreatment antagonized the protective effects of E2 against H2O2 exposure. CONCLUSION: E2 offers mitochondrial protective effects on HUVECs, which is mediated by the estrogen receptors. PMID- 18676251 TI - [BRCA1 regulates progesterone receptors A and B protein expressions in breast cancer cells in vitro]. AB - OBJECTIVE: To study the regulatory role of BRCA1 in the expression of progesterone receptors A and B (PRA and PRB) in breast cancer cells. METHODS: Breast cancer MCF-7 cells were transfected with pFlag-CMV2-BRCA1 wt plasmid containing a full-length BRCA1 cDNA or with BRCA1-specific siRNA via lipofectamine 2000 to induce overexpression or suppressed expression of BRCA1, respectively. Twenty-four hours after the transfection, the cells were incubated in fresh culture medium containing 100 nmol/L progesterone for 24 h. The total RNA extract or whole cell lysate was prepared for detecting BRCA1, PRA and PRB expressions using RT-PCR and Western blotting. RESULTS: The protein expressions of PRA and PRB were significantly decreased whereas their mRNA expressions remained unchanged in MCF-7 cells overexpressing BRCA1. In MCF-7 cells with BRCA1 knock-down, in contrast, the PRA and PRB protein expressions were markedly increased. CONCLUSION: In breast cancer cells, exogenous and endogenous BRCA1 can both down-regulate the expressions of PRA and PRB at the protein level. PMID- 18676252 TI - [Analysis of the allergenicity and immunogenicity of Psilogramma menephron allergen]. AB - OBJECTIVE: To analyze the allergenicity and immunogenicity of Psilogramma menephron allergen so as to provide the basis for preparing recombinant and standardized allergen vaccines of Psilgramma menephorn. METHODS: The extracts of Psilgramma menephorn were analyzed by SDS-PAGE, and the allergenicity and immunogenicity of the extracts were tested with 9 sera from allergic patients by means of immunoblotting. RESULTS: More than 20 allergen proteins were separated from the extract of Psilgramma menephorn by SDS-PAGE, with the relative molecular weight ranging from 12,000 to 128,000. The relative molecular weight of the allergenic proteins were 74,000 (88.9%), 66,000 (22.2%), 49,000 (22.2%), 36,000 (77.8%), or 25,000 (33.3%), and those of the immunogenic proteins were 79,000 (33.3%), 74,000 (66.7%), 66,000 (22.2%), 49,000 (22.2%), 36,000 (44.4%), or 25,000 (55.6%). CONCLUSION: The relative molecular weight of the major allergenic proteins of Psilgramma menephorn are 74,000 and 36,000, and 74,000 and 25,000 for the major immunogenic proteins. These proteins constitute the major allergenic components for diagnosis and specific treatment of Psilgramma menephorn allergy. PMID- 18676253 TI - [Expression of osteopontin mRNA in oral squamous cell carcinoma and normal oral mucosa]. AB - OBJECTIVE: To investigate osteopontin (OPN) mRNA expression in oral squamous cell carcinoma (OSCC) and normal oral mucosa tissues. METHODS: Differential OPN gene expression were detected in 30 cancerous tissues and their paired normal tissues using real-time reverse transcription-PCR (real-time RT-PCR), and the data were analyzed statistically. RESULTS: Real-time RT-PCR results demonstrated that the relative expression level of OPN mRNA in the cancerous tissues were significantly higher than that in paired normal samples (4.17-/+0.51 vs 0.97-/+0.12, P<0.001), showing a 4.3-fold up-regulation. In the 30 OSCC specimens, OPN mRNA expression in the OSCC of histological grades I showed a 3.1-fold down-regulation, significantly lower than the expression in grade II/III tumors (2.16-/+0.17 vs 6.80-/+0.72, P<0.05); its expression was significantly lower in early stage than in advanced stage OSCCs (2.34-/+0.17 vs 4.73-/+0.35, P<0.05). In cases of cervical lymph node metastasis, the expression was significantly higher than that in cases without lymphatic metastasis (6.38-/+0.56 vs 2.89-/+0.32, P<0.05). CONCLUSION: OPN mRNA overexpression may play an important role in OSCC carcinogenesis and can be a potential target for OSCC therapy. PMID- 18676254 TI - [Immunological activity of Streptomyces polysaccharide in normal and immunosuppressed mice]. AB - OBJECTIVE: To investigate the immunological activity of Streptomyces polysaccharide (SMP) on normal and immunosuppressed mice. METHODS: The effect of SMP on the proliferating activity of normal mouse splenocytes was tested in the mixed lymphocyte culture, and the changes of peripheral blood T lymphocytes were evaluated with acid a-naphthyl acetate esterase (ANAE) method. The ratio of Lyt2+ and L3T4+ T cell subsets was measured by flow cytometry. RESULTS: SMP stimulated obvious proliferation of mixed lymphocytes, showed protective effects on T lymphocyte and increased the ratio of Lyt2+ and L3T4+ cell subsets to nearly normal level in immunosuppressed mice. CONCLUSIONS: SMP can regulate the immune function in mice. PMID- 18676255 TI - [Effect of vascular endothelial growth factor gene transfer on proliferation and metabolism of human bone marrow stromal cells in vitro]. AB - OBJECTIVE: To investigate the effect of vascular endothelial growth factor 165 (VEGF165) gene transfer on the proliferation and metabolism of human bone marrow stromal cells (hBMSCs) in vitro. METHODS: hBMSCs were divided into 3 groups and subjected to adenovirus mediated VEGF165 gene transfection, transfection with empty adenoviral vector, or left untreated (control). MTT assay and flow cytometry were performed to analyze the proliferation of the cells after corresponding treatments. The third passage of hBMSCs (2x10(4)/ml), after corresponding transfection procedures, were cultured in conditional medium and tested for ALP content 2, 4 and 6 days after the transfection. Also at 3, 5 and 7 days after the transfection, the cells were examined for osteocalcin (C) and laminin (LN) contents. RESULTS: The number of cells in each group increased with the culture time without obvious differences in the optical density. No significant differences were noted between the 3 groups in the percentage of G1 phase cells or in the proliferation index (PrI) (P>0.05), but compared with the nontransfected and the empty vector-transfected cells, the cells with VEGF165 gene transfection had significantly higher ALP, OC and LN contents (P<0.05). CONCLUSION: VEGf165 gene transfer does not obviously affect the proliferation of cultured hBMSCs, but can increase the cellular secretion of AIP, C and LN, suggesting that VEGF165 promotes the differentiation of hBMSCs into osteoblasts in vitro. PMID- 18676256 TI - [Expression of p38MAPK in the hippocampal CA1 region of rats with Abeta25-35 induced Alzheimer disease]. AB - OBJECTIVE: To investigate the changes of p38MAPK expression in a rat model of Alzheimer disease (AD). METHODS: Seventy-two adult SD rats were randomized equally into 4 groups, and a single-dose injection of Abeta25-35 (dementia group), normal saline (saline group), SB203580 (inhibitor group), or DMSO (inhibitor control group) was administered into the lateral cerebral ventricle. Y maze tast was performed to evaluate the behavioral changes of the rats after the injections, and on days 4, 7 and 14 after the injection, p38MAPK expression in the hippocampal CA1 area was measured by means of immunohistochemistry. RESULTS: On days 7 and 14 following Abeta25-35 injection, the training times, error number and total reaction time were significantly higher in dementia group than in saline group (P<0.05), but all these indices were significantly lowered in the inhibitor group as compared with the dementia group (P<0.05). Immunohistochemistry revealed obvious p38 expression in the dementia group 4 days after Abeta25-35 injection, which increased significantly with the passage of time (P<0.01). The gray scale in the inhibitor group was significantly higher than that in the dementia group (P<0.01). CONCLUSION: p38MAPK activation in the hippocampal CA1 area is an event that persists during the entire course of Abeta25-35-induced AD in rats, and the inhibitor SB203580 prevents p38MAPK expression and improves the learning and memory abilities of the rats. PMID- 18676257 TI - [Development of the recombinant SAG1 antigen of Toxoplasma gondii by high-density fermentation and identification of its immunoreactivity]. AB - OBJECTIVE: To develop a technology for production of recombinant SAG1 of Toxoplasma gondii (T.g) in batches. METHODS: The rSAG1 of T.g was expressed in E.coli by high-density fermentation and purified by Sephadex G-75 column chromatography after Ni-NTA agarose at native condition. The activity of rSAG1 and its efficacy in T.g diagnosis were identified by Western blotting and ELISA, respectively. RESULTS: The optical density (OD) of the bacteria reached 20.21 after induction, and 300 g bacteria were harvested from 11.5 L broth. The rSAG1 was highly expressed in E.coli as a fusion protein, accounting for about 25.82% of the total bacterial protein. The purity of rSAG1 reached 98.54% after purification by Ni-NTA combined with Sephadex G-75 column chromatography. Western blotting revealed a distinct band reacting with the sera of rabbits vaccinated by T.g. Twenty-four of the 25 sera of mice infected with T.g and 36 of the 38 sera of human subjects with IgG antibody against T.g were detected by rSAG1-ELISA. CONCLUSION: A large-scale production of immunoreactive SAG1 of T.g is developed by high-density fermentation and purification with Ni-NTA combined with Sephadex G-75 column chromatography. PMID- 18676258 TI - [Correlations between preS1-antigen, HBV-DNA and HBV serum markers in patients with chronic hepatitis B]. AB - OBJECTIVE: To study the correlations between preS1 antigen, HBV-DNA and hepatitis B virus (HBV) serum markers in patients with chronic hepatitis B. METHODS: The HBV markers, preS1 antigen and HBV-DNA were determined using enzyme- linked immunosorbent assay and quantitative PCR in 1158 patients with chronic hepatitis B. RESULTS: In these patients, the HBV-DNA positivity rate was 68.9%, significantly higher than preS1 antigen positivity (54.8%, chi2=53.24, P<0.005). The positivity rates of both HBV-DNA and PreS1-antigen were significantly higher in HBeAg-positive patients than in HBeAg-negative patients (P<0.005). The coincident rates of preS1-antigen and HBeAg with HBV-DNA were 56.9% and 63.3%, respectively. PreS1 antigen had higher sensitivity but lower specificity than HBeAg. The detection rates of preS1 antigen and HBeAg increased with the level of HBV-DNA, and preS1 antigen positivity was higher than that of HBeAg in patients with low HBV-DNA levels. CONCLUSION: Detection of HBV serum markers along with preS1 antigen and HBV-DNA may help assess the status of viral replication and therapeutic efficacy in patients with chronic hepatitis B. PreS1 antigen may serve as an auxiliary indicator in HBeAg-negative cases or when HBV-DNA detection is impossible. PMID- 18676259 TI - [Familial aggregation and sibling heritability in Kashin-Beck disease]. AB - OBJECTIVE: To explore the family aggregation and the role of hereditary factors in the pathogenesis of Kashin-Beck disease (KBD). METHODS: With a stratified sampling method, the general population of 14 villages of Linyou County were studied, from whom 225 KBD probands were selected using systematic sampling at the rate of (1/2). A total of 304 siblings of the probands were ascertained, and in these sibling pairs, the segregation ratio, heritability in different age groups and weighted mean heritability of the siblings were estimated using the methods of Li-Mantel-Grart and Falconer. RESULTS: The KBD distribution scope in the KBD families exceeded the scope of binomial distribution (P<0.001), suggesting obvious family aggregation. The prevalence rate in the siblings of the KBD pedigree was 19.41% (59/304), significantly higher than that in the 14 KBD villages [10.90% (1180/10823), chi2=21.62, P<0.001]. The segregation ratio and heritability in the siblings of the KBD pedigrees were 0.061 and 28.61%, respectively. CONCLUSION: As a polygenetic inheritance disease, KBD exhibits obvious familial aggregation, and genetic susceptibility accounts for (1/4) of the risk factors for KBD. PMID- 18676260 TI - [Adenovirus-mediated stromal cell-derived factor-1 alpha gene transfer promotes mesenchymal stem cell migration]. AB - OBJECTIVE: To explore the role of stromal-derived factor-1 (SDF-1) in the migration of mesenchymal stem cells (MSCs) and the underlying signal transduction mechanism. METHODS: Rat bone marrow-derived MSCs were infected with 100 ml recombinant adenovirus containing human SDF-1alpha gene (Ad-hSDF-1alpha), and the cell migration changes were observed at 1, 2, and 3 days after the infection. Twelve hours after Ad-hSDF-1alpha transfection, the MSCs in separate cultures were treated with wortmannin (50 nmol/L), LY294002 (10 mmol/L), PD98059 (50 mmol/L), U73122 (10 mmol/L), AMD3100 (0.1 g/L), or verapamil (50 nmol/L), respectively, and the signal transduction pathways involved in MSC migration were analyzed. RESULTS: The MSCs grew in colonies after transfection with Ad-hSDF 1alpha, but this growth pattern was substantially attenuated after treatment with wortmannin, LY294002, PD98059, U73122, AMD3100 and verapamil, among which U73122 and AMD3100 treatments resulted in the most conspicuous inhibitory effects. CONCLUSION: The effect of SDF-1 in promoting MSC migration is related to mitogen activated protein kinase, phosphatidylinositol phospholipase C, and protein kinase signal pathways. PMID- 18676261 TI - [Effect of chronic enhanced external counterpulastion on gene expression profiles of arterial endothelial cells of pigs fed with high-cholesterol diet]. AB - OBJECTIVE: To investigate the effect of chronic enhanced external counterpulastion (EECP) on gene expression profiles of arterial endothelial cells (ECs) of pigs fed with high-cholesterol diet. METHODS: Eight male pigs were fed with high-cholesterol diet for 12 weeks to induce arteriosclerosis and subjected to EECP for accumulative 36 h (2 h every other day for 18 sessions). Another 8 pigs on cholesterol-enriched diet and 6 normally fed pigs served as the arteriosclerosis model group and normal control group, respectively, and the high cholesterol diet was maintained until the end of EECP treatment. The coronary artery was then isolated for transmission electro microscopy, and the abdominal aorta was observed using Sudan III staining. The gene expression profiles in ECs from the thoracic aorta using cDNA microarrays. RESULTS: Macrophages and foam cells were detected beneath the ECs in the coronary artery of pigs in the model group, but not in the other two groups. The ratios of Sudan III-positive area in the celiac aorta were significantly lower in normal control and EECP groups than in the model control group (P<0.05). Compared with the normal control group, the gene expressions of integrins-beta1 and CTGF were up-regulated in the model group. Compared with the model group, the expressions of integrins-beta1, CTGF and VCAM-1 were down-regulated and eNOS up-regulated in EECP group. CONCLUSION: Chronic EECP may reduce endothelial injury, down-regulate the gene expression level of integrin-beta1, CTGF and VCAM-1, lower cholesterol uptake and attenuate arterial endothelial inflammation to protect the pigs fed with high-cholesterol diet from arteriosclerosis. PMID- 18676262 TI - [Mechanism of the effect of Tongluo Recipe against glomerular sclerosis in rats]. AB - OBJECTIVE: To investigate the effects of Tongluo Recipe on the expression of collagen IV (Col IV), fibronectin (FN), laminin (LN), transforming growth factor beta1 (TGF-beta1) in rat renal tissues and explore the mechanism underlying these effects in rats with glomerular sclerosis. METHODS: The pathological changes in the renal tissues of rats with glomerular sclerosis were observed microscopically, and the expressions of Col IV, FN, LN, and TGF-beta1 were detected using immunohistochemical staining and image analysis system. RESULTS: Tongluo Recipe significantly decreased the expressions of Col IV, FN, LN and TGF beta1 in the renal tissue of rats with glomerular sclerosis (P<0.05 or P<0.01) and obviously alleviated the renal pathologies (P<0.01). CONCLUSION: The therapeutic effects of Tongluo Recipe are probably mediated by lowered expressions of Col IV, FN, LN and TGF-beta1. PMID- 18676263 TI - [Cloning and secretory expression of islet neogenesis-associated protein in Pichia pastoris]. AB - OBJECTIVE: To clone the recombinant human islet neogenesis-associated protein (rhINGAP) gene for its secretory expression in Pichia pastoris. METHODS: INGAP gene was amplified with PCR and inserted between Xho I and EcoR I downstream sites of the alpha factor of the recombinant plasmid alpha/pUC18. The fusion gene of alpha factor and INGAP was subsequently inserted between BamH I and EcoR I sites of the plasmid pPIC9K of P. pastoris. After confirmation with restriction enzyme digestion and sequencing, the positive recombinant plasmid that integrated INGAP gene was linearized with Sal I digestion and transformed into the yeast host strain GS115 through electroporation. The yeast transformants that harbored the INGAP gene with high copies were selected with the auxotroph medium and G418, followed then by PCR verification of the positive transformants, from which the expression of recombinant human INGAP was induced with methanol as the only carbone source. The antigenic activity of the desired protein was then detected using Western blotting and enzyme-linked immunosorbent assay (ELISA). RESULTS AND CONCLUSION: The recombinant expression plasmid INGAP/pPIC9K was successfully constructed, and 3 positive transformants were obtained. The expressed protein showed good antigenic activity as confirmed by Western blotting and ELISA. PMID- 18676264 TI - [Autologous bone marrow stem cell transplantation for treatment terminal liver diseases]. AB - OBJECTIVE: To evaluate the effect of autologous bone marrow-derived stem cell (BMSCs) transplantation in the treatment of liver failure and decompensated hepatic cirrhosis. METHODS: Bone marrow was harvested (65-95 ml) from 24 patients in the transplantation group. The BMSCs were isolated and infused into liver or spleen of patients via hepatic or splenic artery. At different time points after the transplantation, the patients' liver function and prothrombin time (PT) were evaluated, and the survival rate and symptoms of the patients were recorded. RESULTS: All the serum biochemical indexes remained stable 2 weeks after the transplantation, and at 4 weeks after transplantation, albumin level increased significantly in comparison with the preoperative level (P<0.05). At 12 weeks, the albumin level further increased (P<0.01) along with Pre-ALB (P<0.01), while total bilirubin, tolal bile acid, PT and fibrinogen were all significantly lowered (P<0.05), and globulin, ALT, and AST remained unchanged (P>0.05). One week after the transplantation, improved appetite was observed in 22 cases (91.67%), and 21 cases (87.5%) showed better physical strength; at 2 weeks, hepatic face improved in 15 cases (62.5%), and spider telangiectasia was significantly reduced in one case; at 12 weeks, the survival rate of the patients was 62.5%, and 9 died or gave up treatment due to chronic liver failure complicated by spontaneous bacterial peritonitis, hepatorenal syndrome, or DIC. No complications associated with the transplantation occurred in these patients. CONCLUSION: BMSC transplantation can significantly improve the liver function of patients with terminal liver disease with good safety and effectiveness. PMID- 18676265 TI - [Cell-free fetal DNA detection in maternal plasma using real-time PCR and cycling probe technology for prenatal screening beta-thalassaemia major]. AB - OBJECTIVE: To analyze cell-free fetal DNA in maternal plasma for prenatal screening of beta-thalassaemia major. METHODS: Six couples undergoing prenatal diagnosis of beta-thalassaemia (gestational age range 23-26 weeks) were enrolled in this study. The husbands were all carriers of the CD17 (A-->T) mutation, and the wives carried another beta-thalassaemia mutation. The allele-specific primers and two fluorescent cycling probes were synthesized for the detection of the CD17 (A-->T) mutation, using FAM and HEX fluorescence labeling, respectively. The cell free fetal DNA in the maternal plasma was detected using real-time PCR, and the fetal genotype was confirmed by cord blood conventional prenatal diagnosis. RESULTS: In the 6 pregnancies, FAM and HEX fluorescent signals were detected in 3 maternal plasma samples; in the other 3 samples, only FAM fluorescent signals were detected, suggesting the absence of paternally derived CD17 (A-->T) mutation. CONCLUSION: Examination of cell-free fetal DNA in maternal plasma using real-time PCR and cycling probe technology can be effective means for prenatal screening of beta-thalassaemia major. PMID- 18676266 TI - [Evaluation of resultant cardiac vector based on voltage distribution of electrocardial field around ex vivo guinea pig heart]. AB - OBJECTIVE: To evaluate the possibility of resultant cardiac vector of ex vivo guinea pig heart. METHODS: ECGs were recorded in 4 directions on every plane, and in each direction 4 points at different distances from the anterior, posterior, right and left of isolated guinea pig ventricles immersed in normal saline and distilled water. RESULTS: Main upward waves of ECGs were recorded at all points in 4 directions at 3 levels in the two media. The voltage distribution of normal electrocardial field of the QRS wave was of all-round outward shape, which did not match the resultant cardiac vectors between the left and right and between the anterior and posterior ventricular walls. CONCLUSION: There are some confusion and contradiction in the resultant cardiac vector based on the voltage distribution of electrocardial field around ex vivo guinea pig heart. PMID- 18676267 TI - [Correlation of HBV genotypes to the therapeutic effect of PEG-interferon and the pathological changes of the liver of chronic hepatitis B patients]. AB - OBJECTIVE: To study the correlation of HBV genotypes to the response to PEG interferon alpha (PEG-IFN) in chronic hepatitis B (CHB) patients. METHODS: Real time fluorescent PCR was used for HBV genotyping in 48 CHB patients, and the therapeutic effects of PEG-IFN and hepatic pathological changes were observed. RESULTS: No obvious differences were noted in ALT and HBV DNA levels or negative rate for HBeAg between patients with genotypes B and C (P>0.05). The sustained response rate was significantly higher in genotype B than in genotype C patients 48 weeks o after the therapy. CONCLUSION: HBV genotype is the main factor for predicting PEG-IFN therapy response in CHB patients, and the sustained response rate is significantly higher in genotype B than in genotype C patients. PMID- 18676268 TI - [Role of bile in rat gastric mucosal injury due to duodenogastric reflux]. AB - OBJECTIVE: To explore the effect of bile in inducing gastric mucosal injury in rats. METHODS: SD rats were divided into 4 groups, namely bile duct ligation group, duodenogastric reflux (DGR) group, DGR plus bile duct ligation group and normal control group. The pathological changes in the gastric mucosa and tight junction 3 months after gastrojejunostomy were observed and compared with the findings in the normal control rats. RESULTS: Compared with the rats in DGR plus bile duct ligation group, the rats in DGR group showed obvious gastric mucosal hyperemia, foveolar hyperplasia and severely impaired tight junction between the gastric mucosal cells. CONCLUSION: Bile plays an important role in gastric mucosal injury due to DGR. PMID- 18676269 TI - [Effects of paroxetine on protein kinase PKA, PKC and CaMKII activity in different brain regions in a rat depression model]. AB - OBJECTIVE: To evaluate the effects of paroxetine on protein kinase PKA, PKC and CaMKII activities in different brain regions in a rat model of depression. METHODS: Thirty-six adult male SD rats were randomized into 6 groups, including one control group (I) and 5 groups of depression model established by forcing the rats to swim for 4 weeks. The 5 depression groups received no treatment (II) or were treated with paroxetine at a single dose (III), for a week (IV), 2 weeks (V) or 4 weeks (VI). The radioactivity of PKA, PKC and CaMKII in the hippocampus and prefrontal cortex was quantitatively measured using a liquid scintillation counter. RESULTS: In the rat hippocampus, PKA and CaMKII activities were significantly lower in groups II, III, IV, and V than in groups I and VI (P<0.01 or P<0.05), but comparable between groups VI and I (P>0.05). PKC activity was significantly lower in group II than in group I (P<0.01), but showed no significant difference between the paroxetine-treated groups and group I (P>0.05). In the prefrontal cortex, the activity of PKA in groups I, II, III, and IV was similar (P>0.05), but all significantly lower than that in groups V and VI (P<0.01). PKC activity was significantly higher in groups II and III than that in group I and other paroxetine-treated groups (P<0.01), and similar between groups IV and I (P>0.05); groups V and VI had significantly lower PKC activity than group I (P<0.01). Group I had the highest CaMKII activity among the groups (P<0.01). CONCLUSION: Chronic administration of paroxetine can reverse chronic stress-induced inhibition of PKA, PKC and CaMKII activity in rat hippocampus, while the effects of paroxetine on the protein kinases can be more complex in prefrontal cortex. PMID- 18676270 TI - [Changes of plasma cross-linked D-dimer and neuron-specific enolase in patients with cerebral infarction]. AB - OBJECTIVE: To explore the effect of plasma cross-linked D-dimer (XDP) and neuron specific enolase (NSE) on the infarct volume and neurological function deficit in patients with cerebral infarction (CI). METHODS: Plasma XDP and NSE levels were measured in 66 CI patients on the different days after onset and also in 46 normal individuals, and the changes in XDP and NSE levels were analyzed in the CI patients with different infarct volume and neurological function deficit scores. RESULTS: Within 48 h following CI onset, plasma XDP and NSE levels increased significantly (P<0.01) and reached the highest levels on day 5 (P<0.001), recovering the normal level till day 18. Plasma XDP and NSE levels were significantly higher in patients with moderate to large infarct volume and in those with moderate to severe neurological deficits than in those with small infarct volume and mild neurological deficits (P<0.001). CONCLUSION: Increment of XDP and NSE levels is an important pathological process in CI in close relation to the infarct volume and neurological deficits. XDP and NSE may serve as reliable indices for early diagnosis and evaluation of CI. PMID- 18676271 TI - [Refined mapping of the loss of heterozygosity on chromosome 1q21 in gastric carcinoma]. AB - OBJECTIVE: To obtain allelic loss mapping and define the minimal lost region on chromosome 1q21 in gastric carcinomas, and explore role of 1q21 loss of heterozygosity (LOH) in the development and progression of gastric carcinogenesis. METHODS: Using 7 high-density microsatellite markers and PCR method, lq21 LOH was analyzed in 30 paired specimens of fresh gastric carcinoma, and the relation between 1q21 LOH and the clinicopathological features of the malignancy was tested. RESULTS: The LOH frequency on chromosome 1q21 from these gastric carcinoma tissues reached 60% (18/30). The LOH frequencies of the microsatellite markers D1S514, D1S2696, D1S498, D1S305, D1S2624, D1S2635 and D1S2702 were 13.3%, 10%, 20%, 23.3%, 33.3%, 40% and 23.3%, respectively. The minimal lost region on 1q21 LOH in the gastric carcinoma tissues was located in the region between D1S2624 and D1S2707, in the vicinity of D1S2635. No significant relations of 1q21 LOH to the patients' age, gender, location of the primary foci, clinical staging, or the tumor differentiation were noted (P>0.05), but 1q21 LOH was correlated to lymph node metastases of the malignancy (P<0.05). CONCLUSION: Higher frequency of 1q21 LOH occurs in gastric carcinoma cells, suggesting the presence of potential tumor suppressor genes closely associated with gastric carcinogenesis near the region of D1S2635. PMID- 18676272 TI - [Inhibitory effects of immunotargeting of Chinese cobra cytotoxin and iodine-131 against nasopharyngeal carcinoma cells in vitro]. AB - OBJECTIVE: To explore a new specific therapy of nasopharyngeal carcinoma (NPC) using an anti-nasopharyngeal carcinoma (NPC) monoclonal antibody BAC5 conjugate with Chinese cobra (CT) and iodine-131(131I). METHODS: BAC5 was labeled with 131I by chloramine-T method, CT was labeled with 125I using iodogen method, and BAC5 and 125I-CT were conjugated by SPDP method. The inhibitory effect of the conjugate on cultured human NPC CNE2 cells was observed using MTT assay. RESULTS: The IC50 of 125I-CT-BAC5 conjugate was 9.17x10(-8) mol/L, and that of 131I-BAC5 was 5.83x10(8) Bq/L, and their combined administration showed obvious inhibitory effect on the NPC cells. CONCLUSION: Both 125I-CT-BAC5 and 131I-BAC5 have obvious inhibition effects against NPC cells, but their combined use shows stronger effects. PMID- 18676273 TI - [Variation of serum soluble CD14 levels in patients with chronic heart failure]. AB - OBJECTIVE: To analyze the patterns of serum soluble CD14 (sCD14) and C-reactive protein (CRP) alterations in patients with chronic heart failure (CHF) and investigate the correlations of sCD14 variation to the etiology, clinical symptoms, and the number of mononuclear cells in these patients. METHODS: This study involved 246 CHF patients stratified according to their etiology and clinical symptoms, with 107 normal individuals serving as the control group. Blood samples were collected from these patients the next day after admission and also from the control subjects for measuring serum sCD14 and CPR levels using enzyme-linked immunosorbent assay (ELISA) and rate nephelometry, respectively. RESULTS: The CHF patients had significantly higher serum levels of sCD14 and CRP than the control subjects (P<0.01). In the CHF patients, serum sCD14 and CRP levels differed significantly in the patients with clinical symptoms of different severities (F=3.787, P=0.024), and those with moderate and severe symptoms had significantly higher levels than the asymptomatic patients (P<0.05). The difference in etiologies also resulted in significant difference in sCD14 levels (P<0.05), which were significantly lower in coronary artery disease group than in hypertension group (P<0.05). Significant positive correlations were found between sCD14 and the CRP levels in the CHF patients (r=0.227, P=0.018) and between sCD14 level and the clinical symptoms (r=0.206, P=0.001), but sCD14 level was not correlated to the absolute or relative number of mononuclear cells. CONCLUSIONS: Serum sCD14 and CRP levels are significantly elevated in CHF patients, but this condition may vary as the etiologies and clinical symptoms differ. Increased mononuclear cells do not contribute to the elevation of serum sCD14. PMID- 18676274 TI - [Effects of ulinastatin on gut mucosal apoptosis and bacterial translocation in rats with sepsis]. AB - OBJECTIVE: To investigate the effects of ulinastatin on gut mucosal apoptosis and bacterium translocation in a rat model of sepsis. METHODS: Fifty rats were randomly assigned into 4 groups, namely the control (n=5, no operation or drugs), ulinastatin pretreatment (n=15, treated with 25,000 U/kg ulinastatin 2 h before operation), ulinastatin treatment (n=15, treated with 25,000 U/kg ulinastatin 2 h after operation) and sepsis model (n=15, without drug treatment) groups. The rats in the later 3 groups were subjected to cecal ligation and puncture (CLP). At 3, 6 and 12 h after CLP, the rats were sacrificed and the ileum was removed to examine the pathology and apoptosis of the mucosa. The DNA of Bacillus coli in the whole blood was detected using PCR. RESULTS: Sepsis caused of epithelial cell loss in the ileal villi, ulceration and blebbing of the lamina propria. Ulinastatin treatment administered before and after the operation both significantly alleviated these morphological anomalies. The sepsis rats showed significantly increased intestinal mucosal apoptotic index as compared with the other 3 groups (P<0.05). Ulinastatin pretreatment, in comparison ulinastatin treatment 12 h after CLP, significantly increased the intestinal mucosal apoptotic index (P<0.05). Bacillus coli DNA was positive in sepsis and postoperative ulinastatin treatment groups but negative in the control and pretreated groups. CONCLUSION: Increased intestinal musocal apoptosis and gut bacterial translocation occur in rats following sepsis, and ulinastatin can effectively decrease intestinal mucosal apoptosis and inhibit bacterial translocation. PMID- 18676275 TI - [Preoperative intra-arterial chemotherapy for progressive lower rectal cancer]. AB - OBJECTIVE: To evaluate the therapeutic effect of preoperative regional intra arterial chemotherapy (PRAC) on progressive lower rectal cancer. METHODS: Forty five patients with progressive lower rectal cancer were divided into groups A (23 cases) and B (22 cases) for treatment with PRAC 1 to 2 weeks prior to surgical tumor resection or with surgical resection only, respectively. RESULTS: PRAC caused obvious tissue degeneration and necrosis of rectal cancer with a total effective rate of 95.65%. The rates of radical resection in groups A and B were 91.3% and 72.27%, respectively. The 1-year postoperative survival rates of the two groups were 95.65% and 86.36%, with 3-year survival of 89.96% and 68.18%, and 3-year postoperative recurrence rates of 8.69% and 27.27%, respectively. The anal preservation rates of the two groups were 78.26% and 59.09%. CONCLUSION: PRAC can increase radical resection rates, promote the postoperative survival and anal preservation rate, and lower the recurrence rate in patients with lower rectal cancer. PMID- 18676276 TI - [Phloretin induces apoptosis of BEL-7402 cells in vitro]. AB - OBJECTIVE: To examine the effect of phloretin on apoptosis of BEL-7402 cells. METHODS: The viability changes of BEL- 7402 cells as a result of phloretin induced toxicity were analyzed using MTT assay, and the cell morphology changes were observed with fluorescence microscope. Flow cytometry was used to analyze the cell cycle and mitochondrial membrane potential changes, and chromogenic substrate assay performed to detect caspase activity. RESULTS: Phloretin induced obvious cytotoxicity against BEL-7402 cells with IC50 of 89.23 microg/mL. The growth curve demonstrated decreased growth of the cells as phloretin concentration increased. Cell apoptosis occurred 24 h after treatment with 40-160 microg/mL phloretin. Morphological, the cells exposed to phloretin exhibited nuclear chromatin condensation and increased fluorescence intensity. The activity of caspase-9 reached the peak level 12 h after phloretin exposure, and leak levels of caspase-6 and caspase-3 activities occurred 18 and 24 h after the exposure, respectively. CONCLUSION: Phloretin can induce BEL-7402 cell apoptosis though the mitochondrial pathway. PMID- 18676277 TI - [Correlations of S100A4 and MMP9 expressions to infiltration, metastasis and prognosis of non-small cell lung cancer]. AB - OBJECTIVE: To observe the expressions of metastasis-associated protein (S100A4) and matrix metalloproteinase 9 (MMP9) in human non-small cell lung cancer (NSCLC) and investigate their correlations to the infiltration, metastasis and prognosis of NSCLC. METHODS: The expressions of S100A4 and MMP9 were detected in 41 NSCLC specimens and 6 normal lung tissue specimens using immunohistochemistry with SP method. Univariate and multivariate survival analysis were used to analyze the correlations of S100A4 and MMP9 to the clinicopathological characteristics and progrnosis of NSCLC. RESULTS: Compared with normal lung tissues, NSCLC showed significantly increased positivity for S100A4 and MMP9 expression (P<0.05); their expression were significantly higher in adenocarcinoma than in squamous cell carcinoma (P<0.01), and higher in metastatic NSCLC than in that without lymphatic metastasis (P<0.01). The positive expression rates of S100A4 and MMP9 were significantly higher in tumors in TNM stages III +IV than in stages II+I (P<0.05). S100A4 expression was positively correlated to tumor size (P<0.001), while MMP9 was inversely correlated to tumor differentiation (P<0.05). The expressions of S100A4 and MMP9 were both correlated to lymphatic metastasis, TNM stages and pathological types (P<0.05), and they also showed a mutual correlation (P<0.01). Univariate survival analysis confirmed the effects of histological types, lymphatic metastasis, clinical TNM stages and expressions of S100A4 and MMP9 on the survival time of NSCLC patients (P<0.001). Multivariate survival analysis identified clinical TNM stages and expressions of S100A4 and MMP9 as the independent factors affecting the prognosis of NSCLC (P<0.05). CONCLUSION: The expressions of S100A4 and MMP9 are up-regulated in NSCLC and have significant correlations to the clinical and biological behaviors of NSCLC. S100A4 and MMP9 status are independent prognostic predictors of NSCLC, and detection of their expressions may help evaluate the prognosis of NSCLC. PMID- 18676278 TI - [Effect of peptide YY on hepatic fibrosis in rats]. AB - OBJECTIVE: To investigate the therapeutic effects of peptide YY against hepatic fibrosis in rats and explore the possible mechanism. METHOD: Rat models of hepatic fibrosis were established with a subcutaneous injection of carbon tetrachloride and randomized into normal control group, model group, peptide YY (PYY)-treated group, octreotide-treated group, and interferon gamma-treated group. Serum levels of the hepatic function indices and hepatic fibrotic index were detected, and the hepatic fibrosis grade was assessed using HE staining. The expression of transforming growth factor beta1 (TGFbeta1) were determined with immunohistochemical staining method. RESULTS: The rats in PYY-treated group showed significantly different serum levels of TBIL, HA and LN from the rats in the model group (P<0.05). PYY significantly reduced hepatic fibrosis scores and lowered TGFbeta1 expression as compared with the model group. CONCLUSIONS: PYY can down-regulate TGFbeta1 expression to inhibit the development of hepatic fibrosis with comparable efficacy with interferon gamma and octreotide. PMID- 18676279 TI - [Construction and identification of RNA interference lentiviral expression vector of galectin-3 gene]. AB - OBJECTIVE: To construct a recombinant lentiviral U6 plasmids for RNA interference (RNAi) of galectin-3 gene and select the optimal target sequence of galectin-3 gene for RNAi. METHODS: Double-stranded oligo DNAs were designed and synthesized according to the sequence of galectin-3 gene, and ligated into linearized pGCL GFP/U6 plasmid followed by transformation into competent DH5alpha cells. After PCR and sequence analysis for verification of the positive clones, the plasmid pGCL-GFP/U6 Gal-3shRNA-1 was extracted and transfected into CaCl2-treated 293T cells to obtain the viral vectors containing the RNAi sequence. MCF-7 cells were infected with pGCL-GFP/U6 Gal-3shDNA-1, and at the infection rate over 50%, the cells were harvested to extract the RNA. Real time-PCR was performed to determine the expression level of galectin-3 mRNA in the infected cells. RESULTS: The recombinant vector was successfully constructed as confirmed by sequence analysis. High titer of the virus was obtained, and after infection of MCF-7 cells, RNAi targeting the 1# and 3# sequences in galectin-3 gene resulted in suppression of galectin-3 mRNA expression by 95% and 85%, respectively. CONCLUSION: The recombinant lentiviral U6 plasmid for RNAi of Galectin-3 gene has been successfully constructed, which provides the basis for further study of the role of galectin-3 gene in tumor cells. PMID- 18676280 TI - [Protective effects of Shenkangwan against podocyte injury in rats with early diabetic nephropathy]. AB - OBJECTIVE: To investigate the morphological changes and expressions of desmin and podocin in podocytes of rats with diabetic nephropathy (DN) rats and renal protection mechanism of Shenkangwan. METHODS: DN model was established in rats by a single injection of streptozotocin. The rats were then randomly divided into model group, Shenkangwan treatment group, irbesartan treatment group, and Shenkangwan plus irbesartan treatment group, with normal rats as the control group. All the rats received daily gavage for 8 weeks. The urinary protein quantity in 24 h were detected, and the morphological changes of the kidneys were observed with optic and transmission electron microscopes. The expressions of desmin and podocin in the podocytes were detected by immunohistochemistry. RESULTS: Shenkangwan and irbesartan reduced the urinary protein quantity in 24 h and alleviated the renal damage in DN rats, and the expression of desmin was significantly attenuated while podocin expression increased in the podocytes. CONCLUSIONS: Shenkangwan can provide renal protection against DN in rats and alleviate the structural and functional damages of podocytes possibly by reducing desmin expression and increasing podocin expression in the podocytes. PMID- 18676281 TI - [Circumferential pulmonary vein ostial isolation for atrial fibrillation guided by EnSite NavX three-dimensional electrophysiological mapping]. AB - OBJECTIVE: To investigate the efficacy and safety of circumferential pulmonary vein ostial isolation guided by EnSite NavX three-dimensional electrophysiological mapping in patients with atrial fibrillation (AF). METHODS: Thirty-eight patients with drug refractory paroxysmal or persistent AF underwent circumferential pulmonary vein ostial isolation and were followed up to investigate the efficacy and safety of the treatment. RESULTS: All cases reached the endpoint of the ablation, and both sides of the pulmonary vein were completely isolated, with an average procedure time of 200.4-/+37.0 min, X-ray exposure time of 54.7-/+9.7 min, and three-dimensional left atrial geometry reconstruction time of 27.5-/+7.5 min. During the follow-up for 9-/+3 months, the success rate of initial ablation was 89.5%, and the incidence of procedure related complications were 7.9%. CONCLUSIONS: Circumferential pulmonary vein ostial isolation guided by EnSite NavX three-dimensional electrophysiological mapping can be effective and safe for AF treatment. PMID- 18676282 TI - [Clinical analysis of coronary artery disease in elderly patients with sleep disordered breathing]. AB - OBJECTIVE: To understand the prevalence of sleep disordered breathing (SDB) in elderly patients with coronary artery disease (CAD) and explore the relations between SDB and CAD. METHODS: Sixty-two elderly patients with and 18 without CAD identified by coronary angiography underwent examinations by polysomnography (PSG). Left ventricular ejection fraction (LVEF) was measured by 99Tc equilibrium radionuclide angiography. RESULTS: In the 62 elderly patients with CAD, 53.2% had SDB, a rate significantly higher that (22.2%) in the 18 non-CAD patients. The CAD patients with SDB had higher respiratory disturbance index (RDI) and body mass index (BMI) and lower arterial saturation of oxygen (SaO2) during sleep, with longer duration of low SPO2 (less that 90%). The incidence of hypertension was higher in CAD patients with SDB than in those without SDB. No significant correlation was found between the severity of coronary artery disease and RDI (r= 0.16, P>0.05). CONCLUSION: The elderly patients with CAD have higher incidence of SDB, and appropriate interventions should be administered in those with severe SDB. PMID- 18676283 TI - [Preliminary study of the value of contrast-enhanced ultrasound with high frequency linear-array transducer in the diagnosis of carotid stenosis]. AB - OBJECTIVE: To study the value of contrast-enhanced ultrasound with high-frequency linear-array transducer in the diagnosis of carotid stenosis. METHODS: The carotid arteries of 31 patients was examined with color Doppler ultrasound (CDU), harmonic contrast ultrasound (HCU) and digital subtractive angiography (DSA), respectively. The result of DSA was used as the golden standard for evaluating the accuracy, sensitivity and specificity of CDU and HCU in the diagnosis of carotid stenosis. RESULTS: Satisfactory images of 62 carotid arteries were obtained from the patients using HCU with high-frequency linear-array transducer. After intravenous injection of contrast agent, densely dotted hyperechogenic signals were found in the vascular lumen, with stronger intensity than that of the vascular walls. The filling defect areas in the lumen indicated vascular stenosis. Thirty carotid stenoses were detected by DSA, and 25 by CDU. The accuracy, sensitivity and specificity of CDU was 83.3%, 82.0% and 84.9%, respectively. HCU identified 29 carotid stenoses with accuracy, sensitivity and specificity of 96.7%, 96.7% and 97.0%, respectively, significantly higher than those of CDU (P<0.05). CONCLUSION: HCU with high frequency linear-array transducer has better the accuracy than CDU and may serve as a good alternative for diagnosis of carotid stenosis. PMID- 18676284 TI - [Allogeneic leukocyte immunization combined with IVF-ET for treatment of infertility induced by recurrent spontaneous abortion]. AB - OBJECTIVE: To investigate the effect of allogeneic leukocyte immunization combined with in vitro fertilization-embryo transfer (IVF-ET) for treatment of infertility induced by habitual abortion. METHODS: Allogeneic leukocyte immunization was performed in 9 patients with infertility induced by habitual abortion, with another 9 patients undergoing IVF-ET without habitual abortion as the control group. All the patients were treated with long GnRH-a protocols. The infertility patients with recurrent spontaneous abortion history were immunized with lymphocytes from the husband for before IVF-ET and after clinical pregnancy. RESULTS: The fertilization rates of the immunotherapy group and control group were 81.3% and 82.2%, respectively, showing no significant difference (P>0.05). Five patients in each group had clinical pregnancy, and a twin pregnancy occurred in the control group. The embryo implantation rates were also comparable between the two groups (22.7% vs 28.6%, P>0.05). All the fetuses resulted from IVF-ET developed normally and were healthily delivered. CONCLUSION: Allogeneic leukocyte immunotherapy along with IVF-ET is effective for treatment of infertility resulting from recurrent spontaneous abortion. PMID- 18676285 TI - [Effect of small interfering RNA targeting multidrug resistance-related protein and bcl-2 on drug resistance and apoptosis of K562 and K562/ADM cells]. AB - OBJECTIVE: To observe the effect of small interfering RNA (siRNA) targeting multidrug resistance-related protein (MRP) and bcl-2 genes in modulating drug resistance and apoptosis of K562 and K562/ADM cells. METHODS: Two siRNA constructs targeting respectively bcl-2 and MRP genes, were synthesized and transfected either alone or in combination into K562 and K562/ADM cells via lipofectamine2000. MTT assay was used to evaluate the viability of the transfected cells at 24, 48 and 72 h Post-fransfection, and RT-PCR was performed to determine the mRNA levels of bcl-2 and MRP. The effects of MRP siRNA and bcl2 siRNA on the apoptosis and the protein expression of Bcl-2 and MRP were evaluated with flow cytometry. RESULTS: In K562/ADM cells, the IC (50) decreased from 12.81 microg/ml (ADM group) to 3.74 microg/ml (ADM+MRP siRNA group), 6.82 microg/ml (ADM+bcl2 siRNA group) and 2.51 microg/ml (ADM+MRP siRNA+bcl2 siRNA). Similarly, in K562 cells, the IC50 decreased significantly from 6.75 microg/ml (ADM) to 3.22 microg/ml (ADM+MRP siRNA), 3.56 microg/ml (ADM+bcl2 siRNA) and 1.84 microg/ml (ADM+MRP siRNA+bcl2 siRNA) (P<0.05). Flow cytometry demonstrated significantly increased apoptosis of the cells following MRP siRNA and bcl2 siRNA transfection, which also resulted in significantly decreased expressions of MRP and bcl-2 proteins (P<0.05). CONCLUSION: Treatment with both MRP and bcl-2 siRNAs inhibits the target gene expression, and increases the drug sensitivity and apoptosis of K562 and K562/ADM cells. PMID- 18676286 TI - [Early diagnosis of female genital tuberculosis by phage amplified biological assay]. AB - OBJECTIVE: To establish an early diagnostic method for detecting female genital tuberculosis. METHODS: Eighty-six women with genital tuberculosis during January 2005-September 2007 were examined by phage amplified biological assay, and the results were compared with those from leucorrhea culture, smear and PCR. RESULTS: Forty-five patients were tuberculosis positive with 100% of specificity identified by phage amplified biological assay. Twenty patients were tuberculosis positive by PCR. Five patients were culture-positive tuberculosis and no case had smear-positive tuberculosis. CONCLUSION: Phage amplified biologically assay is sensitive and specific, which could be used for the early diagnosis of female genital tuberculosis. PMID- 18676287 TI - Right coronary artery originating from distal circumflex artery in a patient with single coronary artery. PMID- 18676288 TI - Warfarin-induced benign acral cutaneous lesions in two cardiac patients with decreased protein C and S activity. PMID- 18676289 TI - Successful reimplantation of prematurely displaced stent to the target lesion without balloon inflation during percutaneous intervention to the right coronary artery. PMID- 18676290 TI - Myocardial blushing during coronary angiography due to high pressure applied by an inexperienced operator. PMID- 18676291 TI - Right ventricular lipoma. PMID- 18676292 TI - Ball thrombus in a ligated left atrial appendage. PMID- 18676293 TI - Giant metastatic left atrial tumor leading to pulmonary edema. PMID- 18676294 TI - Management of an enlarging pericardial cyst. PMID- 18676295 TI - A prominent Chiari network prolapsing into right ventricle. PMID- 18676296 TI - Giant aneurysm of the ductus arteriosus. PMID- 18676297 TI - [On SSS, Bratislava-III, doctors who make history, publisher's new advancements, books and once more painful loss]. PMID- 18676298 TI - The protective effect of melatonin on nicotine-induced myocardial injury in newborn rats whose mothers received nicotine. AB - OBJECTIVE: Nicotine, one of the most dangerous substances in tobacco, can pass the placenta and affect the fetal hemodynamics. The aim of this study was to evaluate the protective effects of melatonin on hearts of nicotine exposed newborn rats whose mothers received nicotine. METHODS: This is an experimental, randomized, controlled study. Study groups were composed of five groups of rats; high-dose nicotine (HDN), HDN+melatonin (HDNM), low-dose nicotine (LDN), LDN+melatonin (LDNM), control. Myocardial and plasma malondialdehyde (MDA), nitric oxide(NO), glutathione peroxidase (GSHPx) and superoxide dismutase (SOD) were analyzed and myocardial tissue was examined histopathologically. Comparisons of groups were done with Kruskal-Wallis one-way analysis test. All pairwise multiple comparisons and the comparisons between control and other groups were done with Dunn's nonparametric multiple comparison test. RESULTS: Plasma and tissue MDA levels among groups were different (p=0.001 for plasma MDA and p=0.001 for tissue MDA). Plasma MDA levels of HDN, HDNM, LDN, and tissue MDA levels of HDN and LDN were significantly higher than in control group (p<0.05 for plasma MDA and for tissue MDA). Plasma and tissue NO levels among groups were also different (p=0.011 for plasma NO and p=0.001 for tissue NO). Plasma NO of LDN group was higher than of LDNM group, and plasma NO of LDNM group was lower than in control group (p<0.05). Tissue NO levels of HDN and LDN groups were higher than of control group (p<0.05). There was no difference between plasma GSHPx levels among groups (p=0.221) but statistically significant different was detected between tissue GSHPx levels among groups (p=0.001). Tissue GSHPx level was found lower in HDN group than in control group (p<0.05). Tissue GSHPx level of LDNM group was higher than of LDN group, and tissue GSHPx level of HDNM group was higher than of HDN group (p<0.05). A difference was found between plasma and tissue SOD among groups (p=0.005 for plasma SOD and p=0.001 for tissue SOD). Plasma SOD of LDN group was significantly lower than of HDNM and LDNM groups (p<0.05). Tissue SOD analyzes revealed lower levels in HDN and LDN groups than in control group (p<0.05). Severe cardiomyopathy was determined in HDN and LDN groups (p<0.05). CONCLUSION: Nicotine exposure depletes myocardial antioxidant enzymes and increases free radicals and lipid peroxidation products. Melatonin particularly prevents the nicotine-induced cardiac injury as an antioxidant. PMID- 18676299 TI - Predicting coronary artery disease using different artificial neural network models. AB - OBJECTIVE: Eight different learning algorithms used for creating artificial neural network (ANN) models and the different ANN models in the prediction of coronary artery disease (CAD) are introduced. METHODS: This work was carried out as a retrospective case-control study. Overall, 124 consecutive patients who had been diagnosed with CAD by coronary angiography (at least 1 coronary stenosis > 50% in major epicardial arteries) were enrolled in the work. Angiographically, the 113 people (group 2) with normal coronary arteries were taken as control subjects. Multi-layered perceptrons ANN architecture were applied. The ANN models trained with different learning algorithms were performed in 237 records, divided into training (n=171) and testing (n=66) data sets. The performance of prediction was evaluated by sensitivity, specificity and accuracy values based on standard definitions. RESULTS: The results have demonstrated that ANN models trained with eight different learning algorithms are promising because of high (greater than 71%) sensitivity, specificity and accuracy values in the prediction of CAD. Accuracy, sensitivity and specificity values varied between 83.63%-100%, 86.46% 100% and 74.67%-100% for training, respectively. For testing, the values were more than 71% for sensitivity, 76% for specificity and 81% for accuracy. CONCLUSIONS: It may be proposed that the use of different learning algorithms other than backpropagation and larger sample sizes can improve the performance of prediction. The proposed ANN models trained with these learning algorithms could be used a promising approach for predicting CAD without the need for invasive diagnostic methods and could help in the prognostic clinical decision. PMID- 18676300 TI - [The determination of the factors impacting on in-hospital mortality in patients with acute heart failure in a tertiary referral center]. AB - OBJECTIVE: Despite impressive advances in therapeutics in the last years, acute heart failure (AHF) remains a major cause of cardiovascular morbidity and mortality. Patients hospitalized because of heart failure (HF), irrespective of left ventricular systolic function, represent a high-risk population with limited short-term prognosis. A substantial component of HF-related mortality occurs during a hospital stay. In this study, we aimed to determine the factors impacting on in-hospital mortality in patients with AHF. METHODS: During a 15 month period (December 2005-March 2007), 85 consecutive patients with (mean age: 64+/-8 years, male: 54%) an episode of AHF were included in this study. The effect of demographic, clinical, electrocardiographic, and echocardiographic characteristics, laboratory findings on in-hospital mortality were evaluated retrospectively. RESULTS: Of 85 patients 24.7% of patients had new-onset HF. Coronary artery disease (61%) was the most common underlying disease. The 44.7% of patients had hypertension, 37.6% had diabetes mellitus, 21% had chronic renal failure and 16.4% had chronic obstructive pulmonary disease. Left ventricular ejection fraction was 35+/-7%. In-hospital mortality rate was found as 11.7% (10 patients). The major cause of mortality was the progression of HF to cardiogenic shock in 60% of deaths. In comparison with surviving patients in terms of the clinical, demographic, electrocardiographic, and laboratory characteristics and left and right ventricular functions, patients died during hospitalization had higher blood urea nitrogen (45+/-20 mg/dl vs. 36+/-12 mg/dl, p=0.04), higher creatinine level (2.2+/-0.8 mg/dl vs. 1.1+/-0.5 mg/dl, p=0.001), and wider QRS duration (130+/-13 ms vs. 116+/-18 ms, p=0.04) whereas they had lower plasma sodium level (128+/-5 mmol/l vs. 135+/-9 mmol/l, p=0.02) and systolic blood pressure (p=0.01). Logistic regression analysis revealed that plasma creatinine level (OR 1.5, 95% CI 1.2 to 2.1, p=0.01), blood urea nitrogen (OR 2.1, 95% CI 1.8 to 3.1, p=0.001), plasma sodium level (OR 1.3, 95% CI 1.1 to 1.7, p=0.02), and systolic blood pressure (OR 2.2, 95% CI 1.9 to 2.8, p=0.01) were the independent predictors of in-hospital mortality. CONCLUSION: In-hospital mortality increases in patients who had lower systolic blood pressure, lower plasma sodium level, and renal dysfunction on admission. PMID- 18676301 TI - Drug-induced torsades de pointes in patients aged 80 years or more. AB - OBJECTIVE: We studied all English-written peer-reviewed reports on drug-induced torsades de pointes (TdP) in patients aged 80 years or more in order to characterize the clinical circumstances leading to this serious complication. METHODS: Our literature search yielded 24 reports on 25 patients aged 80-95 years with drug-induced TdP. We systematically reviewed each report and recorded the non-modifiable risk factors for drug-induced TdP (i.e., female sex and structural heart disease) as well as preventable clinical circumstances, which might have been associated with drug-induced TdP. RESULTS: The most prevalent risk factors for drug-induced TdP were non-modifiable risk factors: 22 (88%) patients were female patients and 19 (76%) patients had structural heart disease. Overall, 16 (64%) patients were female patients with structural heart disease. The literature did not report any elderly male patients without structural heart disease. Among the preventable clinical circumstances, which might have been associated with drug-induced TdP, the most prevalent were: administrating QT prolonging agents despite long QT interval (n=11; 44%) and co-administration of two or more QT prolonging agents (n=9; 36%). The most prevalent QT prolonging agents found to trigger TdP were macrolides and quinolones (n=9; 36%). All but three patients had at least one or more preventable clinical circumstances, which might have been associated with drug-induced TdP. CONCLUSION: Physicians should be more aware of the risk for drug-induced TdP in patients aged 80 years or more while administrating QT prolonging agents despite long QT interval and co- administrating two or more QT prolonging agents, specifically in elderly female patients with structural heart disease. PMID- 18676302 TI - QT dispersion during hypobaric hypoxia. AB - OBJECTIVE: Hypoxia is one of the major concerns in aviation. Clinical hypoxia has been shown to increase QT dispersion (QTd). We aimed to examine QTd during hypobaric chamber training to observe the effect of hypobaric hypoxia on QT dispersion. METHODS: A total of 38 healthy male aviators volunteered to take part in this longitudinal study. Subjects' electrocardiograms were recorded by 12-lead digital Holter device before, during, and after hypobaric exposure at simulated altitude of 30,000 ft. Data from 23 of the subjects, aged 27.91+/-6.02 years (range 22-39) was used. QT intervals were measured manually. QT dispersion and heart rate adjusted QTd (QTcd) were calculated for each subject. Statistical significance of changes in parameters was analyzed using the Friedman test. Comparison of pre-post exposure clusters was made using Dunn's test. RESULTS: QT dispersion values were as following: prehypoxic 64.09+/-8.39 ms, hypoxic 50.35+/ 11.06 ms and posthypoxic 59.83+/-9.06 ms (Median: 64, 50, 60; Mean rank: 2.65, 1.28, 2.07) (p=0.0001 for prehypoxic-hypoxic, p=0.046-prehypoxic-posthypoxic, and p=0.002 for posthypoxic-hypoxic). Heart rate values were as following: prehypoxic 74.09+/-6.43 beats/min, hypoxic 127.1+/-17.39 beats/min, and posthypoxic 95.17+/ 11.35 beats/min (Median: 75, 122, 92; Mean rank: 1, 3, 2) (p=0.0001 for prehypoxic-hypoxic, prehypoxic-posthypoxic, and posthypoxic-hypoxic). The change in QTd and HR during hypobaric chamber exposure was statistically significant but, the change in QTcd was not (p<0.001, p<0.001, p>0.1, respectively). CONCLUSION: From the findings of present study, it is not possible to directly comment on the validity of QTd in revealing arrhythmogenic predisposition of healthy subjects exposed to hypobaric hypoxia. The relationship between QT dispersion and hypobaric hypoxic exposure is not clear, particularly when QTd is corrected for the increased heart rate. QT dispersion measurement has not been proven a reliable and practical method to show arrhythmia predisposition during a hypobaric hypoxic exposure in healthy individuals. PMID- 18676303 TI - Assessment of aortic stiffness and ventricular functions in familial Mediterranean fever. AB - OBJECTIVE: To investigate systolic and diastolic ventricular functions, aortic elastic properties and the presence of pericardial effusion in familial Mediterranean fever (FMF) patients. METHODS: A case-controlled, cross-sectional study was performed on 44 FMF patients and 27 controls. Subjects with hypertension, diabetes mellitus and hyperlipidemia were excluded. Left and right ventricular functions were measured using echocardiography including two dimensional, M-mode, and conventional Doppler as well as pulsed wave tissue Doppler imaging (TDI). Aortic elasticity was analyzed using M-mode tracing guided by the two-dimensional echocardiography. Statistical analysis was performed using Mann Whitney U, Spearman rho correlation and Fisher's exact tests. RESULTS: Age, sex, body mass index, smoking status and lipids were comparable in patients and controls (p>0.05). None of the subjects had pericarditis and/or pericardial effusion. Aortic wall properties were similar between groups (p>0.05). The TDI parameters of mitral lateral annulus revealed significantly lower Em/Am ratios in patients compared to controls [1.77 (0.6-3.4) vs. 1.79 (0.9-4.8), p=0.02]. Mitral flow propagation velocity was significantly lower in patients than healthy subjects [63 (39-100) vs. 74 (40-94) cm/s, p=0.008]. Tricuspid annular plane systolic excursion (TAPSE) was significantly reduced in FMF group than in controls [2 (1.3-2.5) vs. 2.5 (1.7-3.2) cm; p<0.001]. Eight of the patients and one control had impaired TAPSE (<2 cm; p=0.025). There was no difference regarding right ventricular diastolic dysfunction (RVDD) as assessed by using standard Doppler echocardiography (p>0.05). However, pronounced RVDD was observed in FMF patients documented by TDI (Em/Am<1; 19 patients vs. 0 controls, p<0.001). CONCLUSION: Subclinical myocardial involvement is present in a cohort of relatively young FMF patients who were also free of classical cardiovascular risk factors. Pericardium and aorta seem to be spared during attack free periods of FMF. PMID- 18676304 TI - [Seven years of experience in patients with pulmonary arterial hypertension in Ege University Hospital: diagnostic approach of a single center]. AB - OBJECTIVE: Pulmonary arterial hypertension (PAH) is a term used to define a variety of progressive conditions that have in common, increased pulmonary vascular resistance leading to right heart failure and death. There has been considerable decrease in mortality and morbidity with the advances in PAH treatment over the past decade. However, since there is no epidemiologic study in Turkey, the prevalence of PAH and its importance is not known yet. This study aimed to evaluate the diagnostic clinical experience of Ege University Medical School Cardiology Department with PAH patients. METHODS: We evaluated the diagnostic approach to patients referred to our department with the diagnosis of PAH since 2000 by retrospective analysis method. RESULTS: The diagnosis of pulmonary hypertension was definite in 70 patients (mean age 47+/-16 years, 61% women). Etiology from most prevalent to least was as following: congenital heart diseases (27%), chronic thromboembolic pulmonary hypertension (24%), connective tissue diseases-scleroderma (14%), idiopathic PAH (8%), diastolic dysfunction (3%), pulmonary disease (3%), pulmonary veno-occlusive disease (2%), hepatopulmonary hypertension (1%), and HIV-infection associated PAH (1%). At diagnosis, 68% of patients were in NYHA functional class-III or IV. Six-minute walk test was 263+/-127 m. Mean pulmonary artery pressure was 65+/-20 mmHg. The prognostic marker pro-BNP (brain natriuretic peptide) level was 3208+/-4145 pg/ml. CONCLUSION: Our practice shows that PAH is diagnosed late in the course of the disease in Turkey. This can be overcome with structured management in designated centers with multidisciplinary team-working in a shared care approach. There is also an urgent need for an epidemiological registry in order to determine the burden of PAH in Turkey and increase the awareness of doctors. PMID- 18676305 TI - Surgical experiences in acute spontaneous dissection of the infrarenal abdominal aorta. AB - OBJECTIVE: Spontaneous dissection of the abdominal aorta should be considered as a vascular surgical emergency. We report here our experience with this rare pathology. METHODS: At our hospital, we operated on 200 patients for abdominal aortic aneurysm over a 4-year period, and aortic dissection was the underlying pathology in eight of them. Diagnosis was based on history, imaging study findings and visualization at surgery. RESULTS: The mean size of the aorta at the time of diagnosis was 5.5 +/- 0.4 cm. Aortic dissection was complicated by acute ischemia of lower limb and paraplegia in 1 patient and four patients were admitted to hospital with severe abdominal and back pain. Two patients with contained rupture were admitted with hemodynamic collapse and shock. One of the patients who presented with abdominal pain experienced also sudden onset paraplegia. The diagnosis of dissecting abdominal aortic aneurysm was established incidentally in the remaining 2 patients. All patients were treated using open techniques with paramedian transperitoneal incision. We performed aorto-bifemoral bypass operation procedure in all patients; there was no post-operative mortality in our series. One patient had to be reoperated because of bleeding during early postoperative period. One patient required transient dialysis at early postoperative period because of transient rise of serum creatinine level. CONCLUSION: The surgical treatment of spontaneous abdominal aortic dissection is usually associated with low morbidity and mortality rates. However, the clinical form of presentation of acute abdominal aortic dissection is usually nonspecific, and diagnosis requires a high index of suspicion. PMID- 18676306 TI - [The role of multislice tomography to prevent postoperative strokes related with atherosclerotic plaques in the ascending aorta after coronary artery bypass]. AB - OBJECTIVE: Atheromatous plaques in the ascending aorta are major risk factors for strokes caused by macroembolization after coronary artery surgery. Detection of plaque formations and changes in the surgical strategy are very important. This study was planned to compare value direct palpation and multislice computerized tomography to establish aortic plaques and to establish clinical predictors of aortic calcification. METHODS: Fifty-four patients who underwent coronary bypass surgery were included in this prospective and diagnostic study. Proximal portion of each patient's aorta was evaluated with multislice computerized tomography and was compared with direct palpation. The efficacy of intraoperative palpation to predict calcifications was studied with ROC analysis and the predictors of aortic plaque formation were analyzed using logistic regression analysis. RESULTS: Atheromatous plaques were detected with palpation in four patients (7.4%), and with multislice tomography in six patients (11.1%). The aortic instrumentation was changed in two patients (3.7%) and strategy was changed in one patient 1.8%). According to the ROC analysis, when multislice computerized tomography was taken as the reference, intraoperative direct palpation was 67% sensitive and 100% specific to predict aortic plaques. Logistic regression analysis of the risk factors showed that the older age was the only significant risk factor (OR-1.3, 95% CI -1.114-1.568, p=0.001) for plaque formation in the aorta. Neither stroke nor other neurological disorders have been observed during the study. CONCLUSIONS: It can be stated that multislice computerized tomography is more effective to show aortic plaques, but it is not sufficient. Multislice tomography may give additional information about the ascending aorta and the opportunity to visualize the aortic arch. It can be preferred in patients with aortic aneurysm or dissection. PMID- 18676308 TI - [Suggestions for definition and classification of double right coronary artery, which is frequently seen in Turkey despite its rarity in the literature]. PMID- 18676307 TI - Biotechnology and stem cell research: a glance into the future. AB - The present review addresses the issues related to innovative contributions in biotechnology and their potential role in stem cell research at present and in the future. We can expect that future developments and applications in biotechnological sciences and industry will effect the direction of emerging cellular therapies. The use of these advances may offer a unique opportunity to investigate the mechanisms related to the journey from embryonic cells or bone marrow derived stem/progenitor cells to cardiomyocytes or endothelial cells and the molecular regulators of cell differentiation. PMID- 18676309 TI - Amebic pericarditis: a summary of Thai cases. PMID- 18676310 TI - [Thrombus fragmentation using iliac balloon angioplasty in a patient with massive pulmonary embolism]. PMID- 18676311 TI - Isolated left pulmonary artery agenesis. PMID- 18676312 TI - Fibroelastoma of the posterior mitral leaflet. PMID- 18676313 TI - Perventricular closure of muscular ventricular septal defect in an infant. PMID- 18676314 TI - Bilateral common peroneal nerve palsy following cardiac surgery. PMID- 18676315 TI - A rare pathogen causing endocarditis: Streptococcus constellatus. PMID- 18676316 TI - May positive U waves in V1-V3 leads predict left main coronary artery occlusion? PMID- 18676317 TI - A rare cause of dilated cardiomyopathy; Alstrom syndrome. PMID- 18676318 TI - The use of renal stents in percutaneous treatment of very large coronary arteries. PMID- 18676319 TI - Continuous international network building to promote scientific publication: Scientific Summer School, Pezinok, Slovakia, May 25-30, 2008. PMID- 18676320 TI - Becoming an oncology nurse through osmosis. PMID- 18676321 TI - Wounded healers. PMID- 18676322 TI - Three little words no one wants to hear. PMID- 18676323 TI - At an increased risk: tumor lysis syndrome. PMID- 18676324 TI - Severe fatigue: could it be aplastic anemia? PMID- 18676325 TI - Managing pain from advanced cancer in the palliative care setting. AB - Managing complex pain at the end of life is an essential aspect of palliative care. Such care is best guided by a comprehensive evaluation of the physiologic sources of pain to determine appropriate analgesia. Using the case of Mrs. J, a woman with advanced ovarian cancer, key principles of complex pain management at the end of life are reviewed, including optimum use of opioids and co-analgesics. In addition to physical assessment, total care of the patient and family facing imminent death should be based on an assessment of psychological, social, and spiritual factors. The assessment and management of pain and suffering are guided by an interdisciplinary team focused on goals of comfort and facilitating a death that respects the life of the patient who is dying. PMID- 18676326 TI - Updates in non-small cell lung cancer. AB - Lung cancer is the leading cause of cancer death in both men and women. Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, representing more than 80% of lung cancer diagnoses. Five-year survival remains at 15%, but new strategies for prevention, early detection, and treatment may improve survival rates. This article provides an overview of updates in NSCLC, with an emphasis on advances in treatment strategies. Newer targeted therapies, as well as advances in genetic blueprinting, will be discussed. Nurses play a pivotal role in the assessment and management of patients with NSCLC and, therefore, must remain abreast of the most current prevention, screening, and treatment options. PMID- 18676327 TI - Evidence-based guidelines for adjuvant therapy for resected adenocarcinoma of the pancreas. AB - Pancreatic cancer, the fourth most common cause of cancer deaths, has a five-year survival rate of 5% or less. Surgical removal of the tumor may improve survival, but survival remains poor even in optimally resected patients. The best adjuvant therapy for patients with resected pancreatic cancer is not clear. Surgical resection followed by chemoradiation and maintenance chemotherapy has been considered the most beneficial treatment for improving survival, but more recent studies have suggested that chemotherapy alone is more effective. The purpose of this article is to review randomized controlled studies of adjuvant chemoradiation or chemotherapy alone in the treatment of resected pancreatic cancer and to determine the optimal adjuvant therapy after curative resection with negative or microscopically positive margins. The outcomes of interest were overall survival and disease-free survival. The results indicate that chemoradiation is an acceptable option for adjuvant treatment. Three of the four randomized controlled trials suggest that adjuvant chemoradiation for resected pancreatic cancer improves overall survival. Adding gemcitabine to the chemoradiation regimen also confers increased disease-free survival. Providers counseling patients regarding treatment options for resected pancreatic cancer should continue to recommend adjuvant therapy--a combination of chemotherapy including gemcitabine and radiotherapy--for appropriately selected patients. PMID- 18676328 TI - Respectful death: a model for end-of-life care. AB - The Respectful Death Model (RDM) is a research-based, holistic, and practical model developed to improve end-of-life care. A respectful death is one which supports dying patients, their families, and professionals in the completion of life cycles and can be used by all members of the healthcare team. The model is a process method commencing with the establishment of a therapeutic relationship with the dying patient and his or her family and, as a result, their stories are heard and incorporated into the care plan. This article demonstrates that hospice and palliative care nurses have been practicing this model since the origination of care of the dying. Other topics addressed are the current culture toward death in the United States, the roles of nurses in the RDM, and the barriers and benefits of the RDM. Recommendations for future research in end-of-life care also are addressed. PMID- 18676329 TI - Cardiovascular toxicity associated with cancer treatment. AB - Cardiotoxicity is a well-described and potentially lethal side effect of certain chemotherapeutic agents. Cardiotoxicity is a broad term used to depict conditions ranging from benign forms of arrhythmias to potentially fatal conditions, such as myocardial ischemia or infarction and heart failure. Anthracyclines (daunorubicin, doxorubicin, and epirubicin), mitomycin, and monoclonal antibodies such as trastuzumab have been associated with cardiotoxicities, but other chemotherapeutic agents, such as fluorouracil, cyclophosphamide, interferons, and interleukin-2 and other targeted agents, also can cause this side effect. Although several theories exist about the process that leads to cardiotoxicity from some chemotherapeutic agents, the exact mechanism of action is unknown. Oncology nurses should know the agents associated with cardiotoxicity, including newer targeted therapy drugs. Knowledge of the potential mechanism of action, as well as the possible reversibility of cardiotoxicity with specific agents, is important. PMID- 18676330 TI - Temsirolimus, an mTOR inhibitor for treatment of patients with advanced renal cell carcinoma. AB - Temsirolimus is a targeted therapy that inhibits mammalian target of rapamycin (mTOR), a central regulator of tumor cell responses to growth stimuli. Temsirolimus has a broad anticancer activity profile that impacts tumor cell growth, proliferation, and survival through its specific inhibition of mTOR. In a randomized phase III trial that enrolled previously untreated patients with advanced renal cell carcinoma (RCC) and poor prognostic features, temsirolimus significantly prolonged overall survival compared with interferon-alpha, a standard therapy (p = 0.008). Because of the results, temsirolimus was approved by the U. S. Food and Drug Administration for treatment and is considered a first line treatment for patients with advanced RCC with poor prognostic features. Temsirolimus is administered at a flat weekly IV dose of 25 mg given over 30-60 minutes. Gastrointestinal disorders (stomatitis, anorexia, nausea, diarrhea, and vomiting), rash, fatigue, edema, infections, and dyspnea, as well as hematologic and metabolic laboratory abnormalities occur in patients receiving temsirolimus. Metabolic side effects include hyperglycemia, hypercholesterolemia, hypertriglyceridemia, and hypophosphatemia. Most adverse reactions associated with temsirolimus can be managed medically or addressed by supportive measures. Nurses can improve patient outcomes through early recognition of side effects and prompt interventions. PMID- 18676331 TI - Understanding urinary incontinence after radical prostatectomy: a nursing framework. AB - Prostate cancer is one of the most prevalent malignancies diagnosed in North American men. Typically, men diagnosed with localized prostate cancer have two options for curative treatment: radiation therapy or radical prostatectomy (RP). Many men choose RP to remove the cancer; however, the intervention has two possible side effects that patients dread: erectile dysfunction and urinary incontinence (UI). At least 50% of men who undergo RP suffer from UI, which can lead to embarrassment, loss of a sense of control, depression, and decreased social interactions. The Human Response to Illness Model provides a framework to gain a comprehensive understanding of the physiologic, pathophysiologic, behavioral, and experiential perspectives as well as personal and environmental factors related to UI following RP. Knowledge gained from these perspectives will help nurses design strategies that facilitate coping and improve outcomes in men with UI following RP. PMID- 18676332 TI - Vitamin D in older patients with cancer. AB - Vitamin D insufficiency has been found to be as high as 75% among community dwelling adults aged 65 and older. The purpose of this article is to provide a review of published literature focused on the benefits of vitamin D and calcium supplementation in older patients with cancer. Insufficient vitamin D levels may have considerable effects on cancer-related diagnosis and treatment. Patient education should include information concerning vitamin D and calcium administration to reduce falls and injury from falls and enhance functional status. This article will consider the issue of vitamin D levels related to cancer prevention, diagnosis, treatment, functional status, and falls in older patients with cancer. Dosing and serum measurement information will be discussed. PMID- 18676333 TI - Managing chronic pain in patients with cancer who have a history of substance abuse. AB - Oncology nurses may encounter patients recovering from substance abuse who will need acute or chronic pain management. Knowing how to assess, treat, and manage that pain is a benefit to the nurse and patient. In addition, understanding and overcoming bias toward patients with a history of substance abuse can lead to a trusting relationship and more effective pain management. A thorough assessment and documentation of the patient's pain during each visit provide a solid basis for prescribing opioids to patients with a history of substance abuse. The use of long-acting and higher-dose opioids in this population will be discussed. Functional improvement versus absence of pain may be a more realistic goal for patients recovering from substance abuse, and complementary and alternative therapy may be considered. Setting standards to deal with lost prescriptions or medication, missed appointments, and the use of contracts for all patients receiving opioids establishes unbiased treatment. PMID- 18676334 TI - Relieving the pain of sentinel lymph node biopsy tracer injection. AB - Sentinel lymph node biopsy (SLNB) has been reliably accurate as a minimally invasive surgical alternative for identifying lymphatic breast metastasis. During mapping, the injection of a radioactive tracer or isosulfan blue dye to differentiate the SLN is acutely painful. The use of the eutectic mixture of lidocaine and prilocaine (EMLA) cream to reduce tracer injection pain has been reported anecdotally. A retrospective study compared injection discomfort of 20 women who had undergone SLNB without EMLA and 20 women who had undergone SLNB with the EMLA protocol. Results indicated a significant difference in mean pain rating. Standards of care should include the use of EMLA prior to intradermal SLN tracer injection unless contraindicated. PMID- 18676335 TI - Does the use of cell phones cause brain tumors? PMID- 18676336 TI - Chest mass mimicking an implanted port. PMID- 18676337 TI - Assessing pain in today's global society. PMID- 18676338 TI - Cognitive impairment after breast cancer treatment. PMID- 18676339 TI - Pharmacy information systems: the experience and user satisfaction within a chain of Dutch pharmacies. AB - This paper proposes two models for evaluating Pharmacy Information Systems (PIS) on their user satisfaction, thereby exploring one of the major requirements in designing PIS for pharmacy networks. The first model is developed to measure pharmacies' satisfaction with their PIS and the second model is developed to specify the determinants of PIS satisfaction. Both models were validated by data from 142 members of a Dutch pharmacy chain. Based on the user satisfaction model, the explanatory model showed that Information Technology (IT) experience and knowledge are the most important drivers for pharmacies' PIS satisfaction. The implications for the evolution of PIS in pharmacy chains are discussed. PMID- 18676340 TI - Investigation of enablers of knowledge transfer in the medical industry. AB - This paper presents a research model for investigating the relationship between organisational enablers and the Knowledge Transfer (KT) Performance (KTP) in the medical industry. The enablers include leadership, organisational culture, Information Technology (IT) and individual performance measurement, and KTP is determined by individual capability, organisational capability and product/service innovation. This paper chose professional medical personnel as the research subject to determine whether or not these enablers affect KT. The findings show that only leadership directly affects the KTP, with IT also impacting both organisational capability and product/service innovation. The implications of these findings are discussed based on interviews with experts and practitioners. PMID- 18676341 TI - Personal identity verification for regional emergency workers. AB - South Central Texas emergency planners have organised a regional Personal Identity Verification (PIV) project for hospital and emergency workers. This paper describes the development and administrative challenges in designing and implementing an electronic PIV system across multiple organisations in a 22 county region. PMID- 18676342 TI - The critical factors influencing the successful implementation of mobile nursing stations: a case study in Taiwan. AB - The purpose of this study is to conduct a comprehensive review of the factors affecting the successful implementation of Mobile Nursing Stations (MNS) by case study. A thorough validation process was used to identify the nine critical factors which influence the implementation of MNS, including the degree of peer competition, governmental and insurance policies, the Information Technology (IT) infrastructure, vendor selection, a clinical champion, top management support, task communications, user participation and training issues. The results of this study can enhance managements' understanding of the complete possibilities for the utilisation of MNS. PMID- 18676343 TI - An analysis of the healthcare informatics and systems in Southeast Asia: a current perspective from seven countries. AB - This paper presents an overview of the healthcare systems in Southeast Asia, with a focus on the healthcare informatics development and deployment in seven countries, namely, Singapore, Cambodia, Malaysia, Thailand, Laos, the Philippines and Vietnam. Brief geographic and demographic information is provided for each country, followed by a historical review of the national strategies for healthcare informatics development. An analysis of the state-of-the-art healthcare infrastructure is also given, along with a critical appraisal of national healthcare provisions. PMID- 18676344 TI - Mobile healthcare: the opportunities and challenges. AB - Mobile healthcare is an important extension of electronic healthcare. It enables the caregivers to have an ubiquitous and uninterrupted access to patients' clinical data and the latest medical knowledge; concurrently, it allows patients with chronic conditions to remain under constant observation without needing to be physically present at the clinic. The critical challenges to a full-scale implementation include establishing interoperability among electronic health records, developing better display technologies and security controls for mobile devices and developing smart algorithms to detect clinically significant events before notifying caregivers. As a consequence of mobile healthcare, new opportunities for physician-patient joint decision-making and personalized healthcare are beginning to take shape. Accompanying them are the challenges of mindset adjustment, the empowerment of patients with medical knowledge in everyday language and ensuring the confidentiality of patient data. PMID- 18676345 TI - Collapsing glomerulopathy and haemophagocytic syndrome related to malaria: a case report. AB - Authors report a 37-year-old Senegalese woman with no known history of nephropathy who was admitted for fever related to malaria, severe acute renal failure requiring dialysis with nephrotic syndrome. Biological examinations and bone marrow aspiration showed hemophagocytic syndrome. A kidney biopsy found a 'collapsing glomerulopathy' (CG). A protracted course of steroids yielded a complete, unexpected remission of the nephrotic syndrome and renal function was normal at 18 months. PMID- 18676346 TI - Abdominal aortic calcification in dialysis patients: results of the CORD study. AB - BACKGROUND: Patients with chronic kidney disease stage 5 have a high prevalence of vascular calcification, but the specific anatomical distribution and severity of abdominal aortic calcification (AAC), in contrast to coronary calcification, is less well documented. AAC may be recorded using plain radiographs. The present report is an analysis of baseline data on AAC in patients enrolled in the CORD (Calcification Outcome in Renal Disease) study. METHODS: A total of 47 centres in six European countries participated in this cross-sectional study. Inclusion criteria were age >or=18 years and duration of dialysis >or=3 months. Lateral lumbar radiography of the abdominal aorta was used to determine the overall AAC score, which is related to the severity of calcific deposits at lumbar vertebral segments L1-L4. The reliability of the method was tested by double reading of 64 radiographs (coefficient of correlation 0.9). RESULTS: A lateral lumbar radiograph was obtained in 933 patients. Calcification (AAC score >or= 1) was present in 81% of the patients; its severity increased significantly from L1 to L4 (P < 0.0001) and affected all of these segments in 51% of patients. Independent predictors for the presence and severity of calcification were age (odds ratio [OR] 1.103/year; P < 0.0001), duration of dialysis (OR 1.110/year; P = 0.002) and history of cardiovascular disease (OR 3.247; P < 0.0001). CONCLUSIONS: AAC detected by lateral lumbar radiograph is associated with several risk factors of uraemic calcification. This semi-quantitative method is more widely available and less expensive than the current procedures for studying calcification and could form part of a pre-transplant workup and cardiovascular risk stratification. PMID- 18676347 TI - PTH, FGF-23 and early CKD. PMID- 18676348 TI - Oral calcium load and fractional intestinal calcium absorption. PMID- 18676349 TI - A positive effect of AII inhibitors on peritoneal membrane function in long-term PD patients. AB - BACKGROUND: Experimental studies showed that inhibition of AII effects attenuates the development of peritoneal membrane fibrosis and neoangiogenesis. The latter leads to an increase of peritoneal solute transport and ultrafiltration failure. The results of a single-centre study showed that use of ACEI/ARB can prevent the increase of small solute transport in long-term PD patients. Our aim was to investigate whether these results would also be present in a larger population and influence patient and technique survival in long-term PD. METHODS: We analysed data from 217 long-term CAPD patients, participating in the Netherlands Cooperative Study on Adequacy of Dialysis (NECOSAD). Included patients underwent CAPD therapy for at least 2 years; 120 of them were treated with the ACE/AII inhibitors-ACEI/ARB group. The control group consisted of 87 patients who received none of the above drugs and 10 patients who had them for <25% of their time on PD. RESULTS: A significant difference in the time course of peritoneal transport was found between the two groups. The value of 24-h D/P creatinine was associated with the PD duration (P = 0.01) and its time course was influenced by use of ACEI/ARB (P = 0.05). We found no effect of ACEI/ARB on patient survival, but some benefit was found for the technique survival: in a multivariate model the hazard ratio for the group with the longest use of ACEI/ARB was 0.5 (CI 0.22 1.4), P = 0.19. CONCLUSIONS: We conclude that AII inhibition prevents the increase in small solute transport in long-term PD. These drugs may also have some positive influence on PD technique survival. PMID- 18676350 TI - A comparison of haemoglobin levels and doses in haemodialysis patients treated with subcutaneous or intravenous darbepoetin alfa: a German prospective, randomized, multicentre study. AB - BACKGROUND: The different efficacy of subcutaneous and intravenous rHuEPO results in higher doses and costs in intravenously treated patients. Darbepoetin alfa has a different pharmacokinetic profile compared to rHuEPO, and previous clinical experience suggests that subcutaneous and intravenous darbepoetin alfa may have similar efficacy. Objective. The aim of this study was to compare the efficacy of intravenous and subcutaneous darbepoetin alfa regarding haemoglobin levels and doses. METHODS: Patients treated with subcutaneous darbepoetin alfa for at least 6 months were randomized 1:1 to continue with subcutaneous treatment of darbepoetin alfa or to switch to the intravenous administration route. The application frequency was not altered. Darbepoetin alfa dose as well as haemoglobin concentrations were evaluated as per patient average at baseline (Week -3 +/- 1), Week 24 +/- 3 and Week 48 +/- 3. RESULTS: One hundred fourteen patients in 9 German dialysis centres were included. Fifty-three patients were treated intravenously and 61 patients continued the subcutaneous therapy. Mean haemoglobin levels and mean weekly darbepoetin alfa dose did not change significantly in either treatment group. CONCLUSIONS: Our data suggest that the darbepoetin alfa dose can be kept constant if patients are switched from subcutaneous to intravenous treatment. PMID- 18676351 TI - Correlation of enhanced thrombospondin-1 expression, TGF-beta signalling and proteinuria in human type-2 diabetic nephropathy. AB - BACKGROUND: Activation of the thrombospondin-1 (TSP-1)-TGF-beta pathway by glucose and the relevance of TSP-1-dependent activation of TGF-beta for renal matrix expansion, renal fibrosis and sclerosis have previously been demonstrated by our group in in vivo and in vitro studies. Design and methods. We investigated renal biopsies (n = 40) and clinical data (n = 30) of patients with diabetic nephropathy. Ten kidneys without evidence of renal disease served as controls. Glomerular and cortical expression of TSP-1, p-smad2/3, fibrosis and glomerular sclerosis (PAS) were assessed by immunhistochemical staining and related with clinical data. RESULTS: Glomerular (g) and cortical (c) TSP-1 were increased during diabetic nephropathy (g: 2.62 +/- 2.65; c: 4.5 +/- 4.2) compared to controls (g: 0.67 +/- 0.7; c: 1.5 +/- 1.2). P-smad2/3 was significantly increased (g: 16.7 +/- 12.9; c: 148.7 +/- 92.8) compared to controls (g: 7.1 +/- 3.6; c: 55 +/- 25; P < 0.05). TSP-1 was coexpressed with p-smad2/3 as an indicator of TGF beta activation. TSP-1 correlated with enhanced tubulointerstitial p-smad2/3 positivity (r = 0.39 and r = 0.4, P < 0.05) and glomerular p-smad2/3 correlated with proteinuria (r = 0.35, P < 0.05). CONCLUSIONS: In summary, the present study suggests a functional activity of the TSP-1/TGF-beta axis, especially in the tubulointerstitium of patients with diabetic nephropathy. The positive correlation of glomerular p-smad2/3 positivity with proteinuria further supports the importance of the TSP-1/TGF-beta system as a relevant mechanism for progression of human type-2 diabetic nephropathy. PMID- 18676353 TI - Down-regulation of asymmetric arginine methylation during replicative and H2O2 induced premature senescence in WI-38 human diploid fibroblasts. AB - Protein arginine methylation is one of the post-translational modifications which yield monomethyl and dimethyl (asymmetric or symmetric) arginines in proteins. In the present study, we investigated the status of protein arginine methylation during human diploid fibroblast senescence. When the expression of protein arginine methyltransferases (PRMTs), namely PRMT1, PRMT4, PRMT5 and PRMT6 was examined, a significant reduction was found in replicatively senescent cells as well as their catalytic activities against histone mixtures compared with the young cells. Furthermore, when the endogenous level of arginine-dimethylated proteins was determined, asymmetric modification (the product of type I PRMTs including PRMT1, PRMT4 and PRMT6) was markedly down-regulated. In contrast, both up- and down-regulations of symmetrically arginine-methylated proteins (the product of type II PRMTs including PRMT5) during replicative senescence were found. Furthermore, when young fibroblasts were induced to premature senescence by sub-cytotoxic H2O2 treatment, results similar to replicative senescence were obtained. Finally, we found that SV40-mediated immortalized WI-38 and HeLa cell lines maintained a higher level of asymmetrically modified proteins as well as type I PRMTs than young fibroblasts. These results suggest that the maintenance of asymmetric modification in the expressed target proteins of type I PRMTs might be critical for cellular proliferation. PMID- 18676352 TI - Endothelium-specific interference with peroxisome proliferator activated receptor gamma causes cerebral vascular dysfunction in response to a high-fat diet. AB - The ligand-activated transcription factor peroxisome proliferator activated receptor gamma (PPARgamma) is expressed in vascular endothelium where it exerts anti-inflammatory and antioxidant effects. However, its role in regulating vascular function remains undefined. We examined endothelial function in transgenic mice expressing dominant-negative mutants of PPARgamma under the control of an endothelial-specific promoter to test the hypothesis that endothelial PPARgamma plays a protective role in the vasculature. Under baseline conditions, responses to the endothelium-dependent agonist acetylcholine were not affected in either aorta or the basilar artery in vitro. In response to feeding a high-fat diet for 12 weeks, acetylcholine produced dilation that was markedly impaired in the basilar artery of mice expressing dominant-negative mutants, but not in mice expressing wild-type PPARgamma controlled by the same promoter. Unlike basilar artery, 12 weeks of a high-fat diet was not sufficient to cause endothelial dysfunction in the aorta of mice expressing dominant-negative PPARgamma, although aortic dysfunction became evident after 25 weeks. The responses to acetylcholine in basilar artery were restored to normal after treatment with a scavenger of superoxide. Baseline blood pressure was only slightly elevated in the transgenic mice, but the pressor response to angiotensin II was augmented. Thus, interference with PPARgamma in the endothelium produces endothelial dysfunction in the cerebral circulation through a mechanism involving oxidative stress. Consistent with its role as a fatty acid sensor, these findings provide genetic evidence that endothelial PPARgamma plays a critical role in protecting blood vessels in response to a high-fat diet. PMID- 18676354 TI - The effects of LAMP1 and LAMP3 on M180 amelogenin uptake, localization and amelogenin mRNA induction by amelogenin protein. AB - We previously demonstrated that the uptake of M180 amelogenin protein in dental epithelial cells (HAT-7) results in increased levels of amelogenin mRNA through enhanced mRNA stabilization. To determine the processes involved in the uptake of extracellular M180 amelogenin by cells and in amelogenin intracellular trafficking in the amelogenin protein-mediated amelogenin mRNA expression pathway, we investigated the effects of LAMP1 and LAMP3, which are candidate M180 amelogenin receptors, on M180 amelogenin uptake, localization and amelogenin mRNA induction by amelogenin protein, using anti-LAMP-1 and anti-LAMP-3 antibodies and siRNA analysis. The results indicate that LAMP3 blocking by anti-LAMP-3 decreases M180 amelogenin uptake, but does not affect amelogenin mRNA induction by amelogenin protein, suggesting that LAMP3 is related to amelogenin degradation. Down-regulation by siRNA of LAMP1, which is the receptor for small amelogenin protein (LRAP), does not affect M180 amelogenin uptake, localization or amelogenin mRNA induction by amelogenin protein. Thus, while LAMP1 is the specific receptor for LRAP, it is not a receptor for M180 amelogenin. These findings will aid further research into the understanding of M180 amelogenin function and expression. PMID- 18676355 TI - A novel tool to analyze MRI recurrence patterns in glioblastoma. AB - At least 10% of glioblastoma relapses occur at distant and even contralateral locations. This disseminated growth limits surgical intervention and contributes to neurological morbidity. Preclinical data pointed toward a role for temozolomide (TMZ) in reducing radiotherapy-induced glioma cell invasiveness. Our objective was to develop and validate a new analysis tool of MRI data to examine the clinical recurrence pattern of glioblastomas. MRIcro software was used to map the location and extent of initial preoperative and recurrent tumors on MRI of 63 patients in the European Organisation for Research and Treatment of Cancer (EORTC) 26981/22981/National Cancer Institute of Canada (NCIC) CE.3 study into the same stereotaxic space. This allowed us to examine changes of site and distance between the initial and the recurrent tumor on the group level. Thirty of the 63 patients were treated using radiotherapy, while the other patients completed a radiotherapy-plus-TMZ treatment. Baseline characteristics (median age, KPS) and outcome data (progression-free survival, overall survival) of the patients included in this analysis resemble those of the general study cohort. The patient groups did not differ in the promoter methylation status of methyl guanine methyltransferase (MGMT). Overall frequency of distant recurrences was 20%. Analysis of recurrence patterns revealed no difference between the groups in the size of the recurrent tumor or in the differential effect on the distance of the recurrences from the preoperative tumor location. The data show the feasibility of groupwise recurrence pattern analysis. An effect of TMZ treatment on the recurrence pattern in the EORTC 26981/22981/NCIC CE.3 study could not be demonstrated. PMID- 18676358 TI - In reference to Maschio et Al. (Neuro-oncology. 2008;10:106-107). PMID- 18676359 TI - In vivo gene delivery by embryonic-stem-cell-derived astrocytes for malignant gliomas. AB - The treatment of malignant gliomas with current therapies remains a challenge in neurooncology. Our recent work showed that embryonic stem cell (ESC)-derived astrocytes conditionally expressing genes can be used to induce apoptosis in malignant glioma cells in vitro. The tumor necrosis factor-related apoptosis inducing ligand (TRAIL) gene has been shown to induce apoptosis in a variety of tumor cells, including gliomas. The aim of this study was to assess the proapoptotic effects of transgenic TRAIL delivered by ESC-derived astrocytes on malignant gliomas in vivo. Malignant glioma A172 cells were used to induce heterotopic xenografts in nude mice. ESC-derived astrocytes conditionally expressing TRAIL were injected into the xenografts. TRAIL expression was documented in the malignant glioma xenografts by reverse transcription PCR and immunohistochemistry after external gene induction. A significant reduction in tumor volume occurred 48 h after a single injection (14%) and double injections (31%) in the experimental groups. Terminal dUTP nick end labeling (TUNEL) revealed abundant apoptotic tumor cells in the experimental groups. Seven days after injection, the tumor had undergone severe necrosis, with only scattered residual tumor cells at the periphery. Death receptor DR4 expression increased significantly in the experimental groups compared with controls. Our data suggest that ESC-derived astrocytes conditionally expressing TRAIL should be considered as vectors to deliver gene therapy for malignant gliomas. PMID- 18676360 TI - Retarded differentiation of Leydig cells and increased apoptosis of germ cells in the initial round of spermatogenesis of rats with lethal dwarf and epilepsy (lde/lde) phenotypes. AB - The lde/lde rats show a severe dwarf phenotype with early postnatal lethality and a high incidence of epileptic seizure. Seizures are first detected in this model between 16 and 63 days of age, and mostly begin as wild running and progress to generalized tonic-clonic convulsions. Because our histological examination detected many extracellular vacuoles in the hippocampus and amygdaloid bodies of these animals at 28 days of age, these pathological alterations may be related to the epileptogenesis in lde/lde rats. In addition to these defects, male lde/lde rats have apparently smaller testes with reduced number of germ cells and poorly matured adult-type Leydig cells in comparison with wild-type controls. In the present study, we performed anatomical, histological, and endocrinologic examinations to characterize the testicular phenotype of lde/lde rats at 21, 28, 35, and 56 days of age. Male lde/lde rats showed severely retarded growth of the testes and accessory sex organs. Their seminiferous tubules were significantly smaller and contained markedly fewer germ cells at all time points examined as compared with controls. Significantly fewer Sertoli cells at 21 and 28 days of age, markedly decreased spermatocyte number at 28 days of age, and delayed appearance of spermatids at 56 days of age were observed in the testes of lde/lde rats. More TUNEL (T&T-mediated duTP-biotin nick-end labeling)-positive cells were detected in lde/lde seminiferous tubules, and the largest number of apoptotic cells was recorded at 28 days of age. The increases in 3beta-hydroxysteroid dehydrogenase-positive adult-type Leydig cells and 11beta-hydroxysteroid dehydrogenase-positive mature adult-type Leydig cells were also severely retarded in the testes of lde/lde rats. Consistent with these defects, significantly lower plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone concentrations were detected in lde/lde males at 28 days of age, and weak immunostaining for FSH and smaller cytoplasm of LH-positive cells were detected in the anterior pituitary lobes of lde/lde males. Despite a normal level of plasma LH after 35 days of age, a significantly lower level of plasma testosterone was detected at 56 days of age. These results indicate that the normal lde allele is related to prepubertal elevations of gonadotropins and normal development of adult-type Leydig cells. Because lde/lde rats experience epileptic seizures during the period when the hypothalamus-pituitary-testicular axis is established, lde/lde rats would be useful as a model for reproductive disorder with pediatric epilepsy. PMID- 18676356 TI - Biological background of pediatric medulloblastoma and ependymoma: a review from a translational research perspective. AB - Survival rates of pediatric brain tumor patients have significantly improved over the years due to developments in diagnostic techniques, neurosurgery, chemotherapy, radiotherapy, and supportive care. However, brain tumors are still an important cause of cancer-related deaths in children. Prognosis is still highly dependent on clinical characteristics, such as the age of the patient, tumor type, stage, and localization, but increased knowledge about the genetic and biological features of these tumors is being obtained and might be useful to further improve outcome for these patients. It has become clear that the deregulation of signaling pathways essential in brain development, for example, sonic hedgehog (SHH), Wnt, and Notch pathways, plays an important role in pathogenesis and biological behavior, especially for medulloblastomas. More recently, data have become available about the cells of origin of brain tumors and the possible existence of brain tumor stem cells. Newly developed array-based techniques for studying gene expression, protein expression, copy number aberrations, and epigenetic events have led to the identification of other potentially important biological abnormalities in pediatric medulloblastomas and ependymomas. PMID- 18676361 TI - Curcumin blocks the activation of androgen and interlukin-6 on prostate-specific antigen expression in human prostatic carcinoma cells. AB - Curcumin, a naturally occurring compound, exhibits anticancer chemopreventive effects. We evaluated the effects and mechanisms of curcumin on the gene expression of prostate-specific antigen (PSA) in human androgen-sensitive prostatic carcinoma cells. LNCaP cells were used to determine the effect of curcumin on PSA expression. Quantitative PSA expression was assessed by reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and immunoblot assay. The modulation of androgen, interlukin-6 (IL 6), and prostate-derived Ets factor (PDEF) on the PSA gene was identified by transient gene expression assay with the use of a PSA reporter vector. The effect of curcumin on the activity of androgen receptors was evaluated by electrophoretic mobility shift assay (EMSA). Immunoblot assays, RT-PCR, and ELISA indicated that curcumin treatments blocked the stimulation of methyltrienolone (R1881) and IL-6 on PSA gene expression in LNCaP cells. The effects of curcumin appear to be mediated via the androgen response element of PSA gene. Results from immunoblot assay and EMSA revealed the modulation of curcumin on the expression of androgen receptor and androgen receptor binding activity on androgen response element of PSA gene. Although overexpression of PDEF dramatically enhanced PSA gene expression, the results of immunoblot assays and transient reporter assays indicated that curcumin treatments did not affect the gene expression of PDEF. Curcumin inhibits R1881- and IL-6-mediated PSA gene expression in LNCaP cells through down-regulation of the expression and activity of androgen receptors. PMID- 18676362 TI - Genetic screening in infertile Mexican men: chromosomal abnormalities, Y chromosome deletions, and androgen receptor CAG repeat length. AB - In our study, we analyzed chromosomal abnormalities, Y chromosome deletions, androgen receptor CAG repeat length and their association with defective spermatogenesis in infertile Mexican men. Eighty-two infertile patients and 40 controls were screened for karyotypic abnormalities, Y chromosome microdeletions, and CAG repeats. Nine infertile males (11%) carried chromosomal abnormalities and 10 (12.2%) presented Y chromosome microdeletions. The mean CAG repeat length was 21.6 and 20.88 base pairs in idiopathic infertile males and controls, respectively. Our results suggest that chromosomal aberrations and Y-chromosomal microdeletions are related to male infertility in Mexican men. In addition, expansion of the CAG repeat segments of the androgen receptor is not correlated with male idiopathic infertility. PMID- 18676363 TI - Transketolase and 2',3'-cyclic-nucleotide 3'-phosphodiesterase type I isoforms are specifically recognized by IgG autoantibodies in multiple sclerosis patients. AB - The presence of autoantibodies in multiple sclerosis (MuS) is well known, but their target antigens have not been clearly identified. In the present study, IgG autoreactivity to neural antigens of normal human white matter separated by bidimensional electrophoresis was assessed in serum and cerebrospinal fluid of 18 MuS and 20 control patients. Broad IgG autoreactivity was detected by two dimensional immunoblotting in all cases to neural antigens, most of which were identified by mass spectrometry. The comparative analysis of MuS and non-MuS reactive spots showed that a restricted number of neural protein isoforms were specifically recognized by MuS IgG. Almost all MuS patients had cerebrospinal fluid IgG directed to isoforms of one of the oligodendroglial molecules, transketolase, 2',3'-cyclic-nucleotide 3'-phosphodiesterase type I, collapsin response mediator protein 2, and tubulin beta 4. Interestingly 50% of MuS IgG recognized transketolase, which was mostly localized on oligodendrocytes in human white matter from normal and MuS samples. IgG autoreactivity to cytoskeletal proteins (radixin, sirtuin 2, and actin-interacting protein 1) was prevalent in secondary progressive MuS patients. Among the proteins recognized by serum IgG, almost all MuS patients specifically recognized a restricted number of neuronal/cytoskeletal proteins, whereas 2',3'-cyclic-nucleotide 3' phosphodiesterase type I was the oligodendroglial antigen most frequently recognized (44%) by MuS seric IgG. Our immunomics approach shed new light on the autoimmune repertoire present in MuS patients revealing novel oligodendroglial and/or neuronal putative autoantigens with potential important pathogenic and diagnostic implications. PMID- 18676364 TI - Banking of biological fluids for studies of disease-associated protein biomarkers. AB - With the increasing demand of providing personalized medicine the need for biobanking of biological material from individual patients has increased. Such samples are essential for molecular research aimed at characterizing diseases at several levels ranging from epidemiology and diagnostic and prognostic classification to prediction of response to therapy. Clinically validated biomarkers may provide information to be used for diagnosis, screening, evaluation of risk/predisposition, assessment of prognosis, monitoring (recurrence of disease), and prediction of response to treatment and as a surrogate response marker. Many types of biological fluids or tissues can be collected and stored in biorepositories. Samples of blood can be further processed into plasma and serum, and tissue pieces can be either frozen or fixed in formalin and then embedded into paraffin. The present review focuses on biological fluids, especially serum and plasma, intended for study of protein biomarkers. In biomarker studies the process from the decision to take a sample from an individual to the moment the sample is safely placed in the biobank consists of several phases including collection of samples, transport of the samples, and handling and storage of samples. Critical points in each step important for high quality biomarker studies are described in this review. Failure to develop and adhere to robust standardized protocols may have significant consequences as the quality of the material stored in the biobank as well as conclusions and clinical recommendations based on analysis of such material may be severely affected. PMID- 18676365 TI - Chemical and pathway proteomics: powerful tools for oncology drug discovery and personalized health care. AB - In recent years mass spectrometry-based proteomics has moved beyond a mere quantitative description of protein expression levels and their possible correlation with disease or drug action. Impressive progress in LC-MS instrumentation together with the availability of new enabling tools and methods for quantitative proteome analysis and for identification of posttranslational modifications has triggered a surge of chemical and functional proteomics studies dissecting mechanisms of action of cancer drugs and molecular mechanisms that modulate signal transduction pathways. Despite the tremendous progress that has been made in the field, major challenges, relating to sensitivity, dynamic range, and throughput of the described methods, remain. In this review we summarize recent advances in LC-MS-based approaches and their application to cancer drug discovery and to studies of cancer-related pathways in cell culture models with particular emphasis on mechanistic studies of drug action in these systems. Moreover we highlight the emerging utility of pathway and chemical proteomics techniques for translational research in patient tissue. PMID- 18676366 TI - Adaptation of connexin 43-hemichannel prostaglandin release to mechanical loading. AB - Bone tissues respond to mechanical loading/unloading regimens to accommodate (re)modeling requirements; however, the underlying molecular mechanism responsible for these responses is largely unknown. Previously, we reported that connexin (Cx) 43 hemichannels in mechanosensing osteocytes mediate the release of prostaglandin, PGE(2), a crucial factor for bone formation in response to anabolic loading. We show here that the opening of hemichannels and release of PGE(2) by shear stress were significantly inhibited by a potent antibody we developed that specifically blocks Cx43-hemichannels, but not gap junctions or other channels. The opening of hemichannels and release of PGE(2) are magnitude dependent on the level of shear stress. Insertion of a rest period between stress enhances this response. Hemichannels gradually close after 24 h of continuous shear stress corresponding with reduced Cx43 expression on the cell surface, thereby reducing any potential negative effects of channels staying open for extended periods. These data suggest that Cx43-hemichannel activity associated with PGE(2) release is adaptively regulated by mechanical loading to provide an effective means of regulating levels of extracellular signaling molecules responsible for initiation of bone (re)modeling. PMID- 18676367 TI - Regulation of cholesterologenesis by the oxysterol receptor, LXRalpha. AB - Cholesterol is required for normal cellular and physiological function, yet dysregulation of cholesterol metabolism is associated with diseases such as atherosclerosis. Cholesterol biosynthesis is regulated by end product negative feedback inhibition where the levels of sterols and oxysterols regulate the expression of cholesterologenic enzymes. Sterol regulatory element-binding protein-2 is responsive to both sterols and oxysterols and has been shown to mediate the transcriptional response of the cholesterologenic enzymes to these lipids. Here, we show that the nuclear hormone receptor for oxysterols, the liver X receptor alpha (LXRalpha), regulates cholesterol biosynthesis by directly silencing the expression of two key cholesterologenic enzymes (lanosterol 14alpha demethylase (CYP51A1), and squalene synthase (farnesyl diphosphate farnesyl transferase 1)) via novel negative LXR DNA response elements (nLXREs) located in each of these genes. Examination of the CYP51A1 gene revealed that both the SRE and nLXRE are required for normal oxysterol-dependent repression of this gene. Thus, these data suggest that LXRalpha plays an important role in the regulation of cholesterol biosynthesis. PMID- 18676368 TI - Multiple ATP binding is required to stabilize the "activated" (clamp open) clamp loader of the T4 DNA replication complex. AB - Most DNA replication systems include a sliding clamp that encircles the genomic DNA and links the polymerase to the template to control polymerase processivity. A loading complex is required to open the clamp and place it onto the DNA. In phage T4 this complex consists of a trimeric clamp of gp45 subunits and a pentameric loader assembly of four gp44 and one gp62 subunit(s), with clamp loading driven by ATP binding. We measure this binding as a function of input ligand concentration and show that four ATPs bind to the gp44/62 complex with equal affinity. In contrast, the ATPase rate profile of the clamp-clamp loader complex exhibits a marked peak at an input ATP concentration close to the overall Kd (approximately 30 microm), with further increases in bound ATP decreasing the ATPase rate to a much lower level. Thus the progressive binding of the four ATPs triggers a conformational change in the complex that markedly inhibits ATPase activity. This inhibition is related to ring opening by using a clamp that is covalently cross-linked across its subunit interfaces and thus rendered incapable of opening. Binding of this clamp abolishes substrate inhibition of the ATPase but leaves ATP binding unchanged. We show that four ATP ligands must bind to the T4 clamp loader before the loader can be fully "activated" and the clamp opened, and that ATP hydrolysis is required only for release of the loader complex after clamp loading onto the replication fork has been completed. PMID- 18676369 TI - Nonspecific interaction of prefibrillar amyloid aggregates with glutamatergic receptors results in Ca2+ increase in primary neuronal cells. AB - It is widely reported that the Ca(2+) increase following nonspecific cell membrane permeabilization is among the earliest biochemical modifications in cells exposed to toxic amyloid aggregates. However, more recently receptors with Ca(2+) channel activity such as alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA), N-methyl D-aspartate (NMDA), ryanodine, and inositol 1,4,5 trisphosphate receptors have been proposed as mediators of the Ca(2+) increase in neuronal cells challenged with beta-amyloid peptides. We previously showed that prefibrillar aggregates of proteins not associated with amyloid diseases are toxic to exposed cells similarly to comparable aggregates of disease-associated proteins. In particular, prefibrillar aggregates of the prokaryotic HypF-N were shown to be toxic to different cultured cell lines by eliciting Ca(2+) and reactive oxygen species increases. This study was aimed at assessing whether NMDA and AMPA receptor activations could be considered a generic feature of cell interaction with amyloid aggregates rather than a specific effect of some aggregated protein. Therefore, we investigated whether NMDA and AMPA receptors were involved in the Ca(2+) increase following exposure of rat cerebellar granule cells to HypF-N prefibrillar aggregates. We found that the intracellular Ca(2+) increase was associated with the early activation of NMDA and AMPA receptors, although some nonspecific membrane permeabilization was also observed at longer times of exposure. This result matched a significant co-localization of the aggregates with both receptors on the plasma membrane. Our data support the possibility that glutamatergic channels are generic sites of interaction with the cell membrane of prefibrillar aggregates of different peptides and proteins as well as the key structures responsible for the resulting early membrane permeabilization to Ca(2+). PMID- 18676370 TI - Re-evaluating the roles of proposed modulators of mammalian target of rapamycin complex 1 (mTORC1) signaling. AB - Signaling through mammalian target of rapamycin complex 1 (mTORC1) is stimulated by amino acids and insulin. Insulin inactivates TSC1/2, the GTPase-activator complex for Rheb, and Rheb.GTP activates mTORC1. It is not clear how amino acids regulate mTORC1. FKBP38 (immunophilin FK506-binding protein, 38 kDa), was recently reported to exert a negative effect on mTORC1 function that is relieved by its binding to Rheb.GTP. We confirm that Rheb binds wild type FKBP38, but inactive Rheb mutants showed contrasting abilities to bind FKBP38. We were unable to observe any regulation of FKBP38/mTOR binding by amino acids or insulin. Furthermore, FKBP38 did not inhibit mTORC1 signaling. The translationally controlled tumor protein (TCTP) in Drosophila was recently reported to act as the guanine nucleotide-exchange factor for Rheb. We have studied the role of TCTP in mammalian TORC1 signaling and its control by amino acids. Reducing TCTP levels did not reproducibly affect mTORC1 signaling in amino acid-replete/insulin stimulated cells. Moreover, overexpressing TCTP did not rescue mTORC1 signaling in amino acid-starved cells. In addition, we were unable to see any stable interaction between TCTP and Rheb or mTORC1. Accumulation of uncharged tRNA has been previously proposed to be involved in the inhibition of mTORC1 signaling during amino acid starvation. To test this hypothesis, we used a Chinese hamster ovary cell line containing a temperature-sensitive mutation in leucyl-tRNA synthetase. Leucine deprivation markedly inhibited mTORC1 signaling in these cells, but shifting the cells to the nonpermissive temperature for the synthetase did not. These data indicate that uncharged tRNA(Leu) does not switch off mTORC1 signaling and suggest that mTORC1 is controlled by a distinct pathway that senses the availability of amino acids. Our data also indicate that, in the mammalian cell lines tested here, neither TCTP nor FKBP38 regulates mTORC1 signaling. PMID- 18676371 TI - Demonstration of the iron-regulated surface determinant (Isd) heme transfer pathway in Staphylococcus aureus. AB - In this study, we report experimental results that provide the first complete challenge of a proposed model for heme acquisition by Staphylococcus aureus via the Isd pathway first put forth by Mazmanian, S. K., Skaar, E. P., Gaspar, A. H., Humayun, M., Gornicki, P., Jelenska, J., Joachmiak, A., Missiakas, D. M., and Schneewind, O. (2003) Science 299, 906-909. The heme-binding NEAT domains of Isd proteins IsdA, IsdB (domain 2), IsdC, and HarA/IsdH (domain 3), and the heme binding IsdE protein, were overexpressed and purified in apo (heme-free) form. Absorption and magnetic circular dichroism spectral data, together with electrospray ionization mass spectrometry were used to unambiguously identify that heme transfers from NEAT-A through NEAT-C to IsdE. Heme transfer was demonstrated to occur in a unidirectional fashion in the sequence NEAT-B2 --> NEAT-A --> NEAT-C --> IsdE or, alternatively, initiating from NEAT-H3 instead of NEAT-B2: NEAT-H3 --> NEAT-A --> NEAT-C --> IsdE. Under the conditions of our experiments, only NEAT-H3 and NEAT-B2 could transfer bidirectionally, which is in the reverse direction as well, and only with each other. Whereas apo-IsdE readily accepted heme from holo-NEAT-C, it would not accept heme from holo-NEAT-A. Heme transfer to IsdE requires the presence of holo-NEAT-C, in agreement with the proposal that IsdC serves as the central conduit of the heme transfer pathway. These experimental findings corroborate the heme transfer model first proposed by the Schneewind group. Our data show that heme transport from the wall-anchored IsdH/IsdB proteins proceeds directly to IsdE at the membrane and, for this to occur, we propose that specific protein-protein interactions must take place. PMID- 18676373 TI - Hurp deficiency in mice leads to female infertility caused by an implantation defect. AB - First revealed in cancer studies, HURP (hepatoma up-regulated protein) is a cell cycle-associated gene with elevated expression in the G(2)/M phase. Cell culture studies have revealed that HURP is an essential factor required for spindle formation and chromosome congression during mitosis. However, the function of HURP in an in vivo context has not been explored. We generated a Hurp knock-out (Hurp(-/-)) mouse to investigate the role of HURP in development under normal physiological conditions. Hurp(-/-) mice develop normally and are indistinguishable from their wild-type littermates. Interestingly, breeding experiments revealed that Hurp(-/-) females are completely infertile, whereas the males reproduce normally. Ovulation, fertilization, and pre-implantation embryo development are normal; however, the Hurp(-/-) females are unable to form implantation sites due to an inability to undergo the decidual reaction. This is caused by a defect in endometrial stromal proliferation that leads to implantation failure. Additionally, HURP expression in the uterus coincides with the implantation stage and can be induced by estrogen treatment. Our results demonstrate for the first time that HURP affects endometrial stromal proliferation during implantation but is dispensable during normal development in mice; specifically, HURP has an essential function in uterine biology. PMID- 18676372 TI - Ceramide generated by sphingomyelin hydrolysis and the salvage pathway is involved in hypoxia/reoxygenation-induced Bax redistribution to mitochondria in NT-2 cells. AB - Ceramide functions as an important second messenger in apoptosis signaling pathways. In this report, we show that treatment of NT-2 neuronal precursor cells with hypoxia/reoxygenation (H/R) resulted in ceramide up-regulation. This elevation in ceramide was primarily due to the actions of acid sphingomyelinase and ceramide synthase LASS 5, demonstrating the action of the salvage pathway. Hypoxia/reoxygenation treatment led to Bax translocation from the cytoplasm to mitochondria and cytochrome c release from mitochondria. Down-regulation of either acid sphingomyelinase or LASS 5-attenuated ceramide accumulation and H/R induced Bax translocation to mitochondria. Overall, we have demonstrated that ceramide up-regulation following H/R is pertinent to Bax activation to promote cell death. PMID- 18676374 TI - cAMP-responding element-binding protein and c-Ets1 interact in the regulation of ATP-dependent MUC5AC gene expression. AB - Exogenous ATP activates purinoreceptors on the cell surface that regulate diverse cellular functions, including mucous cell secretion in the respiratory epithelium. In this study, ATP increased MUC5AC mRNA in primary human nasal epithelial cells and in NCI-H292 pulmonary adenocarcinoma cells in vitro. ATP induced MUC5AC mRNA was mediated by phospholipase Cbeta3. A dominant-negative mutation in the PDZ binding domain of PLCbeta3 inhibited ATP-induced MUC5AC gene expression. ATP sequentially activated the phosphorylation of Akt, ERK1/2, p38, RSK1, and cAMP-responding element-binding protein (CREB) in a protein kinase C independent manner. ATP-induced MUC5AC mRNA levels were regulated by CREB via direct interaction with c-Ets1 on the MUC5AC gene promoter (located -938 to 930). Effects of CREB and c-Ets1 were additive. Inhibition of either CREB or c Ets1 inhibited ATP-induced MUC5AC gene expression. Stimulation with ATP caused the direct binding of CREB and c-Ets1 to the MUC5AC promoter, increasing the phosphorylation of c-Ets1. Chromatin immunoprecipitation assays demonstrated that in the presence of ATP, both c-Ets1 and CREB bound to the MUC5AC promoter. The effects of exogenous ATP on MUC5AC gene expression are mediated by a complex regulatory cascade controlling interactions between CREB and c-Ets1 that bind to a promoter element in the MUC5AC gene enhancing MUC5AC gene transcription. ATP dependent activation of MUC5AC gene expression via CREB-c-Ets1 may contribute to mucous cell hypersecretion associated with common respiratory disorders. PMID- 18676375 TI - Calmodulin association with connexin32-derived peptides suggests trans-domain interaction in chemical gating of gap junction channels. AB - Calmodulin plays a key role in the chemical gating of gap junction channels. Two calmodulin-binding regions have previously been identified in connexin32 gap junction protein, one in the N-terminal and another in the C-terminal cytoplasmic tail of the molecule. The aim of this study was to better understand how calmodulin interacts with the connexin32-binding domains. Lobe-specific interactions of calmodulin with connexin32 peptides were studied by stopped flow kinetics, using Ca(2+) binding-deficient mutants. Peptides corresponding to the N terminal tail (residues 1-22) of connexin32 engaged both the N- and C-terminal lobes (N- and C-lobes) of calmodulin, binding with higher affinity to the C-lobe of calmodulin (Ca(2+) dissociation rate constants k(3,4), 1.7+/-0.5 s(-1)) than to the N-lobe (k(1,2), 10.8+/-1.3 s(-1)). In contrast, peptides representing the C-terminal tail domain (residues 208-227) of connexin32 bound either the C- or the N-lobe but only one calmodulin lobe at a time (k(3,4), 2.6+/-0.1 s(-1) or k(1), 13.8+/-0.5 s(-1) and k(2), 1000 s(-1)). The calmodulin-binding domains of the N- and C-terminal tails of connexin32 were best defined as residues 1-21 and 216-227, respectively. Our data, showing separate functions of the N- and C-lobes of calmodulin in the interactions with connexin32, suggest trans-domain or trans subunit bridging by calmodulin as a possible mechanism of gap junction gating. PMID- 18676376 TI - Cooperative synergy between NFAT and MyoD regulates myogenin expression and myogenesis. AB - Calcineurin/NFAT signaling is involved in multiple aspects of skeletal muscle development and disease. The myogenic basic helix-loop-helix transcription factors, MyoD, myogenin, Myf5, and MRF4 specify the myogenic lineage. Here we show that calcineurin/NFAT (nuclear factor of activated T cells) signaling is required for primary myogenesis by transcriptional cooperation with the basic helix-loop-helix transcription factor MyoD. Calcineurin/NFAT signaling is involved in myogenin expression in differentiating myoblasts, where the myogenic regulatory factor MyoD synergistically cooperates with NFATc2/c3 at the myogenin promoter. Using gel shift and chromatin immunoprecipitation assays, we identified two conserved NFAT binding sites in the myogenin promoter that were occupied by NFATc3 upon skeletal muscle differentiation. The transcriptional integration between NFATc3 and MyoD is crucial for primary myogenesis in vivo, as myogenin expression is weak in myod:nfatc3 double null embryos, whereas myogenin expression is unaffected in embryos with null mutations for either factor alone. Thus, the combined findings provide a novel transcriptional paradigm for the first steps of myogenesis, where a calcineurin/NFATc3 pathway regulates myogenin induction in cooperation with MyoD during myogenesis. PMID- 18676377 TI - Sialylation of beta1 integrins blocks cell adhesion to galectin-3 and protects cells against galectin-3-induced apoptosis. AB - In previous studies, we determined that beta1 integrins from human colon tumors have elevated levels of alpha2-6 sialylation, a modification added by beta galactosamide alpha-2,6-sialyltranferase I (ST6Gal-I). Intriguingly, the beta1 integrin is thought to be a ligand for galectin-3 (gal-3), a tumor-associated lectin. The effects of gal-3 are complex; intracellular forms typically protect cells against apoptosis through carbohydrate-independent mechanisms, whereas secreted forms bind to cell surface oligosaccharides and induce apoptosis. In the current study, we tested whether alpha2-6 sialylation of the beta1 integrin modulates binding to extracellular gal-3. Herein we report that SW48 colonocytes lacking alpha2-6 sialylation exhibit beta1 integrin-dependent binding to gal-3 coated tissue culture plates; however, binding is attenuated upon forced expression of ST6Gal-I. Removal of alpha2-6 sialic acids from ST6Gal-I expressors by neuraminidase treatment restores gal-3 binding. Additionally, using a blot overlay approach, we determined that gal-3 binds directly and preferentially to unsialylated, as compared with alpha2-6-sialylated, beta1 integrins. To understand the physiologic consequences of gal-3 binding, cells were treated with gal-3 and monitored for apoptosis. Galectin-3 was found to induce apoptosis in parental SW48 colonocytes (unsialylated), whereas ST6Gal-I expressors were protected. Importantly, gal-3-induced apoptosis was inhibited by function blocking antibodies against the beta1 subunit, suggesting that beta1 integrins are critical transducers of gal-3-mediated effects on cell survival. Collectively, our results suggest that the coordinate up-regulation of gal-3 and ST6Gal-I, a feature that is characteristic of colon carcinoma, may confer tumor cells with a selective advantage by providing a mechanism for blockade of the pro apoptotic effects of secreted gal-3. PMID- 18676378 TI - Effect of low root temperature on hydraulic conductivity of rice plants and the possible role of aquaporins. AB - The role of root temperature T(R) in regulating the water-uptake capability of rice roots and the possible relationship with aquaporins were investigated. The root hydraulic conductivity Lp(r) decreased with decreasing T(R) in a measured temperature range between 10 degrees C and 35 degrees C. A single break point (T(RC) = 15 degrees C) was detected in the Arrhenius plot for steady-state Lp(r). The temperature dependency of Lp(r) represented by activation energy was low (28 kJ mol(-1)) above T(RC), but the value is slightly higher than that for the water viscosity. Addition of an aquaporin inhibitor, HgCl(2), into root medium reduced osmotic exudation by 97% at 25 degrees C, signifying that aquaporins play a major role in regulating water uptake. Below T(RC), Lp(r) declined precipitously with decreasing T(R) (E(a) = 204 kJ mol(-1)). When T(R) is higher than T(RC), the transient time for reaching the steady-state of Lp(r) after the immediate change in T(R) (from 25 degrees C) was estimated as 10 min, while it was prolonged up to 2-3 h when T(R) < T(RC). The Lp(r) was completely recovered to the initial levels when T(R) was returned back to 25 degrees C. Immunoblot analysis using specific antibodies for the major aquaporin members of PIPs and TIPs in rice roots revealed that there were no significant changes in the abundance of aquaporins during 5 h of low temperature treatment. Considering this result and the significant inhibition of water-uptake by the aquaporin inhibitor, we hypothesize that the decrease in Lp(r) when T(R) < T(RC) was regulated by the activity of aquaporins rather than their abundance. PMID- 18676379 TI - Identification of maize silicon influx transporters. AB - Maize (Zea mays L.) shows a high accumulation of silicon (Si), but transporters involved in the uptake and distribution have not been identified. In the present study, we isolated two genes (ZmLsi1 and ZmLsi6), which are homologous to rice influx Si transporter OsLsi1. Heterologous expression in Xenopus laevis oocytes showed that both ZmLsi1 and ZmLsi6 are permeable to silicic acid. ZmLsi1 was mainly expressed in the roots. By contrast, ZmLsi6 was expressed more in the leaf sheaths and blades. Different from OsLsi1, the expression level of both ZmLsi1 and ZmLsi6 was unaffected by Si supply. Immunostaining showed that ZmLsi1 was localized on the plasma membrane of the distal side of root epidermal and hypodermal cells in the seminal and crown roots, and also in cortex cells in lateral roots. In the shoots, ZmLsi6 was found in the xylem parenchyma cells that are adjacent to the vessels in both leaf sheaths and leaf blades. ZmLsi6 in the leaf sheaths and blades also exhibited polar localization on the side facing towards the vessel. Taken together, it can be concluded that ZmLsi1 is an influx transporter of Si, which is responsible for the transport of Si from the external solution to the root cells and that ZmLsi6 mainly functions as a Si transporter for xylem unloading. PMID- 18676380 TI - On "a four-week, task-specific neuroprosthesis program..." Dunning K, et al. Phys Ther. 2008;88:397-405. PMID- 18676381 TI - On "breathing: a sign of life..." Ekerholt and Bergland. Phys Ther. 2008;88:832 840. PMID- 18676382 TI - First, do no harm: balancing the risks and benefits of regional anesthesia in patients with underlying neurological disease. PMID- 18676383 TI - Consent in the crossfire. PMID- 18676384 TI - Effects of baricity of 0.5% or 0.75% levobupivacaine on the onset time of spinal anesthesia: a randomized trial. AB - PURPOSE: Levobupivacaine has been extensively evaluated for spinal anesthesia. Its pharmacologic properties are similar to those of bupivacaine, but differences in density between two commercially-available preparations of plain levobupivacaine might lead to different block profiles. We examined the characteristics of spinal anesthesia using the same dose of 0.5% and 0.75% plain levobupivacaine. METHODS: In this randomized, controlled, double-blind clinical trial, 60 ASA I-III patients, scheduled for hip fracture surgery, were randomly allocated to receive spinal anesthesia with 15 mg of either 0.5% (group Levo 0.5), or 0.75% (group Levo-0.75) plain levobupivacaine. Onset time (primary outcome), sensory block height level by pinprick, motor block using a modified Bromage scale, and hemodynamic variables were recorded. RESULTS: In group Levo 0.5 (n = 29), surgical anesthesia was obtained within 15 (10-20) min, compared to 12 (7-21) min in group Levo-0.75 (n = 31) (p = 0.409). The highest sensory block level was T11 (T6-T12) in group Levo-0.5, and T10 (T8 - T12) in group Levo-0.75 (p = 0.759). Mean duration of anesthesia was 285 (224-303) min in group Levo-0.5, and 318 (243-375) min in group Levo-0.75 (p = 0.117). The groups were similar in regards to the number of failed blocks requiring general anesthesia, and the number of patients requiring vasopressors. CONCLUSIONS: In this population of elderly patients, spinal anesthesia with 15 mg of either 0.5% or 0.75% plain levobupiva-caine resulted in similar onset of pharmacological actions, and outcomes. Potential differences in baricity did not appear to be clinically relevant in this setting. PMID- 18676385 TI - Hemodilution does not alter the coronary vasodilating effects of endogenous or exogenous nitric oxide. AB - INTRODUCTION: It is well known that hemoglobin is a scavenger of nitric oxide (NO). The present study used a canine model to test the hypothesis that acute normovolemic hemodilution (ANH) affects NO-mediated coronary vasodilation. METHODS: Studies were performed in 18 open-chest, anesthetized dogs. In Series 1, the contribution of endogenous NO to coronary vasodilatation during ANH with 5% dextran-40 (reduction in hematocrit by 50%) was assessed. This was accomplished by comparing myocardial blood flow (MBF; radioactive microspheres) in the left anterior descending (LAD) region, which was treated with the NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), to that in the circumflex (control) region. In Series 2, the LAD was perfused via a controlled-pressure extracorporeal system with coronary blood flow (CBF) measured with an ultrasonic, transit-time flow transducer. The dose-dependent increases in CBF caused by acetylcholine (ACh), which releases endogenous NO from the vascular endothelium, and sodium nitroprusside (SNP), which provides exogenous NO, were compared before and during ANH. RESULTS: Acute normovolemic hemodilution caused similar (approximately twofold) increases in MBF (P < 0.01) in the absence and presence of L-NAME, and it did not affect the dose-related increases in CBF caused by ACh and SNP. CONCLUSIONS: Series 1: under baseline conditions, hemoglobin in red blood cells does not limit the coronary vasodilatation resulting from tonic release of NO; NO does not mediate coronary vasodilation during ANH. Series 2: ANH does not influence the coronary vasodilating effects of increased levels of NO, whether due to endogenous release (ACh) or infusion of an NO donor (SNP). PMID- 18676386 TI - Ultrasound-guided peripheral regional blockade in patients with Charcot-Marie Tooth disease: a review of three cases. AB - PURPOSE: To describe the clinical presentation of three patients with Charcot Marie-Tooth disease, who underwent uneventful upper limb surgery following successful peripheral nerve blockade, and to review the anesthetic implications in patients with Charcot-Marie-Tooth disease. CLINICAL FEATURES: In three patients with Charcot-Marie-Tooth disease presenting for surgery of the upper limb, the motor response, following nerve stimulation, was suboptimal. However, ultrasound guidance was effective in visualizing the needle-nerve interaction, and local anesthetic was injected around the nerves. Good block ensued and surgery proceeded in all patients without complications. No exacerbation of the neurological condition was observed in any patient. CONCLUSIONS: Charcot-Marie Tooth disease is a demyelinating, hereditary, motor and sensory neuropathy characterized by abnormalities of nerve conduction. Regional anesthesia of the upper limb is feasible in these patients, and these cases show that ultrasound guidance makes peripheral nerve block possible in patients for whom traditional methods of nerve localization fail. PMID- 18676387 TI - Serotonin toxicity caused by an interaction between fentanyl and paroxetine. AB - PURPOSE: To report a case of serotonin toxicity, presenting in the postoperative period, caused by an interaction between paroxetine (a selective serotonin reuptake inhibitor, SSRI) and fentanyl (a phenylpiperidine opioid). Serotonin toxicity precipitated by fentanyl is unusual and has not previously been described in combination with SSRIs in the perioperative setting. CLINICAL FEATURES: A 60-yr-old woman, established on paroxetine for depression, underwent excision of a chest wall myxofibrosarcoma and chest wall reconstruction. Fentanyl was administered for intraoperative and postoperative analgesia (1 mg intraoperatively, and 2.5 mg by infusion in the first 36 hr, postoperatively). She developed a vague affectation, intermittent agitation, bilateral hyper reflexia, inducible clonus, and a period of hypertension, suggestive of serotonin toxicity. There was complete resolution after cessation of fentanyl and paroxetine. CONCLUSION: The co-administration of SSRIs and fentanyl may precipitate serotonin toxicity. There must be consideration of this unusual interaction when administering fentanyl to patients established on SSRIs. Physicians should be vigilant of the features of serotonin toxicity developing in such patients. PMID- 18676388 TI - General anesthesia using remifentanil for Cesarean delivery in a parturient with Marfan's syndrome. AB - PURPOSE: Due to cardiovascular and skeletal abnormalities, anesthetic management of parturients with Marfan's syndrome can be particularly challenging. Parturients with aortic root dilatation are at risk for aortic dissection. We describe the anesthetic management of a parturient with Marfan's syndrome and aortic root dilatation, who required general anesthesia for Cesarean delivery. CLINICAL FEATURES: At 26 weeks gestation, a nulliparous woman with Marfan's syndrome presented to the Anesthesia Clinic. Her history revealed asymptomatic aortic root dilatation of 41 mm, and partial correction of scoliosis with Harrington rods. Her cardiologist advised metoprolol, serial echocardiograms, and Cesarean delivery to decrease the risk of aortic dissection. At a multidisciplinary conference, a decision was made to proceed with Cesarean delivery, at term, at the cardiac surgery centre. After placement of arterial and central lines, general anesthesia was induced with remifentanil, propofol, and succinylcholine. Anesthesia was maintained with N(2)O, sevoflurane, and remifentanil (0.02-0.08 microg x kg(-1) x min(-1)). Transesophageal echocardiography examination confirmed stable aortic root dilatation. The patient remained hemodynamically stable. The baby's Apgars were 4 and 8, at one and five minutes, respectively. At the end of the procedure, the patient's trachea was extubated when she was awake. Initial postoperative care was in the intensive care unit. Both mother and baby recovered uneventfully. CONCLUSIONS: Peripartum hemodynamic changes can be life-threatening to the parturient with Marfan's syndrome and aortic dilatation. Anesthetic goals for delivery included preparation for possible aortic dissection, and avoidance of increased aortic root shear stress, through careful hemodynamic monitoring, and general anesthesia using remifentanil. PMID- 18676389 TI - A critical review of thromboembolic complications associated with central venous catheters. AB - PURPOSE: Central venous catheters (CVC) are commonly used in critical care. While thrombosis is a well-recognized and frequent complication associated with their use, CVC-related thromboembolic complications, including pulmonary embolism (PE) and right heart thromboembolism (RHTE), occur less frequently and often evade diagnosis. Little information exists to guide clinicians in the diagnosis and management of CVC-related thromboembolic complications. SOURCE: We critically review and synthesize the literature highlighting the incidence of CVC-related thrombosis. We highlight the risk for developing thromboembolic complications and provide approaches to diagnosing and managing RHTE. PRINCIPLE FINDINGS: The incidence of CVC-related thrombosis varies depending on patient, site, instrument, and infusate-related factors. Central venous catheters-related thrombosis represents an important source of morbidity and mortality for affected patients. Pulmonary embolism occurs in approximately 15% of patients with CVC related upper extremity deep venous thrombosis (UEDVT). More frequent use of transesophageal echocardiography, in patients with suspected and confirmed PE, has resulted in increased detection of RHTE. While it is recognized that the occurrence of RHTE, in association with PE, increases mortality, the optimal strategy for their management has not been established in a clinical trial. CONCLUSION: Central venous catheter-related thrombosis occurs frequently and represents an important source of morbidity and mortality for affected patients. Our review supports that surgery and thrombolysis have both been demonstrated to enhance survival in patients with RHTE and PE. However, important patient, clot, and institutional considerations mandate that treatment for patients with RHTE and PE be individualized. PMID- 18676390 TI - The unresolved issues with risk stratification and management of patients with coronary artery disease undergoing major vascular surgery. AB - PURPOSE: The purpose of this article, with a specific focus on patients undergoing vascular surgery, is to review controversial issues related to mechanisms of perioperative myocardial infarction (MI), coronary artery disease detection, and strategies to reduce perioperative complications. We propose explanations for the many conflicting results that have recently emerged in the literature.Source documents: We searched MEDLINE and reviewed all relevant manuscripts and scientific statements regarding management of patients undergoing non-cardiac surgery. PRINCIPAL FINDINGS: Identification and prevention of ischemia in patients undergoing vascular surgery remains controversial. While the identification of preoperative ischemia is a marker of a higher perioperative risk, the value of identifying such ischemia has been questioned. We believe this may be, at least in part, due to our limited understanding of perioperative MI. Appropriate management of patients, based on the results of such testing, is likely the key to improving outcomes, and deserves further investigation. Efforts aimed at reducing the ischemic consequences of severe coronary plaques (by revascularization or beta-blocker therapy) have yielded conflicting results. The use of high doses of preoperative beta-blocker therapy may be harmful. Some studies suggest a promising role for statin therapy. Benefits of acetylsalicylic acid must be weighted against the risk of bleeding. CONCLUSION: Many questions remain unanswered about the impact of detecting inducible ischemia, and the role of revascularization or beta-blockers in patients undergoing vascular surgery. A better understanding of the pathophysiology of perioperative MI is critical, in order to identify the best approach to improve cardiac outcomes in these patients. PMID- 18676391 TI - Echocardiography images of inferior vena cava tumour thrombus in patient with renal cell carcinoma. PMID- 18676392 TI - Perioperative glycemic control and postoperative cognitive dysfunction: an expression of caution. PMID- 18676393 TI - Safeguarding against errors with free flow protection devices of patient controlled analgesia systems. PMID- 18676394 TI - Use of the Airtraq with a fibreoptic bronchoscope in a difficult intubation outside the operating room. PMID- 18676395 TI - Mitral valve repair for ischemic mitral regurgitation. AB - Our aim was to evaluate midterm results in patients who underwent mitral valve repair (MVR) for ischemic mitral regurgitation (IMR) in our most recent experience. From March 2006 to March 2008, 105 patients underwent MVR for IMR. Mean IMR grade was 2.6 +/- 1.1, with 46 patients having or=3/4. Five patients (4.8%) died within first month; Two-year freedom from death any cause was 85.5% +/- 3.8, freedom from cardiac death was 88.7% +/- 3.4. NYHA Class of the survivors was 1.3 +/- 0.6, with 3 patients in NYHA Class III. Freedom from death any cause and NYHA Class III-IV was 78.6% +/- 4.6. IMR grade decreased from 2.6 +/- 1.1 to 0.1 +/- 0.3 at the discharge and to 0.5 +/- 0.3 after a mean of 7 +/- 4 months, with no patient with IMR grade 3/4 or 4/4. MVR for IMR should be performed in patients with moderate-or-more IMR grade or when the MV is excessively dilated, to achieve good midterm results. PMID- 18676396 TI - Granulocyte colony-stimulating factor exacerbates cardiac fibrosis after myocardial infarction in a rat model of permanent occlusion. AB - AIMS: Controversy exists regarding the effects of granulocyte colony-stimulating factor (G-CSF) on post-infarction remodelling, which is regulated by matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). The aim of this study was to investigate the impact of G-CSF administration on cardiac MMP/TIMP ratios and long-term remodelling in a rat model of acute myocardial infarction (MI). METHODS AND RESULTS: Sprague-Dawley rats underwent coronary ligation to produce MI. Rats surviving the MI for 3 h were randomized to receive G-CSF (50 microg/kg/day for 5 consecutive days, n = 16) or saline (n = 10). Sham-operated animals received no treatment (n = 10). G-CSF injection significantly increased circulating white blood cells, neutrophils, and monocytes. Western blotting revealed that the ratios of MMP-2/TIMP-1 and MMP 9/TIMP-1 were significantly decreased in the infarcted myocardium. At 3 months, echocardiographic and haemodynamic examinations showed that the G-CSF treatment induced left ventricular (LV) enlargement and dysfunction. Histological analysis revealed that the extent of myocardial fibrosis and infarct size were larger in the G-CSF group than in the Saline group. Furthermore, G-CSF treated animals showed a significantly lower post-MI survival during the study period. CONCLUSION: Decrease of cardiac MMP/TIMP ratios by G-CSF after infarction may be important as a mechanism in promotion of myocardial fibrosis, which further facilitates infarct expansion and LV dysfunction. PMID- 18676397 TI - Articular cartilage: structure, injuries and review of management. AB - INTRODUCTION: Chondral and osteochondral injuries are commonly seen in today's clinical practice. Articular cartilage provides an ultimate low-friction gliding surface, which none of the artificial constructs have been able to replace successfully. Retrospective review of the knee arthroscopies has revealed an underestimated incidence of this complex problem. Cartilage injuries in the knee joint if left untreated lead to pre-mature early arthritis and affect the activities of daily living. Various different treatment methods of cartilage regeneration have shown encouraging results, but unfortunately none has proved to be the ultimate solution. SOURCES OF DATA: This article re-visits the intricate structure of articular cartilage and reviews the different methods of regeneration described in the literature, based on evidence-based effectiveness. The methods described by their originators and their results are considered gold standards for those methods, as being the best available evidence. AREAS OF AGREEMENT: Majority of the authors agree that cartilage injuries are complex and difficult to treat. If untreated, cartilage defects lead to early osteoarthritis. Great debate still persists about the best available treatment for symptomatic chondral or osteochondral defect(s). AREAS OF CONTROVERSY: The controversy about the management outplays its aetiological theories. Several authors have reported good results with different techniques; however none has proved to be the solution for the problem. GROWING POINTS: Up until 1990, marrow stimulation techniques were routine form of management for chondral defects. However, ever since autologous chondrocyte implantation was successfully introduced in humans, it has provided a new dimension for the treatment of chondral defects. AREAS TIMELY FOR DEVELOPING RESEARCH: The success of any treatment lies in its longevity. The new minimally invasive techniques are being invented. However, timely research, on the basis of randomized controlled trial comparing different methods of cartilage reconstruction is necessary for decision-making in today's evidence-based medical world. PMID- 18676398 TI - Still without impact on adverse post-operative outcomes: pre-operative statin therapy in patients undergoing cardiac surgery. PMID- 18676399 TI - Atrial fibrillation is associated with impaired cognitive function and hippocampal atrophy: silent cerebral ischaemia vs. Alzheimer's disease? PMID- 18676401 TI - The histone subcode: poly(ADP-ribose) polymerase-1 (Parp-1) and Parp-2 control cell differentiation by regulating the transcriptional intermediary factor TIF1beta and the heterochromatin protein HP1alpha. AB - Recent advances reveal emerging unique functions of poly(ADP-ribose) polymerase-1 (Parp-1) and Parp-2 in heterochromatin integrity and cell differentiation. However, the chromatin-mediated molecular and cellular events involved remain elusive. Here we describe specific physical and functional interactions of Parp-1 and Parp-2 with the transcriptional intermediary factor (TIF1beta) and the heterochromatin proteins (HP1) that affect endodermal differentiation. We show that Parp-2 binds to TIF1beta with high affinity both directly and through HP1alpha. Both partners colocalize at pericentric heterochromatin in primitive endoderm-like cells. Parp-2 also binds to HP1beta but not to HP1gamma. In contrast Parp-1 binds weakly to TIF1beta and HP1beta only. Both Parps selectively poly(ADP-ribosyl)ate HP1alpha. Using shRNA approaches, we provide evidence for distinct participation of both Parps in endodermal differentiation. Whereas Parp 2 and its activity are required for the relocation of TIF1beta to heterochromatic foci during primitive endodermal differentiation, Parp-1 and its activity modulate TIF1beta-HP1alpha association with consequences on parietal endodermal differentiation. Both Parps control TIF1beta transcriptional activity. In addition, this work identifies both Parps as new modulators of the HP1-mediated subcode histone.-Quenet, D., Gasser, V., Fouillen, L., Cammas, F., Sanglier Cianferani, S., Losson, R., Dantzer, F. The histone subcode: poly(ADP-ribose) polymerase-1 (Parp-1) and Parp-2 control cell differentiation by regulating the transcriptional intermediary factor TIF1beta and the heterochromatin protein HP1alpha. PMID- 18676402 TI - Vitamin A depletion causes oxidative stress, mitochondrial dysfunction, and PARP 1-dependent energy deprivation. AB - A significant unresolved question is how vitamin A deprivation causes, and why retinoic acid fails to reverse, immunodeficiency. When depleted of vitamin A, T cells undergo programmed cell death (PCD), which is enhanced by the natural competitor of retinol, anhydroretinol. PCD does not happen by apoptosis, despite the occurrence of shared early events, including mitochondrial membrane depolarization, permeability transition pore opening, and cytochrome c release. It also lacks caspase-3 activation, chromatin condensation, and endonuclease mediated DNA degradation, hallmarks of apoptosis. PCD following vitamin A deprivation exhibits increased production of reactive oxygen species (ROS), drastic reductions in ATP and NAD(+) levels, and activation of poly-(ADP-ribose) polymerase (PARP) -1. These latter steps are causative because neutralizing ROS, imposing hypoxic conditions, or inhibiting PARP-1 by genetic or pharmacologic approaches prevents energy depletion and PCD. The data highlight a novel regulatory role of vitamin A in mitochondrial energy homeostasis. PMID- 18676403 TI - Depletion of phytosterols from the plant plasma membrane provides evidence for disruption of lipid rafts. AB - Involvement of sterols in membrane structural properties has been extensively studied in model systems but rarely assessed in natural membranes and never investigated for the plant plasma membrane (PM). Here, we address the question of the role of phytosterols in the organization of the plant PM. The sterol composition of tobacco BY-2 cell PM was determined by gas chromatography. The cyclic oligosaccharide methyl-beta-cyclodextrin, commonly used in animal cells to decrease cholesterol levels, caused a drastic reduction (50%) in the PM total free sterol content of the plant material, without modification in amounts of steryl-conjugates. Fluorescence spectroscopy experiments using DPH, TMA-DPH, Laurdan, and di-4-ANEPPDHQ indicated that such a depletion in sterol content increased lipid acyl chain disorder and reduced the overall liquid-phase heterogeneity in correlation with the disruption of phytosterol-rich domains. Methyl-beta-cyclodextrin also prevented isolation of a PM fraction resistant to solubilization by nonionic detergents, previously characterized in tobacco, and induced redistribution of the proteic marker of this fraction, NtrbohD, within the membrane. Altogether, our results support the role of phytosterols in the lateral structuring of the PM of higher plant cells and suggest that they are key compounds for the formation of plant PM microdomains. PMID- 18676405 TI - Writing science: the abstract is poetry, the paper is prose. PMID- 18676404 TI - The transcription factor ST18 regulates proapoptotic and proinflammatory gene expression in fibroblasts. AB - Suppression of tumorigenicity 18 (ST18) and the homologues neural zinc-finger protein-3 (NZF3) and myelin transcription factor 3 (Myt3) are transcription factors with unknown function. Previous studies have established that they repress transcription of a synthetic reporter construct consisting of the consensus sequence AAAGTTT linked to the thymidine kinase promoter. In addition, ST18 exhibits significantly reduced expression in breast cancer and breast cancer cell lines. We report here for the first time evidence that ST18 mediates tumor necrosis factor (TNF) -alpha induced mRNA levels of proapoptotic and proinflammatory genes in fibroblasts by mRNA profiling and silencing with ST18 small interfering RNA (siRNA). Gene set enrichment analysis and mRNA profiling support this conclusion by identifying several apoptotic and inflammatory pathways that are down-regulated by ST18 siRNA. In addition, ST18 siRNA reduces TNF-induced fibroblast apoptosis and caspase-3/7 activity. Fibroblasts that overexpress ST18 by transient transfection exhibit significantly increased apoptosis and increased expression of TNF-alpha, interleukin (IL) -1alpha, and IL 6. In addition, cotransfection of ST18 and a TNF-alpha or IL-1alpha reporter construct demonstrates that ST18 overexpression in fibroblasts significantly enhanced promoter activity of these genes. Taken together, these studies demonstrate that the transcription factor ST18/NZF3 regulates the mRNA levels of proapoptotic and proinflammatory genes in revealing a previously unrecognized function. PMID- 18676406 TI - Serum homocysteine levels in relation to clinical progression in multiple sclerosis. AB - BACKGROUND: Elevated homocysteine levels are associated with various neurodegenerative diseases and have even been identified as a risk factor for some of these. Homocysteine levels may be elevated in patients with multiple sclerosis (MS) but large studies are lacking and the relation with disease progression remains to be determined. AIM: The aim of the study was to investigate homocysteine levels in patients with MS and in controls, and to study the relationship between homocysteine levels and clinical progression in MS. METHODS: Serum homocysteine levels were compared between MS subtypes (n = 219) and controls (n = 152). Homocysteine levels were associated with baseline and follow-up clinical severity scores. RESULTS: The results showed that serum homocysteine values were similar in patients with MS and controls. Baseline scores on the Expanded Disability Status Scale were higher in patients with secondary progressive MS (SPMS) in the top compared with the bottom quartile of homocysteine levels (p = 0.02). The baseline scores on the Multiple Sclerosis Functional Composite (MSFC) and Paced Auditory Serial Addition Test (PASAT), which measures cognitive functioning, were lower in patients with SPMS in the top compared with the bottom quartile of homocysteine levels (MSFC, p = 0.02; PASAT, p = 0.02). High homocysteine levels were associated with a decline in PASAT scores during follow-up in patients with primary progressive MS (p = 0.009). CONCLUSION: Serum total homocysteine levels are associated with several measures of disease progression in MS but are not elevated in patients with MS compared with controls. The association of homocysteine levels with cognition in patients with progressive MS raises the question of whether homocysteine directly impacts on MS or reflects a more general neurodegenerative process. PMID- 18676407 TI - Visual sensorial preference delays balance control compensation after vestibular schwannoma surgery. AB - BACKGROUND: Balance control performance after vestibular schwannoma surgical removal follows a course that is characterised by a deterioration in postural performance immediately after unilateral vestibular deafferentation (uVD) and a recovery process (vestibular compensation). However, sensory strategies for balance vary during tumoral growth, which could lead to differences in the preferential use of sensory afferences. This longitudinal study aimed to assess the post-operative sensorimotor strategies of postural regulation according to sensory preference of balance control before surgery. METHODS: Twenty-two patients with vestibular schwannoma (11 relying less on vision (G1), 11 relying more on vision (G2), to control balance before surgery), underwent vestibular, subjective visual vertical (SVV), static posturography and sensory organisation (SOT) tests, before and 8 days, 1 and 3 months after surgery. RESULTS: In G1 patients, little static posturographic and SOT performance deterioration after uVD was observed, despite vestibular test and SVV modifications. In G2 patients, uVD-related modifications followed a time-course characterised by a degradation in posturographic and SOT, vestibular and SVV performances immediately after uVD and a progressive restoration and even improvement 1 month and particularly 3 months after surgery. CONCLUSIONS: High preference for vision before surgery intervenes in postural degradation immediately after surgery, thus delaying the short-term effects of vestibular compensation on postural control. Long-term performance being similar whatever the visual status before surgery, the time dependent implementation of the central adaptive mechanisms due to neuroplasticity leads to a modification of neurosensory information hierarchy, allowing reliance on appropriate information, the gain varying according to the postural task to be performed. PMID- 18676408 TI - Extremely preterm births: recommendations for treatment in European countries. PMID- 18676409 TI - Random safety audits in the neonatal unit. AB - BACKGROUND: Random safety audits have been shown to be effective in improving standards of practice in high-risk industries. They are process audits rapidly performed during real-time clinical activity, with immediate feedback, allowing for immediate change of practice. AIM: Based on a concept described by the Vermont-Oxford Network, we aimed to introduce random safety audits to our unit to improve infection control and routine neonatal care. METHOD: We designed simple data collection tables to audit 11 infection control and four routine care standards. Audits were undertaken during the weekly grand round. Immediate feedback was given. RESULTS: In 6 months we completed three cycles of 15 audits each. Complete results were available for 14 audits. The compliance with the infection control standards improved from a median of 70% (range 20%-100%) to 95% (range 66%-100%). The results of the routine care standards were more variable. CONCLUSION: We have shown that this innovative method of random safety audits is effective in quickly improving practice. We believe this to be due to the instant feedback, continued emphasis on infection control and good clinical practice, and improved teamwork. PMID- 18676410 TI - Bronchopulmonary dysplasia and inflammatory biomarkers in the premature neonate. AB - Bronchopulmonary dysplasia (BPD) is the most common, serious sequela of premature birth. Inflammation is a major contributor to the pathogenesis of BPD. Often initiated by a pulmonary fetal inflammatory response, lung inflammation is exacerbated by mechanical ventilation and exposure to supplemental oxygen. In response to these initiators of injury, a complex interaction occurs between proteins that attract inflammatory cells (ie, chemokines), proteins that facilitate the transendothelial migration of inflammatory cells from blood vessels (ie, adhesion molecules), proteins that promote tissue damage (ie, pro inflammatory cytokines and proteases), and proteins that modulate the process (eg, anti-inflammatory cytokines, binding proteins and receptor antagonists). In addition, during recovery from inflammatory injury, growth factors and other substances that control normal lung growth and mediate repair influence subsequent lung structure. In this review, we discuss the role of each aspect of the inflammatory process in the development of BPD. This discussion will include data from measurements of biomarkers in samples of fluid aspirated from the airways of human infants relevant to each phase of inflammation. Despite their limitations, these measurements provide some insight into the role of inflammation in the development of BPD and may be useful in identifying infants at risk for the disease. PMID- 18676411 TI - Mask ventilation in the early management of congenital central hypoventilation syndrome. PMID- 18676412 TI - The effect of two levels of pressure support ventilation on tidal volume delivery and minute ventilation in preterm infants. AB - OBJECTIVE: To study the effect of different levels of pressure support ventilation (PSV) on respiratory parameters in preterm infants during the weaning phase of mechanical ventilation. DESIGN/METHODS: In this quasi-experimental crossover study, a total of 19 154 breaths were analysed from 10 ventilated infants of <32 weeks' gestation. Breath-to-breath data on minute ventilation, tidal volume, respiratory rate, peak inspiratory pressure and mean airway pressure were collected during three study epochs: synchronised intermittent mandatory ventilation (SIMV) alone, SIMV with partial PSV (PS(min)), and SIMV with full PSV (PS(max)). PS(min) was set to provide an exhaled tidal volume (V(Te)) between 2.5-4 ml/kg and PS(max) 5-8 ml/kg V(Te). Statistical analyses were performed using analysis of variance (ANOVA) for repeated measures. RESULTS: The addition of full PSV (PS(max)) was associated with a significant increase in total minute ventilation as compared with SIMV alone (392 ml/kg/min vs 270 ml/kg/min, respectively; p<0.05). This difference in minute ventilation was still present when PS(min) was used (332 ml/kg/min as compared with 270 ml/kg/min in SIMV; p<0.05). There was also a concomitant decrease in the respiratory rate with both PS(max) (59 breaths per minute) and PS(min) (65 breaths per minute) compared with SIMV alone (72 breaths per min) (p<0.05). CONCLUSIONS: Pressure support ventilation increases total minute ventilation and stabilises breathing in proportion to the level of pressure support used. This may be advantageous and provide a useful ventilation strategy for use during weaning stages of mechanical ventilation in preterm infants. PMID- 18676413 TI - Association of gastric fluid microbes at birth with severe bronchopulmonary dysplasia. AB - OBJECTIVE: Gastric fluid microbes were examined in preterm infants at birth to assess their influence on the postnatal outcome. STUDY DESIGN: Prospective cohort study. SETTING: Level III neonatal intensive care unit. PATIENTS: A total of 103 premature neonates with a gestational age of less than 32 weeks. MAIN OUTCOME MEASURE: Gastric fluid microbes were identified by analysis of bacterial 16S ribosomal RNA gene. Additionally, the urease gene of Ureaplasma species was detected by polymerase chain reaction of gastric fluid obtained at birth and/or tracheal aspirate from ventilated preterm infants. The association between detection of microbes and bronchopulmonary dysplasia was investigated through assessment from clinical features and by a lung injury marker (KL-6). RESULTS: Forty-two of 103 gastric fluid specimens were positive for microbes. Ureaplasma species were detected in 23 of the 42 (55%) gastric fluid specimens. All infants with Ureaplasma species in tracheal aspirate fluid also had positive gastric fluid specimens. Compared to infants negative for gastric fluid microbes, infants positive for microbes had higher rates of maternal chorioamnionitis (18% vs 78%), premature rupture of membranes (11% vs 55%), severe bronchopulmonary dysplasia (1.6% vs 14%) and showed higher plasma KL-6 levels during the initial 4 weeks of life. CONCLUSION: Detection of gastric fluid microbes was correlated well with antenatal infection and severe bronchopulmonary dysplasia. Detection of Ureaplasma species in gastric fluid was associated with subsequent respiratory colonisation. These results suggest that antenatal exposure of the immature fetus to microbes may cause lung injury and promote the onset of bronchopulmonary dysplasia. PMID- 18676414 TI - Size matters: just how big is BIG?: Quantifying realistic sample size requirements for human genome epidemiology. AB - BACKGROUND: Despite earlier doubts, a string of recent successes indicates that if sample sizes are large enough, it is possible-both in theory and in practice to identify and replicate genetic associations with common complex diseases. But human genome epidemiology is expensive and, from a strategic perspective, it is still unclear what 'large enough' really means. This question has critical implications for governments, funding agencies, bioscientists and the tax-paying public. Difficult strategic decisions with imposing price tags and important opportunity costs must be taken. METHODS: Conventional power calculations for case-control studies disregard many basic elements of analytic complexity-e.g. errors in clinical assessment, and the impact of unmeasured aetiological determinants-and can seriously underestimate true sample size requirements. This article describes, and applies, a rigorous simulation-based approach to power calculation that deals more comprehensively with analytic complexity and has been implemented on the web as ESPRESSO: (www.p3gobservatory.org/powercalculator.htm). RESULTS: Using this approach, the article explores the realistic power profile of stand-alone and nested case-control studies in a variety of settings and provides a robust quantitative foundation for determining the required sample size both of individual biobanks and of large disease-based consortia. Despite universal acknowledgment of the importance of large sample sizes, our results suggest that contemporary initiatives are still, at best, at the lower end of the range of desirable sample size. Insufficient power remains particularly problematic for studies exploring gene-gene or gene-environment interactions. Discussion Sample size calculation must be both accurate and realistic, and we must continue to strengthen national and international cooperation in the design, conduct, harmonization and integration of studies in human genome epidemiology. PMID- 18676415 TI - Protein-ligand interaction prediction: an improved chemogenomics approach. AB - MOTIVATION: Predicting interactions between small molecules and proteins is a crucial step to decipher many biological processes, and plays a critical role in drug discovery. When no detailed 3D structure of the protein target is available, ligand-based virtual screening allows the construction of predictive models by learning to discriminate known ligands from non-ligands. However, the accuracy of ligand-based models quickly degrades when the number of known ligands decreases, and in particular the approach is not applicable for orphan receptors with no known ligand. RESULTS: We propose a systematic method to predict ligand-protein interactions, even for targets with no known 3D structure and few or no known ligands. Following the recent chemogenomics trend, we adopt a cross-target view and attempt to screen the chemical space against whole families of proteins simultaneously. The lack of known ligand for a given target can then be compensated by the availability of known ligands for similar targets. We test this strategy on three important classes of drug targets, namely enzymes, G protein-coupled receptors (GPCR) and ion channels, and report dramatic improvements in prediction accuracy over classical ligand-based virtual screening, in particular for targets with few or no known ligands. AVAILABILITY: All data and algorithms are available as Supplementary Material. PMID- 18676416 TI - Genome-scale classification of metabolic reactions and assignment of EC numbers with self-organizing maps. AB - MOTIVATION: The automatic perception of chemical similarities between metabolic reactions is required for a variety of applications ranging from the computer aided validation of classification systems, to genome-scale reconstruction (or comparison) of metabolic pathways, to the classification of enzymatic mechanisms. Comparison of metabolic reactions has been mostly based on Enzyme Commission (EC) numbers, which are extremely useful and widespread, but not always straightforward to apply, and often problematic when an enzyme catalyzes several reactions, when the same reaction is catalyzed by different enzymes, when official full EC numbers are unavailable or when reactions are not catalyzed by enzymes. Different methods should be available to compare metabolic reactions. Simultaneously, methods are required for the automatic assignment of EC numbers to reactions still not officially classified. RESULTS: We have proposed the MOLMAP reaction descriptors to numerically encode the structural transformations resulting from a chemical reaction. Here, such descriptors are applied to the mapping of a genome-scale database of almost 4000 metabolic reactions by Kohonen self-organizing maps (SOMs), and its screening for inconsistencies in EC numbers. This approach allowed for the SOMs to assign EC numbers at the class, subclass and sub-subclass levels for reactions of independent test sets with accuracies up to 92, 80 and 70%, respectively. Different levels of similarity between training and test sets were explored. The approach also led to the identification of a number of similar reactions bearing differences at the EC class level. AVAILABILITY: The programs to generate MOLMAP descriptors from atomic properties included in SDF files are available upon request for evaluation. PMID- 18676417 TI - Large-scale computation of elementary flux modes with bit pattern trees. AB - MOTIVATION: Elementary flux modes (EFMs)--non-decomposable minimal pathways--are commonly accepted tools for metabolic network analysis under steady state conditions. Valid states of the network are linear superpositions of elementary modes shaping a polyhedral cone (the flux cone), which is a well-studied convex set in computational geometry. Computing EFMs is thus basically equivalent to extreme ray enumeration of polyhedral cones. This is a combinatorial problem with poorly scaling algorithms, preventing the large-scale analysis of metabolic networks so far. RESULTS: Here, we introduce new algorithmic concepts that enable large-scale computation of EFMs. Distinguishing extreme rays from normal (composite) vectors is one critical aspect of the algorithm. We present a new recursive enumeration strategy with bit pattern trees for adjacent rays--the ancestors of extreme rays--that is roughly one order of magnitude faster than previous methods. Additionally, we introduce a rank updating method that is particularly well suited for parallel computation and a residue arithmetic method for matrix rank computations, which circumvents potential numerical instability problems. Multi-core architectures of modern CPUs can be exploited for further performance improvements. The methods are applied to a central metabolism network of Escherichia coli, resulting in approximately 26 Mio. EFMs. Within the top 2% modes considering biomass production, most of the gain in flux variability is achieved. In addition, we compute approximately 5 Mio. EFMs for the production of non-essential amino acids for a genome-scale metabolic network of Helicobacter pylori. Only large-scale EFM analysis reveals the >85% of modes that generate several amino acids simultaneously. AVAILABILITY: An implementation in Java, with integration into MATLAB and support of various input formats, including SBML, is available at http://www.csb.ethz.ch in the tools section; sources are available from the authors upon request. PMID- 18676418 TI - Video-assisted instruction improves the success rate for tracheal intubation by novices. AB - BACKGROUND: Tracheal intubation via laryngoscopy is a fundamental skill, particularly for anaesthesiologists. However, teaching this skill is difficult since direct laryngoscopy allows only one individual to view the larynx during the procedure. The purpose of this study was to determine if video-assisted laryngoscopy improves the effectiveness of tracheal intubation training. METHODS: In this prospective, randomized, crossover study, 37 novices with less than six prior intubation attempts were randomized into two groups, video-assisted followed by traditional instruction (Group V/T) and traditional instruction followed by video-assisted instruction (Group T/V). Novices performed intubations on three patients, switched groups, and performed three more intubations. All trainees received feedback during the procedure from an attending anaesthesiologist based on standard cues. Additionally, during the video-assisted part of the study, the supervising anaesthesiologist incorporated feedback based on the video images obtained from the fibreoptic camera located in the laryngoscope. RESULTS: During video-assisted instruction, novices were successful at 69% of their intubation attempts whereas those trained during the non-video assisted portion were successful in 55% of their attempts (P=0.04). Oesophageal intubations occurred in 3% of video-assisted intubation attempts and in 17% of traditional attempts (P<0.01). CONCLUSIONS: The improved rate of successful intubation and the decreased rate of oesophageal intubation support the use of video laryngoscopy for tracheal intubation training. PMID- 18676420 TI - Diagnostic utility of alarm features for colorectal cancer: systematic review and meta-analysis. AB - OBJECTIVE: Colorectal cancer is the second most common cause of cancer death in Europe and North America. Alarm features are used to prioritize access to urgent investigation, but there is little information concerning their utility in the diagnosis of colorectal cancer. METHODS: A systematic review and meta-analysis of the published literature was carried out to assess the diagnostic accuracy of alarm features in predicting colorectal cancer. Primary or secondary care-based studies in unselected cohorts of adult patients with lower gastrointestinal symptoms were identified by searching MEDLINE, EMBASE and CINAHL (up to October 2007). The main outcome measures were accuracy of alarm features or statistical models in predicting the presence of colorectal cancer after investigation. Data were pooled to estimate sensitivity, specificity, and positive and negative likelihood ratios. The quality of the included studies was assessed according to predefined criteria. RESULTS: Of 11 169 studies identified, 205 were retrieved for evaluation. Fifteen studies were eligible for inclusion, evaluating 19 443 patients, with a pooled prevalence of colorectal carcinoma of 6% (95% CI 5% to 8%). Pooled sensitivity of alarm features was poor (5% to 64%) but specificity was >95% for dark red rectal bleeding and abdominal mass, suggesting that the presence of either rules the diagnosis of colorectal cancer in. Statistical models had a sensitivity of 90%, but poor specificity. CONCLUSIONS: Most alarm features had poor sensitivity and specificity for the diagnosis of colorectal carcinoma, whilst statistical models performed better in terms of sensitivity. Future studies should examine the utility of dark red rectal bleeding and abdominal mass, and concentrate on maximising specificity when validating statistical models. PMID- 18676421 TI - In this issue of Occupational Medicine. PMID- 18676419 TI - Adiponectin suppresses colorectal carcinogenesis under the high-fat diet condition. AB - BACKGROUND AND AIMS: The effect of adiponectin on colorectal carcinogenesis has been proposed but not fully investigated. We investigated the effect of adiponectin deficiency on the development of colorectal cancer. METHODS: We generated three types of gene-deficient mice (adiponectin-deficient, adiponectin receptor 1-deficient, and adiponectin receptor 2-deficient) and investigated chemical-induced colon polyp formation and cell proliferation in colon epithelium. Western blot analysis was performed to elucidate the mechanism which affected colorectal carcinogenesis by adiponectin deficiency. RESULTS: The numbers of colon polyps were significantly increased in adiponectin-deficient mice compared with wild-type mice fed a high-fat diet. However, no difference was observed between wild-type and adiponectin-deficient mice fed a basal diet. A significant increase in cell proliferative activity was also observed in the colonic epithelium of the adiponectin-deficient mice when compared with wild-type mice fed a high-fat diet; however, no difference was observed between wild-type and adiponectin-deficient mice fed a basal diet. Similarly, an increase in epithelial cell proliferation was observed in adiponectin receptor 1-deficient mice, but not in adiponectin receptor 2-deficient mice. Western blot analysis revealed activation of mammalian target of rapamycin, p70 S6 kinase, S6 protein and inactivation of AMP-activated protein kinase in the colon epithelium of adiponectin-deficient mice fed with high-fat diet. CONCLUSIONS: Adiponectin suppresses colonic epithelial proliferation via inhibition of the mammalian target of the rapamycin pathway under a high-fat diet, but not under a basal diet. These studies indicate a novel mechanism of suppression of colorectal carcinogenesis induced by a Western-style high-fat diet. PMID- 18676422 TI - We can eliminate occupational cancer from chemicals. PMID- 18676423 TI - Philip Jacques de Loutherbourg, Coalbrookdale at Night (1801): oil on canvas, 68 x 106.5 cm. Science Museum, London. PMID- 18676425 TI - Doctors' health and fitness to practise: the need for a bespoke model of assessment. AB - Doctors' performance and fitness to practise are attracting increased attention. High profile cases have brought into question the assessment of fitness to practise and the monitoring of professional performance. In the UK, the chief medical adviser for England has proposed strengthening systems to improve the performance of doctors which include addressing problems of ill-health. The behaviour of the impaired physician, or the doctor-patient, presents unique challenges and a review of the various issues highlights the need to address how the medical profession and society deal with the occurrence of illness in doctors. Conditions such as mental ill-health and substance abuse may affect doctors' fitness to practise, but other conditions may also be relevant. This paper will discuss the occurrence of ill-health and the need for a bespoke model of assessment. PMID- 18676424 TI - Doctors' health and fitness to practise: assessment models. AB - The assessment of doctors' health and fitness to practise is recognized as a specialist area of occupational medicine practice. This paper will consider the involvement of specialists from a variety of disciplines in the overall assessment process and will discuss some current assessment models. The paper will make recommendations for further developments in this area. PMID- 18676426 TI - Doctors' health and fitness to practise: performance problems in doctors and cognitive impairments. AB - BACKGROUND: As a response to concerns over the safety of patient care and quality of care provided by doctors, there has been an increasing interest in identifying the reasons for medical errors. METHODS: This paper reviews briefly the common neurocognitive causes for performance problems in doctors and provides an updated account of the current literature. Search on Medline and PsychINFO for English language articles between 1956 and September 2006 was performed, as well as a manual search by the authors for other relevant articles. RESULTS: Neuropsychiatric and neuropsychological assessment is increasingly accepted as an accurate evaluation tool to clarify the performance problems in doctors. Furthermore, it seems that neurocognitive difficulties are commonly found to be the cause for such problems. CONCLUSIONS: The performance problems in doctors need to be acknowledged 'better too soon than too late'. Neuropsychiatric and neuropsychological assessment helps to create an accurate treatment and rehabilitation plan for the specific functional tasks of the particular doctor's duties. PMID- 18676427 TI - Doctors' health and fitness to practise: treating addicted doctors. AB - The literature describing the diagnostic process in the addicted doctor is scant. Figures from North America indicate that the prevalence of alcohol problems in doctors may be no higher than in the population as a whole, whereas high rates of prescription drug use have been recognized. This practice of self-treatment with controlled drugs is a 'unique concern' for doctors. The development of substance misuse problems in doctors cannot be reduced to a single factor: Anxiety and depression, personality problems, stress at work, family stress, bereavement, an injury or accident at work, pain and a non-specific drift into drinking have been implicated. Early diagnosis is critical because doctors are often reluctant to seek help and colleagues reluctant to intervene. Medical schools and continuing medical education programmes must give greater emphasis to addiction and substance misuse in doctors with a view to reducing the incidence of 'impaired physicians' and promoting and encouraging early treatment and rehabilitation. The relationship between the addiction psychiatrist and the occupational physician is key given that these problems occur at the interface between occupational health and regulatory systems. The need for individually tailored back to work programmes requires careful coordination and monitoring and may be difficult to implement without their involvement. Generally, the prognosis for doctors' recovery is good and it is possible to predict which doctors will 'make it'. PMID- 18676428 TI - International Occupational Medicine: Croatia. PMID- 18676429 TI - Drs Shanahan and Ahern raised important concerns in their editorial. PMID- 18676430 TI - Post-traumatic Stress Diagnostic Scale (PDS). PMID- 18676432 TI - Retraction. AB - Garcia MLG, Calvo C, Pozo F, et al. Detection of human bocavirus in ill and healthy Spanish children: a 2-year study. This paper has been retracted. PMID- 18676433 TI - Childhood leukaemias and lymphomas in Greece (1996-2006): a nationwide registration study. AB - BACKGROUND: Monitoring time trends in the incidence of childhood leukaemias and lymphomas requires efficient and continuous data collecting systems. In countries without official cancer registries, such as Greece, ad hoc nationwide registration of incident childhood leukaemias and lymphomas could help elucidate the underlying aetiology and monitor socioeconomic differentials in health care delivery. METHODS: We registered all cases and produced age, gender, type and immunophenotype specific figures and overall crude and age adjusted annual incidence rates and secular trends for 863 leukaemia and 311 lymphoma incident cases diagnosed in children <15 years of age across Greece during 1996-2006, namely the first 11 years of the Nationwide Registry for Childhood Hematological Malignancies. RESULTS: The epidemiological profiles of leukaemias/lymphomas in Greece are similar to those in industrialised countries. No secular trends are observed for either malignancy during the studied period. However, the calculated incidence for leukaemia (46.60 cases per 1 million children annually) is among the highest in the EU-27 (19% higher than average; p<0.001), whereas that for lymphoma (16.8 cases per 1 million children annually) is around the EU-27 average. CONCLUSIONS: Minimal secular changes in childhood leukaemias/lymphomas have been noted recently in the EU-27, which cannot be easily explained in countries with small populations. Therefore, centralised EU databases such as the Automated Childhood Cancer Information System (ACCIS) should be enlarged to generate sufficient statistical power for monitoring time trends. It would be interesting to explore whether different lifestyle patterns across the EU might be responsible for the observed excess leukaemia incidence in countries such as Greece. PMID- 18676434 TI - The variation in stool patterns from 1 to 42 months: a population-based observational study. AB - OBJECTIVE: To document the normal stool patterns of young children. DESIGN: Prospective population-based longitudinal study. SETTING: Avon Longitudinal Study of Parents and Children (ALSPAC). SUBJECTS: 12,984 children, whose parents completed questionnaires at 4 weeks, 6, 18, 30 and 42 months on their frequency of bowel movements and the consistency and colour of their stools. RESULTS: Stool frequency declined from a mean of 3.0 times/day (3rd centile 0.6, 97th centile 5.9) at 4 weeks to 1.3 times/day (0.6, 2.7) at 42 months. Stool consistency was soft in most babies with nearly half passing liquid or curdy stool at 4 weeks. 14% of babies usually passed a hard stool at 4 weeks, rising to 30% at 42 months. Stool colour was commonly yellow at 4 weeks and had changed to brown by 6 months. Black stools were extremely unusual at all ages. CONCLUSIONS: These data on the changes with age in the stool patterns of young children will be useful for clinicians. PMID- 18676435 TI - Pilot study of a novel educational programme for 11-16 year olds with type 1 diabetes mellitus: the KICk-OFF course. AB - AIMS: To pilot an educational programme (KICk-OFF) for children and adolescents with type 1 diabetes mellitus (DM). Evaluation included (i) independent assessment of curriculum quality, (ii) acceptability to families and (iii) possible impact on standardised outcome measures (HbA1c, body mass index (BMI), diary reports of hypoglycaemia, quality of life (QoL)). METHODS: 48 children aged 11-16 years (mean age 13.60 (SD 1.36) years) were recruited from three UK centres. Six 5-day outpatient courses on carbohydrate counting and insulin dose adjustment were held. Semi-structured interviews were conducted with children and parents before and after the course. Glycaemic control (HbA1c), BMI and frequency of hypoglycaemia were assessed before the course and at 3 and 6 months after intervention. QoL was assessed before the course and at 2 weeks, 3 and 6 months. RESULTS: Educational evaluation indicated the course format was appropriate and consistent. Parent and child interviews suggested that pre-course expectations were largely met. There were no changes in HbA1c, BMI or episodes of hypoglycaemia, but children and parents reported improved QoL (p<0.05). CONCLUSIONS: KICk-OFF was well-received by children and parents and was associated with improved QoL. In this small pilot study, glycaemic control did not change but findings justify conducting a future randomised controlled trial involving a revised curriculum, a larger study population including a control group and longer follow-up. PMID- 18676436 TI - When to use drugs to help sleep. AB - Paediatric sleep medicine is a relatively new but important and rapidly growing field. It is increasingly recognised that many "adult" sleep disorders begin in childhood where the consequences of missed diagnoses can be devastating. Adequate training of all health care professionals and careful eliciting of symptoms remains the first step in ensuring accurate and timely diagnoses. Although behavioural approaches play a huge role in paediatric sleep medicine, at times severe sleep disorders also require pharmacological treatments. However, the evidence base for these is weak, and currently treatments of conditions including insomnia, narcolepsy and restless legs syndrome in childhood still rely on hopeful extrapolation from adult data. A growth in randomised controlled trials over the last 5 years is encouraging; trial methodologies are also improving. International working groups and multicentre trials will increasingly be needed to evaluate the new technologies and pharmacological treatments that are emerging. PMID- 18676437 TI - A simplified gentamicin dosing chart is quicker and more accurate for nurse verification than the BNFc. AB - BACKGROUND: At least 5% of paediatric admissions are complicated by medication error. Nurses can prevent some errors by correctly verifying prescriptions before administering drugs, which requires adequate drug calculation skills and familiarity with the BNFc. We wished to explore whether a newly devised chart would improve nurses' dose calculation ability and thus potentially reduce doctors' prescription errors. AIMS: To explore nurses' ability to calculate doses of gentamicin for neonates and children using a new simple dosing chart compared with the BNFc. METHODS: Two gentamicin dosing charts (paediatric and neonatal) devised by a multidisciplinary group to simplify dose calculation and selection of frequency were compared with the BNFc using four questions (two neonatal, two paediatric) asking ward nurses to calculate gentamicin doses. Answers were scored for both the correct dosage and correct frequency. RESULTS: 51 nurses participated. 11 nurses (22%) answered all four questions correctly. A higher proportion correctly answered both the dosage and frequency questions simultaneously when using the chart compared with the BNFc: paediatric questions 100% (51/51 chart) versus 80% (41/51 BNFc) (OR 0.20) and neonatal questions 55% (28/51 chart) versus 35% (18/51 BNFc) (OR 0.2). Errors when using the BNFc were due to selection of the incorrect regimen (23%), wrong frequency (17%) and one 10 fold dosing error. When using the chart, there were no dosing errors, only frequency errors for the neonatal regimen. CONCLUSION: The chart was more reliable, quicker and may be useful for patient safety. Revising the format of the BNFc may be beneficial for nurses. PMID- 18676439 TI - Perivascular spaces--MRI marker of inflammatory activity in the brain? AB - The Virchow-Robin spaces (VRS), perivascular compartments surrounding small blood vessels as they penetrate the brain parenchyma, are increasingly recognized for their role in leucocyte trafficking as well as for their potential to modulate immune responses. In the present study, we investigated VRS numbers and volumes in different brain regions in 45 multiple sclerosis patients and 30 healthy controls of similar age and gender distribution, applying three different MRI sequence modalities (T(2)-weighted, T(1)-weighted and FLAIR). VRS were detected in comparable numbers in both multiple sclerosis patients and healthy individuals, indicating that perivascular compartments present on MRI are not a unique feature of multiple sclerosis. However, multiple sclerosis patients had significantly larger VRS volumes than healthy controls (P = 0.004). This finding was not explained by a significantly lower brain parenchymal fraction (BPF), resulting from a higher degree of atrophy, in the patient cohort. In a multiple linear regression analysis, age had a significant influence on VRS volumes in the control group but not in multiple sclerosis patients (P = 0.023 and P = 0.263, respectively). A subsequent prospective longitudinal substudy with monthly follow up MRI over a period of up to 12 months in 18 patients revealed a significant increase in VRS volumes and counts accompanying the occurrence of contrast enhancing lesions (CEL). At time points when blood-brain barrier (BBB) breakdown was indicated by the appearance of CEL, total VRS volumes and counts were significantly higher compared with preceding time points without CEL (P = 0.011 and P = 0.041, respectively), whereas a decrease thereafter was not statistically significant. Thus, our data points to an association of VRS with CEL as a sign for inflammation rather than with factors such as age, observed in healthy controls, and therefore suggests a role of VRS in inflammatory processes of the brain. PMID- 18676440 TI - HIV programmes reduce deaths and new infections, UNAIDS report shows. PMID- 18676441 TI - Advanced glaucomatous visual loss and oral steroids. PMID- 18676442 TI - Epilepsy. PMID- 18676445 TI - Over 60s' use of prescription drugs has doubled in past decade in England. PMID- 18676446 TI - NHS Care for Overseas Patients. Free for all? PMID- 18676447 TI - Determination of the solubility and size distribution of radioactive aerosols in the uranium processing plant at NRCN. AB - Inhalation is the main route of internal exposure to radioactive aerosols in the nuclear industry. To assess the radiation dose from the intake of these aerosols, it is necessary to know their physical (aerodynamic diameter distribution) and chemical (dissolution rate in extracellular lung fluid) characteristics. Air samples were taken from the uranium processing plant at the Nuclear Research Center, Negev. Measurements of aerodynamic diameter distribution using a cascade impactor indicated an average activity median aerodynamic diameter value close to 5 microm, in accordance with the recent recommended values of International Commission on Radiological Protection (ICRP) model. Solubility profiles of these aerosols were determined by performing in vitro solubility tests over 100 d in a simultant solution of the extracellular fluid. The tests indicated that the uranium aerosols should be assigned to an absorption between Types M and S (as defined by the ICRP Publication 66 model). PMID- 18676448 TI - An approach of orthology detection from homologous sequences under minimum evolution. AB - In the field of phylogenetics and comparative genomics, it is important to establish orthologous relationships when comparing homologous sequences. Due to the slight sequence dissimilarity between orthologs and paralogs, it is prone to regarding paralogs as orthologs. For this reason, several methods based on evolutionary distance, phylogeny and BLAST have tried to detect orthologs with more precision. Depending on their algorithmic implementations, each of these methods sometimes has increased false negative or false positive rates. Here, we developed a novel algorithm for orthology detection that uses a distance method based on the phylogenetic criterion of minimum evolution. Our algorithm assumes that sets of sequences exhibiting orthologous relationships are evolutionarily less costly than sets that include one or more paralogous relationships. Calculation of evolutionary cost requires the reconstruction of a neighbor joining (NJ) tree, but calculations are unaffected by the topology of any given NJ tree. Unlike tree reconciliation, our algorithm appears free from the problem of incorrect topologies of species and gene trees. The reliability of the algorithm was tested in a comparative analysis with two other orthology detection methods using 95 manually curated KOG datasets and 21 experimentally verified EXProt datasets. Sensitivity and specificity estimates indicate that the concept of minimum evolution could be valuable for the detection of orthologs. PMID- 18676449 TI - The splice variants of UBF differentially regulate RNA polymerase I transcription elongation in response to ERK phosphorylation. AB - The mammalian architectural HMGB-Box transcription factor UBF is ubiquitously expressed in two variant forms as the result of a differential splicing event, that in the UBF2 deletes 37 amino acid from the second of six HMGB-boxes. Several attempts to define a function for this shorter UBF2 protein have been less than satisfactory. However, since all mammals appear to display similar levels of the longer and shorter UBF variants, it is unlikely that UBF2 is simply nonfunctional. Previously we showed that phosphorylation of UBF by the MAP-kinase ERK regulates chromatin folding and transcription elongation, explaining the rapid response of the ribosomal RNA genes to growth factors. Here we have investigated the roles the UBF variants play in the response of these genes to ERK activity. We demonstrate that the variant HMGB-box 2 of UBF2 has lost the ability to bind bent DNA and hence to induce chromatin folding. As a result it is significantly less effective than UBF1 at arresting RNAPI elongation but at the same time is more responsive to ERK phosphorylation. Thus, UBF2 functionally simulates a hemi-phosphorylated UBF whose expression may provide a means by which to tune the response of the ribosomal RNA genes to growth factor stimulation. PMID- 18676450 TI - Crystal engineering of HIV-1 reverse transcriptase for structure-based drug design. AB - HIV-1 reverse transcriptase (RT) is a primary target for anti-AIDS drugs. Structures of HIV-1 RT, usually determined at approximately 2.5-3.0 A resolution, are important for understanding enzyme function and mechanisms of drug resistance in addition to being helpful in the design of RT inhibitors. Despite hundreds of attempts, it was not possible to obtain the structure of a complex of HIV-1 RT with TMC278, a nonnucleoside RT inhibitor (NNRTI) in advanced clinical trials. A systematic and iterative protein crystal engineering approach was developed to optimize RT for obtaining crystals in complexes with TMC278 and other NNRTIs that diffract X-rays to 1.8 A resolution. Another form of engineered RT was optimized to produce a high-resolution apo-RT crystal form, reported here at 1.85 A resolution, with a distinct RT conformation. Engineered RTs were mutagenized using a new, flexible and cost effective method called methylated overlap extension ligation independent cloning. Our analysis suggests that reducing the solvent content, increasing lattice contacts, and stabilizing the internal low energy conformations of RT are critical for the growth of crystals that diffract to high resolution. The new RTs enable rapid crystallization and yield high resolution structures that are useful in designing/developing new anti-AIDS drugs. PMID- 18676451 TI - A probabilistic generative model for GO enrichment analysis. AB - The Gene Ontology (GO) is extensively used to analyze all types of high throughput experiments. However, researchers still face several challenges when using GO and other functional annotation databases. One problem is the large number of multiple hypotheses that are being tested for each study. In addition, categories often overlap with both direct parents/descendents and other distant categories in the hierarchical structure. This makes it hard to determine if the identified significant categories represent different functional outcomes or rather a redundant view of the same biological processes. To overcome these problems we developed a generative probabilistic model which identifies a (small) subset of categories that, together, explain the selected gene set. Our model accommodates noise and errors in the selected gene set and GO. Using controlled GO data our method correctly recovered most of the selected categories, leading to dramatic improvements over current methods for GO analysis. When used with microarray expression data and ChIP-chip data from yeast and human our method was able to correctly identify both general and specific enriched categories which were overlooked by other methods. PMID- 18676452 TI - Consolidated strategy for the analysis of microarray spike-in data. AB - As the number of users of microarray technology continues to grow, so does the importance of platform assessments and comparisons. Spike-in experiments have been successfully used for internal technology assessments by microarray manufacturers and for comparisons of competing data analysis approaches. The microarray literature is saturated with statistical assessments based on spike-in experiment data. Unfortunately, the statistical assessments vary widely and are applicable only in specific cases. This has introduced confusion into the debate over best practices with regards to which platform, protocols and data analysis tools are best. Furthermore, cross-platform comparisons have proven difficult because reported concentrations are not comparable. In this article, we introduce two new spike-in experiments, present a novel statistical solution that enables cross-platform comparisons, and propose a comprehensive procedure for assessments based on spike-in experiments. The ideas are implemented in a user friendly Bioconductor package: spkTools. We demonstrated the utility of our tools by presenting the first spike-in-based comparison of the three major platforms- Affymetrix, Agilent and Illumina. PMID- 18676456 TI - Confirmation of deposit, but confirmation of what? PMID- 18676454 TI - Recombineering-mediated tagging of Drosophila genomic constructs for in vivo localization and acute protein inactivation. AB - Studying gene function in the post-genome era requires methods to localize and inactivate proteins in a standardized fashion in model organisms. While genome wide gene disruption and over-expression efforts are well on their way to vastly expand the repertoire of Drosophila tools, a complementary method to efficiently and quickly tag proteins expressed under endogenous control does not exist for fruit flies. Here, we describe the development of an efficient procedure to generate protein fusions at either terminus in an endogenous genomic context using recombineering. We demonstrate that the fluorescent protein tagged constructs, expressed under the proper control of regulatory elements, can rescue the respective mutations and enable the detection of proteins in vivo. Furthermore, we also adapted our method for use of the tetracysteine tag that tightly binds the fluorescent membrane-permeable FlAsH ligand. This technology allows us to acutely inactivate any tagged protein expressed under native control using fluorescein-assisted light inactivation and we provide proof of concept by demonstrating that acute loss of clathrin heavy chain function in the fly eye leads to synaptic transmission defects in photoreceptors. Our tagging technology is efficient and versatile, adaptable to any tag desired and paves the way to genome-wide gene tagging in Drosophila. PMID- 18676457 TI - Prevotella histicola sp. nov., isolated from the human oral cavity. AB - Three strains of anaerobic, variably pigmenting, Gram-negative bacilli isolated from human oral mucosal tissue were subjected to a comprehensive range of phenotypic and genotypic tests and were found to comprise a homogeneous group. 16S rRNA gene sequence analysis and DNA-DNA hybridization revealed that the strains constituted a novel group within the genus Prevotella, being most closely related to Prevotella melaninogenica and Prevotella veroralis. A novel species, Prevotella histicola sp. nov., is proposed to accommodate these strains. Prevotella histicola is saccharolytic and produces acetic acid and succinic acid as major end products of fermentation and trace to minor amounts of isovaleric acid and lactic acid. The G+C content of the DNA of the type strain is 43 mol%. The type strain of Prevotella histicola is T05-04T (=DSM 19854T=CCUG 55407T). PMID- 18676453 TI - The triple helix: 50 years later, the outcome. AB - Triplex-forming oligonucleotides constitute an interesting DNA sequence-specific tool that can be used to target cleaving or cross-linking agents, transcription factors or nucleases to a chosen site on the DNA. They are not only used as biotechnological tools but also to induce modifications on DNA with the aim to control gene expression, such as by site-directed mutagenesis or DNA recombination. Here, we report the state of art of the triplex-based anti-gene strategy 50 years after the discovery of such a structure, and we show the importance of the actual applications and the main challenges that we still have ahead of us. PMID- 18676458 TI - Rhodobacter megalophilus sp. nov., a phototroph from the Indian Himalayas possessing a wide temperature range for growth. AB - Two strains of phototrophic, purple non-sulfur bacteria capable of growing at low temperatures (5 degrees C) were isolated from the Himalayas. The two strains showed positive phototaxis and grew over a relatively wide temperature range (5 40 degrees C). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JA194T clustered with members of the genus Rhodobacter. Strain JA194T showed highest 16S rRNA gene sequence similarity with Rhodobacter sphaeroides DSM 158T (99 %). However, DNA-DNA hybridization experiments between Rba. sphaeroides DSM 158T and strain JA194T revealed a level of relatedness of only 67 %. The DNA base composition of strain JA194T was 66.67 mol% G+C (by HPLC). Based on 16S rRNA gene sequence analysis, morphological, physiological, Fourier transform infrared fingerprinting and DNA-DNA hybridization studies, strain JA194T (=KCTC 5602T =JCM 14598T) is sufficiently different from other Rhodobacter species to merit its description as the type strain of a novel species, for which the name Rhodobacter megalophilus sp. nov. is proposed. PMID- 18676459 TI - Rubellimicrobium mesophilum sp. nov., a mesophilic, pigmented bacterium isolated from soil. AB - A pink- to light-reddish-pigmented, aerobic, pointed, budding, mesophilic strain designated MSL-20T was isolated from soil from Bigeum island, Korea. The isolate was Gram-negative. Cells were motile rods without flagella. The temperature and pH for growth were 20-37 degrees C (optimum 28 degrees C) and pH 7.0-11.0 (optimum pH 7.0+/-0.2). Q-10 was the predominant respiratory ubiquinone. The major fatty acids were C16:0 (37 %), C18: 1 omega7c (36 %) and 11-methyl C18:1 omega7c (12 %). Phylogenetic analysis based on the 16S rRNA gene sequence revealed that Rubellimicrobium thermophilum DSM 16684T was its closest neighbour (95.2 % similarity); the isolate had less than 97.0 % similarity to all species with validly published names. Chemotaxonomic data were determined for this isolate, including peptidoglycan type, diagnostic sugar composition, fatty acid profiles, ubiquinones and polar lipids; the results supported the affiliation of strain MSL-20T to the genus Rubellimicrobium. The DNA G+C content was 72.3 mol%. On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain MSL 20(T) represents a novel species of the genus Rubellimicrobium, for which the name Rubellimicrobium mesophilum sp. nov. is proposed. The type strain is MSL-20T (=KCTC 22012T =DSM 19309T). PMID- 18676460 TI - Sphingobium rhizovicinum sp. nov., isolated from rhizosphere soil of Fortunella hindsii (Champ. ex Benth.) Swingle. AB - The taxonomic status of a pale-yellow-coloured bacterial isolate from rhizosphere soil of Fortunella hindsii (Champ. ex Benth.) Swingle was characterized using a polyphasic taxonomic approach. Comparative analysis of the 16S rRNA gene sequence showed that the isolate constituted a distinct branch within the genus Sphingobium. The generic assignment was confirmed by chemotaxonomic data, which revealed the presence of a fatty acid profile that was characteristic for the genus Sphingobium, consisting of straight-chain saturated and unsaturated as well as 2-OH fatty acids and the lack of 3-OH fatty acids, ubiquinone with ten isoprene units (Q-10) as the predominant respiratory quinone, and a polar lipid pattern that consisted of the predominant compounds phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine, diphosphatidylglycerol, sphingoglycolipid and an unknown glycolipid. Spermidine was the major polyamine component. The genotypic and phenotypic data (physiology and fatty acid and polar lipid profiles) showed that the isolate merits classification as representing a novel species of the genus Sphingobium, for which the name Sphingobium rhizovicinum sp. nov. is proposed. The type strain is CC-FH12-1T (=CCM 7491(T)=BCRC 17770T) [corrected] PMID- 18676461 TI - Complete rpoB gene sequencing as a suitable supplement to DNA-DNA hybridization for bacterial species and genus delineation. AB - DNA-DNA hybridization (DDH), the gold standard for bacterial species delineation, is a laborious method and the alternative, average nucleotide identity (ANI), a genomic sequence-derived parameter, is not applicable to non-sequenced species. A universal cut-off value to delineate bacterial species does not exist, yet a DDH value <70 % and ANI <95+/-0.5 % have proved useful in selected examples. We herein compare published values for DDH and ANI with sequence similarity of rpoB gene sequences retrieved from GenBank for strains of 230 bacterial species representative of 45 genera. Intraspecific rpoB sequence similarity was 98.2-100 %. We observed that an rpoB gene sequence similarity 1 family risk factor, who had a serious injury, who had a child risk factor other than an inconsistent injury, who were black, or who were unfamiliar to the clinician were more likely to be reported. Clinicians who had not reported all suspicious injuries during their career or who had lost families as patients because of previous reports were more likely to report suspicious injuries. CONCLUSIONS: Clinicians had some degree of suspicion that approximately 10% of the injuries they evaluated were caused by child abuse. Clinicians did not report all suspicious injuries to child protective services, even if the level of suspicion was high (likely or very likely caused by child abuse). Child, family, and injury characteristics and clinician previous experiences influenced decisions to report. PMID- 18676508 TI - Child abuse recognition and reporting: supports and resources for changing the paradigm. AB - As shown previously in the Child Abuse Recognition Experience Study (CARES), primary care providers reported that they decided not to report a substantial proportion of injuries that they suspected might have resulted from abuse. The most serious cases result in death. This article provides detailed reports of 2 illustrative cases from the author's experience as a member of a multidisciplinary child fatality review team and discusses several alternative legal mechanisms for improving mandated reporting. Regional centers of excellence in child protection may be an effective way of improving child protection; current proposals to establish such centers are reviewed. PMID- 18676509 TI - Professional education in child abuse and neglect. AB - Physicians have reported feeling that they were not adequately trained to identify and report child abuse. This article reviews the current state of medical education and residency training and the needs of physicians in practice and proposes changes and additions that can be made to improve the ability and confidence of physicians who are faced with the responsibility of keeping children safe. PMID- 18676510 TI - Translating child abuse research into action. AB - The Child Abuse Recognition Experience Study revealed that primary care clinicians did not always follow the legal mandate to report suspected child abuse to child protective services. National child abuse experts representing different disciplines met in a 2-day conference in January 2007 to discuss and develop new strategies that would address the barriers to reporting suspected child abuse and improve the protection of children. This article describes the rationale, structure, and goals of the conference. PMID- 18676511 TI - Doing the right thing: a primary care pediatrician's perspective on child abuse reporting. AB - The Child Abuse Recognition Experience Study (CARES) demonstrated that primary care physicians and nurse practitioners who treat injured children find the decision of whether to report suspected abuse difficult. This commentary briefly summarizes and responds to the papers presented at the Child Abuse Recognition, Research, and Education Translation (CARRET) Conference. The commentator traces her own changing views of child protective services as she became more involved in a multidisciplinary assessment team. Pediatricians are called on to advocate for more effective collaborations to better protect vulnerable maltreated children. PMID- 18676512 TI - In-hospital falls of newborn infants: data from a multihospital health care system. AB - OBJECTIVE: The fall of a newborn infant to the hospital floor is an error that has received little or no attention in medical publications. We sought to analyze the circumstances surrounding all such falls that occurred in an 18-hospital health care system during a 3-year period. METHODS: Information was located by using electronic and risk-management records. Demographic features, circumstances of the fall, and outcomes were tabulated for each event. RESULTS: During the study period, 88774 live births occurred at the Intermountain Healthcare hospitals. Fourteen neonatal in-hospital falls were identified during this period (incidence estimate: 1.6 falls per 10000 births). Seven falls occurred when a parent, holding the infant in a hospital bed or reclining chair, fell asleep and the infant fell to the floor. Six of these 7 falls occurred between 1:30 am and 9:00 am. Four falls occurred in the delivery room, 2 in the hallway while a nurse was wheeling a bassinette, and 1 from an infant swing. No deaths occurred. One patient sustained a depressed skull fracture and was transported to the regional children's hospital. At hospital discharge, 13 of the 14 were reported to have a normal examination. No specific protocols for preventing in-hospital falls of neonates were in place. CONCLUSIONS: If the incidence of a neonatal in-hospital fall in this study is representative, then 600 to 700 such falls occur annually in the United States. Relatively few scenarios explain the majority of falls. We speculate that the prevalence of this error could be reduced significantly by enacting programs aimed at eliminating or monitoring the most common circumstances under which these falls occur. PMID- 18676513 TI - Girls who disclose sexual abuse: urogenital symptoms and signs after genital contact. AB - BACKGROUND: Little information is available about idiosyncratic historical details provided by sexually abused girls, yet this information can help medical professionals diagnose sexual abuse. OBJECTIVES: Our goals were to describe types and frequencies of urogenital symptoms/signs reported by girls who disclosed direct genital contact and to explore factors associated with this reporting. METHODS: We reviewed 161 medical charts of 3- to 18-year-old girls who disclosed sexual abuse by direct genital contact for urogenital symptoms/signs, type of genital contact (oral, object, digital, or genital), time interval between last perpetrator contact and physical examination, age and sexual maturity at the time of last perpetrator contact, genital findings, and other medical diagnoses. Regression analyses were performed to determine factors that were most predictive of symptom/sign reporting. RESULTS: Many of the girls reported multiple types of genital contact; 33% reported oral/object-genital contact, 72% reported digital genital contact, and 55% reported genital-genital contact. Sixty percent of the girls reported experiencing >or=1 symptom/sign; 53% of the total sample had genital pain, 37% had dysuria, and 11% had genital bleeding. Symptoms/signs were highly associated with genital-genital contact: 48% of the girls reporting genital-genital contact had dysuria compared with 25% of girls not reporting genital-genital contact, 72% had genital pain/soreness compared with 32% not reporting genital-genital contact, and 16% had bleeding compared with 4% of those not reporting genital-genital contact. Using regression analysis, the strongest factor predictive of symptom reporting by the girls was genital-genital contact. CONCLUSIONS: Sexually abused girls who experienced direct genital contact frequently reported symptoms related to the abusive episode. These symptoms were reported most frequently with genital-to-genital contact. This information sheds some light on the mechanism of injury leading to symptom reporting and can be used to further study symptoms/signs reported by sexually abused girls compared with the general population. PMID- 18676514 TI - Reduced exercise capacity in children born very preterm. AB - OBJECTIVE: In the past 20 years, there has been an increase in survivors of very preterm birth, but little is known regarding their long-term respiratory and fitness outcomes. We aimed to assess the 10-year lung function and fitness outcomes for children who were born weighing <1000 g and before 32 weeks' gestation in 1992-1994. METHODS: A cross-sectional study was conducted of 126 children (mean age: 10 years) who were born at a mean gestation of 27 weeks and 34 term-born control subjects. Extensive lung function (spirometry, lung volumes, and gas exchange) and fitness (6-minute walk and 20-m shuttle run tests) assessments were conducted at a single visit according to previously validated techniques. RESULTS: The preterm group had significantly lower values for all measured spirometric parameters compared with the control group. In contrast to airflow, the preterm group had significantly higher percentage predicted values in all standard lung-volume parameters and transfer factor than the control group. The exercise capacity of the preterm group was approximately half that of the control group. There was no significant difference in the distance walked in the 6-minute walk test. CONCLUSIONS: In the largest cohort of school-aged children (born very preterm in the 1990s) to undergo extensive lung function and fitness assessments, we demonstrated significant impairment in exercise capacity despite evidence of only mild small-airway obstruction and gas trapping. Additional studies are required to evaluate the cause of this exercise limitation and whether it can be improved with a training program. PMID- 18676515 TI - Adverse childhood experiences predict earlier age of drinking onset: results from a representative US sample of current or former drinkers. AB - OBJECTIVE: Our goal was to determine whether adverse childhood experiences predicted the age at which drinking was initiated and drinking motives in a representative sample of current or former drinkers in the United States. METHODS: In 2006, a probability sample of 3592 US current or former drinkers aged 18 to 39 were surveyed. Multinomial logistic regression examined whether each of 10 adverse childhood experiences was associated with earlier ages of drinking onset, controlling for demographics, parental alcohol use, parental attitudes toward drinking, and peers' drinking in adolescence. We also examined whether there was a graded relationship between the number of adverse childhood experiences and age of drinking onset and whether adverse childhood experiences were related to self-reported motives for drinking during the first year that respondents drank. RESULTS: Sixty-six percent of respondents reported >or=1 adverse childhood experiences, and 19% reported experiencing >or=4. The most commonly reported adverse childhood experiences were parental separation/divorce (41.3%), living with a household member who was a problem drinker (28.7%), mental illness of a household member (24.8%), and sexual abuse (19.1%). Of the 10 specific adverse childhood experiences assessed, 5 were significantly associated with initiating drinking at or=21 years of age) after adjustment for confounders, including physical abuse, sexual abuse, having a mentally ill household member, substance abuse in the home, and parental discord or divorce. Compared with those without adverse childhood experiences, respondents with adverse childhood experiences were substantially more likely to report that they drank to cope during the first year that they used alcohol. CONCLUSIONS: Results suggest that children with particular adverse childhood experiences may initiate drinking earlier than their peers and that they may be more likely to drink to cope with problems (rather than for pleasure or to be social). PMID- 18676516 TI - A video game improves behavioral outcomes in adolescents and young adults with cancer: a randomized trial. AB - OBJECTIVE: Suboptimal adherence to self-administered medications is a common problem. The purpose of this study was to determine the effectiveness of a video game intervention for improving adherence and other behavioral outcomes for adolescents and young adults with malignancies including acute leukemia, lymphoma, and soft-tissue sarcoma. METHODS: A randomized trial with baseline and 1- and 3-month assessments was conducted from 2004 to 2005 at 34 medical centers in the United States, Canada, and Australia. A total of 375 male and female patients who were 13 to 29 years old, had an initial or relapse diagnosis of a malignancy, and currently undergoing treatment and expected to continue treatment for at least 4 months from baseline assessment were randomly assigned to the intervention or control group. The intervention was a video game that addressed issues of cancer treatment and care for teenagers and young adults. Outcome measures included adherence, self-efficacy, knowledge, control, stress, and quality of life. For patients who were prescribed prophylactic antibiotics, adherence to trimethoprim-sulfamethoxazole was tracked by electronic pill monitoring devices (n = 200). Adherence to 6-mercaptopurine was assessed through serum metabolite assays (n = 54). RESULTS: Adherence to trimethoprim sulfamethoxazole and 6-mercaptopurine was greater in the intervention group. Self efficacy and knowledge also increased in the intervention group compared with the control group. The intervention did not affect self-report measures of adherence, stress, control, or quality of life. CONCLUSIONS: The video-game intervention significantly improved treatment adherence and indicators of cancer-related self efficacy and knowledge in adolescents and young adults who were undergoing cancer therapy. The findings support current efforts to develop effective video-game interventions for education and training in health care. PMID- 18676517 TI - Unexpected infant deaths associated with use of cough and cold medications. AB - OBJECTIVE: The objective of this study was to determine whether caregivers had given infants who died unexpectedly over-the-counter cough and cold medications before the infant deaths to identify sociodemographic risk factors for their use. METHODS: The Arizona Child Fatality Review Program reviews the circumstances surrounding every child death that occurs in the state each year. By statute, the multidisciplinary review teams have access to all medical charts, autopsy reports, law enforcement reports, and other records for their review and use these data to determine the cause of death and its preventability. The data on all infants who died unexpectedly in 2006 and had an autopsy and postmortem toxicologic studies were reviewed for this analysis. RESULTS: Ten unexpected infant deaths that were associated with cold-medication use were identified. The infants ranged in age from 17 days to 10 months. Postmortem toxicology testing found evidence of recent administration of pseudoephedrine, antihistamine, dextromethorphan, and/or other cold-medication ingredients in these infants. The families who used these medications were poor and publicly insured, and 50% of them had limited English proficiency. Only 4 of these infants had received medical care for their current illness before their death. The over-the-counter cough and cold medication had been prescribed by a clinician for only 1 of these infants. CONCLUSIONS: Review of these infants' deaths raises concern about the role of the over-the-counter cough and cold medications in these deaths. These findings support the recommendation that such medications not be given to infants. In addition, these findings suggest that warnings on these medications "to consult a clinician" before use are not being followed by parents. Educational campaigns to decrease the use of over-the-counter cough and cold medications in infants need to be increased. PMID- 18676518 TI - Cough and cold medication use by US children, 1999-2006: results from the slone survey. AB - OBJECTIVE: Pediatric cough and cold medications are widely marketed in the United States, but the precise patterns of use among children are not known. Such information is especially important given recent reports suggesting that these medications are responsible for previously underappreciated serious adverse events and deaths among children. We sought to describe the prevalence and patterns of pediatric use of cough and cold medications, with particular attention to use among young children. METHODS: We analyzed data on the use of cough and cold medications, defined as any oral medication that contains >or=1 antitussive, decongestant, expectorant, and/or first-generation antihistamine active ingredients, among 4267 US children who were younger than 18 years and enrolled during 1999-2006 in the Slone Survey, a national random-digit-dial telephone survey of medication use by the US population. RESULTS: In a given week, a cough and cold medication was used by 10.1% of US children. Exposure was highest to decongestants (6.3%; mostly pseudoephedrine) and first-generation antihistamines (6.3%; most common were chlorpheniramine, diphenhydramine, and brompheniramine), followed by antitussives (4.1%; mostly dextromethorphan) and expectorants (1.5%; almost exclusively guaifenesin). Multiple-ingredient products accounted for 64.2% of all cough and cold medications used. Exposure to antitussives, decongestants, and first-generation antihistamines was highest among 2- to 5-year-olds (7.0%, 9.9%, and 10.1%, respectively) followed by children who were younger than 2 years (5.9%, 9.4%, and 7.6%, respectively); expectorant use was low in all age groups. The use of cough and cold medications declined from 1999 through 2006. CONCLUSIONS: Approximately 1 in 10 US children uses a cough and cold medication in a given week. The especially high prevalence of use among children of young age is noteworthy, given concerns about potential adverse effects and the lack of data on the efficacy of cough and cold medications in this age group. PMID- 18676519 TI - Long-term outcome of the ketogenic diet for intractable childhood epilepsy with focal malformation of cortical development. AB - OBJECTIVE: We evaluated the efficacy and long-term outcome of the ketogenic diet in patients with intractable childhood epilepsy as a result of focal malformation of cortical development. METHODS: A retrospective analysis evaluated seizure outcomes of 47 patients who had intractable epilepsy from (and) surgically remediable focal malformation of cortical development and were first treated with the classic ketogenic diet, involving the 4:1 lipid/nonlipid ratio. The long-term prognosis of 21 patients, who became seizure-free 3 months after the ketogenic diet, was followed up with that of 22 patients who eventually underwent epilepsy surgery. RESULTS: Three months after diet initiation, 29 (61.7%) patients showed a reduction in seizure frequency of >50%, including 21 (44.7%) who became seizure free. Of the 21 patients with complete seizure control at 3 months, 16 (76.2%) successfully completed the diet for 2 years without relapse, and 10 (47.6%) have remained seizure-free after cessation of the diet (mean follow-up for 3 years and 10 months), including 1 patient who remained seizure-free with additional medication after a relapse. Of the 22 patients who underwent epilepsy surgery, a seizure-free outcome was obtained for 13 (59.1%). CONCLUSIONS: The ketogenic diet should be considered to be an additional option even in patients with focal malformation of cortical development, and long-term seizure-free outcome can be expected for patients who become seizure-free 3 months after the diet. PMID- 18676520 TI - Growth of children on classical and medium-chain triglyceride ketogenic diets. AB - OBJECTIVE: The objectives of this study were to examine growth in children on classical and medium-chain triglyceride ketogenic diets and to investigate any association between growth and calorie or protein intake. METHODS: Weight, height, and BMI z scores were recorded for children who were initiated on 1 of 2 ketogenic diets at baseline and after 3, 6, and 12 months, if continued. Mean calorie and protein intakes during treatment were calculated for children who completed 12 months on the diet. Changes in growth were compared between the 2 diets, and the association between growth and dietary intake was examined. RESULTS: Seventy-five children provided growth data. Weight z scores decreased significantly between baseline and 3, 6, and 12 months; height z scores showed no change at 3 months but decreased significantly by 6 and 12 months. This was more significant in the younger and ambulatory children. Subdivision according to diet type showed weight z scores to decrease significantly in the medium-chain triglyceride group only at 3 and 6 months and in both groups at 12 months. Height z scores decreased significantly in both groups by 6 and 12 months. Forty children completed 12 months of treatment; in this group, the slopes of best-fit regression lines of serial z-score measures were used to represent growth trend. There were no significant differences in mean slope between classical and medium chain triglyceride diet groups for weight, height, or BMI. There was no significant difference in mean calorie intake during the 12 months between the 2 diets, but the medium-chain triglyceride group had significantly higher protein intake. There was no significant correlation between calorie or protein intakes and the slope of the best-fit line for weight, height, or BMI. CONCLUSIONS: Both weight and height z scores decreased during diet treatment. By 12 months, there was no difference in outcome between classical and medium-chain triglyceride protocols despite the increased protein in the latter diet. PMID- 18676521 TI - Birth weight and cognitive ability in childhood among siblings and nonsiblings. AB - OBJECTIVES: The purpose of this work was to examine whether the positive association between birth weight and childhood cognitive ability is seen within siblings from the same family, as well as between nonsiblings, and to determine whether these associations vary with age. METHODS: We compared the association of birth weight with cognitive ability measured at ages 5 to 6, 7 to 9, and 11 to 12 years among a total of 5402 children from different families with that among 2236 to 3083 sibships from the National Longitudinal Study of Youth 1979-Children. RESULTS: In the whole cohort, there were positive associations between birth weight and cognitive ability at all ages, with the association increasing with age from a 0.81-point increase at ages 5 to 6 years to 1.30 and 1.44 points at ages 7 to 9 and 11 to 12 years, respectively, per 1 SD of gestational age- and gender-adjusted birth weight z score. With adjustment for covariates, there was marked attenuation of these associations. Mean differences were 0.28 points in children aged 5 to 6 years, 0.67 points in those aged 7 to 9 years, and 0.52 points in those aged 11 to 12 years after adjusting for child's gender, race or ethnicity, year of birth, and age at test; maternal age, height, parity, education, smoking during pregnancy, and cognitive ability; and household income. Our family-based analyses that separated within- and between-family effects found that the between-family associations were much stronger than the within-family associations. However, adjustment for potential confounders attenuated the between-family associations, and there was no evidence for a difference in association comparing the between- and within-family associations. CONCLUSIONS: In these data, the positive association between birth weight and childhood cognitive ability at ages 5 to 12 years is explained largely by family characteristics rather than a specific intrauterine effect. PMID- 18676522 TI - Accidental and nonaccidental poisonings as a cause of apparent life-threatening events in infants. AB - BACKGROUND: Apparent life-threatening events are a relatively common event in children for which there may be a number of causes. Previous reports have suggested that poisonings, either accidental or intentional, may be causes of some events. However, this theory has not been systematically studied. METHODS: We conducted a prospective, descriptive study of infants aged <2 years presenting to a pediatric emergency department of a large, urban tertiary care children's hospital with signs and symptoms of an apparent life-threatening event. All of the children presenting with an apparent life-threatening event were to undergo a standardized evaluation, which included obtaining a comprehensive urine toxicology screen. A positive toxicology screen result was defined as follows: a clinically insignificant screen result (identification of a medication that would not cause an apparent life-threatening event) or a clinically significant screen result (identification of a medication that could cause apnea or other event consistent with an apparent life-threatening event, even if it was a medication that the child was known to be taking). RESULTS: During the study period, 596 children presented to the emergency department with an apparent life-threatening event, and 274 (46.0%) had a toxicology screen performed. Of 274 toxicology screen results, 50 were considered truly positive (18.2%), and 23 positive screen results were considered clinically significant (23 of 274 [8.4%]). Thirteen toxicology screen results were positive for an over-the-counter cold preparation (13 of 274 [4.7%]). No parent admitted to having given his or her child an over the-counter cold preparation. CONCLUSIONS: A substantial number of children presenting to the emergency department with an apparent life-threatening event had a positive toxicology screen result. In particular, a number of children were found to have been given an over-the-counter cold preparation. We would recommend that toxicology screens be included as part of the routine evaluation of children who present with an apparent life-threatening event. PMID- 18676523 TI - Clinical effort against secondhand smoke exposure: development of framework and intervention. AB - OBJECTIVE: The purpose of this work was to describe a novel process and present results of formative research to develop a pediatric office intervention that uses available systems of care for addressing parental smoking. METHODS: The scientific development of the intervention occurred in 3 stages. In stage 1, we designed an office system for parental tobacco control in the pediatric outpatient setting on the basis of complementary conceptual frameworks of preventive services delivery, conceptualized for the child health care setting through a process of key interviews with leaders in the field of implementing practice change; existing Public Health Service guidelines that had been shown effective in adult practices; and adaptation of an evidence-based adult office system for tobacco control. This was an iterative process that yielded a theoretically framed intervention prototype. In stage 2, we performed focus-group testing in pediatric practices with pediatricians, nurses, clinical assistants, and key office staff. Using qualitative methods, we adapted the intervention prototype on the basis of this feedback to include 5 key implementation steps for the child health care setting. In stage 3, we presented the intervention to breakout groups at 2 national meetings of pediatric practitioners for additional refinements. RESULTS: The main result was a theoretically grounded intervention that was responsive to the barriers and suggestions raised in the focus groups and at the national meetings. The Clinical Effort Against Secondhand Smoke Exposure intervention was designed to be flexible and adaptable to the particular practices' staffing, resources, and physical configuration. Practice staff can choose materials relevant to their own particular systems of care (www.ceasetobacco.org). CONCLUSIONS: Conceptually grounded and focus-group-tested strategies for parental tobacco control are now available for implementation in the pediatric outpatient setting. The tobacco-control intervention-development process might have particular relevance for other chronic pediatric conditions that have a strong evidence base and have available treatments or resources that are underused. PMID- 18676524 TI - A medical home for children with insulin-dependent diabetes: comanagement by primary and subspecialty physicians--convergence and divergence of opinions. AB - OBJECTIVE: The purpose of this work was to examine pediatricians' and endocrinologists' views about management for routine preventive and acute care, diabetes-specific care, and family education and care coordination for children with insulin-dependent diabetes. METHODS: We conducted a mixed-mode survey of all of the pediatricians in 1 medicaid managed care network and all of the pediatric and adult endocrinologists who treat children with diabetes in North Carolina. RESULTS: Of the 201 pediatricians surveyed, 132 responded (65%). Among the 61 endocrinologists who treat children, 59% replied. Nearly all of the respondents agreed that primary care physicians should have responsibility for routine primary care (eg, well-child checkups, treating minor illnesses or injuries, and immunizations). Likewise, large majorities favored endocrinologists as leads for diabetes-specific care (eg, 94% for training in use of an insulin pump and 82% for training in use of a glucometer). Many generalists and subspecialists reported that specific aspects of diabetes care should be comanaged (eg, 31% for tracking of hemoglobin A1c). However, large proportions of pediatricians and endocrinologists expressed differing opinions about the primary responsibility for family education and care coordination and for specific diabetes services. For example, 80% of endocrinologists saw subspecialists as leads for monitoring blood sugar levels, whereas 52% of pediatricians favored comanagement. CONCLUSIONS: An effective medical home model of care depends on establishing clear lines of responsibility between the primary care physician and subspecialist. Our findings suggest that primary care physicians and subspecialists agree on who should lead most aspects of care for patients with insulin-dependent diabetes and that some aspects of care should be comanaged. However, primary care physicians and subspecialists did not agree either between or within disciplines on who should be more responsible for the basic aspect of monitoring of blood sugar levels. Approaches that recognize the appropriate division of care between primary care physicians and subspecialists, facilitate comanagement when it is needed, and reward the collaboration required to provide medical homes for patients should be investigated as models of care. PMID- 18676525 TI - Diving-related injuries in children <20 years old treated in emergency departments in the United States: 1990-2006. AB - OBJECTIVE: The purpose of this work was to comprehensively examine diving-related injuries in the United States among children and adolescents <20 years of age. METHODS: We conducted a retrospective analysis of diving-related injury data from the National Electronic Injury Surveillance System, including patients aged <20 years old who were seen in an emergency department for a diving-related injury from 1990 through 2006. RESULTS: An estimated 111341 patients aged 6 million hospital stays, costing almost $50 billion annually, hospitalized children represent an important population for which most inpatient quality indicators are not applicable. Our aim was to develop indicators using inpatient administrative data to assess aspects of the quality of inpatient pediatric care and access to quality outpatient care. METHODS: We adapted the Agency for Healthcare Research and Quality quality indicators, a publicly available set of measurement tools refined previously by our team, for a pediatric population. We systematically reviewed the literature for evidence regarding coding and construct validity specific to children. We then convened 4 expert panels to review and discuss the evidence and asked them to rate each indicator through a 2-stage modified Delphi process. From the 2000 and 2003 Agency for Healthcare Research and Quality Healthcare Cost and Utilization Project Kids' Inpatient Database, we generated national estimates for provider level indicators and for area level indicators. RESULTS: Panelists recommended 18 indicators for inclusion in the pediatric quality indicator set based on overall usefulness for quality improvement efforts. The indicators included 13 hospital level indicators, including 11 based on complications, 1 based on mortality, and 1 based on volume, as well as 5 area-level potentially preventable hospitalization indicators. National rates for all 18 of the indicators varied minimally between years. Rates in high-risk strata are notably higher than in the overall groups: in 2003 the decubitus ulcer pediatric quality indicator rate was 3.12 per 1000, whereas patients with limited mobility experienced a rate of 22.83. Trends in rates by age varied across pediatric quality indicators: short term complications of diabetes increased with age, whereas admissions for gastroenteritis decreased with age. CONCLUSIONS: Tracking potentially preventable complications and hospitalizations has the potential to help prioritize quality improvement efforts at both local and national levels, although additional validation research is needed to confirm the accuracy of coding. PMID- 18676530 TI - Cognitive function and brain structure in females with a history of adolescent onset anorexia nervosa. AB - OBJECTIVE: Abnormalities in cognitive function and brain structure have been reported in acutely ill adolescents with anorexia nervosa, but whether these abnormalities persist or are reversible in the context of weight restoration remains unclear. Brain structure and cognitive function in female subjects with adolescent-onset anorexia nervosa assessed at long-term follow-up were studied in comparison with healthy female subjects, and associations with clinical outcome were investigated. PATIENTS AND METHODS: Sixty-six female subjects (aged 21.3 +/- 2.3 years) who had a diagnosis of adolescent-onset anorexia nervosa and treated 6.5 +/- 1.7 years earlier in a tertiary care hospital and 42 healthy female control subjects (aged 20.7 +/- 2.5 years) were assessed. All participants underwent a clinical examination, magnetic resonance brain scan, and cognitive evaluation. Clinical data were analyzed first as a function of weight recovery (n = 14, <85% ideal body weight; n = 52, >or=85% ideal body weight) and as a function of menstrual status (n = 18, absent/irregular menses; n = 29, oral contraceptive pill; n = 19, regular menses). Group comparisons were made across structural brain volumes and cognitive scores. RESULTS: Compared with control subjects, participants with anorexia nervosa who remained at low weight had larger lateral ventricles. Twenty-four-hour urinary free-cortisol levels were positively correlated with volumes of the temporal horns of the lateral ventricles and negatively correlated with volumes of the hippocampi in clinical participants. Participants who were amenorrheic or had irregular menses showed significant cognitive deficits across a broad range of many domains. CONCLUSIONS: Female subjects with adolescent-onset anorexia nervosa showed abnormal cognitive function and brain structure compared with healthy individuals despite an extended period since diagnosis. To our knowledge, this is the first study to report a specific relationship between menstrual function and cognitive function in this patient population. Possible mechanisms underlying neural and cognitive deficits with anorexia nervosa are discussed. Additional examination of the effects of estrogen on cognitive function in female subjects with anorexia nervosa is necessary. PMID- 18676531 TI - Macrophage migration inhibitory factor and autism spectrum disorders. AB - OBJECTIVE: Autistic spectrum disorders are childhood neurodevelopmental disorders characterized by social and communicative impairment and repetitive and stereotypical behavior. Macrophage migration inhibitory factor (MIF) is an upstream regulator of innate immunity that promotes monocyte/macrophage activation responses by increasing the expression of Toll-like receptors and inhibiting activation-induced apoptosis. On the basis of results of previous genetic linkage studies and reported altered innate immune response in autism spectrum disorder, we hypothesized that MIF could represent a candidate gene for autism spectrum disorder or its diagnostic components. METHODS: Genetic association between autism spectrum disorder and MIF was investigated in 2 independent sets of families of probands with autism spectrum disorder, from the United States (527 participants from 152 families) and Holland (532 participants from 183 families). Probands and their siblings, when available, were evaluated with clinical instruments used for autism spectrum disorder diagnoses. Genotyping was performed for 2 polymorphisms in the promoter region of the MIF gene in both samples sequentially. In addition, MIF plasma analyses were conducted in a subset of Dutch patients from whom plasma was available. RESULTS: There were genetic associations between known functional polymorphisms in the promoter for MIF and autism spectrum disorder-related behaviors. Also, probands with autism spectrum disorder exhibited higher circulating MIF levels than did their unaffected siblings, and plasma MIF concentrations correlated with the severity of multiple autism spectrum disorder symptoms. CONCLUSIONS: These results identify MIF as a possible susceptibility gene for autism spectrum disorder. Additional research is warranted on the precise relationship between MIF and the behavioral components of autism spectrum disorder, the mechanism by which MIF contributes to autism spectrum disorder pathogenesis, and the clinical use of MIF genotyping. PMID- 18676532 TI - Perceptions of oral health training and attitudes toward performing oral health screenings among graduating pediatric residents. AB - BACKGROUND: There is a lack of access to oral health services for American children. Some propose that pediatricians be trained to deliver preventive screening and education to families until they access a dental home. However, little is known about the current oral health training provided during pediatric residency. OBJECTIVE. The purpose of this work was to evaluate the oral health training that pediatricians receive during residency and their attitudes toward performing basic oral health screenings. METHODS: The American Academy of Pediatrics Survey of Graduating Residents is an annual, randomly sampled national survey of graduating pediatric residents. The 2006 Survey of Graduating Residents surveyed 611 residents and examined their perception of their oral health training and their attitudes about performing oral health screenings. RESULTS: Thirty-five percent of residents received no oral health training during residency. Of those who did, 73% had <3 hours of training, and only 14% had clinical observation time with a dentist. Seventy-one percent felt that they had too little oral health training, and only 21% felt that their residency was very good or excellent in preparing them to perform oral health-risk assessments. Residents felt confident in their ability to offer anticipatory guidance but not to do the more technical oral health screenings. The majority of residents believed that pediatricians should conduct basic oral health screenings. Multiple regression analysis indicated that residents who received >or=3 hours of oral health training, who applied for jobs in the inner city, and whose career goal was to work in primary practice are those most likely to support this idea. CONCLUSIONS: Pediatric residents currently receive little training in oral health, and the majority wish for more. This study shows that oral health training during residency can increase pediatrician confidence in participating in important oral health promotion tasks, including anticipatory guidance, oral screenings, and oral health-risk assessment. PMID- 18676533 TI - Effect of supplementing pregnant and lactating mothers with n-3 very-long-chain fatty acids on children's IQ and body mass index at 7 years of age. AB - OBJECTIVES: Arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3) are essential for brain growth and cognitive development. We have reported that supplementing pregnant and lactating women with n-3 very-long-chain polyunsaturated fatty acids promotes higher IQ scores at 4 years of age as compared with maternal supplementation with n-6 polyunsaturated fatty acids. In our present study, the children were examined at 7 years of age with the same cognitive tests as at 4 years of age. We also examined the relation between plasma fatty acid pattern and BMI in children, because an association between arachidonic acid and adipose tissue size has been suggested. METHODS: The study was randomized and double-blinded. The mothers took 10 mL of cod liver oil or corn oil from week 18 of pregnancy until 3 months after delivery. Their children were tested with the Kaufman Assessment Battery for Children at 7 years of age, and their height and weight were measured. RESULTS: We did not find any significant differences in scores on the Kaufman Assessment Battery for Children test at 7 years of age between children whose mothers had taken cod liver oil (n = 82) or corn oil (n = 61). We observed, however, that maternal plasma phospholipid concentrations of alpha-linolenic acid (18:3n-3) and docosahexaenoic acid during pregnancy were correlated to sequential processing at 7 years of age. We observed no correlation between fatty acid status at birth or during the first 3 months of life and BMI at 7 years of age. CONCLUSION: This study suggests that maternal concentration of n-3 very-long-chain polyunsaturated fatty acids during pregnancy might be of importance for later cognitive function, such as sequential processing, although we observed no significant effect of n-3 fatty acid intervention on global IQs. Neonatal fatty acid status had no influence on BMI at 7 years of age. PMID- 18676534 TI - Access to and use of paid sick leave among low-income families with children. AB - OBJECTIVE: The ability of employed parents to meet the health needs of their children may depend on their access to sick leave, especially for low-income workers, who may be afforded less flexibility in their work schedules to accommodate these needs yet also more likely to have children in poor health. Our goal was to provide rates of access to paid sick leave and paid vacation leave among low-income families with children and to assess whether access to these benefits is associated with parents' leave taking to care for themselves or others. METHODS: We used a sample of low-income families (<200% of the federal poverty level) with children aged 0 to 17 years in the 2003 and 2004 Medical Expenditure Panel Survey to examine bivariate relationships between access to and use of paid leave and characteristics of children, families, and parents' employer. RESULTS: Access to paid leave was lower among children in low-income families than among those in families with higher income. Within low-income families, children without >or=1 full-time worker in the household were especially likely to lack access to this benefit, as were children whose parents work for small employers. Among children whose parents had access to paid sick leave, parents were more likely to take time away from work to care for themselves or others. This relationship is even more pronounced among families with the highest need, such as children in fair or poor health and children with all parents in full-time employment. CONCLUSIONS: Legislation mandating paid sick leave could dramatically increase access to this benefit among low-income families. It would likely diminish gaps in parents' leave taking to care for others between families with and without the benefit. However, until the health related consequences are better understood, the full impact of such legislation remains unknown. PMID- 18676535 TI - Pediatric injuries from needles discarded in the community: epidemiology and risk of seroconversion. AB - OBJECTIVES: Although anxiety exists concerning the perceived risk of transmission of bloodborne viruses after community-acquired needlestick injuries, seroconversion seems to be rare. The objectives of this study were to describe the epidemiology of pediatric community-acquired needlestick injuries and to estimate the risk of seroconversion for HIV, hepatitis B virus, and hepatitis C virus in these events. METHODS: The study population included all of the children presenting with community-acquired needlestick injuries to the Montreal Children's Hospital between 1988 and 2006 and to Hopital Sainte-Justine between 1995 and 2006. Data were collected prospectively at Hopital Sainte-Justine from 2001 to 2006. All of the other data were reviewed retrospectively by using a standardized case report form. RESULTS: A total of 274 patients were identified over a period of 19 years. Mean age was 7.9 +/- 3.4 years. A total of 176 (64.2%) were boys. Most injuries occurred in streets (29.2%) or parks (24.1%), and 64.6% of children purposely picked up the needle. Only 36 patients (13.1%) noted blood on the device. Among the 230 patients not known to be immune for hepatitis B virus, 189 (82.2%) received hepatitis B immunoglobulin, and 213 (92.6%) received hepatitis B virus vaccine. Prophylactic antiretroviral therapy was offered beginning in 1997. Of the 210 patients who presented thereafter, 82 (39.0%) received chemoprophylaxis, of whom 69 (84.1%) completed a 4-week course of therapy. The use of a protease inhibitor was not associated with a significantly higher risk of adverse effects or early discontinuation of therapy. At 6 months, 189 were tested for HIV, 167 for hepatitis B virus, and 159 for hepatitis C virus. There were no seroconversions. CONCLUSIONS: We observed no seroconversions in 274 pediatric community-acquired needlestick injuries, thereby confirming that the risk of transmission of bloodborne viruses in these events is very low. PMID- 18676536 TI - A critique of criteria for evaluating vaccines for inclusion in mandatory school immunization programs. AB - Several new vaccines for children and young adults have been introduced recently and now appear on the Advisory Committee on Immunization Practices' recommended childhood and adolescent immunization schedule (meningococcal, rotavirus, human papillomavirus). As new vaccines are introduced, states face complex decisions regarding which vaccines to fund and which vaccines to require for school or child care entry. This complexity is evidenced by the current debate surrounding the human papillomavirus vaccine. We present a critique to the approach and criteria for evaluating vaccines for inclusion in mandatory school immunization programs that have been adopted by the Washington State Board of Health by illustrating how these criteria might be applied to the human papillomavirus vaccine. We conclude that these 9 criteria can help ensure a deliberate and informed approach to important public policy decisions, but we argue that several clarifications of the review process are needed along with the addition of a 10th criterion that ensures that a new vaccine mandate relates in some manner to increasing safety in the school environment. PMID- 18676537 TI - Seasonal invasive pneumococcal disease in children: role of preceding respiratory viral infection. AB - OBJECTIVE: Our objective was to demonstrate correlations between invasive pneumococcal disease in children and circulating respiratory viruses. METHODS: This retrospective study included 6 winter respiratory viral seasons (2001-2007) in Intermountain Healthcare, an integrated health system in the Intermountain West, including Primary Children's Medical Center in Salt Lake City, Utah. Children <18 years of age who were hospitalized with either invasive pneumococcal disease in any Intermountain Healthcare facility or culture-confirmed invasive pneumococcal disease at Primary Children's Medical Center were included. We analyzed the correlation between invasive pneumococcal disease and circulating respiratory viruses. RESULTS: A total of 435 children with invasive pneumococcal disease and 203 with culture-confirmed invasive pneumococcal disease were hospitalized in an Intermountain Healthcare facility or Primary Children's Medical Center during the study period. During the same period, 6963 children with respiratory syncytial virus, 1860 with influenza virus, 1459 with parainfluenza virus, and 818 with adenoviruses were evaluated at Primary Children's Medical Center. A total of 253 children with human metapneumovirus were identified during the last 5 months of the study. There were correlations between invasive pneumococcal disease and seasonal respiratory syncytial virus, influenza virus, and human metapneumovirus activity. The correlation with invasive pneumococcal disease was strong up to 4 weeks after respiratory syncytial virus activity. For influenza virus and human metapneumovirus, the correlations were strong at 2 weeks after activity of these viruses. Pneumonia was the most common clinical disease associated with culture-confirmed invasive pneumococcal disease, mostly attributable to serotypes 1, 19A, 3, and 7F. CONCLUSIONS: In the post-pneumococcal conjugate vaccine era, seasonal increases in respiratory syncytial virus, influenza virus, and human metapneumovirus infections in children were associated with increased pediatric admissions with invasive pneumococcal disease, especially pneumonia caused by nonvaccine serotypes. PMID- 18676538 TI - Blood pressure variability and classification of prehypertension and hypertension in adolescence. AB - OBJECTIVE: There is little information in pediatrics on the persistence of the prehypertension and hypertension classifications or on the progression of prehypertension to hypertension. This study aimed to examine those issues. METHODS: An analysis of data from the National Childhood Blood Pressure database was conducted to examine the longitudinal blood pressure outcomes for adolescents classified after a single measurement of blood pressure. Adolescent subjects (N = 8535) for whom serial single blood pressure measurements were obtained at intervals of 2 years were identified. Subjects were stratified according to blood pressure status at the initial measurement, as having normotension, prehypertension, or hypertension. RESULTS: Among subjects designated as having prehypertension (n = 1470), 14% of boys and 12% of girls had hypertension 2 years later. Among subjects designated as having hypertension, 31% of boys and 26% of girls continued to exhibit hypertension, and 47% of boys and 26% of girls had blood pressure values in the prehypertensive range. Regression models showed no significant effect of race on blood pressure changes but significant effects of initial BMI and changes in BMI. CONCLUSIONS: These data indicated that the rate of progression of prehypertension to hypertension was approximately 7% per year. Prehypertension can be predictive of future hypertension and may benefit from preventive interventions, especially lifestyle changes. PMID- 18676539 TI - Pediatric residents' use of jargon during counseling about newborn genetic screening results. AB - OBJECTIVE: The goal was to investigate pediatric residents' usage of jargon during discussions about positive newborn screening test results. METHODS: An explicit-criteria abstraction procedure was used to identify jargon usage and explanations in transcripts of encounters between residents and standardized parents of a fictitious infant found to carry cystic fibrosis or sickle cell hemoglobinopathy. Residents were recruited from a series of educational workshops on how to inform parents about positive newborn screening test results. The time lag from jargon words to explanations was measured by using "statements," each of which contained 1 subject and 1 predicate. RESULTS: Duplicate abstraction revealed reliability kappa of 0.92. The average number of unique jargon words per transcript was 20; the total jargon count was 72.3 words. There was an average of 7.5 jargon explanations per transcript, but the explained/total jargon ratio was only 0.17. When jargon was explained, the average time lag from the first usage to the explanation was 8.2 statements. CONCLUSION: The large number of jargon words and the small number of explanations suggest that physicians' counseling about newborn screening may be too complex for some parents. PMID- 18676540 TI - Effects of the accreditation council for graduate medical education duty hour limits on sleep, work hours, and safety. AB - OBJECTIVE: To mitigate the risks of fatigue-related medical errors, the Accreditation Council for Graduate Medical Education introduced work hour limits for resident physicians in 2003. Our goal was to determine whether work hours, sleep, and safety changed after implementation of the Accreditation Council for Graduate Medical Education standards. METHODS: We conducted a prospective cohort study in which residents from 3 large pediatric training programs provided daily reports of work hours and sleep. In addition, they completed reports of near-miss and actual motor vehicle crashes, occupational exposures, self-reported medical errors, and ratings of educational experience. They were screened for depression and burnout. Concurrently, at 2 of the centers, data on medication errors were collected prospectively by using an established active surveillance method. RESULTS: A total of 220 residents provided 6007 daily reports of their work hours and sleep, and 16 158 medication orders were reviewed. Although scheduling changes were made in each program to accommodate the standards, 24- to 30-hour shifts remained common, and the frequency of residents' call remained largely unchanged. There was no change in residents' measured total work hours or sleep hours. There was no change in the overall rate of medication errors, and there was a borderline increase in the rate of resident physician ordering errors, from 1.06 to 1.38 errors per 100 patient-days. Rates of motor vehicle crashes, occupational exposures, depression, and self-reported medical errors and overall ratings of work and educational experiences did not change. The mean length of extended-duration (on-call) shifts decreased 2.7% to 28.5 hours, and rates of resident burnout decreased significantly (from 75.4% to 57.0%). CONCLUSIONS: Total hours of work and sleep did not change after implementation of the duty hour standards. Although fewer residents were burned out, rates of medication errors, resident depression, and resident injuries and educational ratings did not improve. PMID- 18676541 TI - Clinicians' description of factors influencing their reporting of suspected child abuse: report of the Child Abuse Reporting Experience Study Research Group. AB - OBJECTIVES: Primary care clinicians participating in the Child Abuse Reporting Experience Study did not report all suspected physical child abuse to child protective services. This evaluation of study data seeks (1) to identify factors clinicians weighed when deciding whether to report injuries they suspected might have been caused by child abuse; (2) to describe clinicians' management strategies for children with injuries from suspected child abuse that were not reported; and (3) to describe how clinicians explained not reporting high suspicion injuries. METHODS: From the 434 pediatric primary care clinicians who participated in the Child Abuse Reporting Experience Study and who indicated they had provided care for a child with an injury they perceived as suspicious, a subsample of 75 of 81 clinicians completed a telephone interview. Interviewees included 36 clinicians who suspected child abuse but did not report the injury to child protective services (12 with high suspicion and 24 with some suspicion) and 39 who reported the suspicious injury. Interviews were analyzed for major themes and subthemes, including decision-making regarding reporting of suspected physical child abuse to child protective services and alternative management strategies. RESULTS: Four major themes emerged regarding the clinicians' reporting decisions, that is, familiarity with the family, reference to elements of the case history, use of available resources, and perception of expected outcomes of reporting to child protective services. When they did not report, clinicians planned alternative management strategies, including active or informal case follow-up management. When interviewed, some clinicians modified their original opinion that an injury was likely or very likely caused by abuse, to explain why they did not report to child protective services. CONCLUSIONS: Decisions about reporting to child protective services are guided by injury circumstances and history, knowledge of and experiences with the family, consultation with others, and previous experiences with child protective services. PMID- 18676542 TI - Morbidity of overweight (>or=85th percentile) in the first 2 years of life. AB - OBJECTIVES: Our hypothesis was that morbidity related to overweight/obesity is already evident in infants and young toddlers. The major objectives of this study were (1) to assess the prevalence of overweight in a sample of hospitalized infants and (2) to assess the prevalence of morbidity in overweight infants in a community-based sample. METHODS: The hospital admission study population included 2139 infants, or=85th weight-for height percentile in >or=2 measurements, >or=3 months apart), or=95th percentile had more admissions than expected, as well as a larger number of repeated admissions. In the second part of the study, we found that rates of developmental delays (mainly delayed gross motor skills) and snoring were significantly higher in infants of >or=85th percentile. In addition, although the results were not statistically significant, infants with overweight suffered more frequently from breathing problems, such as asthma and stridor. When the mothers were asked to assess whether their child was overweight, only 31.6% of mothers of overweight children thought that the child was overweight. CONCLUSIONS: The high admission rates for infants of >or=95th percentile and the high incidence rates of respiratory morbidity, snoring, and delayed gross motor skills in overweight infants support our hypothesis regarding early morbidity associated with overweight. PMID- 18676543 TI - Pediatric autoimmune neuropsychiatric disorders associated with streptococcus: comparison of diagnosis and treatment in the community and at a specialty clinic. AB - OBJECTIVES: This study aimed to examine whether pediatric autoimmune neuropsychiatric disorders associated with streptococcus were appropriately diagnosed in the community and to determine subsequent rates of unwarranted use of antibiotic treatment for tics and obsessive-compulsive symptoms without the identification of an infection. METHODS: The design was a retrospective, cross sectional, observational study of 176 children and adolescents who were evaluated in a specialty program for tics, Tourette's disorder, and related problems. Previously published diagnostic criteria were used to establish the diagnosis of pediatric autoimmune neuropsychiatric disorders associated with streptococcus in our clinic. RESULTS: Subjects were significantly less likely to receive a diagnosis of pediatric autoimmune neuropsychiatric disorders associated with streptococcus at the specialty clinic than in the community. In the community, subjects were significantly more likely to be treated with antibiotics or immunosuppressant medication if they received a diagnosis of pediatric autoimmune neuropsychiatric disorders associated with streptococcus. Of the 27 subjects with a community diagnosis of pediatric autoimmune neuropsychiatric disorders associated with streptococcus who were treated with antibiotics, 22 (82%) were treated without laboratory evidence of an infection; 2 were treated with immunomodulatory medications. CONCLUSIONS: Our results support our hypothesis that pediatric autoimmune neuropsychiatric disorders associated with streptococcus are frequently diagnosed in the community without the application of all working diagnostic criteria. This phenomenon has resulted in unwarranted use of antibiotic treatment for tics/obsessive-compulsive disorder without evidence of laboratory infection. PMID- 18676544 TI - Influence of having breakfast on cognitive performance and mood in 13- to 20-year old high school students: results of a crossover trial. AB - OBJECTIVE: The goal was to determine whether breakfast had effects on the cognitive performance and mood of high school students. METHODS: A crossover trial was performed in boarding schools, involving 104 students between 13 and 20 years of age. The participants were randomly assigned to 2 equal-size groups on the morning of the first testing day. One half of the total sample received a standardized breakfast, whereas the other half received no breakfast. Seven days later, the treatment order was reversed. Measurements of cognitive function included standardized tests of attention and concentration, as well as tests of verbal and spatial memory. In addition, mood was rated with a self-administered questionnaire covering the dimensions of positive and negative affect, information uptake, arousal, and alertness. Statistical analysis consisted of repeated-measures analysis of variance. RESULTS: Breakfast had no effect on sustained attention among high school students. Visuospatial memory was improved in male students. Self-reported alertness improved significantly in the entire study population. Male students reported feeling more positive after consuming breakfast, compared with the fasting condition. CONCLUSIONS: This crossover trial demonstrated positive short-term effects of breakfast on cognitive functioning and self-reported alertness in high school students. PMID- 18676545 TI - Impact of cesarean section on neonatal mortality rates among very preterm infants in the United States, 2000-2003. AB - OBJECTIVE: The objective of this analysis was to compare the neonatal mortality rates for infants delivered through primary cesarean section versus vaginal delivery, taking into consideration a number of potentially risk-modifying conditions. METHODS: US linked birth and infant death certificate files for 2000 2003 were used. Demographic, medical, and labor and delivery complications were abstracted from the files with infant information. The primary outcome examined was neonatal death (death at 0-27 days of age). Because of concern regarding misclassification of gestational age, a procedure was used to trim away births for which the birth weight for a specific gestational age was incongruous. Adjusted odds ratios were calculated for the risk of neonatal death relative to the mode of delivery (primary cesarean section versus vaginal delivery), using logistic regression analysis. RESULTS: There were data for 13,733 neonatal deaths and 106,809 survivors available from the trimmed data set for analysis for the 4 year period. More than 80% of pregnancies with delivery between 22 and 31 weeks of gestation experienced >or=1 risk factor. Adjusted odds ratios demonstrated significantly reduced risk of neonatal death for infants delivered through cesarean section at 22 to 25 weeks of gestation (adjusted odds ratios of 0.58, 0.52, 0.72, and 0.81 for 22, 23, 24, and 25 weeks, respectively). CONCLUSION: Cesarean section does seem to provide survival advantages for the most immature infants delivered at 22 to 25 weeks of gestation, independent of maternal risk factors for cesarean section. PMID- 18676546 TI - Supplemental oxygen compromises the use of pulse oximetry for detection of apnea and hypoventilation during sedation in simulated pediatric patients. AB - OBJECTIVE: The goal was to assess the time to recognition of apnea in a simulated pediatric sedation scenario, with and without supplemental oxygen. METHODS: A pediatric human patient simulator mannequin was used to simulate apnea in a 6 year-old patient who received sedation for resetting of a fractured leg. Thirty pediatricians participating in a credentialing course for sedation were randomly assigned to 2 groups. Those in group 1 (N = 15) used supplemental oxygen, and those in group 2 (N = 15) did not use supplemental oxygen. A third group (N = 10), consisting of anesthesiology residents (postgraduate years 2 and 3 equivalent), performed the scenario with oxygen supplementation, to ensure validity and reliability of the simulation. The time interval from simulated apnea to bag-mask ventilation was recorded. Oxygen saturation and Paco(2) values were recorded. All recorded variables and measurements were compared between the groups. RESULTS: The time interval for bag-mask ventilation to occur in group 1 (oxygen supplementation) was significantly longer than that in group 2 (without oxygen supplementation) (173 +/- 130 and 83 +/- 42 seconds, respectively). The time interval for bag-mask ventilation to occur was shorter in group 3 (anesthesiology residents) (24 +/- 6 seconds). Paco(2) reached a higher level in group 1 (75 +/- 26 mmHg), compared with groups 2 and 3 (48 +/- 10 and 42 +/- 3 mmHg, respectively). There was no significant difference between the groups in oxygen saturation values at the time of clinical detection of apnea (93 +/- 5%, 88 +/- 5%, and 94 +/- 7%, respectively). CONCLUSIONS: Hypoventilation and apnea are detected more quickly when patients undergoing sedation breathe only air. Supplemental oxygen not only does not prevent oxygen desaturation but also delays the recognition of apnea. PMID- 18676548 TI - Exposure of US adolescents to extremely violent movies. AB - OBJECTIVE: Despite concerns about exposure to violent media, there are few data on youth exposure to violent movies. In this study we examined such exposure among young US adolescents. METHODS: We used a random-digit-dial survey of 6522 US adolescents aged 10 to 14 years fielded in 2003. Using previously validated methods, we determined the percentage and number of US adolescents who had seen each of 534 recently released movies. We report results for the 40 that were rated R for violence by the Motion Picture Association of America, UK 18 by the British Board of Film Classification and coded for extreme violence by trained content coders. RESULTS: The 40 violent movies were seen by a median of 12.5% of an estimated 22 million US adolescents aged 10 to 14 years. The most popular violent movie, Scary Movie, was seen by >10 million (48.1%) children, 1 million of whom were 10 years of age. Watching extremely violent movies was associated with being male, older, nonwhite, having less-educated parents, and doing poorly in school. Black male adolescents were at particularly high risk for seeing these movies; for example Blade, Training Day, and Scary Movie were seen, respectively, by 37.4%, 27.3%, and 48.1% of the sample overall versus 82.0%, 81.0%, and 80.8% of black male adolescents. Violent movie exposure was also associated with measures of media parenting, with high-exposure adolescents being significantly more likely to have a television in their bedroom and to report that their parents allowed them to watch R-rated movies. CONCLUSIONS: This study documents widespread exposure of young US adolescents to movies with extreme graphic violence from movies rated R for violence and raises important questions about the effectiveness of the current movie-rating system. PMID- 18676547 TI - Recurrent postnatal infections are associated with progressive white matter injury in premature infants. AB - OBJECTIVE: Our objective was to identify clinical predictors of progressive white matter injury. METHODS: We evaluated 133 infants of <34 weeks of gestation at birth from 2 university hospitals. Infants underwent MRI twice, initially when in stable condition for transport and again at term-equivalent age or before transfer or discharge. Two neuroradiologists who were blinded to the clinical course graded MRI white matter injury severity by using a validated scale. Potential risk factors were extracted from medical charts. RESULTS: Twelve neonates (9.0%) had progressive white matter injury. In the unadjusted analysis of 10 newborns without Candida meningoencephalitis, recurrent culture-positive postnatal infection and chronic lung disease were associated with progressive white matter injury. Exposure to multiple episodes of culture-positive infection significantly increased the risk of progressive white matter injury. Of the 11 neonates with >1 infection, 36.4% (4 infants) had progressive injury, compared with 5.0% (6 infants) of those with 10 days, and 6 with bronchopulmonary dysplasia. Placental growth factor concentrations in tracheal aspirate fluid were measured in 70 samples from 20 preterm infants during the first postnatal week. RESULTS: In immunohistochemical analyses, placental growth factor staining was seen in bronchial epithelium and macrophages in all groups. Distal airway epithelium positivity was observed mostly in fetuses and in preterm infants who died soon after birth. Vascular endothelial growth factor receptor-2 staining was seen in vascular endothelium in all groups and also in lymphatic endothelium in fetuses. Vascular endothelial growth factor receptor-2 staining in arterial endothelium was associated with higher and staining in venous endothelium with lower gestational age. In capillaries, less vascular endothelial growth factor receptor 2 staining was seen in bronchopulmonary dysplasia. The mean placental growth factor protein concentration in tracheal aspirate fluid during the first postnatal week was 0.64 +/- 0.42 pg/mL per IgA-secretory component unit. Concentrations during the first postnatal week were stable. Lower placental growth factor concentrations correlated with chorioamnionitis and lactosyl ceramide positivity. CONCLUSIONS: The vascular endothelial growth factor receptor 2 staining pattern seems to reflect ongoing differentiation and activity of different endothelia. Lower vascular endothelial growth factor receptor-2 expression in capillary endothelium in bronchopulmonary dysplasia might be a reflection of the dysregulation of vascular development that is characteristic of bronchopulmonary dysplasia. PMID- 18676551 TI - CD4+/CD8+ T cell ratio for diagnosis of HIV-1 infection in infants: Women and Infants Transmission Study. AB - OBJECTIVE: In this study, we tested the hypothesis that the CD4(+)/CD8(+) T cell ratio could predict HIV infection status in HIV-exposed infants. METHODS: CD4(+)/CD8(+) T cell ratios were determined from data for live-born singleton infants who had been prospectively enrolled in the Women and Infants Transmission Study. Data for 2208 infants with known HIV infection status (179 HIV-infected and 2029 uninfected infants) were analyzed. RESULTS: Receiver operating characteristic curves indicated that the CD4(+)/CD8(+) T cell ratio performed better than the proportion of CD4(+) T cells for diagnosis of HIV infection as early as 2 months of age, and this relationship was unaffected by adjustment for maternal race/ethnicity, infant birth weight, gestational age, and gender. At 4 months of age, 90% specificity for HIV diagnosis was associated with 60% sensitivity. For ease of use, graphical estimates based on cubic splines for the time-dependent parameters in a Box-Cox transformation (L), the median (M), and the coefficient of variation (S) were used to create LMS centile curves to show the sensitivity and specificity of CD4(+)/CD8(+) T cell ratios in HIV-infected and uninfected infants until 12 months of age. At 6 months of age, a simplified equation that incorporated sequential CD4(+)/CD8(+) T cell ratios and hematocrit values resulted in improved receiver operating characteristic curves, with 94% positive predictive value and 98% negative predictive value. The positive and negative predictive values remained above 90% in simulated infant populations over a wide range of HIV infection prevalence values. CONCLUSIONS: In the absence of virological diagnosis, a presumptive diagnosis of HIV infection status can be made on the basis of CD4(+)/CD8(+) T cell ratios in HIV-1-exposed infants after 2 months of age; sensitivity and specificity can be improved at 6 months by using a discriminant analysis equation. PMID- 18676553 TI - Who is at risk for special health care needs: findings from the National Survey of Children's Health. AB - OBJECTIVE: A conceptual model of risk factors for special health care needs in childhood was presented previously. This article uses that conceptual model to identify candidate variables for an exploratory empirical examination of the effects of factors that may increase or decrease the risk of developing a special health care need. METHODS: The National Survey of Children's Health was used for our analysis (N = 102 353). We used multilevel and multivariate analysis methods. We examined risk factors for special health care needs generally and for specific physical, developmental, behavioral, and emotional conditions cooccurring with special health care needs. Risk factors were grouped into 6 major domains, namely, predisposing characteristics, genetic endowment, physical environment, social environment, health-influencing behavior, and health care system characteristics. We examined preschool-aged and school-aged children separately. RESULTS: Significant associations were found in 5 of 6 domains studied (no variables in the health care systems characteristics were significant). Individual variables found to decrease or to increase significantly the odds of experiencing special health care needs were expressed at the child level (eg, age and gender), family level (eg, family structure and family conflict), and neighborhood level (eg, perception of supportiveness of the neighborhood). CONCLUSIONS: This analysis is the first to consider empirically a range of risk factors for special health care needs, using a population health model. Although provisional, the results of our analysis can help us to begin thinking about which characteristics of the child, family, and community are worthy of further exploration. Some of the variables we found to be significantly associated with special health care needs, such as age and ethnicity, are immutable. However, we found a number of significant correlates (ie, possible risk factors) that may be amenable to public health interventions, including breastfeeding practices, exposure to secondhand smoke, family closeness, and neighborhood cohesion. PMID- 18676554 TI - Growth characteristics of infantile hemangiomas: implications for management. AB - OBJECTIVES: Infantile hemangiomas often are inapparent at birth and have a period of rapid growth during early infancy followed by gradual involution. More precise information on growth could help predict short-term outcomes and make decisions about when referral or intervention, if needed, should be initiated. The objective of this study was to describe growth characteristics of infantile hemangioma and compare growth with infantile hemangioma referral patterns. METHODS: A prospective cohort study involving 7 tertiary care pediatric dermatology practices was conducted. Growth data were available for a subset of 526 infantile hemangiomas in 433 patients from a cohort study of 1096 children. Inclusion criteria were age younger than 18 months at time of enrollment and presence of at least 1 infantile hemangioma. Growth stage and rate were compared with clinical characteristics and timing of referrals. RESULTS: Eighty percent of hemangioma size was reached during the early proliferative stage at a mean age of 3 months. Differences in growth between hemangioma subtypes included that deep hemangiomas tend to grow later and longer than superficial hemangiomas and that segmental hemangiomas tended to exhibit more continued growth after 3 months of age. The mean age of first visit was 5 months. Factors that predicted need for follow-up included ongoing proliferation, larger size, deep component, and segmental and indeterminate morphologic subtypes. CONCLUSIONS: Most infantile hemangioma growth occurs before 5 months, yet 5 months was also the mean age at first visit to a specialist. Recognition of growth characteristics and factors that predict the need for follow-up could help aid in clinical decision-making. The first few weeks to months of life are a critical time in hemangioma growth. Infants with hemangiomas need close observation during this period, and those who need specialty care should be referred and seen as early as possible within this critical growth period. PMID- 18676555 TI - Does connection to primary care matter for children with attention deficit/hyperactivity disorder? AB - OBJECTIVE: Whether high-quality primary care in the form of a medical home effectively meets the health care needs of children with attention deficit/hyperactivity disorder is unknown. The objectives of this study were to (1) describe the percentage who report unmet health care need, (2) evaluate whether having a medical home is associated with lower risk for having unmet needs, and (3) compare the impact of having a medical home on unmet need for children with attention-deficit/hyperactivity disorder with those with asthma. METHODS: Cross-sectional analysis was conducted of the National Survey of Children's Health, 2003, a nationally representative sample. The primary outcome variable was parent-reported unmet health care need. Multivariate logistic regression tested the impact of having a medical home on unmet needs for children with attention-deficit/hyperactivity disorder and asthma. RESULTS: The National Survey of Children's Health interviewed parents of 6030 children who had attention-deficit/hyperactivity disorder and 6133 children who had asthma and were between the ages of 6 and 17 years. A total of 16.8% of children with attention-deficit/hyperactivity disorder had at least 1 unmet need compared with 6.7% of children with asthma. Although the proportion of children with a medical home was comparable, children with attention-deficit/hyperactivity disorder were 3.5 times more likely to have an unmet need than were children with asthma. Children with asthma who have a medical home have less than half the likelihood of reporting an unmet need in comparison with those without a medical home; however, among children with attention-deficit/hyperactivity disorder, having a medical home was not associated with decreased likelihood of reporting an unmet need. CONCLUSIONS: Having a medical home is not associated with fewer unmet needs for children with attention-deficit/hyperactivity disorder. Our results suggest that high-quality primary care may not be as successful at meeting the needs of children with behavioral health conditions compared with those with physical conditions. PMID- 18676557 TI - A phase I/II trial of high-dose erythropoietin in extremely low birth weight infants: pharmacokinetics and safety. AB - OBJECTIVES: High-dose recombinant erythropoietin is neuroprotective in animal models of neonatal brain injury. Extremely low birth weight infants are at high risk for brain injury and neurodevelopmental problems and might benefit from recombinant erythropoietin. We designed a phase I/II trial to test the safety and determine the pharmacokinetics of high-dose recombinant erythropoietin in extremely low birth weight infants. METHODS: In a prospective, dose-escalation, open-label trial, we compared 30 infants who were treated with high-dose recombinant erythropoietin with 30 concurrent control subjects. Eligible infants were <24 hours old, A (Leiden) were completely concordant by snapback genotyping. We distinguished 7 genotypes in 2 regions of CFTR exon 10 with symmetric PCR using 2 snapback primers followed by product dilution to favor intramolecular hybridization. CONCLUSIONS: Snapback primer genotyping with saturating dyes provides the specificity of a probe with only 2 primers that are free of special covalent labels in a closed-tube system. PMID- 18676585 TI - Clinical evaluation of bionime rightest GM310 biosensors with a simplified electrode fabrication for alternative-site blood glucose tests. AB - BACKGROUND: Most processes for fabricating biosensors applied to screen-printed carbon electrodes (SPCEs) are complex. This study presents a novel one-step process for manufacturing electrodes for injection-molding biosensors. METHODS: During the sensor-fabrication process, barrel-plated gold electrodes were inserted into an injection-molded base. The electrode directly touched the electrical contact of a meter. We analyzed technical measurements for this biosensor, including tests of the measurement range, within-run imprecision, and between-meter imprecision. In clinical trials, experienced technicians tested 3 alternative sites (fingertip, palm, and arm). The results were simultaneously compared with plasma values obtained with the hexokinase method on the Olympus AU640 instrument. Analytical results were evaluated according to International Standards Organization 15197 (ISO 15197:2003) criteria and by Clarke error grid analysis (EGA), and CVs were calculated to evaluate within-run imprecision. RESULTS: The glucose measurement range was 0.6- 33.3 mmol/L (y = 0.96x + 0.07 mmol/L; r(2) = 0.9977). The CVs in the within-run imprecision test were 1.7% 3.5%, and the overall CV was 2.1%, indicating good reproducibility of results. The Student t-tests of mean values from 5 meters revealed statistically insignificant differences (P > 0.05). In clinical trials, the agreement of the Rightest GM310 meter results with those of a laboratory method complied with ISO 15197:2003 criteria. In the EGA, 100% of the values were within the acceptable zones (A + B), and the proportion of values within zone A exceeded 95%. CONCLUSIONS: The Bionime Rightest GM310 meter applied a simplified process for biosensor fabrication and displayed acceptable performance for monitoring glucose concentrations at alternative test sites. PMID- 18676586 TI - Key issues in the developing synergism between cardiovascular imaging and biomarkers. AB - BACKGROUND: Sophisticated methods of cardiac imaging have the potential to revolutionize the care of patients with cardiovascular disease. The benefits of these state-of-the art imaging techniques can be enhanced by their use in combination with new cardiac biomarkers. This review addresses potentially useful interactions between imaging and biomarkers. CONTENT: Areas were defined in which the combined use of novel imaging techniques and biomarkers would be most beneficial. This review addresses multiple cardiovascular conditions for which the useful aspects of imaging and biomarkers are likely to be positively synergistic, including acute and chronic ischemic heart disease, heart failure, myocarditis, hypertension, and atherosclerosis. CONCLUSIONS: The synergistic use of imaging techniques and biomarkers will enhance the investigation of many key issues and questions and will be an important resource in the future. PMID- 18676587 TI - New automated multiplex assay for bone turnover markers in osteoporosis. AB - BACKGROUND: Serum C-terminal cross-linked telopeptide of type I collagen (CTX-I), N-terminal propeptide of type I collagen (PINP), and osteocalcin (OC) are among the most sensitive bone turnover markers for evaluating osteoporosis. Each marker is currently measured individually by manual or automated immunoassays that are time consuming and require substantial sample volume. We evaluated the performance characteristics of a novel, fully automated, protein-array chip system that allows the simultaneous measurement of CTX-I, PINP, OC, and intact parathyroid hormone (PTH) in 20 microL of serum. METHODS: We measured CTX-I, PINP, OC, and PTH using multiplex and corresponding automated single assays in 157 healthy premenopausal women, 74 healthy men, and 56 postmenopausal osteoporotic women before and 6 months after treatment with oral ibandronate (150 mg/month). RESULTS: Within- and between-run CVs of the multiplex assay were similar to those of single measurement assays (<10% for all markers), whereas the limit of quantification was lower, except for OC. Multiplex values highly correlated (r > 0.93, P < 0.0001 for all markers) with the corresponding single assays, and measured concentrations were comparable. After 6 months of ibandronate, CTX-I, PINP, and OC decreased by a median of 48%, 63%, and 52%, respectively (P < 0.0001 for all 3 markers), magnitudes similar to those of the corresponding single assays. CONCLUSIONS: The automated protein-array chip demonstrated similar analytical precision, improved analytical sensitivity, and comparable measured concentrations to those of single assays. The multiplex assay should be useful for assessing bone metabolism in large clinical studies, particularly when sample volume is limited. PMID- 18676588 TI - Use of colorimetric test strips for monitoring the effect of hemodialysis on salivary nitrite and uric acid in patients with end-stage renal disease: a proof of principle. AB - BACKGROUND: Initial screening of potential biomarkers for monitoring dialysis was performed with saliva samples collected from patients with end-stage renal disease (ESRD). A more thorough analysis of the most promising markers identified in the initial screening was conducted with saliva samples acquired at hourly intervals throughout dialysis to monitor analyte concentrations as dialysis progressed. We observed that salivary nitrite (NO(2)(-)) and uric acid (UA) concentrations consistently decreased as dialysis proceeded. METHODS: Solution based colorimetric-detection chemistries for NO(2)(-) and UA were converted to a test strip format to produce a simple method for semiquantitatively measuring NO(2)(-) and UA concentrations in the clinic or at the patient's home. We assessed the test strips with saliva samples collected from both ESRD patients undergoing dialysis and healthy control volunteers to qualitatively monitor the effect of dialysis on salivary NO(2)(-) and UA. We used computer software to analyze digital images of the resulting test strip color intensities. RESULTS: Test strip measurements showed that mean salivary concentrations of NO(2)(-) and UA were decreased in ESRD patients by 86% and 39%, respectively, compared with 15% and 9% for time-matched controls. Comparison of test strip results with calibrated solution-based assays suggests that the test strips can semiquantitatively measure salivary concentrations of NO(2)(-) and UA. CONCLUSIONS: The colorimetric test strips monitored changes in salivary NO(2)(-) and UA concentrations that occurred in ESRD patients during dialysis. The test strips may prove useful for noninvasively evaluating dialysis progress and may also be useful for monitoring renal disease status. PMID- 18676589 TI - Parental smoking and adolescent problem behavior: an adoption study of general and specific effects. AB - OBJECTIVE: It is essential to understand the effect of parental smoking on offspring tobacco use. In biologically related families, parents who smoke may transmit a nonspecific genetic risk for offspring disinhibited behavior, including tobacco use. Studying adoptive families allows one to control for genetic confounding when examining the environmental effect of exposure to parental smoking. The purpose of this study was to examine the genetic and environmental contributions to the risk represented by exposure to parental smoking and to assess the specificity of that risk. METHODS: Adolescents adopted in infancy were systematically ascertained from records of three private Minnesota adoption agencies; nonadopted adolescents were ascertained from Minnesota birth records. Adolescents and their rearing parents participated in all assessments in person. The main outcome measures were self-reports of behavioral deviance, substance use, and personality, as well as DSM-IV clinical assessments of childhood disruptive disorders. RESULTS: The data from adoptive families suggest that exposure to parental smoking represents an environmental risk for substance use in adolescent offspring. In biologically related families, the effect of exposure to parental smoking is larger and more diverse, including substance use, disruptive behavior disorders, delinquency, deviant peer affiliations, aggressive attitudes, and preference for risk taking. CONCLUSIONS: This study provides evidence for an environmentally mediated pathway by which parental smoking increases risk specifically for substance use in adolescent offspring. The data are also consistent with a genetically mediated pathway by which nonadoptive parents who smoke may also transmit a nonspecific genetic risk to their offspring for disinhibited behavior. PMID- 18676590 TI - Overcrowding in hospital wards as a predictor of antidepressant treatment among hospital staff. AB - OBJECTIVE: This report assessed whether hospital ward overcrowding predicts antidepressant use among hospital staff. METHOD: The extent of hospital ward overcrowding was determined using administrative records of monthly bed occupancy rates between 2000 and 2004 in 203 somatic illness wards in 16 Finnish hospitals providing specialized health care. Information on job contracts for personnel was obtained from the employers' registers. Comprehensive daily data on purchased antidepressant prescriptions (World Health Organization's Anatomical Therapeutic Chemical classification code N06A) for nurses (N=6,699) and physicians (N=641) was derived from national registers. Cox proportional hazards models were used to examine the association between bed occupancy rate and subsequent antidepressant treatment. Monthly bed occupancy rates were used as a time-dependent exposure that could change in value over the course of observation. Hazard ratios were adjusted for sex, age, occupation, type and length of employment contract, hospital district, specialty, and calendar year. RESULTS: Exposure over 6 months to an average bed occupancy rate over 10% in excess of the recommended limit was associated with new antidepressant treatment. This association followed a dose response pattern, with increasing bed occupancy associated with an increasing likelihood of antidepressant use. There was no evidence of reverse causality; antidepressant treatment among employees did not predict subsequent excess bed occupancy. CONCLUSIONS: The increased risk of antidepressant use observed in this study suggests that overcrowding in hospital wards may have an adverse effect on the mental health of staff. PMID- 18676591 TI - Mentalization: ontogeny, assessment, and application in the treatment of borderline personality disorder. AB - This article aims to review the development of the concept of mentalization, its applications in the understanding and treatment of borderline personality disorder, and the issue of its assessment. While conceptually derivative of theory of mind, Fonagy's concept of mentalization concerns more affectively and interpersonally complex understandings of oneself and others, reflecting abilities that enable an individual not only to navigate the social world effectively but also to develop an enriched, stable sense of self. The components of mentalization can be organized around self-/other-oriented, implicit/explicit, and cognitive/affective dimensions. Concepts of mindfulness, psychological mindedness, empathy, and affect consciousness are shown to partially overlap with mentalization within these three dimensions. Mentalization is assessed by the measure of reflective function, a scale to be used adjunctively on semistructured narrative interviews such as the Adult Attachment Interview. Its validity has not been fully tested, and its usage has been hampered by the time and expense it requires. Although the concept of mentalization is a useful heuristic that enables clinicians to adopt a coherent treatment approach, it may be too broad and multifaceted to be operationalized as a marker for specific forms of psychopathology such as borderline personality disorder. Research elucidating the relationship between reflective function, overlapping concepts, and features of borderline psychopathology is needed. PMID- 18676592 TI - The posttraumatic stress disorder diagnosis in preschool- and elementary school age children exposed to motor vehicle accidents. AB - OBJECTIVE: Increasingly, children are being diagnosed with psychiatric disorders, including preschool-age children. These diagnoses in young children raise questions pertaining to 1) how diagnostic algorithms for individual disorders should be modified for young age groups, 2) how psychopathology is best detected at an early stage, and 3) how to make use of multiple informants. The authors examined these issues in a prospective longitudinal assessment of preschool- and elementary school-age children who were exposed to a traumatic event. METHOD: Participants were 114 children (age range: 2-10 years) who had experienced a motor vehicle accident. Parents and older children (age range: 7-10 years) completed structured interviews 2-4 weeks (initial assessment) and 6 months (6 month follow-up) after the traumatic event. A recently proposed alternative symptom algorithm for diagnosing posttraumatic stress disorder (PTSD) was utilized and compared with the standard DSM-IV algorithms for diagnosing PTSD and acute stress disorder. RESULTS: At the 2- to 4-week assessment, 11.5% of the children met conditions for a diagnosis of PTSD based on the alternative algorithm criteria per parent report, and 13.9% met criteria for this diagnosis at the 6-month follow-up. These percentages were much higher than those for DSM IV diagnoses of acute stress disorder and PTSD. Among 7- to 10-year-old subjects, the use of combined parent- and child-reported symptoms to derive a diagnosis resulted in an increased number of children in this age group who were identified with psychiatric illness relative to the use of parent report alone. Agreement between parent and child on symptoms for 1) a diagnosis of PTSD based on the alternative algorithm criteria and 2) diagnoses of DSM-IV acute stress disorder and PTSD in this age group was poor. Among 2- to 6-year-old subjects, the alternative algorithm PTSD diagnosis per parent report was a more sensitive predictor of later onset psychopathology relative to a diagnosis of DSM-IV acute stress disorder or PTSD per parent report. However, among 7- to 10-year-old subjects, a combined symptom report (from both parent and child) was optimal in predicting posttraumatic psychopathology. CONCLUSIONS: These findings support the use of the proposed alternative algorithm for assessing PTSD in young children and suggest that the diagnosis of PTSD based on the alternative algorithm criteria is stable from the acute phase onward. When both parent- and child reported symptoms are utilized for the assessment of PTSD among 7- to 10-year-old children, the alternative algorithm and DSM-IV criteria have broad comparable validity. However, in the absence of child-reported symptoms, the alternative algorithm criteria per parent report appears to be an optimal diagnostic measure of PTSD among children in this age group, relative to the standard DSM-IV algorithm for diagnosing the disorder. PMID- 18676593 TI - Cholinergic agonists as novel treatments for schizophrenia: the promise of rational drug development for psychiatry. PMID- 18676594 TI - Neural networks in schizophrenia. PMID- 18676595 TI - New insights into clinical response in schizophrenia: from dopamine D2 receptor occupancy to patients' quality of life. PMID- 18676596 TI - Divergent approaches converge on frontal lobe dysfunction in schizophrenia. PMID- 18676597 TI - Accelerating new knowledge in schizophrenia. PMID- 18676598 TI - Clinically responsible genetic testing in neuropsychiatric patients: a bridge too far and too soon. PMID- 18676599 TI - Eight ball. PMID- 18676600 TI - Asperger's syndrome: diagnosis and treatment. PMID- 18676601 TI - Schizophrenia in a patient with spinocerebellar ataxia 2: coincidence of two disorders or a neurodegenerative disease presenting with psychosis? PMID- 18676602 TI - Franco Basaglia, 1924-1980. PMID- 18676603 TI - Antidepressant prescribing and rapid cycling. PMID- 18676604 TI - Mixed depression and rapid cycling. PMID- 18676605 TI - Maimonides and depression. PMID- 18676606 TI - D-cycloserine augmentation for behavioral therapy. PMID- 18676607 TI - Father-to-son transmission of 6;17 translocation in Tourette's syndrome. PMID- 18676608 TI - The use of iconic symbols to represent our experience. PMID- 18676609 TI - Takotsubo cardiomyopathy in a patient with emotional stress. PMID- 18676612 TI - Novel role of calpain-3 in the triad-associated protein complex regulating calcium release in skeletal muscle. AB - Calpain-3 (CAPN3) is a non-lysosomal cysteine protease that is necessary for normal muscle function, as mutations in CAPN3 result in an autosomal recessive form of limb girdle muscular dystrophy type 2A. To elucidate the biological roles of CAPN3 in skeletal muscle, we performed a search for potential substrates and interacting partners. By yeast-two-hybrid analysis we identified the glycolytic enzyme aldolase A (AldoA) as a binding partner of CAPN3. In co-expression studies CAPN3 degraded AldoA; however, no accumulation of AldoA was observed in total extracts from CAPN3-deficient muscles suggesting that AldoA is not an in vivo substrate of CAPN3. Instead, we found CAPN3 to be necessary for recruitment of AldoA to one specific location, namely the triads, which are structural components of muscle responsible for calcium transport and excitation-contraction coupling. Both aldolase and CAPN3 are present in the triad-enriched fraction and are able to interact with ryanodine receptors (RyR) that form major calcium release channels. Levels of triad-associated AldoA and RyR were decreased in CAPN3-deficient muscles compared with wild-type. Consistent with these observations we found calcium release to be significantly reduced in fibers from CAPN3-deficient muscles. Together, these data suggest that CAPN3 is necessary for the structural integrity of the triad-associated protein complex and that impairment of calcium transport is a phenotypic feature of CAPN3-deficient muscle. PMID- 18676613 TI - The intracellular region of Notch ligands Dll1 and Dll3 regulates their trafficking and signaling activity. AB - Genetic studies have shown that ubiquitination and endocytosis of the Drosophila ligand Delta in signal-sending cells are required for activation of Notch signaling, but how these events promote Notch activation remains poorly understood. Here, we show that an ubiquitination-defective mutant of the murine Delta-homologue Dll1 is endocytosed but, in contrast to the wild-type Dll1, is unable to subsequently recycle back to the cell surface or to bind Notch1 efficiently. These results demonstrate that ubiquitination, although not required for endocytosis, is essential for Dll1 recycling and that recycling is required to acquire affinity for the receptor. On the other hand, a chimeric molecule encompassing the extracellular domain of Dll1 and the transmembrane/intracellular domain of Dll3, which contains no lysine, is endocytosed, recycled, and interacts with Notch1 but is unable to induce transendocytosis of the extracellular region of Notch1 or to signal. These observations suggest that the chimera uses an ubiquitination-independent signal to recycle, allowing it to acquire affinity for Notch1. Our results support the idea that ligand recycling determines its competence to bind efficiently to the receptor but that this is insufficient to allow it to perform transendocytosis, an event required for activation of Notch signaling. Finally, the present study indicates that Dll1 partially localizes to lipid microdomains, whereas both ubiquitination-defective Dll1 and the Dll1-3 chimera are excluded from these compartments, suggesting that these microdomains provide the environment necessary for Dll1 to activate Notch signaling. PMID- 18676614 TI - Nonerythropoietic, tissue-protective peptides derived from the tertiary structure of erythropoietin. AB - Erythropoietin (EPO), a member of the type 1 cytokine superfamily, plays a critical hormonal role regulating erythrocyte production as well as a paracrine/autocrine role in which locally produced EPO protects a wide variety of tissues from diverse injuries. Significantly, these functions are mediated by distinct receptors: hematopoiesis via the EPO receptor homodimer and tissue protection via a heterocomplex composed of the EPO receptor and CD131, the beta common receptor. In the present work, we have delimited tissue-protective domains within EPO to short peptide sequences. We demonstrate that helix B (amino acid residues 58-82) of EPO, which faces the aqueous medium when EPO is bound to the receptor homodimer, is both neuroprotective in vitro and tissue protective in vivo in a variety of models, including ischemic stroke, diabetes-induced retinal edema, and peripheral nerve trauma. Remarkably, an 11-aa peptide composed of adjacent amino acids forming the aqueous face of helix B is also tissue protective, as confirmed by its therapeutic benefit in models of ischemic stroke and renal ischemia-reperfusion. Further, this peptide simulating the aqueous surface of helix B also exhibits EPO's trophic effects by accelerating wound healing and augmenting cognitive function in rodents. As anticipated, neither helix B nor the 11-aa peptide is erythropoietic in vitro or in vivo. Thus, the tissue-protective activities of EPO are mimicked by small, nonerythropoietic peptides that simulate a portion of EPO's three-dimensional structure. PMID- 18676615 TI - Orientation dependence in fluorescent energy transfer between Cy3 and Cy5 terminally attached to double-stranded nucleic acids. AB - We have found that the efficiency of fluorescence resonance energy transfer between Cy3 and Cy5 terminally attached to the 5' ends of a DNA duplex is significantly affected by the relative orientation of the two fluorophores. The cyanine fluorophores are predominantly stacked on the ends of the helix in the manner of an additional base pair, and thus their relative orientation depends on the length of the helix. Observed fluorescence resonance energy transfer (FRET) efficiency depends on the length of the helix, as well as its helical periodicity. By changing the helical geometry from B form double-stranded DNA to A form hybrid RNA/DNA, a marked phase shift occurs in the modulation of FRET efficiency with helix length. Both curves are well explained by the standard geometry of B and A form helices. The observed modulation for both polymers is less than that calculated for a fully rigid attachment of the fluorophores. However, a model involving lateral mobility of the fluorophores on the ends of the helix explains the observed experimental data. This has been further modified to take account of a minor fraction of unstacked fluorophore observed by fluorescent lifetime measurements. Our data unequivocally establish that Forster transfer obeys the orientation dependence as expected for a dipole-dipole interaction. PMID- 18676616 TI - Potent activation of FGF-2 IRES-dependent mechanism of translation during brain development. AB - Fibroblast growth factor-2 (FGF-2) plays a fundamental role in brain functions. This role may be partly achieved through the control of its expression at the translational level via an internal ribosome entry site (IRES)-dependent mechanism. Transgenic mice expressing a bicistronic mRNA allowed us to study in vivo and ex vivo where this translational mechanism operates. Along brain development, we identified a stringent spatiotemporal regulation of FGF-2 IRES activity showing a peak at post-natal day 7 in most brain regions, which is concomitant with neuronal maturation. At adult age, this activity remained relatively high in forebrain regions. By the enrichment of this activity in forebrain synaptoneurosomes and by the use of primary cultures of cortical neurons or cocultures with astrocytes, we showed that this activity is indeed localized in neurons, is dependent on their maturation, and correlates with endogenous FGF-2 protein expression. In addition, this activity was regulated by astrocyte factors, including FGF-2, and spontaneous electrical activity. Thus, neuronal IRES-driven translation of the FGF-2 mRNA may be involved in synapse formation and maturation. PMID- 18676617 TI - Upf1 potentially serves as a RING-related E3 ubiquitin ligase via its association with Upf3 in yeast. AB - Three Upf proteins are essential to the nonsense-mediated mRNA decay (NMD) pathway. Although these proteins assemble on polysomes for recognition of aberrant mRNAs containing premature termination codons, the significance of this assembly remains to be elucidated. The Cys- and His-rich repeated N terminus (CH domain) of Upf1 has been implicated in its binding to Upf2. Here, we show that CH domain also plays a RING-related role for Upf1 to exhibit E3 ubiquitin ligase activity in yeast. Despite the sequence divergence from typical E3-RING fingers, the CH domain of yeast Upf1 specifically and directly interacted with the yeast E2 Ubc3. Interestingly, Upf1 served as a substrate for the in vitro self ubiquitination, and the modification required its association with Upf3 rather than Upf2. Substitution of the coordinated Cys and His residues in the CH domain impaired not only self-ubiquitination of Upf1 but also rapid decay of aberrant mRNAs. These results suggest that Upf1 may serve as an E3 ubiquitin ligase upon its association with Upf3 and play an important role in signaling to the NMD pathway. PMID- 18676618 TI - Group II introns in eubacteria and archaea: ORF-less introns and new varieties. AB - Group II introns are a major class of ribozymes found in bacteria, mitochondria, and plastids. Many introns contain reverse transcriptase open reading frames (ORFs) that confer mobility to the introns and allow them to persist as selfish DNAs. Here, we report an updated compilation of group II introns in Eubacteria and Archaea comprising 234 introns. One new phylogenetic class is identified, as well as several specialized lineages. In addition, we undertake a detailed search for ORF-less group II introns in bacterial genomes in order to find undiscovered introns that either entirely lack an ORF or encode a novel ORF. Unlike organellar group II introns, we find only a handful of ORF-less introns in bacteria, suggesting that if a substantial number exist, they must be divergent from known introns. Together, these results highlight the retroelement character of bacterial group II introns, and suggest that their long-term survival is dependent upon retromobility. PMID- 18676619 TI - The anandamide analog, Met-F-AEA, controls human breast cancer cell migration via the RHOA/RHO kinase signaling pathway. AB - The endocannabinoid system regulates cell proliferation and migration in human breast cancer cells. In this study, we showed that a metabolically stable analog of anandamide, 2-methyl-2'-F-anandamide (Met-F-AEA), inhibited the RHOA activity and caused a RHOA delocalization from the cell membrane to cytosol determining a decrease in actin stress fibers. Overexpression of a dominant negative of RHOA activity and treatment of these cells with a RHO-associated protein kinase (ROCK) inhibitor, Y 27632, mimicked Met-F-AEA effects on actin organization and cell migration. We suggest that the inhibitory effect of Met-F-AEA on tumor cell migration could be related to RHOA-ROCK-dependent signaling pathway. PMID- 18676620 TI - Tigecycline for the treatment of multidrug-resistant Enterobacteriaceae: a systematic review of the evidence from microbiological and clinical studies. AB - OBJECTIVES: Antimicrobial drug resistance is spreading among Enterobacteriaceae, limiting the utility of traditionally used agents. We sought to systematically review the microbiological activity and clinical effectiveness of tigecycline for multidrug-resistant (MDR) Enterobacteriaceae, including those resistant to broad spectrum beta-lactams due to the expression of extended-spectrum beta-lactamases (ESBLs), AmpC enzymes and carbapenemases (including metallo-beta-lactamases). METHODS: PubMed was searched for articles including relevant data. RESULTS: Twenty-six microbiological and 10 clinical studies were identified. Tigecycline was active against more than 99% of 1936 Escherichia coli isolates characterized by any of the above resistance patterns (including 1636 ESBL-producing isolates) using the US Food and Drug Administration (FDA) breakpoint of susceptibility (MIC < or = 2 mg/L). Findings were not different using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoint (< or = 1 mg/L). Susceptibility rates for Klebsiella spp. with any of the above resistance patterns were 91.2% for 2627 isolates by the FDA criteria and 72.3% for 1504 isolates by the EUCAST criteria (92.3% for 2030 and 72.3% for 1284 ESBL-producing isolates, by the FDA and EUCAST criteria, respectively). The degree of microbiological activity of tigecycline against 576 MDR Enterobacter spp. isolates was moderate. In clinical studies, 69.7% of the 33 reported patients treated with tigecycline achieved resolution of an infection caused by a carbapenem-resistant or ESBL-producing or MDR Enterobacteriaceae. CONCLUSIONS: Tigecycline is microbiologically active against almost all of the ESBL or MDR E. coli isolates and the great majority of ESBL or MDR Klebsiella spp. isolates. Further evaluation of its clinical utility against such resistant Enterobacteriaceae, particularly regarding non-labelled indications, is warranted. PMID- 18676621 TI - Gene expression and sensitivity in response to copper stress in rice leaves. AB - Gene expression in response to Cu stress in rice leaves was quantified using DNA microarray (Agilent 22K Rice Oligo Microarray) and real-time PCR technology. Rice plants were grown in hydroponic solutions containing 0.3 (control), 10, 45, or 130 microM of CuCl(2), and Cu accumulation and photosynthesis inhibition were observed in leaves within 1 d of the start of treatment. Microarray analysis flagged 305 Cu-responsive genes, and their expression profile showed that a large proportion of general and defence stress response genes are up-regulated under excess Cu conditions, whereas photosynthesis and transport-related genes are down regulated. The Cu sensitivity of each Cu-responsive gene was estimated by the median effective concentration value (EC50) and the range of fold-changes (F) under the highest (130 microM) Cu conditions (|log(2)F|(130)). Our results indicate that defence-related genes involved in phytoalexin and lignin biosynthesis were the most sensitive to Cu, and that plant management of abiotic and pathogen stresses has overlapping components, possibly including signal transduction. PMID- 18676622 TI - Regulation of pigment epithelium-derived factor production and release by retinal glial (Muller) cells under hypoxia. AB - PURPOSE: To assess the regulation of pigment epithelium-derived factor (PEDF) production by retinal Muller glial cells, especially under ischemic or hypoxic conditions. METHODS: PEDF was determined in surgically excised retinal tissue originating from patients with ischemic diabetic retinopathy and in primary guinea pig Muller cell cultures exposed to the protein synthesis inhibitor cycloheximide (CHX) and to hypoxia. PEDF production and secretion were studied by immunohistochemistry, quantitative RT-PCR, ELISA, fluorescence-activated cell sorter analysis, and Western blotting. RESULTS: Gliotic Muller cells displayed decreased PEDF immunoreactivity in fibrovascular tissue from patients with diabetes compared with tissue from subjects with pathologic myopia. In Muller cell cultures, CHX treatment resulted in an increase, whereas mild hypoxia (2.5% 10% O(2)) induced a decrease, of PEDF mRNA and protein levels. However, strong hypoxia (0.2% O(2)) induced an upregulation of PEDF mRNA expression and resulted in only slightly reduced PEDF levels after 24 hours, detected as either a released, soluble, or cell surface-linked protein. CONCLUSIONS: These results suggest that under certain conditions including mild hypoxia, Muller cells synthesize a protein factor that downregulates PEDF expression or its turnover. Generally, the cells appear to generate a biphasic response to hypoxia. In moderate hypoxia, PEDF is downregulated such that the VEGF-to-PEDF ratio increases (and angiogenesis is facilitated). During severe (or chronic) oxygen deficiency, however, the PEDF decline is arrested or even reversed; thus, the neurotrophic effects of PEDF remain available. PMID- 18676623 TI - Documenting fixation at an extrafoveal locus with a modified slit lamp in age related macular degeneration. AB - PURPOSE: To develop a simple and clinically useful technique for observing fixation at an extrafoveal locus (preferred retinal locus [PRL]) with different targets and texts in age-related macular degeneration (AMD). METHODS: A standard slit lamp was modified by adding several fixation targets in the illumination pathway for direct observation and documentation of fixation during fundus examination. Fixation patterns were analyzed in 30 subjects with AMD. RESULTS: The location and stability of fixation with various stimuli was possible to record in each subject. In 23 subjects, there was no difference between the fixations at star and wagon wheel stimuli; in seven subjects, they were in clearly different retinal locations. Fixation was unstable in three subjects. The PRL for reading words was detectable in all subjects. CONCLUSIONS: The present assessment technique seems to offer a simple, clinically available technique to record fixation patterns to different targets and texts. PMID- 18676624 TI - Localization and functional mapping of AMPA receptor subunits in the developing rabbit retina. AB - PURPOSE: Glutamate has been suggested to regulate the development of retinal neurons, but ontogenic expression of ionotropic glutamate receptors has only recently been characterized in the rat retina. The purpose of this study was to examine the expression patterns of AMPA receptors and to functionally map glutamatergic drive in the developing rabbit retina. METHODS: The retinas from New Zealand White rabbits of different developmental stages (embryonic days [E]21 and 26, postnatal days [P]0-10, and adult) were isolated and cryosectioned into vertical slices. Antibodies against GluR1, -R2/3, and -R4 were used to examine the postnatal expression patterns of the AMPA receptor subunits. To further map the glutamatergic drive in the developing rabbit retina, an agmatine (AGB) activation assay was also used. RESULTS: All AMPA receptor subunits, including GluR1, -R2/3, and -R4, were expressed in the inner plexiform layer as early as E26 and were convincingly labeled in the outer plexiform layer at P2. These AMPA subunits showed different spatial distribution and temporal expression patterns across the postnatal stages examined. The immunoreactivity of the AMPA subunits was weak at P0 to P2 and then showed a striking increase at P4 to P6. The AGB activation assay revealed that some amacrine and ganglion cells were activated with 2 microM AMPA as early as E26 and, in the presence of an increased concentration of AMPA (20 microM), some potential horizontal cells were activated at the same stage. CONCLUSIONS: AMPA glutamate receptors express and function during the early stages of the developing rabbit retina, indicating that AMPA receptors are functional before synapse formation. The period of increasing expression pattern of AMPA subunits also coincides with the switch of the glutamatergic drive of the retinal wave and thus may contribute to the synaptic maturation in the retinal circuits. PMID- 18676625 TI - Constant volume of the human lens and decrease in surface area of the capsular bag during accommodation: an MRI and Scheimpflug study. AB - PURPOSE: A change in surface area of the capsular bag and a change in volume of the lens can indicate whether a change in the shape of the lens during accommodation is due to the compressibility or the elasticity of the lens material. METHODS: 3D magnetic resonance imaging (MRI) was used to image the complete shape of the lens in a group of five healthy subjects between 18 and 35 years of age. A parametric representation of the cross-sectional shape was fitted to the edges of the lens, which were determined with a Canny edge filter. Based on a partition of the lens into eight parts, the parametric shape makes it possible to calculate the mean cross-sectional area, the volume, and the surface area as a function of accommodation. Corrected Scheimpflug imaging was used to validate the results obtained with MRI. RESULTS: No significant difference in central anterior and posterior radius of curvature and thickness was found between the MRI and Scheimpflug measurements. In accordance with the Helmholtz accommodation theory, a decrease in the anterior and posterior radius of curvature and equatorial diameter and an increase in lens thickness occurred with accommodation. During accommodation, the mean cross-sectional area increased and the surface area decreased. However, no significant change in lens volume was found. CONCLUSIONS: The preservation of lens volume implies that the internal human lens material can be assumed to be incompressible and is undergoing elastic deformation. Furthermore, the change in surface area indicates that the capsular bag also undergoes elastic deformation. PMID- 18676627 TI - Lutein and zeaxanthin measured separately in the living human retina with fundus reflectometry. AB - PURPOSE: To separately measure the optical densities of lutein (L) and zeaxanthin (Z) in the human retina in vivo. L and Z are the basic constituents of the macular pigment (MP). METHODS: Spectral fundus reflectance was measured in 23 subjects (group 1) at 0 degrees , 1 degrees , 2 degrees , 4 degrees , and 8 degrees eccentricity with a modified macular pigment reflectometer. A model generated the optical densities of L (LOD) and Z (ZOD), using their slightly different absorption spectra. Three other subjects (group 2) took 20 mg/d zeaxanthin for 6 months; they were measured approximately monthly for 18 months. RESULTS: Mean LOD for group 1 at the central fovea was 0.200 +/- 0.061 (range, 0.085-0.305), mean ZOD was 0.494 +/- 0.169 (range, 0.169-0.806), resulting in a mean Z fraction [ZOD/(LOD + ZOD)] of 0.71. ZOD dropped faster toward the periphery than LOD, measuring 0.044 and 0.010 (Z fraction 0.18) at 8 degrees , respectively. Zeaxanthin supplementation in group 2 caused a significant increase in ZOD, and no or minor changes in LOD. ZOD further increased over a 10-month period after supplementation in all subjects. CONCLUSIONS: LOD and ZOD had different spatial profiles that, apart from scaling factors, showed similarity to in vitro literature data. Supplementation with Z caused LOD to decrease and ZOD to increase. These results strongly suggest that the optical densities of L and Z can be assessed in vivo by fundus reflectometry, opening new ways of investigating the putative protective roles of L and Z in retinal disease. PMID- 18676626 TI - Cell cycle arrest by kynurenine in lens epithelial cells. AB - PURPOSE: Indolemine 2,3-dioxygenase (IDO)-mediated oxidation of tryptophan produces kynurenines (KYNs), which may play a role in cataract formation. The molecular mechanisms by which KYNs cause cellular changes are poorly understood. The effects of KYNs on mouse lens epithelial cells by overexpression of human IDO were investigated. METHODS: Lens epithelial cells (mLECs) derived from human IDO overexpressing hemizygous transgenic (hemTg) and wild-type (Wt) mice were used. IDO activity was measured by quantifying kynurenine (KYN) by HPLC. KYN-mediated protein modifications were detected by immunocytochemistry and measured by ELISA. Cell proliferation and apoptosis were measured with commercially available kits. Cell distribution between cell cycle phases was examined with flow cytometric analysis. Immunoprecipitation followed by LC/MS was used to identify kynurenine modified proteins. RESULTS: mLECs derived from hemTg animals exhibited considerable IDO immunoreactivity and enzyme activity, which were barely detectable in Wt mLECs. KYN and KYN-mediated protein modification were detected in hemTg but not in Wt mLECs; the modified proteins were myosin II and alpha/gamma-actin. HemTg mLECs displayed reduced viability and proliferation. Cell cycle analysis of hemTg mLEC cultures showed approximately a twofold increase in cells at G(2)/M or in both phases, relative to Wt mLECs. Blocking IDO activity with 1-methyl-d,l-tryptophan in hemTg mLECs prevented KYN formation, KYN mediated protein modification, and G(2)/M arrest. CONCLUSIONS: Excess IDO activity in mLECs results in KYN production, KYN-mediated modification of myosin II and alpha/gamma-actin, and cell cycle perturbation. Modification of myosin II and gamma-actin by KYN may interfere with cytokinesis, leading to defective epithelial cell division and thus a decreased number of fiber cells. PMID- 18676628 TI - Inhibition by a selective IkappaB kinase-2 inhibitor of interleukin-1-induced collagen degradation by corneal fibroblasts in three-dimensional culture. AB - PURPOSE: Corneal ulcer results from excessive collagen degradation in the corneal stroma. Interleukin (IL)-1 promotes this process by activating signaling molecules that include nuclear factor (NF)-kappaB and stimulating the synthesis of matrix metalloproteinases (MMPs) in corneal fibroblasts. NF-kappaB activation is mediated by phosphorylation of the inhibitor IkappaB by IkappaB kinase (IKK)-2 and consequent IkappaB degradation. The authors investigated the effects of the IKK-2 inhibitor [5-(p-fluorophenyl)-2-ureido]thiophene-3-carboxamide (TPCA-1) on collagen degradation by corneal fibroblasts. METHODS: Rabbit corneal fibroblasts were cultured in three-dimensional collagen gels. Collagen degradation was evaluated by spectrophotometric quantitation of hydroxyproline in culture supernatants subjected to acid-heat hydrolysis. Expression of MMPs was evaluated by immunoblot analysis, gelatin zymography, and real-time reverse transcription polymerase chain reaction analysis. The phosphorylation and degradation of IkappaBalpha and the subcellular localization of NF-kappaB were examined by immunoblot and immunofluorescence analyses, respectively. RESULTS: IL-1beta induced collagen degradation by corneal fibroblasts was inhibited by TPCA-1 in a concentration- and time-dependent manner. TPCA-1 inhibited the IL-1beta-induced expression of MMP-1, -3, and -9 in these cells at both the mRNA and protein levels and the IL-1beta-induced activation of pro-MMP-2. In contrast to dexamethasone, TPCA-1 inhibited the phosphorylation and degradation of IkappaBalpha and the nuclear translocation of NF-kappaB induced by IL-1beta. CONCLUSIONS: An IKK-2 inhibitor blocked IL-1beta-induced collagen degradation by corneal fibroblasts by inhibiting the activation of the NF-kappaB signaling pathway and the upregulation of MMPs. IKK-2 inhibitors are thus potential alternatives to dexamethasone for the treatment of corneal ulcer. PMID- 18676629 TI - Visualization of sub-retinal pigment epithelium morphologies of exudative macular diseases by high-penetration optical coherence tomography. AB - PURPOSE: To evaluate the clinical significance of the newly developed long wavelength probe optical coherence tomography (LP-OCT) for the diagnosis of exudative macular diseases. METHODS: Fourteen eyes of 13 participants were prospectively enrolled in the study. There were seven type I and five type II choroidal neovascularization (CNV) cases associated with age-related macular degeneration and idiopathic neovascularization and one case of polypoidal choroidal vasculopathy (PCV). A custom-built LP-OCT based on swept-source OCT (SS OCT) technology was used. This new OCT uses a probe beam with a wavelength of 1060 nm that provides deeper penetration into the choroid and higher image contrast to the structures beneath the retinal pigment epithelium (RPE) and pathologic tissues than does conventional OCT. The depth resolution is 10.4 microm in tissue and the measurement speed is 28,000 depth scans/s. All the eyes were also examined by standard short wavelength probe OCT (SP-OCT). The image contrasts of the LP- and SP-OCT were qualitatively evaluated and analyzed by Wilcoxon's paired signed rank test and Spearman's rank correlation test. RESULTS: In 10 of 14 eyes, high-contrast visualization of the diseases beneath the RPE, CNV, or fibrin was attained. These diseases were almost invisible in the SP-OCT images. The LP-OCT of the remaining eyes also revealed significant improvement in the image contrasts beneath the RPE and CNV. Qualitative evaluation of the image contrasts and subsequent statistical test indicated statistically significant improvement in the image penetration to the choroid of LP-OCT to that of SP-OCT. CONCLUSIONS: LP-OCT provided significant improvement in the image contrast of exudative macular diseases. PMID- 18676630 TI - Reproducibility of nerve fiber layer thickness measurements using 3D fourier domain OCT. AB - PURPOSE: Conventional time-domain optical coherence tomography (OCT) has been shown to provide reproducible retinal nerve fiber layer (RNFL) measurements. Recently, high-speed, high-resolution Fourier-domain 3D-OCT has been introduced to improve OCT quality. It can provide 6-mm(2) high-density scans to provide RNFL thickness measurements. The purpose of this study was to test the reproducibility of 3D-OCT RNFL thickness measurements in healthy volunteers. METHODS: Thirty eight eyes were included in the study. High-density 6-mm(2) 3D scans were registered by two independent operators. RNFL thickness was calculated for eight areas corresponding to the ETDRS areas and for two ring areas. The ETDRS grid was centered on the optic disc. Intraclass correlation coefficients (ICC) and coefficients of variation (COV) were calculated. Interobserver reproducibility was visualized by using Bland-Altman analysis. RESULTS: Intrasession reproducibility was good with a mean ICC of 0.90. The mean COV for operator 1 and 2 was 4.2% and 4%, respectively (range, 1.9%-6.7%). Highest reproducibility was found for the two ring areas and the superior and inferior quadrants. Mean differences in RNFL thickness measurements for ring 1 and 2 between operator 1 and 2 were 0.9 microm (limits of agreement, -11.4 to +9.6 microm) and 0.1 microm (limits of agreement -4.1 to +3.9 microm), respectively. CONCLUSIONS: 3D-OCT RNFL thickness measurements in healthy volunteers showed good intra- and interobserver reproducibility. 3D-OCT provides more RNFL thickness information compared to conventional time-domain OCT measurements and may be useful for the management of glaucoma and other optic neuropathies. PMID- 18676631 TI - Segmentation error in Stratus optical coherence tomography for neovascular age related macular degeneration. AB - PURPOSE: To describe the rate of automated segmentation error in Stratus optical coherence tomography (OCT) scans in consecutive patients with neovascular age related macular degeneration (nAMD) receiving treatment and to investigate the effect of the segmentation error on automated retinal thickness measures and whether further imaging reduces the rate of segmentation error. METHODS: A retrospective analysis of fast macular thickness map (FMTM) protocol OCT scans of 50 eyes of 50 consecutive patients with nAMD. Each line scan was analyzed for segmentation error with manual measurement of the center-point retinal thickness allowing calculation of the percentage error in automated thickness. OCT scanning was repeated to overcome segmentation error. RESULTS: Segmentation error was detected in 45 (90%) of the 50 patients with 37 (74%) patients having an error affecting the central 1-mm subfield. Scan sets with a high central segmentation error score (two or more line scans affected of six) had a significantly greater error in automated center-point retinal thickness than scan sets with a low error score (20% compared with 3%, P < 0.000005). Central subfield segmentation error persisted in 30 (60%) patients despite repeat scanning. CONCLUSIONS: There is a high rate of segmentation error in OCT scans of patients with nAMD who are undergoing treatment, leading to errors in automated central retinal thickness measurement. The authors recommend manual measurement of central macular thickness when two or more line scans are affected by segmentation error in the central 1-mm subfield. Repeated scanning reduced the rate of error but did not eliminate the problem. PMID- 18676632 TI - Sporadic bilateral optic neuropathy in children: the role of mitochondrial abnormalities. AB - PURPOSE: To evaluate a group of patients with isolated, early-onset, bilateral optic neuropathy for genetic and biochemical evidence of mitochondrial diseases. METHODS: This case-control study involved 21 patients, 159 control subjects for mitochondrial (mt)DNA sequencing, and 40 control subjects for relative mtDNA content. Patients were identified who had had decreased vision since childhood due to bilateral optic neuropathy characterized by central visual loss with no other major neurologic or ocular abnormality and no clinical evidence of a mitochondrial syndrome. Clinical examination, electroretinograms, and neuroimaging were performed; the entire mtDNA coding region was sequenced in leukocytes of all patients; relative mtDNA content was assessed; and OPA1 and OPA3 nuclear genes associated with dominant and recessive optic atrophy, respectively, were sequenced. Main outcome measures were clinical description, nonsynonymous (NS) mtDNA nucleotide changes, relative mtDNA content, and OPA1 and OPA3 nucleotide changes. RESULTS: Twenty-one unrelated patients (16 male and 5 female; mean age at first examination 13.6 years) had bilateral moderate, relatively symmetric optic neuropathies and normal neurologic examinations other than strabismus in 11 and congenital nystagmus in 9. Four patients had optic nerve hypoplasia. One patient had the nt 11778 primary Leber hereditary optic neuropathy (LHON) mutation, and three others had mtDNA nucleotide changes predicted to be pathologic. The entire group had a small increase (6.7%) in relative mtDNA content of indeterminate statistical significance. No patient had a polymorphism or mutation of OPA1 or OPA3. CONCLUSIONS: A minority of these young patients with sporadic bilateral optic neuropathy had abnormalities of the mitochondrial parameters evaluated. This bilateral optic neuropathy may be due to other genetic, epigenetic, or environmental injury to the optic nerve or to mitochondrial defects not studied. PMID- 18676633 TI - Advanced glycation end product (AGE) accumulation on Bruch's membrane: links to age-related RPE dysfunction. AB - PURPOSE: Advanced glycation end products (AGEs) accumulate during aging and have been observed in postmortem eyes within the retinal pigment epithelium (RPE), Bruch's membrane, and subcellular deposits (drusen). AGEs have been associated with age-related dysfunction of the RPE-in particular with development and progression to age-related macular degeneration (AMD). In the present study the impact of AGEs at the RPE-Bruch's membrane interface was evaluated, to establish how these modifications may contribute to age-related disease. METHODS: AGEs on Bruch's membrane were evaluated using immunohistochemistry. A clinically relevant in vitro model of substrate AGE accumulation was established to mimic Bruch's membrane ageing. Responses of ARPE-19 growing on AGE-modified basement membrane (AGE-BM) for 1 month were investigated by using a microarray approach and validated by quantitative (q)RT-PCR. In addition to identified AGE-related mRNA alterations, lysosomal enzyme activity and lipofuscin accumulation were also studied in ARPE-19 grown on AGE-BM. RESULTS: Autofluorescent and glycolaldehyde derived AGEs were observed in clinical specimens on Bruch's membrane and choroidal extracellular matrix. In vitro analysis identified a range of dysregulated mRNAs in ARPE-19 exposed to AGE-BM. Altered ARPE-19 degradative enzyme mRNA expression was observed on exposure to AGE-BM. AGE-BM caused a significant reduction in cathepsin-D activity in ARPE-19 (P < 0.05) and an increase in lipofuscin accumulation (P < 0.01). CONCLUSIONS: AGEs influence ARPE 19 mRNA expression profiles and may contribute to reduced lysosomal enzyme degradative capacity and enhanced accumulation of lipofuscin. Formation of AGEs on Bruch's membrane may have important consequences for age-related dysfunction of the RPE, perhaps leading to age-related outer retinal disease. PMID- 18676634 TI - Effect of systemic nitric oxide synthase inhibition on optic disc oxygen partial pressure in normoxia and in hypercapnia. AB - PURPOSE: To investigate the effect of systemic nitric oxide synthase (NOS) inhibition on optic disc oxygen partial pressure (PO(2)) in normoxia and hypercapnia. METHODS: Intervascular optic disc PO(2) was measured in 12 anesthetized minipigs by using oxygen-sensitive microelectrodes placed <50 microm from the optic disc. PO(2) was measured continuously during 10 minutes under normoxia, hyperoxia (100% O(2)), carbogen breathing (95% O(2), 5% CO(2)), and hypercapnia (increased inhaled CO(2)). Measurements were repeated after intravenous injection of N(omega)-nitro-L-arginine methyl ester (L-NAME) 100 mg/kg. Intravenous L-arginine 100 mg/kg was subsequently given to three animals. RESULTS: Before L-NAME injection, an increase was observed in optic disc PO(2) during hypercapnia (DeltaPO(2) = 3.2 +/- 1.7 mm Hg; 18%; P = 0.001) and carbogen breathing (DeltaPO(2) = 12.8 +/- 5.1 mm Hg; 69%; P < 0.001). Optic disc PO(2) in normoxia remained stable for 30 minutes after L-NAME injection (4% decrease from baseline; P > 0.1), despite a 21% increase of mean arterial pressure. Optic disc PO(2) increase under hypercapnia was blunted after L-NAME injection (DeltaPO(2) = 0.6 +/- 1.1 mm Hg; 3%; P > 0.1), and this effect was reversible by L-arginine. Moreover, L-NAME reduced the response to carbogen by 29% (DeltaPO(2) = 9.1 +/- 4.4 mm Hg; 49%; P = 0.01 versus before L-NAME). The response to hyperoxia was not affected. CONCLUSIONS: Whereas systemic NOS inhibition did not affect optic disc PO(2) in normoxia, a blunting effect was noted on the CO(2)-induced optic disc PO(2) increase. Nitric oxide appears to mediate the hypercapnic optic disc PO(2) increase. PMID- 18676635 TI - Granulocyte colony-stimulating factor protects retinal photoreceptor cells against light-induced damage. AB - PURPOSE: Granulocyte colony stimulating factor (G-CSF) has been shown to have neuroprotective and anti-inflammatory effects in cerebral damage models. In addition, bone-marrow-derived hematopoietic cells, which can be mobilized with G CSF, have a neuroprotective effect in hereditary retinal cell death. The present study was conducted to investigate whether G-CSF protects photoreceptors from light-induced cell death. METHODS: G-CSF or vehicle was systemically injected before the light exposure and for four consecutive days after the exposure. Morphologic and electrophysiologic examinations were performed 1 week after the exposure to light. Gamma ray irradiation (6.5 Gy) was used to examine the involvement of bone marrow-derived cells increased by G-CSF injection. The expression of G-CSF receptor in the retina was analyzed by immunohistochemistry and quantitative RT-PCR. RESULTS: The outer nuclear layer thickness was partially preserved in G-CSF-treated mice (measured at 300 microm superior from the optic disc, G-CSF: 14.9 +/- 6.3 microm versus control: 6.7 +/- 2.5 microm), and an electroretinogram confirmed the preservation of wave amplitudes (maximum scotopic a-wave G-CSF: 97.7 +/- 48.0 microV versus control: 14.4 +/- 21.9 microV, maximum scotopic b-wave G-CSF: 298.1 +/- 145.3 microV versus control: 33.2 +/- 50.1 microV). The effect was not lost, even with leukocyte depletion by irradiation. G CSF receptor was expressed in retinal cells and upregulated by the light exposure (1.8-fold upregulation 2 hours after light exposure). CONCLUSIONS: G-CSF protects photoreceptor cells against light-induced damage, possibly via G-CSF receptor expressed on retinal cells. These findings may lead to a novel treatment strategy for neural degenerating diseases of the retina. PMID- 18676637 TI - Myosin isoform expression in dog rectus muscles: patterns in global and orbital layers and among single fibers. AB - PURPOSE: To quantitate the distribution of myosin heavy chain (MyHC) isoforms along the global and orbital layers of dog rectus muscles and determine MyHC and myosin light chain (MLC) isoform patterns among single fibers from both layers. METHODS: Serial samples of both layers of rectus muscles were prepared for gel electrophoresis. Relative amounts of each MyHC isoform in each sample were determined with scanning densitometry. Single fibers were isolated from each layer for analyses of MyHC and MLC isoforms. RESULTS: Nine MyHC isoforms were detected. Four prominent MyHC isoforms, and an additional MyHC isoform at very low levels, are expressed in the global layer. Evidence suggests that all nine MyHC isoforms are expressed in the orbital layer. There are marked gradients in the levels of some MyHC isoforms along the length of both layers. Complex patterns of coexpression of multiple MyHC isoforms exist in single fibers from both layers. Most fibers express conventional slow or fast MLC isoforms, in accordance with the type (slow or fast) of MyHC isoform(s) in a given fiber, with the exception that slow fibers in the orbital layer express the atrial/embryonic isoform of MLC1. CONCLUSIONS: MyHC isoform expression patterns differ markedly between and along global and orbital layers of dog rectus muscles, with greater complexity in the orbital layer. Heterogeneity in MyHC isoform expression in rectus muscles is much greater than in limb muscles and presumably is the basis for the broad spectrum of extraocular muscle (EOM) contractile properties in driving oculomotor functions. PMID- 18676636 TI - Human p32 is a novel FOXC1-interacting protein that regulates FOXC1 transcriptional activity in ocular cells. AB - PURPOSE: Mutations in the human forkhead box C1 gene (FOXC1) cause Axenfeld Rieger (AR) malformations, often leading to glaucoma. Understanding the function of FOXC1 necessitates characterizing the proteins that interact with FOXC1. This study was undertaken to isolate FOXC1-interacting proteins and determine their effects on FOXC1. METHODS: To identify FOXC1-interacting proteins, a human trabecular meshwork (HTM) yeast two-hybrid (Y2H) cDNA library was screened. The interaction and colocalization between FOXC1 and its putative protein partner were confirmed by Ni(2+) pull-down assays, immunoprecipitation, and immunofluorescence, respectively. The electrophoretic mobility shift assay (EMSA) was used to study the effect of the interacting protein on FOXC1 DNA-binding ability. Dual luciferase assays using FOXC1 reporter plasmids in HTM cells were performed to determine the effect of the interaction on FOXC1 transcription activity. RESULTS: The human p32 protein was isolated as a putative FOXC1 interacting protein from a Y2H screen. The interaction of FOXC1 with p32 was confirmed by Ni-pull-down assays and immunoprecipitation. Although p32 is predominantly cytoplasmic, the portion of p32 that is within the nucleus colocalizes with FOXC1. The FOXC1 forkhead domain (FHD) was identified as the p32 interaction domain. p32 significantly inhibited FOXC1-mediated transcription activation in a dose-dependent manner but did not affect FOXC1 DNA-binding ability. Of interest, a FOXC1 mutation F112S displayed an impaired interaction with p32. CONCLUSIONS: In the study, the human p32 protein as a novel regulator of FOXC1-mediated transcription activation. Failure of p32 to interact with FOXC1 containing the disease-causing F112S mutation indicates that impaired protein interaction may be a disease mechanism for AR malformations. PMID- 18676639 TI - Mechanisms for ultrafast nonradiative relaxation in electronically excited eumelanin constituents. AB - We investigate the relaxation dynamics of melanin model constituents including monomers, dimers, and tetramers, upon excitation, using state-of-the-art, time dependent, density functional theory calculations. The results explain the ability of these molecules to transform photon energy into thermal energy in a remarkably short timescale of approximately 100 fs. We find that after electronic excitation by light absorption, ultrafast energy conversion takes place through two novel mechanisms: proton transfer on a timescale of 110 fs and state mixing upon oligomerization on a timescale of <50 fs. These results are in good agreement with available experiments and help elucidate melanin's role in photoprotection against ultraviolet radiation. PMID- 18676638 TI - The cardiac Ca2+-sensitive regulatory switch, a system in dynamic equilibrium. AB - The Ca(2+)-sensitive regulatory switch of cardiac muscle is a paradigmatic example of protein assemblies that communicate ligand binding through allosteric change. The switch is a dimeric complex of troponin C (TnC), an allosteric sensor for Ca(2+), and troponin I (TnI), an allosteric reporter. Time-resolved equilibrium Forster resonance energy transfer (FRET) measurements suggest that the switch activates in two steps: a TnI-independent Ca(2+)-priming step followed by TnI-dependent opening. To resolve the mechanistic role of TnI in activation we performed stopped-flow FRET measurements of activation after rapid addition of a lacking component (Ca(2+) or TnI) and deactivation after rapid chelation of Ca(2+). Time-resolved measurements, stopped-flow measurements, and Ca(2+) titration measurements were globally analyzed in terms of a new quantitative dynamic model of TnC-TnI allostery. The analysis provided a mesoscopic parameterization of distance changes, free energy changes, and transition rates among the accessible coarse-grained states of the system. The results reveal that 1), the Ca(2+)-induced priming step, which precedes opening, is the rate-limiting step in activation; 2), closing is the rate-limiting step in de-activation; 3), TnI induces opening; 4), there is an incompletely deactivated population when regulatory Ca(2+) is not bound, which generates an accessory pathway of activation; and 5), there is incomplete activation by Ca(2+)-when regulatory Ca(2+) is bound, a 3:2 mixture of dynamically interconverting open (active) and primed-closed (partially active) conformers is observed (15 degrees C). Temperature-dependent stopped-flow FRET experiments provide a near complete thermokinetic parameterization of opening: the enthalpy change (DeltaH = -33.4 kJ/mol), entropy change (DeltaS = -0.110 kJ/mol/K), heat capacity change (DeltaC(p) = -7.6 kJ/mol/K), the enthalpy of activation (delta(double dagger) = 10.6 kJ/mol) and the effective barrier crossing attempt frequency (nu(adj) = 1.8 x 10(4) s(-1)). PMID- 18676640 TI - Cartilage aggrecan can undergo self-adhesion. AB - Here it is reported that aggrecan, the highly negatively charged macromolecule in the cartilage extracellular matrix, undergoes Ca(2+)-mediated self-adhesion after static compression even in the presence of strong electrostatic repulsion in physiological-like solution conditions. Aggrecan was chemically end-attached onto gold-coated planar silicon substrates and gold-coated microspherical atomic force microscope probe tips (end radius R approximately 2.5 mum) at a density ( approximately 40 mg/mL) that simulates physiological conditions in the tissue ( approximately 20-80 mg/mL). Colloidal force spectroscopy was employed to measure the adhesion between opposing aggrecan monolayers in NaCl (0.001-1.0 M) and NaCl + CaCl(2) ([Cl(-)] = 0.15 M, [Ca(2+)] = 0 - 75 mM) aqueous electrolyte solutions. Aggrecan self-adhesion was found to increase with increasing surface equilibration time upon compression (0-30 s). Hydrogen bonding and physical entanglements between the chondroitin sulfate-glycosaminoglycan side chains are proposed as important factors contributing to aggrecan self-adhesion. Self adhesion was found to significantly increase with decreasing bath ionic strength (and hence, electrostatic double-layer repulsion), as well as increasing Ca(2+) concentration due to the additional ion-bridging effects. It is hypothesized that aggrecan self-adhesion, and the macromolecular energy dissipation that results from this self-adhesion, could be important factors contributing to the self assembled architecture and integrity of the cartilage extracellular matrix in vivo. PMID- 18676641 TI - Structural refinement of membrane proteins by restrained molecular dynamics and solvent accessibility data. AB - We present an approach for incorporating solvent accessibility data from electron paramagnetic resonance experiments in the structural refinement of membrane proteins through restrained molecular dynamics simulations. The restraints have been parameterized from oxygen (PiO(2)) and nickel-ethylenediaminediacetic acid (PiNiEdda) collision frequencies, as indicators of lipid or aqueous exposed spin label sites. These are enforced through interactions between a pseudoatom representation of the covalently attached Nitroxide spin-label and virtual "solvent" particles corresponding to O(2) and NiEdda in the surrounding environment. Interactions were computed using an empirical potential function, where the parameters have been optimized to account for the different accessibilities of the spin-label pseudoatoms to the surrounding environment. This approach, "pseudoatom-driven solvent accessibility refinement", was validated by refolding distorted conformations of the Streptomyces lividans potassium channel (KcsA), corresponding to a range of 2-30 A root mean-square deviations away from the native structure. Molecular dynamics simulations based on up to 58 electron paramagnetic resonance restraints derived from spin-label mutants were able to converge toward the native structure within 1-3 A root mean square deviations with minimal computational cost. The use of energy-based ranking and structure similarity clustering as selection criteria helped in the convergence and identification of correctly folded structures from a large number of simulations. This approach can be applied to a variety of integral membrane protein systems, regardless of oligomeric state, and should be particularly useful in calculating conformational changes from a known reference crystal structure. PMID- 18676642 TI - Protein adsorption onto organically modified silica glass leads to a different structure than sol-gel encapsulation. AB - The secondary structures of two proteins were examined by circular dichroism spectroscopy after adsorption onto a series of organically modified silica glasses. The glasses were prepared by the sol-gel technique and were varied in hydrophobicity by incorporation of 5% methyl, propyl, trifluoropropyl, or n-hexyl silane. Both cytochrome c and apomyoglobin were found to lose secondary structure after adsorption onto the modified glasses. In the case of apomyoglobin, the alpha-helical content of the adsorbed protein ranged from 21% to 28%, well below the 62% helix found in solution. In contrast, these same glasses led to a striking increase in apomyoglobin structure when the protein was encapsulated within the pores during sol-gel processing: the helical content of apomyoglobin increased with increasing hydrophobicity from 18% in an unmodified glass to 67% in a 5% hexyl-modified glass. We propose that proteins preferentially adsorb onto unmodified regions of the silica surface, whereas encapsulated proteins are more susceptible to changes in surface hydration due to the proximity of the alkyl chain groups. PMID- 18676643 TI - Ligand escape pathways and (un)binding free energy calculations for the hexameric insulin-phenol complex. AB - Cooperative binding of phenolic species to insulin hexamers is known to stabilize pharmaceutical preparations of the hormone. Phenol dissociation is rapid on hexamer dissolution timescales, and phenol unbinding upon dilution is likely the first step in the conversion of (pharmaceutical) hexameric insulin to the active monomeric form upon injection. However, a clear understanding of the determinants of the rates of phenol unbinding remains obscure, chiefly because residues implicated in phenol exchange as determined by NMR are not all associated with likely unbinding routes suggested by the best-resolved hexamer structures. We apply random acceleration molecular dynamics simulation to identify potential escape routes of phenol from hydrophobic cavities in the hexameric insulin-phenol complex. We find three major pathways, which provide new insights into (un)binding mechanisms for phenol. We identify several residues directly participating in escape events that serve to resolve ambiguities from recent NMR experiments. Reaction coordinates for dissociation of phenol are developed based on these exit pathways. Potentials of mean force along the reaction coordinate for each pathway are resolved using multiple independent steered molecular dynamics simulations with second-order cumulant expansion of Jarzynski's equality. Our results for DeltaF agree reasonably well within the range of known experimental and previous simulation magnitudes of this quantity. Based on structural analysis and energetic barriers for each pathway, we suggest a plausible preferred mechanism of phenolic exchange that differs from previous mechanisms. Several weakly-bound metastable states are also observed for the first time in the phenol dissociation reaction. PMID- 18676644 TI - Thermodynamic redox behavior of the heme centers in A-type heme-copper oxygen reductases: comparison between the two subfamilies. AB - The study of the thermodynamic redox behavior of the hemes from two members of the A family of heme-copper oxygen reductases, Paracoccus denitrificans aa3 (A1 subfamily) and Rhodothermus marinus caa3 (A2 subfamily) enzymes, is presented. At different pH values, midpoint reduction potentials and interaction potentials were obtained in the framework of a pairwise model for two interacting redox centers. In both enzymes, the hemes have different reduction potentials. For the A1-type enzyme, it was shown that heme a has a pH-dependent midpoint reduction potential, whereas that of heme a3 is pH independent. For the A2-type enzyme the opposite was observed. The midpoint reduction potential of heme c from subunit II of the caa3 enzyme was determined by fitting the data with a single-electron Nernst curve, and it was shown to be pH dependent. The results presented here for these A-type enzymes are compared with those previously obtained for representative members of the B and C families. PMID- 18676645 TI - Biophysical studies on the stability of DNA intrastrand cross-links of transplatin. AB - Clinically ineffective transplatin [trans-diamminedichloridoplatinum(II)] is used in the studies of the structure-pharmacological activity relationship of platinum compounds. In addition, a number of transplatin analogs exhibit promising toxic effects in several tumor cell lines including those resistant to conventional antitumor cisplatin. Moreover, transplatin-modified oligonucleotides have been shown to be effective modulators of gene expression. Owing to these facts and because DNA is also considered the major pharmacological target of platinum complexes, interactions between transplatin and DNA are of great interest. We examined, using biophysical and biochemical methods, the stability of 1,3-GNG intrastrand cross-links (CLs) formed by transplatin in short synthetic oligodeoxyribonucleotide duplexes and natural double-helical DNA. We have found that transplatin forms in double-helical DNA 1,3-GNG intrastrand CLs, but their stability depends on the sequence context. In some sequences the 1,3-GNG intrastrand CLs formed by transplatin in double-helical DNA readily rearrange into interstrand CLs. On the other hand, in a number of other sequences these intrastrand CLs are relatively stable. We show that the stability of 1,3-GNG intrastrand CLs of transplatin correlates with the extent of conformational distortion and thermodynamic destabilization induced in double-helical DNA by this adduct. PMID- 18676646 TI - Theoretical conditions for high-frequency hair bundle oscillations in auditory hair cells. AB - Substantial evidence exists for spontaneous oscillations of hair cell stereociliary bundles in the lower vertebrate inner ear. Since the oscillations are larger than expected from Brownian motion, they must result from an active process in the stereociliary bundle suggested to underlie amplification of the sensory input as well as spontaneous otoacoustic emissions. However, their low frequency (<100 Hz) makes them unsuitable for amplification in birds and mammals that hear up to 5 kHz or higher. To examine the possibility of high-frequency oscillations, we used a finite-element model of the outer hair cell bundle incorporating previously measured mechanical parameters. Bundle motion was assumed to activate mechanotransducer channels according to the gating spring hypothesis, and the channels were regulated adaptively by Ca(2+) binding. The model generated oscillations of freestanding bundles at 4 kHz whose sharpness of tuning depended on the mechanotransducer channel number and location, and the Ca(2+) concentration. Entrainment of the oscillations by external stimuli was used to demonstrate nonlinear amplification. The oscillation frequency depended on channel parameters and was increased to 23 kHz principally by accelerating Ca(2+) binding kinetics. Spontaneous oscillations persisted, becoming very narrow band, when the hair bundle was loaded with a tectorial membrane mass. PMID- 18676647 TI - Hydrophobe-water interactions: methane as a model. AB - Recent molecular-dynamics simulations have demonstrated that the use of an empirical hydrophobic potential displaying two minima, i.e., one for hydrophobes in close contact and one for hydrophobes separated by a hydration layer, leads to a marked improvement in protein structure prediction. This potential is supported by experimental data and simulations, but its physical origin and mathematical formulation have not been derived as yet. Here we show that water-mediated attraction (the "wetting regime") between two hydrophobic molecules originates in the interaction between the dipoles induced at the surface of the hydrophobes by the surrounding structured water. As an example, we derive the effective hydrophobic potential that describes the interaction between two methane molecules, a classical model of a double-well energy function. We found an excellent agreement with published results from all-atom, explicit solvent molecular-dynamics simulations of this interaction. The approach presented here provides the theoretical basis for implementing an adequate representation of the wetting regime of the hydrophobic interactions in force fields used for structure prediction. The results are useful for modeling both protein folding and binding. PMID- 18676648 TI - Rate constant and reaction coordinate of Trp-cage folding in explicit water. AB - We report rate constant calculations and a reaction coordinate analysis of the rate-limiting folding and unfolding process of the Trp-cage mini-protein in explicit solvent using transition interface sampling. Previous transition path sampling simulations revealed that in this (un)folding process the protein maintains its compact configuration, while a (de)increase of secondary structure is observed. The calculated folding rate agrees reasonably with experiment, while the unfolding rate is 10 times higher. We discuss possible origins for this mismatch. We recomputed the rates with the forward flux sampling method, and found a discrepancy of four orders of magnitude, probably caused by the method's higher sensitivity to the choice of order parameter with respect to transition interface sampling. Finally, we used the previously computed transition path sampling ensemble to screen combinations of many order parameters for the best model of the reaction coordinate by employing likelihood maximization. We found that a combination of the root mean-square deviation of the helix and of the entire protein was, of the set of tried order parameters, the one that best describes the reaction coordination. PMID- 18676649 TI - DARS (Decoys As the Reference State) potentials for protein-protein docking. AB - Decoys As the Reference State (DARS) is a simple and natural approach to the construction of structure-based intermolecular potentials. The idea is generating a large set of docked conformations with good shape complementarity but without accounting for atom types, and using the frequency of interactions extracted from these decoys as the reference state. In principle, the resulting potential is ideal for finding near-native conformations among structures obtained by docking, and can be combined with other energy terms to be used directly in docking calculations. We investigated the performance of various DARS versions for docking enzyme-inhibitor, antigen-antibody, and other type of complexes. For enzyme-inhibitor pairs, DARS provides both excellent discrimination and docking results, even with very small decoy sets. For antigen-antibody complexes, DARS is slightly better than a number of interaction potentials tested, but results are worse than for enzyme-inhibitor complexes. With a few exceptions, the DARS docking results are also good for the other complexes, despite poor discrimination, and we show that the latter is not a correct test for docking accuracy. The analysis of interactions in antigen-antibody pairs reveals that, in constructing pairwise potentials for such complexes, one should account for the asymmetry of hydrophobic patches on the two sides of the interface. Similar asymmetry does occur in the few other complexes with poor DARS docking results. PMID- 18676650 TI - Chirality-based signatures of local protein environments in two-dimensional optical spectroscopy of two species photosynthetic complexes of green sulfur bacteria: simulation study. AB - Two-dimensional electronic chirality-induced signals of excitons in the photosynthetic Fenna-Matthews-Olson complex from two species of green sulfur bacteria (Chlorobium tepidum and Prosthecochloris aestuarii) are compared. The spectra are predicted to provide sensitive probes of local protein environment of the constituent bacteriochlorophyll a chromophores and reflect electronic structure variations (site energies and couplings) of the two complexes. Pulse polarization configurations are designed that can separate the coherent and incoherent exciton dynamics contributions to the two-dimensional spectra. PMID- 18676651 TI - Free-energy profiles of membrane insertion of the M2 transmembrane peptide from influenza A virus. AB - The insertion of the M2 transmembrane peptide from influenza A virus into a membrane has been studied with molecular-dynamics simulations. This system is modeled by an atomically detailed peptide interacting with a continuum representation of a membrane bilayer in aqueous solution. We performed replica exchange molecular-dynamics simulations with umbrella-sampling techniques to characterize the probability distribution and conformation preference of the peptide in the solution, at the membrane interface, and in the membrane. The minimum in the calculated free-energy surface of peptide insertion corresponds to a fully inserted, helical peptide spanning the membrane. The free-energy profile also shows that there is a significant barrier for the peptide to enter into this minimum in a nonhelical conformation. The sequence of the peptide is such that hydrophilic amino acid residues at the ends of the otherwise primarily hydrophobic peptide create a trapped, U-shaped conformation with the hydrophilic residues associated with the aqueous phase and the hydrophobic residues embedded in the membrane. Analysis of the free energy shows that the barrier to insertion is largely enthalpic in nature, whereas the membrane-spanning global minimum is favored by entropy. PMID- 18676652 TI - Peptide aggregation and pore formation in a lipid bilayer: a combined coarse grained and all atom molecular dynamics study. AB - We present a simulation study where different resolutions, namely coarse-grained (CG) and all-atom (AA) molecular dynamics simulations, are used sequentially to combine the long timescale reachable by CG simulations with the high resolution of AA simulations, to describe the complete processes of peptide aggregation and pore formation by alamethicin peptides in a hydrated lipid bilayer. In the 1 micros CG simulations the peptides spontaneously aggregate in the lipid bilayer and exhibit occasional transitions between the membrane-spanning and the surface bound configurations. One of the CG systems at t = 1 micros is reverted to an AA representation and subjected to AA simulation for 50 ns, during which water molecules penetrate the lipid bilayer through interactions with the peptide aggregates, and the membrane starts leaking water. During the AA simulation significant deviations from the alpha-helical structure of the peptides are observed, however, the size and arrangement of the clusters are not affected within the studied time frame. Solid-state NMR experiments designed to match closely the setup used in the molecular dynamics simulations provide strong support for our finding that alamethicin peptides adopt a diverse set of configurations in a lipid bilayer, which is in sharp contrast to the prevailing view of alamethicin oligomers formed by perfectly aligned helical alamethicin peptides in a lipid bilayer. PMID- 18676653 TI - Modeling transmembrane transport through cell membrane wounds created by acoustic cavitation. AB - Cells exposed to acoustic cavitation and other mechanical stresses can be transiently permeabilized to permit intracellular uptake of molecules, including drugs, proteins, and genes. Microscopic imaging and other studies suggest that intracellular loading occurs through plasma membrane wounds of submicrometer radius that reseal over time through the aggregation and fusion of lipid vesicles trafficked to the wound site. The goal of this study was to 1), determine the size of membrane wounds as a function of time after in vitro sonication of DU145 prostate cancer cells under conditions that caused extensive acoustic cavitation; and 2), theoretically model transport processes leading to intracellular loading. Our overall hypothesis was that intracellular loading is governed by passive diffusion through porous membrane wounds of up to 300-nm radius containing pores that permit entry of molecules up to at least 28-nm radius over a timescale of minutes. Experimental measurements showed intracellular loading of molecules with radii from 0.6 to 28 nm, where most loading occurred after sonication over a timescale up to minutes and where smaller molecules were taken up to a greater extent and over a longer timescale than larger molecules. Theoretical modeling predicted that membrane wounds would have a 300-nm radius initially and then would shrink, with a half life of 20 to 50 s. Uptake was shown to occur predominantly by diffusion and the increasing levels of uptake with decreasing molecular size was explained primarily by differences in molecular diffusivity and, for the largest molecule, geometrical hindrance within the wound. Mathematical modeling was simplified, because transport through porous wounds of possibly complex internal nanostructure was governed largely by transport at the edge of the wound, and depended only weakly on the size, number, and distribution of nanopores within the wound under the conditions relevant to this study. Overall, this study developed a theoretical framework for analysis of transmembrane transport through cell membrane wounds and thereby provided quantitative estimates of their size and lifetime. PMID- 18676654 TI - Statistical prediction and molecular dynamics simulation. AB - We describe a statistical approach to the validation and improvement of molecular dynamics simulations of macromolecules. We emphasize the use of molecular dynamics simulations to calculate thermodynamic quantities that may be compared to experimental measurements, and the use of a common set of energetic parameters across multiple distinct molecules. We briefly review relevant results from the theory of stochastic processes and discuss the monitoring of convergence to equilibrium, the obtaining of confidence intervals for summary statistics corresponding to measured quantities, and an approach to validation and improvement of simulations based on out-of-sample prediction. We apply these methods to replica exchange molecular dynamics simulations of a set of eight helical peptides under the AMBER potential using implicit solvent. We evaluate the ability of these simulations to quantitatively reproduce experimental helicity measurements obtained by circular dichroism. In addition, we introduce notions of statistical predictive estimation for force-field parameter refinement. We perform a sensitivity analysis to identify key parameters of the potential, and introduce Bayesian updating of these parameters. We demonstrate the effect of parameter updating applied to the internal dielectric constant parameter on the out-of-sample prediction accuracy as measured by cross validation. PMID- 18676656 TI - 3'-phosphoinositides regulate the coordination of speed and accuracy during chemotaxis. AB - The PI3K/PTEN pathway, as the regulator of 3'-phosphoinositide (3'-PI) dynamics, has emerged as a key regulator of chemoattractant gradient sensing during chemotaxis in Dictyostelium and other cell types. Previous results have shown 3' PIs to be important for regulating basal cell motility and sensing the direction and strength of the chemoattractant gradient. We examined the chemotaxis of wild type cells and cells lacking PTEN or PI3K1 and 2 using analytical methods that allowed us to quantitatively discern differences between the genotype's ability to sense and efficiently respond to changes in gradient steepness during chemotaxis. We found that cells are capable of increasing their chemotactic accuracy and speed as they approach a micropipette in a manner that is dependent on the increasing strength of the concentration gradient and 3'-PI signaling. Further, our data show that 3'-PI signaling affects a cell's ability to coordinate speed and direction to increase chemotactic efficiency. Using to our knowledge a new measurement of chemotactic efficiency that reveals the degree of coordination between speed and accuracy, we found that cells also have the capacity to increase their chemotactic efficiency as they approach the micropipette. Like directional accuracy and speed, the increase in chemotactic efficiency of cells with increased gradient strength is sensitive to 3'-PI dysregulation. Our evidence suggests that receptor-driven 3'-PI signaling regulates the ability of a cell to capitalize on stronger directional inputs and minimize the effects of inaccurate turns to increase chemotactic efficiency. PMID- 18676655 TI - Calsequestrin-mediated mechanism for cellular calcium transient alternans. AB - Intracellular calcium transient alternans (CTA) has a recognized role in arrhythmogenesis, but its origin is not yet fully understood. Recent models of CTA are based on a steep relationship between calcium release from the sarcoplasmic reticulum (SR) and its calcium load before release. This mechanism alone, however, does not explain recent observations of CTA without diastolic SR calcium content alternations. In addition, nanoscopic imaging of calcium dynamics has revealed that the elementary calcium release units of the SR can become refractory independently of their local calcium content. Here we show using a new physiologically detailed mathematical model of calcium cycling that luminal gating of the calcium release channels (RyRs) mediated by the luminal buffer calsequestrin (CSQN) can cause CTA independently of the steepness of the release load relationship. In this complementary mechanism, CTA is caused by a beat-to beat alternation in the number of refractory RyR channels and can occur with or without diastolic SR calcium content alternans depending on pacing conditions and uptake dynamics. The model has unique features, in that it treats a realistic number of spatially distributed and diffusively coupled dyads, each one with a realistic number of RyR channels, and that luminal CSQN buffering and gating is incorporated based on experimental data that characterizes the effect of the conformational state of CSQN on its buffering properties. In addition to reproducing observed features of CTA, this multiscale model is able to describe recent experiments in which CSQN expression levels were genetically altered as well as to reproduce nanoscopic measurements of spark restitution properties. The ability to link microscopic properties of the calcium release units to whole cell behavior makes this model a powerful tool to investigate the arrhythmogenic role of abnormal calcium handling in many pathological settings. PMID- 18676657 TI - Conformational transition pathways explored by Monte Carlo simulation integrated with collective modes. AB - Conformational transitions between open/closed or free/bound states in proteins possess functional importance. We propose a technique in which the collective modes obtained from an anisotropic network model (ANM) are used in conjunction with a Monte Carlo (MC) simulation approach, to investigate conformational transition pathways and pathway intermediates. The ANM-MC technique is applied to adenylate kinase (AK) and hemoglobin. The iterative method, in which normal modes are continuously updated during the simulation, proves successful in accomplishing the transition between open-closed conformations of AK and tense relaxed forms of hemoglobin (C(alpha)-root mean square deviations between two end structures of 7.13 A and 3.55 A, respectively). Target conformations are reached by root mean-square deviations of 2.27 A and 1.90 A for AK and hemoglobin, respectively. The intermediate conformations overlap with crystal structures from the AK family within a 3.0-A root mean-square deviation. In the case of hemoglobin, the transition of tense-to-relaxed passes through the relaxed state. In both cases, the lowest-frequency modes are effective during transitions. The targeted Monte Carlo approach is used without the application of collective modes. Both the ANM-MC and targeted Monte Carlo techniques can explore sequences of events in transition pathways with an efficient yet realistic conformational search. PMID- 18676658 TI - Influence of liquid-layer thickness on pulmonary surfactant spreading and collapse. AB - Pulmonary surfactant spreads on the thin ( approximately 0.1 microm) liquid layer that lines the alveoli, forming a film that reduces surface tension and allows normal respiration. Pulmonary surfactant deposited in vitro on liquid layers that are several orders of magnitude thicker, however, does not reach the low surface tensions ( approximately 0.001 N/m) achieved in the lungs during exhalation when the surfactant film compresses. This is due to collapse, a surface phase transition during which the surfactant film, rather than decreasing surface tension by increasing its surface density, becomes thicker at constant surface tension ( approximately 0.024 N/m). Formation of the collapse phase requires transport of surfactant to collapse sites, and this transport can be hindered in thinner liquid layers by viscous resistance to motion. Our objective is to determine the effect of the liquid-layer thickness on surfactant transport, which might affect surfactant collapse. To this end, we developed a mathematical model that accounts for the effect of the liquid-layer thickness on surfactant transport, and focused on surfactant spreading and collapse. Model simulations showed a marked decrease in collapse rates for thinner liquid layers, but this decrease was not enough to completely explain differences in surfactant film behavior between in vitro and in situ experiments. PMID- 18676659 TI - Concentration dependence of alpha-synuclein fibril length assessed by quantitative atomic force microscopy and statistical-mechanical theory. AB - The initial concentration of monomeric amyloidogenic proteins is a crucial factor in the in vitro formation of amyloid fibrils. We use quantitative atomic force microscopy to study the effect of the initial concentration of human alpha synuclein on the mean length of mature alpha-synuclein fibrils, which are associated with Parkinson's disease. We determine that the critical initial concentration, below which low-molecular-weight species dominate and above which fibrils are the dominant species, lies at approximately 15 muM, in good agreement with earlier measurements using biochemical methods. In the concentration regime where fibrils dominate, we find that their mean length increases with initial concentration. These results correspond well to the qualitative predictions of a recent statistical-mechanical model of amyloid fibril formation. In addition, good quantitative agreement of the statistical-mechanical model with the measured mean fibril length as a function of initial protein concentration, as well as with the fibril length distributions for several protein concentrations, is found for reasonable values of the relevant model parameters. The comparison between theory and experiment yields, for the first time to our knowledge, an estimate of the magnitude of the free energies associated with the intermolecular interactions that govern alpha-synuclein fibril formation. PMID- 18676660 TI - Singlet oxygen is the major reactive oxygen species involved in photooxidative damage to plants. AB - Reactive oxygen species act as signaling molecules but can also directly provoke cellular damage by rapidly oxidizing cellular components, including lipids. We developed a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry-based quantitative method that allowed us to discriminate between free radical (type I)- and singlet oxygen ((1)O(2); type II)-mediated lipid peroxidation (LPO) signatures by using hydroxy fatty acids as specific reporters. Using this method, we observed that in non-photosynthesizing Arabidopsis (Arabidopsis thaliana) tissues, nonenzymatic LPO was almost exclusively catalyzed by free radicals both under normal and oxidative stress conditions. However, in leaf tissues under optimal growth conditions, (1)O(2) was responsible for more than 80% of the nonenzymatic LPO. In Arabidopsis mutants favoring (1)O(2) production, photooxidative stress led to a dramatic increase of (1)O(2) (type II) LPO that preceded cell death. Furthermore, under all conditions and in mutants that favor the production of superoxide and hydrogen peroxide (two sources for type I LPO reactions), plant cell death was nevertheless always preceded by an increase in (1)O(2)-dependent (type II) LPO. Thus, besides triggering a genetic cell death program, as demonstrated previously with the Arabidopsis fluorescent mutant, (1)O(2) plays a major destructive role during the execution of reactive oxygen species-induced cell death in leaf tissues. PMID- 18676661 TI - Two new clock proteins, LWD1 and LWD2, regulate Arabidopsis photoperiodic flowering. AB - The "light" signal from the environment sets the circadian clock to regulate multiple physiological processes for optimal rhythmic growth and development. One such process is the control of flowering time by photoperiod perception in plants. In Arabidopsis (Arabidopsis thaliana), the flowering time is determined by the correct interconnection of light input and signal output by the circadian clock. The identification of additional clock proteins will help to better dissect the complex nature of the circadian clock in Arabidopsis. Here, we show LIGHT-REGULATED WD1 (LWD1)/LWD2 as new clock proteins involved in photoperiod control. The lwd1lwd2 double mutant has an early-flowering phenotype, contributed by the significant phase shift of CONSTANS (CO), and, therefore, an increased expression of FLOWERING LOCUS T (FT) before dusk. Under entrainment conditions, the expression phase of oscillator (CIRCADIAN CLOCK ASSOCIATED1 [CCA1], LATE ELONGATED HYPOCOTYL [LHY], TIMING OF CAB EXPRESSION1 [TOC1], and EARLY FLOWERING4 [ELF4]) and output (GIGANTEA, FLAVIN-BINDING, KELCH REPEAT, F-BOX1, CYCLING DOF FACTOR1, CO, and FT) genes in the photoperiod pathway shifts approximately 3 h forward in the lwd1lwd2 double mutant. Both the oscillator (CCA1, LHY, TOC1, and ELF4) and output (COLD, CIRCADIAN RHYTHM, AND RNA BINDING2 and CHLOROPHYLL A/B BINDING PROTEIN2) genes have a short period length in the lwd1lwd2 double mutant. Our data imply that LWD1/LWD2 proteins function in close proximity to or within the circadian clock for photoperiodic flowering control. PMID- 18676662 TI - AtCHX13 is a plasma membrane K+ transporter. AB - Potassium (K+) homeostasis is essential for diverse cellular processes, although how various cation transporters collaborate to maintain a suitable K+ required for growth and development is poorly understood. The Arabidopsis (Arabidopsis thaliana) genome contains numerous cation:proton antiporters (CHX), which may mediate K+ transport; however, the vast majority of these transporters remain uncharacterized. Here, we show that AtCHX13 (At2g30240) has a role in K+ acquisition. AtCHX13 suppressed the sensitivity of yeast (Saccharomyces cerevisiae) mutant cells defective in K+ uptake. Uptake experiments using (86)Rb+ as a tracer for K+ demonstrated that AtCHX13 mediated high-affinity K+ uptake in yeast and in plant cells with a K(m) of 136 and 196 microm, respectively. Functional green fluorescent protein-tagged versions localized to the plasma membrane of both yeast and plant. Seedlings of null chx13 mutants were sensitive to K+ deficiency conditions, whereas overexpression of AtCHX13 reduced the sensitivity to K+ deficiency. Collectively, these results suggest that AtCHX13 mediates relatively high-affinity K+ uptake, although the mode of transport is unclear at present. AtCHX13 expression is induced in roots during K+-deficient conditions. These results indicate that one role of AtCHX13 is to promote K+ uptake into plants when K+ is limiting in the environment. PMID- 18676663 TI - Involvement of the pepper antimicrobial protein CaAMP1 gene in broad spectrum disease resistance. AB - Pathogen-inducible antimicrobial defense-related proteins have emerged as key antibiotic peptides and enzymes involved in disease resistance in plants. A novel antimicrobial protein gene, CaAMP1 (for Capsicum annuum ANTIMICROBIAL PROTEIN1), was isolated from pepper (C. annuum) leaves infected with Xanthomonas campestris pv vesicatoria. Expression of the CaAMP1 gene was strongly induced in pepper leaves not only during pathogen infection but also after exposure to abiotic elicitors. The purified recombinant CaAMP1 protein possessed broad-spectrum antimicrobial activity against phytopathogenic bacteria and fungi. CaAMP1:smGFP fusion protein was localized mainly in the external and intercellular regions of onion (Allium cepa) epidermal cells. The virus-induced gene silencing technique and gain-of-function transgenic plants were used to determine the CaAMP1 gene function in plant defense. Silencing of CaAMP1 led to enhanced susceptibility to X. campestris pv vesicatoria and Colletotrichum coccodes infection, accompanied by reduced PATHOGENESIS-RELATED (PR) gene expression. In contrast, overexpression of CaAMP1 in Arabidopsis (Arabidopsis thaliana) conferred broad-spectrum resistance to the hemibiotrophic bacterial pathogen Pseudomonas syringae pv tomato, the biotrophic oomycete Hyaloperonospora parasitica, and the fungal necrotrophic pathogens Fusarium oxysporum f. sp. matthiolae and Alternaria brassicicola. CaAMP1 overexpression induced the salicylic acid pathway-dependent genes PR1 and PR5 but not the jasmonic acid-dependent defense gene PDF1.2 during P. syringae pv tomato infection. Together, these results suggest that the antimicrobial CaAMP1 protein is involved in broad-spectrum resistance to bacterial and fungal pathogen infection. PMID- 18676665 TI - The stable, functional core of DdrA from Deinococcus radiodurans R1 does not restore radioresistance in vivo. AB - DdrA protein binds to and protects 3' DNA ends and is essential for preserving the genome integrity of Deinococcus radiodurans following treatment by gamma radiation in an environment lacking nutrients. Limited proteolysis was used to identify a stable and functional protein core, designated DdrA157, consisting of the first 157 residues of the protein. In vitro, the biochemical differences between wild-type and mutant proteins were modest. DdrA exhibits a strong bias in binding DNA with 3' extensions but not with 5' extensions. The mutant DdrA157 exhibited a greater affinity for 5' DNA ends but still bound to 3' ends more readily. However, when we replaced the wild-type ddrA gene with the mutant gene for ddrA157, the resulting D. radiodurans strain became almost as sensitive to gamma radiation as the ddrA knockout strain. These results suggest that while the stable protein core DdrA157 is functional for DNA binding and protection assays in vitro, the carboxyl terminus is required for important functions in vivo. The C terminus may therefore be required for protein or DNA interactions or possibly as a regulatory region for DNA binding or activities not yet identified. PMID- 18676664 TI - The CorA Mg2+ channel is required for the virulence of Salmonella enterica serovar typhimurium. AB - CorA is the primary Mg(2+) channel in Salmonella enterica serovar Typhimurium. A corA mutant is attenuated in mice and defective for invasion of and replication within epithelial cells. Microarray studies show that several virulence effectors are repressed in a corA mutant strain, which ultimately manifests itself as a decrease in virulence. PMID- 18676666 TI - Regulation of CorA Mg2+ channel function affects the virulence of Salmonella enterica serovar typhimurium. AB - The CorA Mg(2+) channel is the primary source of intracellular Mg(2+) in Salmonella enterica serovar Typhimurium. In another study, we found that a strain lacking corA was attenuated in mice and also defective for invasion and replication within Caco-2 epithelial cells (K. M. Papp-Wallace, M. Nartea, D. G. Kehres, S. Porwollik, M. McClelland, S. J. Libby, F. C. Fang, and M. E. Maguire, J. Bacteriol. 190:6517-6523, 2008). Therefore, we further examined Salmonella interaction with Caco-2 epithelial cells. Inhibiting CorA acutely or chronically with a high concentration of a selective inhibitor, Co(III) hexaammine, had no effect on S. enterica serovar Typhimurium invasion of Caco-2 epithelial cells. Complementing the corA mutation with corA from various species rescued the invasion defect only if the complementing allele was functional and if it was evolutionarily similar to S. enterica serovar Typhimurium CorA. One explanation for these results could be that regulation of CorA function is needed for optimal virulence. Further experiments examining corA transcription, CorA protein content, CorA transport, and cell Mg(2+) content indicated that both CorA expression and CorA function are differentially regulated. Moreover, the rates of Mg(2+) influx via CorA are not closely correlated with either protein levels or Mg(2+) content. We conclude that loss of the CorA protein disrupts a regulatory network(s) with the ultimate phenotype of decreased virulence. This conclusion is compatible with the microarray results in our other study, which showed that loss of corA resulted in changes in transcription (and protein expression) in multiple metabolic pathways (Papp-Wallace et al., J. Bacteriol. 190:6517-6523, 2008). Further study of the regulation of CorA expression and function provides an opportunity to dissect the complexity of Mg(2+) homeostasis and its ties to virulence within the bacterium. PMID- 18676668 TI - X-ray absorption spectroscopy as a probe of microbial sulfur biochemistry: the nature of bacterial sulfur globules revisited. AB - The chemical nature of the sulfur in bacterial sulfur globules has been the subject of controversy for a number of years. Sulfur K-edge X-ray absorption spectroscopy (XAS) is a powerful technique for probing the chemical forms of sulfur in situ, but two groups have used it with very different conclusions. The root of the controversy lies with the different detection strategies used by the two groups, which result in very different spectra. This paper seeks to resolve the controversy. We experimentally demonstrate that the use of transmittance detection for sulfur K-edge XAS measurements is highly prone to spectroscopic distortions and that much of the published work on sulfur bacteria is very likely based on distorted data. We also demonstrate that all three detection methods used for X-ray absorption experiments yield essentially identical spectra when the measurements are carried out under conditions where no experimental distortions are expected. Finally, we turn to the original question--the chemical nature of bacterial sulfur. We examine isolated sulfur globules of Allochromatium vinosum and intact cells of a strain of magnetotactic coccus and show that XAS indicates the presence of a chemical form of sulfur resembling S(8). PMID- 18676667 TI - The vibrio cholerae hybrid sensor kinase VieS contributes to motility and biofilm regulation by altering the cyclic diguanylate level. AB - Phosphorelay systems are important mediators of signal transduction during bacterial adaptation to new environments. Previously we described the vieSAB operon, encoding a putative three-protein component phosphorelay involved in regulating Vibrio cholerae virulence gene expression. At least part of the regulatory activity of VieSAB is exerted through the cyclic diguanylate (c-di GMP)-degrading activity of the putative response regulator VieA. So far no direct evidence that VieSAB encodes a phosphorelay system exists. In addition, the role VieS plays in modulating VieA activity remains unclear. To address these questions, we expressed and purified VieA and a soluble cytoplasmic portion of VieS and used them in autophosphorylation and phosphotransfer assays. These assays showed that VieS has kinase activity in vitro and is able to selectively phosphorylate VieA. A phenotypic comparison revealed that deletion of vieS results in increased biofilm production comparable to that seen for deletion of vieA, whereas motility was decreased only slightly in the DeltavieS mutant compared to the profound defect observed in a DeltavieA mutant. We also found that the DeltavieS strain has a lower level of vieA transcript and, similar to a DeltavieA mutant, an increased intracellular level of c-di-GMP. Further analysis using site-directed vieA mutants showed that some of the phenotypes observed were due to the phosphorylation status of VieA. The evidence presented in this report is the first to link VieS and VieA biochemically and genetically, lending support to the hypothesis that these proteins function together in a signaling system. PMID- 18676669 TI - Isorenieratene biosynthesis in green sulfur bacteria requires the cooperative actions of two carotenoid cyclases. AB - The cyclization of lycopene to gamma- or beta-carotene is a major branch point in the biosynthesis of carotenoids in photosynthetic bacteria. Four families of carotenoid cyclases are known, and each family includes both mono- and dicyclases, which catalyze the formation of gamma- and beta-carotene, respectively. Green sulfur bacteria (GSB) synthesize aromatic carotenoids, of which the most commonly occurring types are the monocyclic chlorobactene and the dicyclic isorenieratene. Recently, the cruA gene, encoding a conserved hypothetical protein found in the genomes of all GSB and some cyanobacteria, was identified as a lycopene cyclase. Further genomic analyses have found that all available fully sequenced genomes of GSB encode an ortholog of cruA. Additionally, the genomes of all isorenieratene-producing species of GSB encode a cruA paralog, now named cruB. The cruA gene from the chlorobactene-producing GSB species Chlorobaculum tepidum and both cruA and cruB from the brown-colored, isorenieratene-producing GSB species Chlorobium phaeobacteroides strain DSM 266(T) were heterologously expressed in lycopene- and neurosporene-producing strains of Escherichia coli, and the cruB gene of Chlorobium clathratiforme strain DSM 5477(T) was also heterologously expressed in C. tepidum by inserting the gene at the bchU locus. The results show that CruA is probably a lycopene monocyclase in all GSB and that CruB is a gamma-carotene cyclase in isorenieratene-producing species. Consequently, the branch point for the synthesis of mono- and dicyclic carotenoids in GSB seems to be the modification of gamma-carotene, rather than the cyclization of lycopene as occurs in cyanobacteria. PMID- 18676670 TI - Phage-associated mutator phenotype in group A streptococcus. AB - Defects in DNA mismatch repair (MMR) occur frequently in natural populations of pathogenic and commensal bacteria, resulting in a mutator phenotype. We identified a unique genetic element in Streptococcus pyogenes strain SF370 that controls MMR via a dynamic process of prophage excision and reintegration in response to growth. In S. pyogenes, mutS and mutL are organized on a polycistronic mRNA under control of a common promoter. Prophage SF370.4 is integrated between the two genes, blocking expression of the downstream gene (mutL) and resulting in a mutator phenotype. However, in rapidly growing cells the prophage excises and replicates as an episome, allowing mutL to be expressed. Excision of prophage SF370.4 and expression of MutL mRNA occur simultaneously during early logarithmic growth when cell densities are low; this brief window of MutL gene expression ends as the cell density increases. However, detectable amounts of MutL protein remain in the cell until the onset of stationary phase. Thus, MMR in S. pyogenes SF370 is functional in exponentially growing cells but defective when resources are limiting. The presence of a prophage integrated into the 5' end of mutL correlates with a mutator phenotype (10(-7) to 10(-8) mutation/generation, an approximately a 100-fold increase in the rate of spontaneous mutation compared with prophage-free strains [10(-9) to 10(-10) mutation/generation]). Such genetic elements may be common in S. pyogenes since 6 of 13 completed genomes have related prophages, and a survey of 100 strains found that about 20% of them are positive for phages occupying the SF370.4 attP site. The dynamic control of a major DNA repair system by a bacteriophage is a novel method for achieving the mutator phenotype and may allow the organism to respond rapidly to a changing environment while minimizing the risks associated with long term hypermutability. PMID- 18676671 TI - Characterization of the traD operon of naphthalene-catabolic plasmid NAH7: a host range modifier in conjugative transfer. AB - Pseudomonas putida G7 carries a naphthalene-catabolic and self-transmissible plasmid, NAH7, which belongs to the IncP-9 incompatibility group. Adjacent to the putative origin of conjugative transfer (oriT) of NAH7 are three genes, traD, traE, and traF, whose functions and roles in conjugation were previously unclear. These three genes were transcribed monocistronically and thus were designated the traD operon. Mutation of the three genes in the traD operon resulted in 10- to 10(5)-fold decreases in the transfer frequencies of the plasmids from Pseudomonas to Pseudomonas and Escherichia coli and from E. coli to E. coli. On the other hand, the traD operon was essential for the transfer of NAH7 from E. coli to Pseudomonas strains. These results indicated that the traD operon is a host-range modifier in the conjugative transfer of NAH7. The TraD, TraE, and TraF proteins were localized in the cytoplasm, periplasm, and membrane, respectively, in strain G7 cells. Our use of a bacterial two-hybrid assay system showed that TraE interacted in vivo with other essential components for conjugative transfer, including TraB (coupling protein), TraC (relaxase), and MpfH (a channel subunit in the mating pair formation system). PMID- 18676672 TI - The pangenome structure of Escherichia coli: comparative genomic analysis of E. coli commensal and pathogenic isolates. AB - Whole-genome sequencing has been skewed toward bacterial pathogens as a consequence of the prioritization of medical and veterinary diseases. However, it is becoming clear that in order to accurately measure genetic variation within and between pathogenic groups, multiple isolates, as well as commensal species, must be sequenced. This study examined the pangenomic content of Escherichia coli. Six distinct E. coli pathovars can be distinguished using molecular or phenotypic markers, but only two of the six pathovars have been subjected to any genome sequencing previously. Thus, this report provides a seminal description of the genomic contents and unique features of three unsequenced pathovars, enterotoxigenic E. coli, enteropathogenic E. coli, and enteroaggregative E. coli. We also determined the first genome sequence of a human commensal E. coli isolate, E. coli HS, which will undoubtedly provide a new baseline from which workers can examine the evolution of pathogenic E. coli. Comparison of 17 E. coli genomes, 8 of which are new, resulted in identification of approximately 2,200 genes conserved in all isolates. We were also able to identify genes that were isolate and pathovar specific. Fewer pathovar-specific genes were identified than anticipated, suggesting that each isolate may have independently developed virulence capabilities. Pangenome calculations indicate that E. coli genomic diversity represents an open pangenome model containing a reservoir of more than 13,000 genes, many of which may be uncharacterized but important virulence factors. This comparative study of the species E. coli, while descriptive, should provide the basis for future functional work on this important group of pathogens. PMID- 18676673 TI - A defect in menadione biosynthesis induces global changes in gene expression in Staphylococcus aureus. AB - Both the high-resolution two-dimensional protein gel electrophoresis technique and full-genome DNA microarrays were used for identification of Staphylococcus aureus genes whose expression was changed by a mutation in menD. Because the electron transport chain is interrupted, the mutant should be unable to use oxygen and nitrate as terminal electron acceptors. Consistent with this, a mutation in menD was found to cause a gene expression pattern typically detected under anaerobic conditions in wild-type cells: proteins involved in glycolytic as well as in fermentation pathways were upregulated, whereas tricarboxylic acid (TCA) cycle enzymes were significantly downregulated. Moreover, the expression of genes encoding enzymes for nitrate respiration and the arginine deiminase pathway was strongly increased in the mutant strain. These results indicate that the menD mutant, just as the site-directed S. aureus hemB mutant, generates ATP from glucose or fructose mainly by substrate phosphorylation and might be defective in utilizing a variety of carbon sources, including TCA cycle intermediates and compounds that generate ATP only via electron transport phosphorylation. Of particular interest is that there are also differences in the gene expression patterns between hemB and menD mutants. While some anaerobically active enzymes were present in equal amounts in both strains (Ldh1, SACOL2535), other classically anaerobic enzymes seem to be present in higher amounts either in the hemB mutant (e.g., PflB, Ald1, IlvA1) or in the menD mutant (arc operon). Only genes involved in nitrate respiration and the ald1 operon seem to be additionally regulated by a depletion of oxygen in the hemB and/or menD mutant. PMID- 18676674 TI - Dual promoters control expression of the Bacillus anthracis virulence factor AtxA. AB - The AtxA virulence regulator of Bacillus anthracis is required for toxin and capsule gene expression. AtxA is a phosphotransferase system regulatory domain containing protein whose activity is regulated by phosphorylation/dephosphorylation of conserved histidine residues. Here we report that transcription of the atxA gene occurs from two independent promoters, P1 (previously described by Dai et al. [Z. Dai, J. C. Sirard, M. Mock, and T. M. Koehler, Mol. Microbiol. 16:1171-1181, 1995]) and P2, whose transcription start sites are separated by 650 bp. Both promoters have -10 and -35 consensus sequences compatible with recognition by sigma(A)-containing RNA polymerase, and neither promoter depends on the sporulation sigma factor SigH. The dual promoter activity and the extended untranslated mRNA suggest that as-yet-unknown regulatory mechanisms may act on this region to influence the level of AtxA in the cell. PMID- 18676675 TI - Transition state regulator AbrB inhibits transcription of Bacillus amyloliquefaciens FZB45 phytase through binding at two distinct sites located within the extended phyC promoter region. AB - We have previously identified the phyC gene of Bacillus amyloliquefaciens FZB45, encoding extracellular phytase, as a member of the PhoP regulon, which is expressed only during phosphate starvation. Its sigma(A)-dependent promoter is positively and negatively regulated by the phosphorylated PhoP response regulator in a phosphate-dependent manner (O. Makarewicz, S. Dubrac, T. Msadek, and R. Borriss, J. Bacteriol. 188:6953-6965, 2006). Here, we provide experimental evidence that the transcription of phyC underlies a second control mechanism exerted by the global transient-phase regulator protein, AbrB, which hinders its expression during exponential growth. Gel mobility shift and DNase I footprinting experiments demonstrated that AbrB binds to two different regions in the phyC promoter region that are separated by about 200 bp. One binding site is near the divergently orientated yodU gene, and the second site is located downstream of the phyC promoter and extends into the coding region of the phyC gene. Cooperative binding to the two distant binding regions is necessary for the AbrB directed repression of phyC transcription. AbrB does not affect the transcription of the neighboring yodU gene. PMID- 18676676 TI - Reactive oxygen species mediate p53 activation and apoptosis induced by sodium nitroprusside in SH-SY5Y cells. AB - Sodium nitroprusside (SNP) is a water-soluble iron nitrosyl complex clinically used as a powerful vasodilator for treatment of hypertension; and, in basic research, it has been used to mainly investigate the cytotoxic effects of nitrosative stress. Although NO is considered a pharmacologically active molecule, not all of the biological effects of SNP are dependent on its NO moiety. To elucidate the molecular executioner(s) responsible for SNP cytotoxicity, this study determines the involvement of oxidative stress in p53 activation and apoptotic induction elicited by SNP in SH-SY5Y neuroblastoma cells. We demonstrate that proapoptotic activity of SNP is independent of NO production, because SNP and its 2-day light-exhausted compound SNP(ex) trigger apoptosis to the same extent. We provide evidence for the occurrence of oxidative stress and oxidative damage during both SNP and SNP(ex) exposure and demonstrate that iron-derived reactive oxygen species (ROS) are the genuine mediators of their cytotoxicity. We show that p53 is equally activated upon both SNP and SNP(ex) treatments. Moreover, as demonstrated by small interfering RNA experiments, we indicate its primary role in the induction of apoptosis, suggesting the ineffectiveness of NO in its engagement. The attenuation of p53 levels, obtained by oxy-radical scavengers, is consistent with the recovery of cell viability and ROS decrease, demonstrate that SNP-mediated p53 activation is an event triggered by ROS and/or ROS-mediated damages. Together, our results suggest that investigations of the physiopathological effects of SNP should consider the role of ROS, other than NO, particularly in some conditions such as apoptotic induction and p53 activation. PMID- 18676677 TI - Anti-inflammatory, antiproliferative, and cytoprotective activity of NO chimera nitrates of use in cancer chemoprevention. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have shown promise in colorectal cancer (CRC), but they are compromised by gastrotoxicity. NO-NSAIDs are hybrid nitrates conjugated to an NSAID designed to exploit the gastroprotective properties of NO bioactivity. The NO chimera ethyl 2-((2,3 bis(nitrooxy)propyl)disulfanyl)benzoate (GT-094), a novel nitrate containing an NSAID and disulfide pharmacophores, is effective in vivo in rat models of CRC and is a lead compound for design of agents of use in CRC. Preferred chemopreventive agents possess 1) antiproliferative and 2) anti-inflammatory actions and 3) the ability to induce cytoprotective phase 2 enzymes. To determine the contribution of each pharmacophore to the biological activity of GT-094, these three biological activities were studied in vitro in compounds that deconstructed the structural elements of the lead GT-094. The anti-inflammatory and antiproliferative actions of GT-094 in vivo were recapitulated in vitro, and GT 094 was seen to induce phase 2 enzymes via the antioxidant responsive element. In the variety of colon, macrophage-like, and liver cell lines studied, the evidence from structure-activity relationships was that the disulfide structural element of GT-094 is the dominant contributor in vitro to the anti-inflammatory activity, antiproliferation, and enzyme induction. The results provide a direction for lead compound refinement. The evidence for a contribution from the NO mimetic activity of nitrates in vitro was equivocal, and combinations of nitrates with acetylsalicylic acid were inactive. PMID- 18676678 TI - Identification of a putative intracellular allosteric antagonist binding-site in the CXC chemokine receptors 1 and 2. AB - The chemokine receptors CXCR1 and CXCR2 are G-protein-coupled receptors (GPCRs) implicated in mediating cellular functions associated with the inflammatory response. Potent CXCR2 receptor antagonists have been discovered, some of which have recently entered clinical development. The aim of this study was to identify key amino acid residue differences between CXCR1 and CXCR2 that influence the relative antagonism by two compounds that have markedly different chemical structures. By investigating the effects of domain switching and point mutations, we found that the second extracellular loop, which contained significant amino acid sequence diversity, was not important for compound antagonism. We were surprised to find that switching the intracellular C-terminal 60 amino acid domains of CXCR1 and CXCR2 caused an apparent reversal of antagonism at these two receptors. Further investigation showed that a single amino acid residue, lysine 320 in CXCR2 and asparagine 311 in CXCR1, plays a predominant role in describing the relative antagonism of the two compounds. Homology modeling studies based on the structure of bovine rhodopsin indicated a potential intracellular antagonist binding pocket involving lysine 320. We conclude that residue 320 in CXCR2 forms part of a potential allosteric binding pocket on the intracellular side of the receptor, a site that is distal to the orthosteric site commonly assumed to be the location of antagonist binding to GPCRs. The existence of a common intracellular allosteric binding site at GPCRs related to CXCR2 may be of value in the design of novel antagonists for therapeutic intervention. PMID- 18676679 TI - Selenite reactivates silenced genes by modifying DNA methylation and histones in prostate cancer cells. AB - DNA hypermethylation is a common epigenetic alteration in human prostate cancer and is considered to contribute to development of this disease. Accumulating data suggest that dietary factors may alter cancer risk by modifications of epigenetic processes in the cell. The present study was designed to investigate whether selenium (Se) would alter epigenetic events to regulate methylation-silenced genes in human prostate cancer cells. DNA methylation, histone modifications and gene expression were studied in LNCaP cells after selenite treatment using polymerase chain reaction, western blot analysis, chromatin immunoprecipitation assay and enzymatic activity assay. Our study shows that selenite treatment caused partial promoter DNA demethylation and reexpression of the pi-class glutathione-S-transferase (GSTP1) in LNCaP cells in a dose- and time-dependent manner. Selenite treatment decreased messenger RNA levels of DNA methyltransferases (DNMTs) 1 and 3A and protein levels of DNMT1. Selenite also decreased histone deacetylase activity and increased levels of acetylated lysine 9 on histone H3 (H3-Lys 9), but decreased levels of methylated H3-Lys 9. Selenite treatment reduced levels of DNMT1 and methylated H3-Lys 9 associated with the GSTP1 promoter, but increased levels of acetylated H3-Lys 9 associated with this promoter. Additionally, selenite treatment decreased general DNA methylation and caused partial promoter demethylation and reexpression of the tumor suppressor adenomatous polyposis coli and cellular stress response 1, a gene involving tumor growth and metastasis. Our study demonstrates that Se can epigenetically modulate DNA and histones to activate methylation-silenced genes. These epigenetic modifications may contribute to cancer prevention by Se. PMID- 18676680 TI - Pathway-based evaluation of 380 candidate genes and lung cancer susceptibility suggests the importance of the cell cycle pathway. AB - Common genetic variation may play an important role in altering lung cancer risk. We conducted a pathway-based candidate gene evaluation to identify genetic variations that may be associated with lung cancer in a population-based case control study in Xuan Wei, China (122 cases and 111 controls). A total of 1260 single-nucleotide polymorphisms (SNPs) in 380 candidate genes for lung cancer were successfully genotyped and assigned to one of 10 pathways based on gene ontology. Logistic regression was used to assess the marginal effect of each SNP on lung cancer susceptibility. The minP test was used to identify statistically significant associations at the gene level. Important pathways were identified using a test of proportions and the rank truncated product methods. The cell cycle pathway was found as the most important pathway (P = 0.044) with four genes significantly associated with lung cancer (PLA2G6 minP = 0.001, CCNA2 minP = 0.006, GSK3 beta minP = 0.007 and EGF minP = 0.013), after adjusting for multiple comparisons. Interestingly, most cell cycle genes that were associated with lung cancer in this analysis were concentrated in the AKT signaling pathway, which is essential for regulation of cell cycle progression and cellular survival, and may be important in lung cancer etiology in Xuan Wei. These results should be viewed as exploratory until they are replicated in a larger study. PMID- 18676682 TI - Cold-hearted--the electrocardiogram in hypothermia. PMID- 18676681 TI - Interactions between SIRT1 and AP-1 reveal a mechanistic insight into the growth promoting properties of alumina (Al2O3) nanoparticles in mouse skin epithelial cells. AB - The physicochemical properties of nanomaterials differ from those of the bulk material of the same composition. However, little is known about the underlying effects of these particles in carcinogenesis. The purpose of this study was to determine the mechanisms involved in the carcinogenic properties of nanoparticles using aluminum oxide (Al(2)O(3)/alumina) nanoparticles as the prototype. Well established mouse epithelial JB6 cells, sensitive to neoplastic transformation, were used as the experimental model. We demonstrate that alumina was internalized and maintained its physicochemical composition inside the cells. Alumina increased cell proliferation (53%), proliferating cell nuclear antigen (PCNA) levels, cell viability and growth in soft agar. The level of manganese superoxide dismutase, a key mitochondrial antioxidant enzyme, was elevated, suggesting a redox signaling event. In addition, the levels of reactive oxygen species and the activities of the redox sensitive transcription factor activator protein-1 (AP-1) and a longevity-related protein, sirtuin 1 (SIRT1), were increased. SIRT1 knockdown reduces DNA synthesis, cell viability, PCNA levels, AP-1 transcriptional activity and protein levels of its targets, JunD, c-Jun and BcL xl, more than controls do. Immunoprecipitation studies revealed that SIRT1 interacts with the AP-1 components c-Jun and JunD but not with c-Fos. The results identify SIRT1 as an AP-1 modulator and suggest a novel mechanism by which alumina nanoparticles may function as a potential carcinogen. PMID- 18676683 TI - Interaction between statins and clopidogrel: is there anything clinically relevant? AB - Since their introduction several years ago, the 3-hydroxy-3-methylglutaryl coenzyme A (HMGCoA) reductase inhibitors-the statins-have been widely used for hyperlipidemia and for the primary/secondary prevention of cardiovascular diseases. They have been shown to be safe as well as efficacious in a number of different clinical trials; however, studies have suggested that they can interact with other co-administered therapies. More recently, the thienopyridines have been successfully integrated with the conventional medical treatment of coronary disease as they showed effectiveness in reducing platelet activity both in stable and unstable settings. They also improve the outcome of patients treated with percutaneous coronary intervention. The potential interaction of statins and thienopyridines is a matter of concern. Despite some preclinical data suggesting an interaction between statins metabolized by the liver cytochrome P3A4-such as atorvastatin, lovastatin and simvastatin-and clopidogrel, there is no compelling clinical evidence to stop their co-administration. PMID- 18676684 TI - Serum matrix metalloproteinase-9 and coronary heart disease: a prospective study in middle-aged men. AB - BACKGROUND: Matrix metalloproteinase-9 (MMP-9) has a potential role in arterial plaque rupture, but its relation to risk of coronary heart disease (CHD) is uncertain. AIM: To determine whether circulating levels of serum MMP-9 are prospectively related to the risk of CHD in the general population. METHODS: We measured baseline MMP-9 levels in stored serum samples of subjects in a case control study nested within a prospective study of 5661 men followed up for 16 years for CHD events (465 cases, 1076 controls). RESULTS: MMP-9 values were associated with cigarette smoking, and with several inflammatory and haemostatic markers, but not with age, body mass index, blood pressure or lipid measurements. Men in the top third of baseline MMP-9 levels had an age-adjusted odds ratio (OR) for CHD of 1.37 (95% CI 1.04-1.82) compared with those in the bottom third. Adjustment for conventional risk factors (smoking in particular) reduced the odds ratio to borderline significance: OR 1.28 (95% CI 0.95-1.74), while additional adjustment for two markers of generalized inflammation, interleukin-6 and C reactive protein, further attenuated the association: OR 1.13 (0.82-1.56). CONCLUSION: Serum MMP-9 has a modest association with incident CHD in the general population, which is not independent of cigarette smoking exposure and circulating markers of generalized inflammation. MMP-9 is unlikely to be a clinically useful biomarker of CHD risk, but may still play a role in the pathogenesis of CHD. PMID- 18676685 TI - Ultrafast sodium channel block by dietary fish oil prevents dofetilide-induced ventricular arrhythmias in rabbit hearts. AB - Several epidemiologic and clinical studies show that following myocardial infarction, dietary supplements of omega-3 polyunsaturated fatty acids (omega3FA) reduce sudden death. Animal data show that omega3FA have antiarrhythmic properties, but their mechanisms of action require further elucidation. The effects of omega3FA supplementation were studied in female rabbits to analyze whether their antiarrhythmic effects are due to a reduction of triangulation, reverse use-dependence, instability, and dispersion (TRIaD) of the cardiac action potential (TRIaD as a measure of proarrhythmic effects). In Langendorff-perfused hearts challenged by a selective rapidly activating delayed rectifier potassium current inhibitor that has been shown to exhibit proarrhythmic effects (dofetilide; 1 to 100 nM), omega3FA pretreatment (30 days; n=6) prolonged the plateau phase of the monophasic action potential; did not slow the terminal fast repolarization; reduced the dofetilide-induced prolongation of the action potential duration; reduced dofetilide-induced triangulation; and reduced dofetilide-induced reverse use-dependence, instability of repolarization, and dispersion. Dofetilide reduced excitability in omega3FA-pretreated hearts but not in control hearts. Whereas torsades de pointes (TdP) were observed in five out of six in control hearts, none were observed in omega3FA-pretreated hearts. Docosahexaenoic acid (DHA) inhibited the sodium current with ultrafast kinetics. Dietary omega3FA supplementation markedly reduced dofetilide-induced TRIaD and abolished dofetilide-induced TdP. Ultrafast sodium channel block by DHA may account for the antiarrhythmic protection of the dietary supplements of omega3FA against dofetilide-induced proarrhythmia observed in this animal model. PMID- 18676686 TI - Mechanisms of atrial fibrillation termination by rapidly unbinding Na+ channel blockers: insights from mathematical models and experimental correlates. AB - Atrial fibrillation (AF) is the most common sustained clinical arrhythmia and is a problem of growing proportions. Recent studies have increased interest in fast unbinding Na(+) channel blockers like vernakalant (RSD1235) and ranolazine for AF therapy, but the mechanism of efficacy is poorly understood. To study how fast unbinding I(Na) blockers affect AF, we developed realistic mathematical models of state-dependent Na(+) channel block, using a lidocaine model as a prototype, and studied the effects on simulated cholinergic AF in two- and three-dimensional atrial substrates. We then compared the results with in vivo effects of lidocaine on vagotonic AF in dogs. Lidocaine action was modeled with the Hondeghem-Katzung modulated-receptor theory and maximum affinity for activated Na(+) channels. Lidocaine produced frequency-dependent Na(+) channel blocking and conduction slowing effects and terminated AF in both two- and three-dimensional models with concentration-dependent efficacy (maximum approximately 89% at 60 microM). AF termination was not related to increases in wavelength, which tended to decrease with the drug, but rather to decreased source Na(+) current in the face of large ACh-sensitive K(+) current-related sinks, leading to the destabilization of primary generator rotors and a great reduction in wavebreak, which caused primary rotor annihilations in the absence of secondary rotors to resume generator activity. Lidocaine also reduced the variability and maximum values of the dominant frequency distribution during AF. Qualitatively similar results were obtained in vivo for lidocaine effects on vagal AF in dogs, with an efficacy of 86% at 2 mg/kg iv, as well as with simulations using the guarded-receptor model of lidocaine action. These results provide new insights into the mechanisms by which rapidly unbinding class I antiarrhythmic agents, a class including several novel compounds of considerable promise, terminate AF. PMID- 18676688 TI - Fasudil inhibits the myogenic response in the fetal pulmonary circulation. AB - In addition to high pulmonary vascular resistance (PVR) and low pulmonary blood flow, the fetal pulmonary circulation is characterized by mechanisms that oppose vasodilation. Past work suggests that high myogenic tone contributes to high PVR and may contribute to autoregulation of blood flow in the fetal lung. Rho-kinase (ROCK) can mediate the myogenic response in the adult systemic circulation, but whether high ROCK activity contributes to the myogenic response and modulates time-dependent vasodilation in the developing lung circulation are unknown. We studied the effects of fasudil, a ROCK inhibitor, on the hemodynamic response during acute compression of the ductus arteriosus (DA) in chronically prepared, late-gestation fetal sheep. Acute DA compression simultaneously induces two opposing responses: 1) blood flow-induced vasodilation through increased shear stress that is mediated by NO release and 2) stretch-induced vasoconstriction (i.e., the myogenic response). The myogenic response was assessed during acute DA compression after treatment with N(omega)-nitro-L-arginine, an inhibitor of nitric oxide synthase, to block flow-induced vasodilation and unmask the myogenic response. Intrapulmonary fasudil infusion (100 microg over 10 min) did not enhance flow-induced vasodilation during brief DA compression but reduced the myogenic response by 90% (P<0.05). During prolonged DA compression, fasudil prevented the time-dependent decline in left pulmonary artery blood flow at 2 h (183+/-29 vs. 110+/-11 ml/min with and without fasudil, respectively; P<0.001). We conclude that high ROCK activity opposes pulmonary vasodilation in utero and that the myogenic response maintains high PVR in the normal fetal lung through ROCK activation. PMID- 18676687 TI - Proteasome inhibition decreases cardiac remodeling after initiation of pressure overload. AB - We tested the possibility that proteasome inhibition may reverse preexisting cardiac hypertrophy and improve remodeling upon pressure overload. Mice were submitted to aortic banding and followed up for 3 wk. The proteasome inhibitor epoxomicin (0.5 mg/kg) or the vehicle was injected daily, starting 2 wk after banding. At the end of the third week, vehicle-treated banded animals showed significant (P<0.05) increase in proteasome activity (PA), left ventricle-to tibial length ratio (LV/TL), myocyte cross-sectional area (MCA), and myocyte apoptosis compared with sham-operated animals and developed signs of heart failure, including increased lung weight-to-TL ratio and decreased ejection fraction. When compared with that group, banded mice treated with epoxomicin showed no increase in PA, a lower LV/TL and MCA, reduced apoptosis, stabilized ejection fraction, and no signs of heart failure. Because overload-mediated cardiac remodeling largely depends on the activation of the proteasome-regulated transcription factor NF-kappaB, we tested whether epoxomicin would prevent this activation. NF-kappaB activity increased significantly upon overload, which was suppressed by epoxomicin. The expression of NF-kappaB-dependent transcripts, encoding collagen types I and III and the matrix metalloprotease-2, increased (P<0.05) after banding, which was abolished by epoxomicin. The accumulation of collagen after overload, as measured by histology, was 75% lower (P<0.05) with epoxomicin compared with vehicle. Myocyte apoptosis increased by fourfold in hearts submitted to aortic banding compared with sham-operated hearts, which was reduced by half upon epoxomicin treatment. Therefore, we propose that proteasome inhibition after the onset of pressure overload rescues ventricular remodeling by stabilizing cardiac function, suppressing further progression of hypertrophy, repressing collagen accumulation, and reducing myocyte apoptosis. PMID- 18676689 TI - Perivascular nitric oxide and superoxide in neonatal cerebral hypoxia-ischemia. AB - Decreased cerebral blood flow (CBF) has been observed following the resuscitation from neonatal hypoxic-ischemic injury, but its mechanism is not known. We address the hypothesis that reduced CBF is due to a change in nitric oxide (NO) and superoxide anion O(2)(-) balance secondary to endothelial NO synthase (eNOS) uncoupling with vascular injury. Wistar rats (7 day old) were subjected to cerebral hypoxia-ischemia by unilateral carotid occlusion under isoflurane anesthesia followed by hypoxia with hyperoxic or normoxic resuscitation. Expired CO(2) was determined during the period of hyperoxic or normoxic resuscitation. Laser-Doppler flowmetry was used with isoflurane anesthesia to monitor CBF, and cerebral perivascular NO and O(2)(-) were determined using fluorescent dyes with fluorescence microscopy. The effect of tetrahydrobiopterin supplementation on each of these measurements and the effect of apocynin and N(omega)-nitro-L arginine methyl ester (L-NAME) administration on NO and O(2)(-) were determined. As a result, CBF in the ischemic cortex declined following the onset of resuscitation with 100% O(2) (hyperoxic resuscitation) but not room air (normoxic resuscitation). Expired CO(2) was decreased at the onset of resuscitation, but recovery was the same in normoxic and hyperoxic resuscitated groups. Perivascular NO-induced fluorescence intensity declined, and O(2)(-)-induced fluorescence increased in the ischemic cortex after hyperoxic resuscitation up to 24 h postischemia. L-NAME treatment reduced O(2)(-) relative to the nonischemic cortex. Apocynin treatment increased NO and reduced O(2)(-) relative to the nonischemic cortex. The administration of tetrahydrobiopterin following the injury increased perivascular NO, reduced perivascular O(2)(-), and increased CBF during hyperoxic resuscitation. These results demonstrate that reduced CBF follows hyperoxic resuscitation but not normoxic resuscitation after neonatal hypoxic-ischemic injury, accompanied by a reduction in perivascular production of NO and an increase in O(2)(-). The finding that tetrahydrobiopterin, apocynin, and L-NAME normalized radical production suggests that the uncoupling of perivascular NOS, probably eNOS, due to acquired relative tetrahydrobiopterin deficiency occurs after neonatal hypoxic-ischemic brain injury. It appears that both NOS uncoupling and the activation of NADPH oxidase participate in the changes of reactive oxygen concentrations seen in cerebral hypoxic-ischemic injury. PMID- 18676690 TI - Cardiac hypertrophy is associated with altered thioredoxin and ASK-1 signaling in a mouse model of menopause. AB - Oxidative stress is implicated in menopause-associated hypertension and cardiovascular disease. The role of antioxidants in this process is unclear. We questioned whether the downregulation of thioredoxin (TRX) is associated with oxidative stress and the development of hypertension and target-organ damage (cardiac hypertrophy) in a menopause model. TRX is an endogenous antioxidant that also interacts with signaling molecules, such as apoptosis signal-regulated kinase 1 (ASK-1), independently of its antioxidant function. Aged female wild type (WT) and follitropin receptor knockout (FORKO) mice (20-24 wk), with hormonal imbalances, were studied. Mice were infused with ANG II (400 ng x kg(-1) x min(-1); 14 days). Systolic blood pressure was increased by ANG II in WT (166+/ 8 vs. 121+/-5 mmHg) and FORKO (176+/-7 vs. 115+/-5 mmHg; P<0.0001; n=9/group) mice. In ANG II-infused FORKO mice, cardiac mass was increased by 42% (P<0.001). This was associated with increased collagen content and augmented ERK1/2 phosphorylation (2-fold). Cardiac TRX expression and activity were decreased by ANG II in FORKO but not in WT (P<0.01) mice. ASK-1 expression, cleaved caspase III content, and Bax/Bcl-2 content were increased in ANG II-infused FORKO (P<0.05). ANG II had no effect on cardiac NAD(P)H oxidase activity or on O(2)(*-) levels in WT or FORKO. Cardiac ANG II type 1 receptor expression was similar in FORKO and WT. These findings indicate that in female FORKO, ANG II-induced cardiac hypertrophy and fibrosis are associated with the TRX downregulation and upregulation of ASK-1/caspase signaling. Our data suggest that in a model of menopause, protective actions of TRX may be blunted, which could contribute to cardiac remodeling independently of oxidative stress and hypertension. PMID- 18676691 TI - Virtual electrodes and the induction of fibrillation in Langendorff-perfused rabbit ventricles: the role of intracellular calcium. AB - A strong premature electrical stimulus (S(2)) induces both virtual anodes and virtual cathodes. The effects of virtual electrodes on intracellular Ca(2+) concentration ([Ca(2+)](i)) transients and ventricular fibrillation thresholds (VFTs) are unclear. We studied 16 isolated, Langendorff-perfused rabbit hearts with simultaneous voltage and [Ca(2+)](i) optical mapping and for vulnerable window determination. After baseline pacing (S(1)), a monophasic (10 ms anodal or cathodal) or biphasic (5 ms-5 ms) S(2) was applied to the left ventricular epicardium. Virtual electrode polarizations and [Ca(2+)](i) varied depending on the S(2) polarity. Relative to the level of [Ca(2+)](i) during the S(1) beat, the [Ca(2+)](i) level 40 ms after the onset of monophasic S(2) increased by 36+/-8% at virtual anodes and 20+/-5% at virtual cathodes (P<0.01), compared with 25+/-5% at both virtual cathode-anode and anode-cathode sites for biphasic S(2). The VFT was significantly higher and the vulnerable window significantly narrower for biphasic S(2) than for either anodal or cathodal S(2) (n=7, P<0.01). Treatment with thapsigargin and ryanodine (n=6) significantly prolonged the action potential duration compared with control (255+/-22 vs. 189+/-6 ms, P<0.05) and eliminated the difference in VFT between monophasic and biphasic S(2), although VFT was lower for both cases. We conclude that virtual anodes caused a greater increase in [Ca(2+)](i) than virtual cathodes. Monophasic S(2) is associated with lower VFT than biphasic S(2), but this difference was eliminated by the inhibition of the sarcoplasmic reticulum function and the prolongation of the action potential duration. However, the inhibition of the sarcoplasmic reticulum function also reduced VFT, indicating that the [Ca(2+)](i) dynamics modulate, but are not essential, to ventricular vulnerability. PMID- 18676692 TI - Mycobacterium bovis bacillus Calmette-Guerin therapy is a potent immunostimulator for management of systemic malignancies. PMID- 18676693 TI - In vitro and clinical investigation of the relationship between CCR5 receptor occupancy and anti-HIV activity of Aplaviroc. AB - Aplaviroc (GW873140) binds specifically to human cellular CC chemokine receptor 5 (CCR5) and demonstrates potent anti-human immunodeficiency virus activity in vitro in the subnanomolar range. In vitro studies show that aplaviroc selectively inhibits the binding of a particular monoclonal antibody, 45531, to CCR5. Based on this observation, a flow cytometry-based assay was developed to determine percentage CCR5 receptor occupancy (RO). CCR5 receptor occupancy was aplaviroc concentration-dependent and related to anti-human immunodeficiency virus activity in vitro. In the clinical setting, CCR5 receptor occupancy in peripheral blood was >98% in all subjects within 2 to 3 hours of dosing, which is consistent with the peak plasma concentrations of drug. Longitudinal analysis in the drug washout period revealed the time to 50% CCR5 receptor occupancy averaged >100 hours, in both human immunodeficiency virus-positive and human immunodeficiency virus negative subjects, substantially longer than the plasma pharmacokinetic half-life of 3 hours. The duration of CCR5 receptor occupancy appeared to be dose-dependent and associated with antiviral activity as measured by plasma human immunodeficiency virus RNA nadir following 10 days of multiple dose administration. These data demonstrate that the analysis of CCR5 receptor occupancy, in addition to conventional plasma-based pharmacokinetic measures, provides an informative tool to assist in evaluating the pharmacodynamic and antiviral effects of cellular CC chemokine receptor antagonists. PMID- 18676694 TI - Assessment of inhibitory effects of fluoride-coated tubes on biofilm formation by using the in vitro dental unit waterline biofilm model. AB - This study aimed to establish an in vitro model to simulate biofilms formed in dental unit waterlines (DUWLs) and to investigate the ability of polyvinylidene fluoride (PVDF)-coated tubes to inhibit biofilm formation using this model. The water and biofilm samples were obtained from DUWLs which had been clinically used for 2.5 years, and the predominant bacteria were identified. A conventional polyurethane tube was incubated for 24 to 96 h in the mixed flora of isolated bacteria, and the optimal incubation conditions to simulate a clinically formed biofilm were determined by observation with a scanning electron microscope. Biofilm formation on a PVDF-coated tube was observed using this in vitro model, and the adherence of different bacterial species to conventional and PVDF-coated tubes was assessed. Sphingomonas paucimobilis, Acinetobacter haemolytics, and Methylobacterium mesophilicum were predominantly isolated from contaminated DUWLs. Incubation of the polyurethane tube with the mixed flora containing these three species for 96 h resulted in the formation of a mature biofilm similar to the one clinically observed. The PVDF-coated tube was significantly less adhesive to all three bacterial species than the polyurethane tube (P < 0.05 by the Mann Whitney U test), and the attachment of small amounts of rods was observed even after incubation with the mixed flora for 96 h. In conclusion, an in vitro biofilm model was obtained by using a mixed flora of bacteria isolated from DUWLs, and the PVDF-coated tube was found to be effective in preventing biofilm formation using this model. PMID- 18676695 TI - Effects of hypercapnic hypoxia on inactivation and elimination of Vibrio campbellii in the Eastern oyster, Crassostrea virginica. AB - The Eastern oyster, Crassostrea virginica, inhabits shallow coastal waters that frequently experience periods of low dissolved oxygen (hypoxia) and elevated CO(2) (hypercapnia) levels. Bacteria are extremely abundant in these environments and accumulate in large numbers in filter-feeding oysters, which can act as passive carriers of human pathogens. Although hypercapnic hypoxia (HH) can affect certain specific immune mechanisms, its direct effect on the inactivation, degradation and elimination of bacteria in oysters is unknown. This research was conducted to determine whether exposure to HH reduces the ability of C. virginica to inactivate and eliminate Vibrio campbellii following its injection into the adductor muscle. Oysters were held in fully air-saturated (normoxic; partial O(2) pressure [P(O2)] = 20.7 kPa, CO(2) < 0.06 kPa, pH 7.8 to 8.0) or HH (P(O2) = 4 kPa, CO(2) = 1.8 kPa, pH 6.5 to 6.8) seawater at 25 degrees C for 4 h before being injected in the adductor muscle with 10(5) live Vibrio campbellii bacteria and remained under these conditions for the remainder of the experiment (up to 24 h postinjection). Real-time PCR was used to quantify the number of intact V. campbellii bacteria, while selective plating was used to quantify the number of injected bacteria remaining culturable in whole-oyster tissues, seawater, and feces/pseudofeces at 0, 1, 4, and 24 h postinjection. We found that oysters maintained under normoxic conditions were very efficient at inactivating and degrading large numbers of injected bacteria within their tissues. Moreover, a small percentage ( approximately 10%) of injected bacteria were passed into the surrounding seawater, while less than 1% were recovered in the feces/pseudofeces. In contrast, HH increased the percentage of culturable bacteria recovered from the tissues of oysters, suggesting an overall decrease in bacteriostasis. We suggest that poor water quality may increase the risk that oysters will harbor and transmit bacterial pathogens hazardous to human and ecosystem health. PMID- 18676696 TI - Transformation of N-phenylpiperazine by mixed cultures from a municipal wastewater treatment plant. AB - Samples from a wastewater treatment plant were used as inocula for mixed cultures dosed with N-phenylpiperazine (NPP), a model compound containing the piperazine ring found in many fluoroquinolones. Chemical analyses showed that NPP (50 mg liter(-1)) disappeared in 12 days, with the appearance of a transient metabolite and two nitrosated compounds. PMID- 18676697 TI - Persistence of Helicobacter pylori in heterotrophic drinking-water biofilms. AB - Although the route of transmission of Helicobacter pylori remains unknown, drinking water has been considered a possible transmission vector. It has been shown previously that, in water, biofilms are a protective niche for several pathogens, protecting them from stressful conditions, such as low carbon concentration, shear stress, and less-than-optimal temperatures. In this work, the influence of these three parameters on the persistence and cultivability of H. pylori in drinking-water biofilms was studied. Autochthonous biofilm consortia were formed in a two-stage chemostat system and then inoculated with the pathogen. Total numbers of H. pylori cells were determined by microscopy using a specific H. pylori 16S rRNA peptide nucleic acid probe, whereas cultivable cells were assessed by standard plating onto selective H. pylori medium. Cultivable H. pylori could not be detected at any time point, but the ability of H. pylori cells to incorporate, undergo morphological transformations, persist, and even agglomerate in biofilms for at least 31 days without a noticeable decrease in the total cell number (on average, the concentration was between 1.54 x 10(6) and 2.25 x 10(6) cells cm(-2)) or in the intracellular rRNA content may indicate that the loss of cultivability was due to entry into a viable but noncultivable state. Unlike previous results obtained for pure-culture H. pylori biofilms, shear stress did not negatively influence the numbers of H. pylori cells attached, suggesting that the autochthonous aquatic bacteria have an important role in retaining this pathogen in the sessile state, possibly by providing suitable microaerophilic environments or linking biomolecules to which the pathogen adheres. Therefore, biofilms appear to provide not only a safe haven for H. pylori but also a concentration mechanism so that subsequent sloughing releases a concentrated bolus of cells that might be infectious and that could escape routine grab sample microbiological analyses and be a cause of concern for public health. PMID- 18676698 TI - Methanogenesis from methanol at low temperatures by a novel psychrophilic methanogen, "Methanolobus psychrophilus" sp. nov., prevalent in Zoige wetland of the Tibetan plateau. AB - The Zoige wetland of the Tibetan plateau is at permanent low temperatures and is a methane emission heartland of the plateau; however, cold-adaptive methanogens in the soil are poorly understood. In this study, a variety of methanogenic enrichments at 15 degrees C and 30 degrees C were obtained from the wetland soil. It was demonstrated that hydrogenotrophic methanogenesis was the most efficient type at 30 degrees C, while methanol supported the highest methanogenesis rate at 15 degrees C. Moreover, methanol was the only substrate to produce methane more efficiently at 15 degrees C than at 30 degrees C. A novel psychrophilic methanogen, strain R15, was isolated from the methanol enrichment at 15 degrees C. Phylogenetic analysis placed strain R15 within the genus Methanolobus, loosely clustered with Methanolobus taylorii (96.7% 16S rRNA similarity). R15 produced methane from methanol, trimethylamine, and methyl sulfide and differed from other Methanolobus species by growing and producing methane optimally at 18 degrees C (specific growth rate of 0.063 +/- 0.001 h(-1)) and even at 0 degrees C. Based on these characteristics, R15 was proposed to be a new species and named "Methanolobus psychrophilus" sp. nov. The K(m) and V(max) of R15 for methanol conversion were determined to be 87.5 +/- 0.4 microM and 0.39 +/- 0.04 mM h(-1) at 18 degrees C, respectively, indicating a high affinity and conversion efficiency for methanol. The proportion of R15 in the soil was determined by quantitative PCR, and it accounted for 17.2% +/- 2.1% of the total archaea, enumerated as 10(7) per gram of soil; the proportion was increased to 42.4% +/- 2.3% in the methanol enrichment at 15 degrees C. This study suggests that the psychrophilic methanogens in the Zoige wetland are likely to be methylotrophic and to play a role in methane emission of the wetland. PMID- 18676699 TI - Genetic diversity of native bradyrhizobia isolated from soybeans (Glycine max L.) in different agricultural-ecological-climatic regions of India. AB - Fifty isolates from root nodules of soybean plants sampled in five agricultural ecological-climatic regions of India were analyzed by PCR-restriction fragment length polymorphism analysis of the 16S rRNA gene, the intergenic spacer region between the 16S and 23S rRNA genes (IGS), and the nifH and nodC genes. Eight haplotypes assigned to the Bradyrhizobium genus were identified, and the genetic diversity was conserved across regions. Sequence analyses of the IGS and the dnaK, glnII, recA, and nifH genes revealed three groups. One of them (26% of isolates) was assigned to Bradyrhizobium liaoningense. A second group (36% of isolates) was identified as B. yuanmingense but likely forms a new biovar able to nodulate soybean plants. The third lineage (38% of isolates) was different from all described Bradyrhizobium species but showed the same symbiotic genotype as B. liaoningense and B. japonicum bv. glycinearum. PMID- 18676700 TI - Enhancement of recombinant hemoglobin production in Escherichia coli BL21(DE3) containing the Plesiomonas shigelloides heme transport system. AB - To produce recombinant hemoglobin in Escherichia coli, sufficient intracellular heme must be present, or the protein folds improperly and is degraded. In this study, coexpression of human hemoglobin genes and Plesiomonas shigelloides heme transport genes enhanced recombinant hemoglobin production in E. coli BL21(DE3) grown in medium containing heme. PMID- 18676701 TI - Monitoring abundance and expression of "Dehalococcoides" species chloroethene reductive dehalogenases in a tetrachloroethene-dechlorinating flow column. AB - We investigated the distribution and activity of chloroethene-degrading microorganisms and associated functional genes during reductive dehalogenation of tetrachloroethene to ethene in a laboratory continuous-flow column. Using real time PCR, we quantified "Dehalococcoides" species 16S rRNA and chloroethene reductive dehalogenase (RDase) genes (pceA, tceA, vcrA, and bvcA) in nucleic acid extracts from different sections of the column. Dehalococcoides 16S rRNA gene copies were highest at the inflow port [(3.6 +/- 0.6) x 10(6) (mean +/- standard deviation) per gram soil] where the electron donor and acceptor were introduced into the column. The highest transcript numbers for tceA, vcrA, and bvcA were detected 5 to 10 cm from the column inflow. bvcA was the most highly expressed of all RDase genes and the only vinyl chloride reductase-encoding transcript detectable close to the column outflow. Interestingly, no expression of pceA was detected in the column, despite the presence of the genes in the microbial community throughout the column. By comparing the 16S rRNA gene copy numbers to the sum of all four RDase genes, we found that 50% of the Dehalococcoides population in the first part of the column did not contain either one of the known chloroethene RDase genes. Analysis of 16S rRNA gene clone libraries from both ends of the flow column revealed a microbial community dominated by members of Firmicutes and Actinobacteria. Higher clone sequence diversity was observed near the column outflow. The results presented have implications for our understanding of the ecophysiology of reductively dehalogenating Dehalococcoides spp. and their role in bioremediation of chloroethenes. PMID- 18676702 TI - Characterization of an endoglucanase belonging to a new subfamily of glycoside hydrolase family 45 of the basidiomycete Phanerochaete chrysosporium. AB - The wood decay fungus Phanerochaete chrysosporium has served as a model system for the study of lignocellulose conversions, but aspects of its cellulolytic system remain uncertain. Here, we report identifying the gene that encodes the glycoside hydrolase (GH) family 45 endoglucanase (EG) from the fungus, cloning the cDNA, determining its heterologous expression in the methylotrophic yeast Pichia pastoris, and characterizing the recombinant protein. The cDNA consisted of 718 bp, including an open reading frame encoding a 19-amino-acid signal peptide, a 7-amino-acid presequence at the N-terminal region, and a 180-amino acid mature protein, which has no cellulose binding domain. Analysis of the amino acid sequence revealed that the protein has a low similarity (<22%) to known fungal EGs belonging to the GH family 45 (EGVs). No conserved domain of this family was found by a BLAST search, suggesting that the protein should be classified into a new subdivision of this GH family. The recombinant protein has hydrolytic activity toward amorphous cellulose, carboxylmethyl cellulose, lichenan, barley beta-glucan, and glucomannan but not xylan. Moreover, a synergistic effect was observed with the recombinant GH family 6 cellobiohydrolase from the same fungus toward amorphous cellulose as a substrate, indicating that the enzyme may act in concert with other cellulolytic enzymes to hydrolyze cellulosic biomass in nature. PMID- 18676704 TI - Evaluation of a low-cost electrostatic dust fall collector for indoor air endotoxin exposure assessment. AB - Exposure to endotoxin in home environments has become a key issue in asthma and allergy research. Most studies have analyzed floor or mattress dust endotoxin, but its validity as a proxy for airborne exposure is unknown, while active airborne dust sampling is not feasible in large-scale population studies because of logistic and financial limitations. We therefore developed and evaluated a simple passive airborne dust collection method for airborne endotoxin exposure assessment. We explored an electrostatic dust fall collector (EDC), consisting of a 42- by 29.6-cm-sized folder with four electrostatic cloths exposed to the air. The EDC was tested during two 14-day periods in seven nonfarm and nine farm homes and in farm stables. In parallel, active airborne dust sampling was performed with Harvard impactors and floor dust collected by vacuuming, using nylon sampling socks. The endotoxin levels could be measured in all EDC cloth extracts. The levels (in EU/m(2)) between EDCs used simultaneously or in different sampling periods in the same home correlated strongly (r > 0.8). EDC endotoxin also correlated moderately to strongly (r = 0.6 to 0.8) with the endotoxin measured by active airborne dust sampling and living room floor dust sampling and-in farm homes-with the endotoxin captured by the EDC in stables. In contrast, endotoxin levels measured by floor dust sampling showed only a poor correlation with the levels measured by active airborne dust sampling. We therefore conclude that measuring endotoxin levels with the EDC is a valid measure of average airborne endotoxin exposure, while reproducibility over time is at least equivalent to that of reservoir dust analyses. PMID- 18676703 TI - Global occurrence of archaeal amoA genes in terrestrial hot springs. AB - Despite the ubiquity of ammonium in geothermal environments and the thermodynamic favorability of aerobic ammonia oxidation, thermophilic ammonia-oxidizing microorganisms belonging to the crenarchaeota kingdom have only recently been described. In this study, we analyzed microbial mats and surface sediments from 21 hot spring samples (pH 3.4 to 9.0; temperature, 41 to 86 degrees C) from the United States, China, and Russia and obtained 846 putative archaeal ammonia monooxygenase large-subunit (amoA) gene and transcript sequences, representing a total of 41 amoA operational taxonomic units (OTUs) at 2% identity. The amoA gene sequences were highly diverse, yet they clustered within two major clades of archaeal amoA sequences known from water columns, sediments, and soils: clusters A and B. Eighty-four percent (711/846) of the sequences belonged to cluster A, which is typically found in water columns and sediments, whereas 16% (135/846) belonged to cluster B, which is typically found in soils and sediments. Although a few amoA OTUs were present in several geothermal regions, most were specific to a single region. In addition, cluster A amoA genes formed geographic groups, while cluster B sequences did not group geographically. With the exception of only one hot spring, principal-component analysis and UPGMA (unweighted-pair group method using average linkages) based on the UniFrac metric derived from cluster A grouped the springs by location, regardless of temperature or bulk water pH, suggesting that geography may play a role in structuring communities of putative ammonia-oxidizing archaea (AOA). The amoA genes were distinct from those of low-temperature environments; in particular, pair-wise comparisons between hot spring amoA genes and those from sympatric soils showed less than 85% sequence identity, underscoring the distinctness of hot spring archaeal communities from those of the surrounding soil system. Reverse transcription-PCR showed that amoA genes were transcribed in situ in one spring and the transcripts were closely related to the amoA genes amplified from the same spring. Our study demonstrates the global occurrence of putative archaeal amoA genes in a wide variety of terrestrial hot springs and suggests that geography may play an important role in selecting different assemblages of AOA. PMID- 18676705 TI - Attached bacterial populations shared by four species of aquatic angiosperms. AB - Symbiotic relationships between microbes and plants are common and well studied in terrestrial ecosystems, but little is known about such relationships in aquatic environments. We compared the phylogenetic diversities of leaf- and root attached bacteria from four species of aquatic angiosperms using denaturing gradient gel electrophoresis (DGGE) and DNA sequencing of PCR-amplified 16S rRNA genes. Plants were collected from three beds in Chesapeake Bay at sites characterized as freshwater (Vallisneria americana), brackish (Potomogeton perfoliatus and Stuckenia pectinata), and marine (Zostera marina). DGGE analyses showed that bacterial communities were very similar for replicate samples of leaves from canopy-forming plants S. pectinata and P. perfoliatus and less similar for replicate samples of leaves from meadow-forming plants Z. marina and V. americana and of roots of all species. In contrast, bacterial communities differed greatly among plant species and between leaves and roots. DNA sequencing identified 154 bacterial phylotypes, most of which were restricted to single plant species. However, 12 phylotypes were found on more than one plant species, and several of these phylotypes were abundant in clone libraries and represented the darkest bands in DGGE banding patterns. Root-attached phylotypes included relatives of sulfur-oxidizing Gammaproteobacteria and sulfate-reducing Deltaproteobacteria. Leaf-attached phylotypes included relatives of polymer degrading Bacteroidetes and phototrophic Alphaproteobacteria. Also, leaves and roots of three plant species hosted relatives of methylotrophic Betaproteobacteria belonging to the family Methylophilaceae. These results suggest that aquatic angiosperms host specialized communities of bacteria on their surfaces, including several broadly distributed and potentially mutualistic bacterial populations. PMID- 18676706 TI - Molecular detection of fungal communities in the Hawaiian marine sponges Suberites zeteki and Mycale armata. AB - Symbiotic microbes play a variety of fundamental roles in the health and habitat ranges of their hosts. While prokaryotes in marine sponges have been broadly characterized, the diversity of sponge-inhabiting fungi has barely been explored using molecular approaches. Fungi are an important component of many marine and terrestrial ecosystems, and they may be an ecologically significant group in sponge-microbe interactions. This study tested the feasibility of using existing fungal primers for molecular analysis of sponge-associated fungal communities. None of the eight selected primer pairs yielded satisfactory results in fungal rRNA gene or internal transcribed spacer (ITS) clone library constructions. However, 3 of 10 denaturing gradient gel electrophoresis (DGGE) primer sets, which were designed to preferentially amplify fungal rRNA gene or ITS regions from terrestrial environmental samples, were successfully amplified from fungal targets in marine sponges. DGGE analysis indicated that fungal communities differ among different sponge species (Suberites zeteki and Mycale armata) and also vary between sponges and seawater. Sequence analysis of DGGE bands identified 23 and 21 fungal species from each of the two sponge species S. zeteki and M. armata, respectively. These species were representatives of 11 taxonomic orders and belonged to the phyla of Ascomycota (seven orders) and Basidiomycota (four orders). Five of these taxonomic orders (Malasseziales, Corticiales, Polyporales, Agaricales, and Dothideomycetes et Chaetothyriomcetes incertae sedis) have now been identified for the first time in marine sponges. Seven and six fungal species from S. zeteki and M. armata, respectively, are potentially new species because of their low sequence identity (< or =98%) with their references in GenBank. Phylogenetic analysis indicated sponge-derived sequences were clustered into "marine fungus clades" with those from other marine habitats. This is the first report of molecular analysis of fungal communities in marine sponges, adding depth and dimension to our understanding of sponge-associated microbial communities. PMID- 18676707 TI - A new Huanglongbing Species, "Candidatus Liberibacter psyllaurous," found to infect tomato and potato, is vectored by the psyllid Bactericera cockerelli (Sulc). AB - A new huanglongbing (HLB) "Candidatus Liberibacter" species is genetically characterized, and the bacterium is designated "Candidatus Liberibacter psyllaurous." This bacterium infects the psyllid Bactericera cockerelli and its solanaceous host plants potato and tomato, potentially resulting in "psyllid yellowing." Host plant-dependent HLB transmission and variation in psyllid infection frequencies are found. PMID- 18676708 TI - Physiological and transcriptional responses to high concentrations of lactic acid in anaerobic chemostat cultures of Saccharomyces cerevisiae. AB - Based on the high acid tolerance and the simple nutritional requirements of Saccharomyces cerevisiae, engineered strains of this yeast are considered biocatalysts for industrial production of high-purity undissociated lactic acid. However, high concentrations of lactic acid are toxic to S. cerevisiae, thus limiting its growth and product formation. Physiological and transcriptional responses to high concentrations of lactic acid were studied in anaerobic, glucose-limited chemostat cultures grown at different pH values and lactic acid concentrations, resulting in a 50% decrease in the biomass yield. At pH 5, the yield decrease was caused mostly by osmotically induced glycerol production and not by the classic weak-acid action, as was observed at pH 3. Cultures grown at pH 5 with 900 mM lactic acid revealed an upregulation of many genes involved in iron homeostasis, indicating that iron chelation occurred at high concentrations of dissociated lactic acid. Chemostat cultivation at pH 3 with 500 mM lactate, resulting in lower anion concentrations, showed an alleviation of this iron homeostasis response. Six of the 10 known targets of the transcriptional regulator Haa1p were strongly upregulated in lactate-challenged cultures at pH 3 but showed only moderate induction by high lactate concentrations at pH 5. Moreover, the haa1Delta mutant exhibited a growth defect at high lactic acid concentrations at pH 3. These results indicate that iron homeostasis plays a major role in the response of S. cerevisiae to high lactate concentrations, whereas the Haa1p regulon is involved primarily in the response to high concentrations of undissociated lactic acid. PMID- 18676709 TI - Antibacterial activities of naturally occurring compounds against Mycobacterium avium subsp. paratuberculosis. AB - The antibacterial activities of 18 naturally occurring compounds (including essential oils and some of their isolated constituents, apple and green tea polyphenols, and other plant extracts) against three strains of Mycobacterium avium subsp. paratuberculosis (a bovine isolate [NCTC 8578], a raw-milk isolate [806R], and a human isolate [ATCC 43015]) were evaluated using a macrobroth susceptibility testing method. M. avium subsp. paratuberculosis was grown in 4 ml Middlebrook 7H9 broth containing 10% oleic acid-albumin-dextrose-catalase, 0.05% Tween 80 (or 0.2% glycerol), and 2 microg/ml mycobactin J supplemented with five concentrations of each test compound. The changes in the optical densities of the cultures at 600 nm as a measure of CFU were recorded at intervals over an incubation period of 42 days at 37 degrees C. Six of the compounds were found to inhibit the growth of M. avium subsp. paratuberculosis. The most effective compound was trans-cinnamaldehyde, with a MIC of 25.9 microg/ml, followed by cinnamon oil (26.2 microg/ml), oregano oil (68.2 microg/ml), carvacrol (72.2 microg/ml), 2,5-dihydroxybenzaldehyde (74 microg/ml), and 2-hydroxy-5 methoxybenzaldehyde (90.4 microg/ml). With the exception of carvacrol, a phenolic compound, three of the four most active compounds are aldehydes, suggesting that the structure of the phenolic group or the aldehyde group may be important to the antibacterial activity. No difference in compound activity was observed between the three M. avium subsp. paratuberculosis strains studied. Possible mechanisms of the antimicrobial effects are discussed. PMID- 18676710 TI - Regulation of Lactobacillus casei sorbitol utilization genes requires DNA-binding transcriptional activator GutR and the conserved protein GutM. AB - Sequence analysis of the five genes (gutRMCBA) downstream from the previously described sorbitol-6-phosphate dehydrogenase-encoding Lactobacillus casei gutF gene revealed that they constitute a sorbitol (glucitol) utilization operon. The gutRM genes encode putative regulators, while the gutCBA genes encode the EIIC, EIIBC, and EIIA proteins of a phosphoenolpyruvate-dependent sorbitol phosphotransferase system (PTS(Gut)). The gut operon is transcribed as a polycistronic gutFRMCBA messenger, the expression of which is induced by sorbitol and repressed by glucose. gutR encodes a transcriptional regulator with two PTS regulated domains, a galactitol-specific EIIB-like domain (EIIB(Gat) domain) and a mannitol/fructose-specific EIIA-like domain (EIIA(Mtl) domain). Its inactivation abolished gut operon transcription and sorbitol uptake, indicating that it acts as a transcriptional activator. In contrast, cells carrying a gutB mutation expressed the gut operon constitutively, but they failed to transport sorbitol, indicating that EIIBC(Gut) negatively regulates GutR. A footprint analysis showed that GutR binds to a 35-bp sequence upstream from the gut promoter. A sequence comparison with the presumed promoter region of gut operons from various firmicutes revealed a GutR consensus motif that includes an inverted repeat. The regulation mechanism of the L. casei gut operon is therefore likely to be operative in other firmicutes. Finally, gutM codes for a conserved protein of unknown function present in all sequenced gut operons. A gutM mutant, the first constructed in a firmicute, showed drastically reduced gut operon expression and sorbitol uptake, indicating a regulatory role also for GutM. PMID- 18676711 TI - Preferential colonization of Solanum tuberosum L. roots by the fungus Glomus intraradices in arable soil of a potato farming area. AB - The symbiosis between plant roots and arbuscular mycorrhizal (AM) fungi has been shown to affect both the diversity and productivity of agricultural communities. In this study, we characterized the AM fungal communities of Solanum tuberosum L. (potato) roots and of the bulk soil in two nearby areas of northern Italy, in order to verify if land use practices had selected any particular AM fungus with specificity to potato plants. The AM fungal large-subunit (LSU) rRNA genes were subjected to nested PCR, cloning, sequencing, and phylogenetic analyses. One hundred eighty-three LSU rRNA sequences were analyzed, and eight monophyletic ribotypes, belonging to Glomus groups A and B, were identified. AM fungal communities differed between bulk soil and potato roots, as one AM fungal ribotype, corresponding to Glomus intraradices, was much more frequent in potato roots than in soils (accounting for more than 90% of sequences from potato samples and less than 10% of sequences from soil samples). A semiquantitative heminested PCR with specific primers was used to confirm and quantify the AM fungal abundance observed by cloning. Overall results concerning the biodiversity of AM fungal communities in roots and in bulk soils from the two studied areas suggested that potato roots were preferentially colonized by one AM fungal species, G. intraradices. PMID- 18676712 TI - Optimization of metabolic capacity and flux through environmental cues to maximize hydrogen production by the cyanobacterium "Arthrospira (Spirulina) maxima". AB - Environmental and nutritional conditions that optimize the yield of hydrogen (H(2)) from water using a two-step photosynthesis/fermentation (P/F) process are reported for the hypercarbonate-requiring cyanobacterium "Arthrospira maxima." Our observations lead to four main conclusions broadly applicable to fermentative H(2) production by bacteria: (i) anaerobic H(2) production in the dark from whole cells catalyzed by a bidirectional [NiFe] hydrogenase is demonstrated to occur in two temporal phases involving two distinct metabolic processes that are linked to prior light-dependent production of NADPH (photosynthetic) and dark/anaerobic production of NADH (fermentative), respectively; (ii) H(2) evolution from these reductants represents a major pathway for energy production (ATP) during fermentation by regenerating NAD(+) essential for glycolysis of glycogen and catabolism of other substrates; (iii) nitrate removal during fermentative H(2) evolution is shown to produce an immediate and large stimulation of H(2), as nitrate is a competing substrate for consumption of NAD(P)H, which is distinct from its slower effect of stimulating glycogen accumulation; (iv) environmental and nutritional conditions that increase anaerobic ATP production, prior glycogen accumulation (in the light), and the intracellular reduction potential (NADH/NAD(+) ratio) are shown to be the key variables for elevating H(2) evolution. Optimization of these conditions and culture age increases the H(2) yield from a single P/F cycle using concentrated cells to 36 ml of H(2)/g (dry weight) and a maximum 18% H(2) in the headspace. H(2) yield was found to be limited by the hydrogenase-mediated H(2) uptake reaction. PMID- 18676713 TI - Engineering of an Escherichia coli strain for the production of 3-methyl-1 butanol. AB - 3-Methyl-1-butanol is a potential fuel additive or substitute. Previously this compound was identified in small quantities in yeast fermentation as one of the fusel alcohols. In this work, we engineered an Escherichia coli strain to produce 3-methyl-1-butanol from glucose via the host's amino acid biosynthetic pathways. Strain improvement with the removal of feedback inhibition and competing pathways increased the selectivity and productivity of 3-methyl-1-butanol. This work demonstrates the feasibility of production of 3-methyl-1-butanol as a biofuel and shows promise in using E. coli as a host for production. PMID- 18676714 TI - Intraspecific variation in Viola suavis in Europe: parallel evolution of white flowered morphotypes. AB - BACKGROUND AND AIMS: Viola species are commonly grown for their ornamental flowers, but their evolutionary history and taxonomy are often complicated and have been poorly explored so far. This is a study of the polymorphic, typically blue-flowered species Viola suavis, concentrating on the white-flowered populations of uncertain taxonomic assignment that occur in Spain and central and south-eastern Europe. The aim was to resolve their origin and taxonomic status and to study the intraspecific structure and (post)glacial history of this species. METHODS: Viola suavis and five close relatives were sampled from multiple locations and subjected to molecular (AFLP, sequencing of nrDNA ITS) and morphometric analyses. Data on ploidy level and pollen fertility were also obtained, to address an assumed hybrid origin of the white-flowered populations. KEY RESULTS: In V. suavis a strong intraspecific genetic split into two groups was observed, indicating that there has been a long-term isolation and survival in distinct glacial refugia. The white-flowered populations could be placed within the variation range of this species, and it is clear that they evolved independently in two distant areas. Their parallel evolution is supported by both morphological and genetic differentiation. The strongly reduced genetic variation and absence of unique AFLP fragments suggest their derived status and origin from the typical, blue-flowered populations. CONCLUSIONS: These results suggest that intraspecific variation in V. suavis has been largely shaped by population isolations during the last glaciation and subsequent recolonizations, although cultivation and vegetative spread by humans have affected the present picture as well. PMID- 18676715 TI - ASAS Centennial Paper: utilization of pasture and forages by ruminants: a historical perspective. AB - Pastures, forages, and grasslands dominate the landscape across the United States and support a large ruminant population that supplies the nation with value-added animal products. A historical perspective is presented of the innovations as they occurred in the Journal of Animal Science over the past 100 yr in pasture and forage research. Consideration was given to both animal and pasture perspectives. Areas given consideration from the animal perspective were schemes for feedstuff analysis, experimental design and statistics, forage sample preservation, indirect methods of measuring intake and digestion, TDN and energy, nutritive value, harvested forage, and innovations in the grazing environment. Areas given consideration from the forage perspective were a framework for forage-animal interface research, determining pasture yield, choice of stocking method, grazing management, partitioning of forage DM, near-infrared reflectance spectroscopy technology, antiquality constituents, and forage sample preservation. Finally, the importance was discussed of applying research results from the forage-animal interface to general ruminant nutrition research beyond the interface that is focused on altered diets. PMID- 18676716 TI - Odorant production and persistence of Escherichia coli in manure slurries from cattle fed zero, twenty, forty, or sixty percent wet distillers grains with solubles. AB - Corn ethanol production removes starch and concentrates the remaining nutrients, including CP and minerals. When wet distillers grains with solubles (WDGS) are fed to cattle in place of corn, CP and minerals often exceed dietary needs. This may increase N emission, P run-off, and odor production. These variables are evaluated in this study. Crossbred steers (n = 160; 434 +/- 8 kg) were assigned in a completely randomized block design to 9 x 9 m pens with concrete floor (10 animals/pen; 4 pens/treatment). Steers were fed a finishing diet that contained 0, 20, 40, or 60% WDGS on a DM basis, and provided 13.3, 15.5, 20.6, or 24.9% CP, respectively. Two kilograms of manure slurry (14 to 23% DM) were collected from each pen monthly (Aug. 20, Sep. 24, and Oct. 22). Samples were analyzed immediately for odorants, DM, pH, NH(3), total alcohol, l-lactate, and concentrations of generic Escherichia coli. After incubation of the samples at 22 degrees C for 2, 4, 7, 10, 15, 21, and 28 d, samples were analyzed for methane production in addition to the above characteristics. Before incubation, NH(3), H(2)S, indole, phenol, isovalerate, isobutyrate, and acetate increased (P < 0.01) with increasing amounts of WDGS in the diet. Other odorants, including skatole, caproate, valerate, butyrate, and propionate, were greater (P < 0.01) in manure slurries from cattle fed 20 or 40% WDGS, compared to 0% WDGS. The l-lactate was greater (P < 0.01) in slurries from cattle fed 0% WDGS (447 mu mol/g of DM) compared with the other treatment slurries (14 to 15 mu mol/g of DM). After incubation, l-lactate contributed to lowered slurry pH (6.3, 7.1, 7.6, and 8.2, respectively, for 0, 20, 40, and 60% WDGS), which inhibited microbial fermentation, E. coli persistence, and methane production. Because of the favorable, more neutral pH in the 40 and 60% WDGS slurries, many of the odorant compounds were rapidly converted to methane during a 28-d static incubation. Escherichia coli O157:H7 inoculated into subsamples of the manure slurries exhibited behavior similar to that of naturally present generic E. coli, surviving in greater numbers longer (P < 0.05) in 20 and 40% WDGS slurries than in 0% WDGS. These data indicate feeding WDGS can increase odorants in manure slurries and extend the persistence of E. coli. PMID- 18676718 TI - Effects of divergent selection for serum insulin-like growth factor-I concentration on performance, feed efficiency, and ultrasound measures of carcass composition traits in Angus bulls and heifers. AB - Angus bulls and heifers from lines divergently selected for serum IGF-I concentration were used to evaluate the effects of IGF-I selection line on growth performance and feed efficiency in 2 studies. In study 1, bulls (low line, n = 9; high line, n = 8; initial BW = 367.1 +/- 22.9 kg) and heifers (low line, n = 9; high line, n = 13; initial BW = 286.4 +/- 28.6 kg) were adapted to a roughage based diet (ME = 1.95 Mcal/kg of DM) for 24 d and fed individually for 77 d by using Calan gate feeders. In study 2, bulls (low line, n = 15; high line, n = 12; initial BW = 297.5 +/- 34.4 kg) and heifers (low line, n = 9; high line, n = 20; initial BW = 256.0 +/- 25.1 kg) were adapted to a grain-based diet (ME = 2.85 Mcal/kg of DM) for 32 d and fed individually for 70 d by using Calan gate feeders. Blood samples were collected at weaning and at the start and end of each study, and serum IGF-I concentration was determined. Residual feed intake (RFI) was calculated, within study, as the residual from the linear regression of DMI on midtest BW(0.75), ADG, sex, sex by midtest BW(0.75) and sex by ADG. In study 1, calves from the low IGF-I selection line had similar initial and final BW and ADG, compared with calves from the high IGF-I selection line. In addition, DMI and feed conversion ratio were similar between IGF-I selection lines; however, calves from the low IGF-I selection line tended (P < 0.10) to have lesser RFI than calves from the high IGF-I selection line (-0.26 vs. 0.24 +/- 0.31 kg/d). In study 2, IGF-I selection line had no influence on performance or feed efficiency traits. However, there was a tendency (P = 0.15) for an IGF-I selection line x sex interaction for RFI. Bulls from the low IGF-I selection line had numerically lesser RFI than those from the high IGF-I selection line, whereas in heifers, the IGF-I selection line had no effect on RFI. In studies 1 and 2, weaning and initial IGF-I concentrations were not correlated with either feed conversion ratio or RFI. However, regression analysis revealed a sex x IGF-I concentration interaction for initial IGF-I concentration in study 1 and weaning IGF-I concentration in study 2 such that the regression coefficient was positive for bulls and negative for heifers. These data suggest that genetic selection for postweaning serum IGF-I concentration had a minimal effect on RFI in beef cattle. PMID- 18676717 TI - Effects of implants of trenbolone acetate, estradiol, or both, on muscle insulin like growth factor-I, insulin-like growth factor-I receptor, estrogen receptor {alpha}, and androgen receptor messenger ribonucleic acid levels in feedlot steers. AB - We previously showed that a combined trenbolone acetate (TBA)/estradiol-17beta (E2) implant significantly increases IGF-I mRNA levels in the LM of feedlot steers by 28 d after implantation. Here we compare the effects of E2 (25.7 mg), TBA (120 mg), and combined TBA (120 mg)/E2 (24 mg) implants on IGF-I, IGF-I receptor (IGFR-1), estrogen receptor (ER)-alpha and androgen receptor (AR) mRNA levels in the LM of steers. Twenty yearling crossbred steers with an average initial BW of 421.1 +/- 3.6 kg were stratified by BW and randomly assigned to 1 of 4 treatments: 1) nonimplanted, control; 2) implanted with TBA and E2; 3) implanted with E2; or 4) implanted with TBA. Steers were weighed weekly starting on d 0, and muscle biopsy samples were taken from each steer on d 0 (before implantation), 7, 14, and 28. Ribonucleic acid was prepared from each sample and real-time reverse transcription-PCR was used to determine the levels of IGF-I, IGFR-1, ER-alpha, and AR mRNA. Body weight of implanted steers, adjusted by using d-0 BW as a covariant, tended (P = 0.09) to be greater than that of control steers. On d 7 and 28, IGF-I mRNA levels were greater (58 and 78%, respectively; P < 0.009) in E2-implanted animals than in control steers. Similarly, on d 28 the LM IGF-I mRNA level was 65% greater (P = 0.017) in TBA/E2-implanted steers than in control animals. In contrast, the TBA implant did not increase (P = 0.99) LM IGF-I mRNA levels after 28 d of implantation. Muscle IGFR-1, AR, and ER-alpha mRNA levels were not different (P > 0.47) in any of the treated groups compared with the control group. These data suggest that E2 is responsible for the increased muscle IGF-I mRNA level observed in steers implanted with a combined TBA/E2 implant. PMID- 18676720 TI - Docosahexaenoic acid regulates adipogenic genes in myoblasts via porcine peroxisome proliferator-activated receptor gamma. AB - The nuclear transcription factor peroxisome proliferator-activated receptor gamma (PPARgamma) triggers adipocyte differentiation by regulating lipogenic genes. A ligand for PPARgamma is necessary to activate PPARgamma function. Fatty acids are potential ligands for PPARgamma activation. The current experiment was designed to determine the potential for individual fatty acids to activate porcine PPARgamma ectopically expressed in myoblasts. The expression of adipocyte fatty acid binding protein (aP2) and adiponectin in myoblasts stably expressing porcine PPARgamma was increased when docosahexaenoic acid (DHA) was added to the adipogenic medium. The response was positively related to DHA concentration and suggests that DHA may bind to and activate porcine PPARgamma, leading to increased expression of aP2 and adiponectin. The conditioned media collected from myoblasts expressing PPARgamma between d 3 and 6 or between d 6 and 9, but not DHA itself, activated the aP2 gene promoter-driven luciferase activity. These results suggest that a metabolite of DHA is the ligand binding to and activating porcine PPARgamma. The metabolite and pathway for its production are currently unknown. PMID- 18676721 TI - Dietary tryptophan helps to preserve tryptophan homeostasis in pigs suffering from lung inflammation. AB - In pigs, inflammation modifies Trp metabolism and consequently could impact on Trp requirement for growth. In this study, the effects of lung inflammation, induced by the intravenous injection of complete Freund's adjuvant, and dietary Trp content on Trp metabolism and availability were investigated. Two dietary Trp contents, one corresponding to a low-Trp diet (1.5 g of Trp/kg of diet, Basal diet) and the second to an adequate-Trp diet (2 g of Trp/kg of diet, TRP diet), were used. Ten blocks of 4 littermate piglets were selected at 40 d of age. Within each block, piglets were randomly assigned to 1 of the 4 experimental treatments: (1) healthy control and Basal diet, (2) inflammation and Basal diet, (3) inflammation and Basal diet + antioxidant, and (4) inflammation and TRP diet. Inflammation induced an increase in indoleam-ine 2,3 dioxygenase (IDO) activity, an enzyme involved in Trp catabolism, in lung, lymph nodes, heart, and spleen (P < 0.01). Contrary to piglets fed the TRP diet, pigs suffering from inflammation did not maintain their plasma Trp concentrations when they were fed the Basal diet. Furthermore, pigs fed the TRP diet had decreased plasma haptoglobin concentrations, IDO activity, and lung weight than those fed the Basal diet, indicating that the inflammatory response was moderated with the greater Trp supply. Antioxidant addition in the Basal diet decreased the effects of inflammation on plasma Trp concentrations and IDO activity. These results indicated that inflammation increases Trp catabolism and thus may decrease Trp availability for growth. PMID- 18676722 TI - Promoter region of the bovine growth hormone receptor gene: single nucleotide polymorphism discovery in cattle and association with performance in Brangus bulls. AB - Expression of the GH receptor (GHR) gene and its binding with GH is essential for growth and fat metabolism. A GT microsatellite exists in the promoter of bovine GHR segregating short (11 bp) and long (16 to 20 bp) allele sequences. To detect SNP and complete an association study of genotype to phenotype, we resequenced a 1,195-bp fragment of DNA including the GT microsatellite and exon 1A. Resequencing was completed in 48 familialy unrelated Holstein, Jersey, Brown Swiss, Simmental, Angus, Brahman, and Brangus cattle. Nine SNP were identified. Phylogeny analyses revealed minor distance (i.e., <5%) in DNA sequence among the 5 Bos taurus breeds; however, sequence from Brahman cattle averaged 27.4 +/- 0.07% divergence from the Bos taurus breeds, whereas divergence of Brangus was intermediate. An association study of genotype to phenotype was completed with data from growing Brangus bulls (n = 553 from 96 sires) and data from 4 of the SNP flanking the GT microsatellite. These SNP were found to be in Hardy-Weinberg equilibrium and in phase based on linkage disequilibrium analyses (r(2) = 0.84 and D'= 0.92). An A/G tag SNP was identified (ss86273136) and was located in exon 1A, which began 88 bp downstream from the GT microsatellite. Minor allele frequency of the tag SNP was greater than 10%, and Mendelian segregation was verified in 3 generation pedigrees. The A allele was derived from Brahman, and the G allele was derived from Angus. This tag SNP genotype was a significant effect in analyses of rib fat data collected with ultrasound when bulls were ~365 d of age. Specifically, bulls of the GG genotype had 6.1% more (P = 0.0204) rib fat than bulls of the AA and AG genotypes, respectively. Tag SNP (ss86273136), located in the promoter of GHR, appears to be associated with a measure of corporal fat in Bos taurus x Bos indicus composite cattle. PMID- 18676723 TI - Fasting regulates the expression of adiponectin receptors in young growing pigs. AB - Adiponectin is an adipocyte-derived hormone that can improve insulin sensitivity. Its functions in regulating glucose utilization and fatty acid metabolism in mammals are mediated by 2 subtypes of adiponectin receptors (AdipoR1 and AdipoR2). This study was conducted to determine the effect of fasting on the expression of adiponectin and its receptors. The expression of adiponectin was not affected in s.c. adipose tissue, but adiponectin expression increased in visceral adipose tissue after fasting. In contrast, expression of both AdipoR mRNA was increased in the liver and s.c. adipose tissue of 24-h-fasted pigs compared with fed pigs, but the mRNA in muscle and visceral adipose tissue was not affected by fasting. A third putative adiponectin receptor, T-cadherin, was cloned and the mRNA expression was determined. T-Cadherin has been recognized to act as a vascular adiponectin receptor in vascular endothelial and smooth muscle cells. Our data showed that the expression of T-cadherin was decreased in the muscle of fasted pigs, suggesting that the expression of T-cadherin can be regulated by feeding status. In summary, in young pigs, adiponectin mRNA was up regulated by fasting in visceral, but not s.c., adipose tissue, whereas AdipoR1 and AdipoR2 mRNA were increased in s.c., but not visceral, adipose tissue. The adiponectin receptor, T-cadherin, was expressed in s.c. and visceral adipose tissue and in muscle, but only muscle mRNA expression was decreased by fasting. PMID- 18676724 TI - The effect of vitamin A supplementation on postnatal adipose tissue development of lambs. AB - Vitamin A (retinoic acid) is known to be an adipogenic factor influencing both in vitro and in vivo cell development. This study aimed to determine its effect on lamb adipose tissue development during the early phase of postnatal development until 100 d of age. Male lambs (n = 24) of the Rasa Aragonesa breed were used. At birth, lambs were assigned to 1 of 2 experimental groups: 1) the control (C) group, which received feed without vitamin A supplementation, and 2) the vitamin A (V) group, which received a supplement of 500,000 IU/animal twice per week from birth to slaughter. The effect of vitamin A supplementation was studied at 16.8 +/- 0.35 kg of BW (58 +/- 0.7 d of age) and at 27.8 +/- 0.78 kg of BW (101 +/- 6.5 d of age). The variables of lamb growth, carcass, LM area, and lipid content were analyzed. To study adipose tissue development, the amount of adipose tissue accumulated, the size and number of adipocytes, and lipogenic enzyme activities (glycerol 3-phosphate dehydrogenase, fatty acid synthase, and glucose 6-phosphate dehydrogenase) of the omental, perirenal, and s.c. depots were quantified. Results showed that vitamin A supplementation had no influence on growth, carcass variables, LM area, and lipid content during lamb growth but that the number of adipocytes in the perirenal depot was 30% greater in lambs of the V group (P < 0.05) and that these lambs had smaller adipocytes in the omental and perirenal depots (P = 0.06) at 28 kg of BW (101 d of age). These results suggest that the intake of this level of vitamin A during the whole period of growth of the lambs influenced the processes of hyperplasia and hypertrophy in the different adipose depots, depending on their degree of maturity. PMID- 18676725 TI - Effects of multiple concurrent stressors on rectal temperature, blood acid-base status, and longissimus muscle glycolytic potential in market-weight pigs. AB - Sixty-four market-weight (130.0 +/- 0.65 kg) barrows (n = 16) and gilts (n = 48) were used in a split-plot design with a 2 x 2 x 2 factorial arrangement of treatments: 1) handling intensity (gentle vs. aggressive), 2) transport floor space (0.39 vs. 0.49 m(2)/pig), and 3) distance moved during handling (25 vs. 125 m) to determine the effects of multiple concurrent stressors on metabolic responses. For the handling intensity treatment, pigs were moved individually approximately 50 m through a handling course with either 0 (gentle) or 8 (aggressive) shocks from an electric goad. Pigs were loaded onto a trailer and transported for approximately 1 h at floor spaces of either 0.39 or 0.49 m(2)/pig. After transport, pigs were unloaded, and the distance moved treatment was applied; pigs were moved 25 or 125 m through a handling course using livestock paddles. Rectal temperature was measured, and blood samples (to measure blood acid-base status) were collected 2 h before the handling intensity treatment was applied and immediately after the distance moved treatment was applied. A LM sample to measure glycolytic potential was collected after the distance moved treatments on a subset of 32 pigs. There were handling intensity x distance moved interactions (P < 0.05) for several blood acid-base measurements. In general, there was no effect of distance moved on these traits when pigs were previously handled gently. However, when pigs were previously handled aggressively, pigs moved 125 compared with 25 m had greater (P < 0.05) blood lactate and less (P < 0.05) blood pH, bicarbonate, and base-excess. Pigs transported at 0.39 compared with 0.49 m(2)/pig had a greater (P < 0.01) increase in creatine kinase values; however, transport floor space did not affect any other measurements. Data were analyzed by the number of stressors (the aggressive handling, restricted transport floor space, and 125-m distance moved treatments) experienced by each pig (0, 1, 2, or 3). As the number of stressors experienced by the pig increased, rectal temperature, blood lactate, and LM lactate increased linearly (P 0.05), but females had greater kidney knob and pelvic fat proportion than males (P < 0.01). Lambs had greater proportions of the highly valued leg cut and lower proportions of shoulder, anterior rib, and neck cuts than kids. Dissection results indicated that kid carcasses had greater muscle content and lower dissected fat and bone than lambs. Kids had greater (P < 0.001) muscle ultimate pH value than lambs (5.8 +/- 0.02 vs. 5.6 +/- 0.02). Males had greater (P < 0.05) muscle ultimate pH value than females (5.7 +/- 0.02 vs. 5.6 +/- 0.02). The kid meat was significantly lighter (P < 0.05) and less yellow (P < 0.001) than the lamb meat. Kids presented less cooking losses (P < 0.001) than lambs, and shear force value was significantly greater (P < 0.01) in lamb meat. The kid meat had significantly more moisture (P < 0.001) and less intramuscular fat content (P < 0.001) than lambs. At this maturity stage, there were significant differences on both carcass and meat quality attributes of suckling kids and lambs, possibly due to inherent differences between species. PMID- 18676734 TI - Moving toward individualized cancer therapies. AB - In vivo analysis of the gene expression profiles of cancer cells before and after treatment in patients may define mechanisms of sensitivity and resistance to specific drugs and ultimately allow for the selection of optimal individualized therapy to improve outcome in cancer. PMID- 18676735 TI - Erythropoiesis-stimulating agent use in cancer: preclinical and clinical perspectives. AB - Erythropoiesis-stimulating agents (ESA) used for the treatment of chemotherapy induced anemia in cancer patients have been associated with adverse outcomes of enhanced tumor progression and impaired survival in a series of recent clinical trials. As clinical practice guidelines for ESA administration in cancer patients have evolved to improve safety, the mechanisms underlying the adverse outcomes and whether ESAs exert direct and/or indirect effects in primary tumors to modulate tumor cell growth, survival, and chemoradiotherapy responses remain uncertain. Erythropoietin receptor (EpoR) expression in tumor cells has raised the simplistic possibility that Epo signaling mediated via a functional cellular receptor may contribute to tumor progression in a direct manner. However, Epo biology in cancer is likely to be complex and an interplay of multiple factors is potentially involved in the overall tumor response to exogenous Epo. Optimization of ESA use as an important supportive therapy modality in cancer patients, and further investigation of the role of Epo-EpoR in cancer biology will require a combination of carefully designed preclinical and clinical studies designed to ascertain not only the effect of ESA therapy on clinical outcomes such as tumor response, progression-free, and overall survival but also to investigate the potential effects of Epo on biomarkers of EpoR activation and factors related to tumor biology and chemoradiation responsiveness. PMID- 18676736 TI - The genetics of castration-resistant prostate cancer: what can the germline tell us? AB - Androgen deprivation therapy (ADT) is the cornerstone treatment for advanced prostate cancer. Despite frequent responses, the majority of metastatic tumors will progress to castration-resistant prostate cancer. Numerous molecular and genetic perturbations have been described in castration-resistant prostate cancer, which are attributable for gain-of-function changes in the androgen receptor, allowing for cell survival and proliferation with castrate levels of testosterone. The utility of these somatic perturbations, which are selected for in the tumor after ADT, for prognostication of response and response duration in metastatic prostate cancer, is problematic. Here, we discuss recent studies that describe germline polymorphisms that determine the response to ADT. Coding and noncoding germline polymorphisms in genes involved in the androgen pathway affect the response to ADT. These polymorphisms require further study and validation. However, they have the potential to be useful for prognosticating the response to ADT, designing clinical trials for patients who have poor germline prognostic features and designing novel therapies targeted against genes that influence the response to ADT. PMID- 18676737 TI - Loss of protein inhibitors of activated STAT-3 expression in glioblastoma multiforme tumors: implications for STAT-3 activation and gene expression. AB - PURPOSE: STATs activate transcription in response to numerous cytokines, controlling proliferation, gene expression, and apoptosis. Aberrant activation of STAT proteins, particularly STAT-3, is implicated in the pathogenesis of many cancers, including GBM, by promoting cell cycle progression, stimulating angiogenesis, and impairing tumor immune surveillance. Little is known about the endogenous STAT inhibitors, the PIAS proteins, in human malignancies. The objective of this study was to examine the expression of STAT-3 and its negative regulator, PIAS3, in human tissue samples from control and GBM brains. EXPERIMENTAL DESIGN: Control and GBM human tissues were analyzed by immunoblotting and immunohistochemistry to determine the activation status of STAT-3 and expression of the PIAS3 protein. The functional consequence of PIAS3 inhibition by small interfering RNA or PIAS3 overexpression in GBM cells was determined by examining cell proliferation, STAT-3 transcriptional activity, and STAT-3 target gene expression. This was accomplished using [(3)H]TdR incorporation, STAT-3 dominant-negative constructs, reverse transcription-PCR, and immunoblotting. RESULTS AND CONCLUSIONS: STAT-3 activation, as assessed by tyrosine and serine phosphorylation, was elevated in GBM tissue compared with control tissue. Interestingly, we observed expression of PIAS3 in control tissue, whereas PIAS3 protein expression in GBM tissue was greatly reduced. Inhibition of PIAS3 resulted in enhanced glioblastoma cellular proliferation. Conversely, PIAS3 overexpression inhibited STAT-3 transcriptional activity, expression of STAT-3 regulated genes, and cell proliferation. We propose that the loss of PIAS3 in GBM contributes to enhanced STAT-3 transcriptional activity and subsequent cell proliferation. PMID- 18676738 TI - Human papillomavirus type 16/18 up-regulates the expression of interleukin-6 and antiapoptotic Mcl-1 in non-small cell lung cancer. AB - PURPOSE: Human papillomavirus (HPV) 16/18 infection is reported to be associated with nonsmoking Taiwanese female lung cancer. In this study, we attempted to further reveal the association between HPV infection with Mcl-1 and interleukin (IL)-6 expressions and to elucidate the roles of HPV infection in lung tumorigenesis. EXPERIMENTAL DESIGN: IL-6 and Mcl-1 expressions were investigated in 79 tumor tissues from lung cancer patients by immunohistochemistry. Secreting IL-6 levels and Mcl-1 expressions were examined by ELISA and Western blot, respectively, in HPV 16/18 E6- and E7-transfected A549 human lung cancer cells, as well as in the HPV16-infected TL-1 lung cancer cells established from lung cancer patients. RESULTS: Lung tumors (70.9% and 57.0%) had positive IL-6 and Mcl 1 immunostainings, respectively. Significant correlation between IL-6 and Mcl-1 expression were observed (P < 0.0001). Both IL-6 and Mcl-1 expression were significantly associated with HPV 16/18 infection (P = 0.014 and P = 0.004, respectively). IL-6 and Mcl-1 protein levels were not only elevated in HPV 16/18 E6- and E7-transfected A549 cells but also in TL-1 cells. Phosphatidylinositol-3 OH kinase pathway was the major pathway contributing to the up-regulation of Mcl 1 by IL-6 in HPV-infected lung cancer cells. CONCLUSIONS: The up-regulating effects of HPV 16/18 E6 and E7 to IL-6 and Mcl-1 expressions were observed in E6- and E7-transfected A549 cells and in HPV16-infected TL-1 cells, mainly through the phosphatidylinositol-3-OH kinase pathway. The involvement of HPV infection in lung tumorigenesis may be partly through a concomitant increased expression of autocrine and/or paracrine IL-6 and the downstream Mcl-1. PMID- 18676739 TI - OPG is regulated by beta-catenin and mediates resistance to TRAIL-induced apoptosis in colon cancer. AB - PURPOSE: Resistance to apoptosis is a hallmark of cancer and correlates with aggressiveness of tumors and poor prognosis. The Wnt/beta-catenin pathway plays a pivotal role in the genesis of colorectal cancer by mechanisms not fully elucidated yet. Previous studies have linked regulation of osteoprotegerin (OPG) in bone to Wnt/beta-catenin signaling. As OPG also serves as a decoy receptor for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), we hypothesized that OPG might play a role in mediating resistance to apoptosis in colorectal cancer cells. EXPERIMENTAL DESIGN: Expression analysis and functional studies in human colorectal cancer cell lines and determination of expression in primary tumors and sera from patients with colorectal cancer. RESULTS: We found production of OPG in colorectal cancer cells to be regulated by beta-catenin/Tcf 4. Addition of exogenous OPG to colorectal cancer cells caused resistance to TRAIL. Similarly, accumulation of OPG in medium of cultivated cells caused resistance to TRAIL, and this could be reverted by removal of OPG. Furthermore, OPG levels were significantly increased in serum of patients with advanced disease. CONCLUSIONS: We conclude that the Wnt/beta-catenin pathway contributes to carcinogenesis and cancer cell survival by driving expression of OPG. Expression of the survival factor OPG might provide colorectal cancer cells with an essential growth advantage and contribute to cell invasion and metastasis. Inhibition of OPG expression might offer a new therapeutic approach for the treatment of patients with colorectal tumors overexpressing OPG and make these tumors sensitive to TRAIL-induced apoptosis. PMID- 18676740 TI - Delineation of TMPRSS2-ERG splice variants in prostate cancer. AB - PURPOSE: The expression of the ETS-related gene (ERG) is low or undetectable in benign prostate epithelial cells. High prevalence of ERG overexpression in prostate cancer cells due to TMPRSS2-ERG fusions suggest for causal roles of ERG protein in the neoplastic process. TMPRSS2-ERG fusion junctions have been extensively studied in prostate cancer. However, virtually nothing is known about the nature of full-length transcripts and encoded proteins. This study focuses on qualitative and quantitative features of full-length TMPRSS2-ERG transcripts in prostate cancer. EXPERIMENTAL DESIGN: Full-length TMPRSS2-ERG transcripts were cloned and sequenced from a cDNA library generated from pooled RNA of six TMPRSS2 ERG fusion-positive prostate tumors. The encoded ERG proteins were analyzed in HEK293 cells. Copy numbers of TMPRSS2-ERG splice variants were determined by quantitative reverse transcription-PCR in laser capture microdissected prostate cancer cells. RESULTS: Two types of TMPRSS2-ERG cDNAs were identified: type I, which encodes full-length prototypical ERG protein (ERG1, ERG2, ERG3), and type II, encoding truncated ERG proteins lacking the ETS domain (ERG8 and a new variant, TEPC). In microdissected prostate tumor cells from 122 patients, relative abundance of these variants was in the following order: ERG8 > TEPC > ERG 3 > ERG1/2 with combined overexpression rate of 62.3% in prostate cancer. Increased ratio of type I over type II splice forms showed a trend of correlation with less favorable pathology and outcome. CONCLUSIONS: Qualitative and quantitative features of specific ERG splice variants defined here promise to enhance the utility of ERG as a biomarker and therapeutic target in prostate cancer. PMID- 18676741 TI - Improved identification of von Hippel-Lindau gene alterations in clear cell renal tumors. AB - PURPOSE: To provide a comprehensive, thorough analysis of somatic mutation and promoter hypermethylation of the von Hippel-Lindau (VHL) gene in the cancer genome, unique to clear cell renal cancer (ccRCC). Identify relationships between the prevalence of VHL gene alterations and alteration subtypes with patient and tumor characteristics. EXPERIMENTAL DESIGN: As part of a large kidney cancer case control study conducted in Central Europe, we analyzed VHL mutations and promoter methylation in 205 well-characterized, histologically confirmed patient tumor biopsies using a combination of sensitive, high-throughput methods (endonuclease scanning and Sanger sequencing) and analysis of 11 CpG sites in the VHL promoter. RESULTS: We identified mutations in 82.4% of cases, the highest VHL gene mutation prevalence reported to date. Analysis of 11 VHL promoter CpG sites revealed that 8.3% of tumors were hypermethylated and all were mutation negative. In total, 91% of ccRCCs exhibited alteration of the gene through genetic or epigenetic mechanisms. Analysis of patient and tumor characteristics revealed that certain mutation subtypes were significantly associated with Fuhrman nuclear grade, metastasis, node positivity, and self-reported family history of RCC. CONCLUSION: Detection of VHL gene alterations using these accurate, sensitive, and practical methods provides evidence that the vast majority of histologically confirmed ccRCC tumors possess genetic or epigenetic alteration of the VHL gene and support the hypothesis that VHL alteration is an early event in ccRCC carcinogenesis. These findings also indicate that VHL molecular subtypes can provide a sensitive marker of tumor heterogeneity among histologically similar ccRCC cases for etiologic, prognostic, and translational studies. PMID- 18676742 TI - Gene expression profiling of papillary thyroid carcinoma identifies transcripts correlated with BRAF mutational status and lymph node metastasis. AB - PURPOSE: To identify papillary thyroid carcinoma (PTC)-associated transcripts, we compared the gene expression profiles of three Serial Analysis of Gene Expression libraries generated from thyroid tumors and a normal thyroid tissue. EXPERIMENTAL DESIGN: Selected transcripts were validated in a panel of 57 thyroid tumors using quantitative PCR (qPCR). An independent set of 71 paraffin-embedded sections was used for validation using immunohistochemical analysis. To determine if PTC associated gene expression could predict lymph node involvement, a separate cohort of 130 primary PTC (54 metastatic and 76 nonmetastatic) was investigated. The BRAF(V600E) mutational status was compared with qPCR data to identify genes that might be regulated by abnormal BRAF/MEK/extracellular signal-regulated kinase signaling. RESULTS: We identified and validated new PTC-associated transcripts. Three genes (CST6, CXCL14, and DHRS3) are strongly associated with PTC. Immunohistochemical analysis of CXCL14 confirmed the qPCR data and showed protein expression in PTC epithelial cells. We also observed that CST6, CXCL14, DHRS3, and SPP1 were associated with PTC lymph node metastasis, with CST6, CXCL14, and SPP1 being positively correlated with metastasis and DHRS3 being negatively correlated. Finally, we found a strong correlation between CST6 and CXCL14 expression and BRAF(V600E) mutational status, suggesting that these genes may be induced subsequently to BRAF activation and therefore may be downstream in the BRAF/MEK/extracellular signal-regulated kinase signaling pathway. CONCLUSION: CST6, CXCL14, DHRS3, and SPP1 may play a role in PTC pathogenesis and progression and are possible molecular targets for PTC therapy. PMID- 18676743 TI - Epithelial-mesenchymal transition in cervical cancer: correlation with tumor progression, epidermal growth factor receptor overexpression, and snail up regulation. AB - PURPOSE: Acquisition of epithelial-mesenchymal transition (EMT) by primary carcinoma cells is associated with disrupted epithelial integrity, local invasion, and ultimately metastasis. Little is known about the existence and function of EMT in cervical cancer. This study aims to investigate the regulation of EMT in cervical squamous cell carcinoma. EXPERIMENTAL DESIGN: We investigated the molecular events of EMT in surgical specimens, which present the progression of cervical carcinoma. Two cervical cancer cell lines and the primary culture of normal cervical epithelia were used to study the regulatory mechanisms of EMT. RESULTS: The chronic epidermal growth factor (EGF) treatment induces the elongation of cell shape, increases cell scattering, and enhances cell invasion. EGF treatment down-regulates E-cadherin and up-regulates vimentin in cervical cancer cells. These characteristics are consistent with the morphologic changes, molecular events, and functional significance of EMT. EGF receptor (EGFR) signaling inactivates glycogen synthase kinase-3beta, which results in the nuclear accumulation of up-regulated Snail and then leads to EMT program. alpha(5)beta(1) integrin signaling and extracellular matrix fibronectin can modulate EGF-induced EMT. Importantly, the immunofluorescent stainings of surgical specimens indicate that cervical carcinoma progression is accompanied by EGFR overexpression, which is in parallel with decreased E-cadherin and increased vimentin. Up-regulation and nuclear accumulation of Snail correlate with EMT program in tumor tissues. CONCLUSION: EGF cooperates with alpha(5)beta(1) integrin signaling to induce EMT in cervical cancer cells via up-regulated Snail. Blockade of EGFR activity or expression may provide a potential target for the treatment of cervical cancer progression. PMID- 18676744 TI - Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer. AB - PURPOSE: Epidermal growth factor receptor (EGFR) mutations, especially in-frame deletions in exon 19 (DEL) and a point mutation in exon 21 (L858R), predict gefitinib sensitivity in patients with non-small cell lung cancer (NSCLC). In this study, we verified the accuracy of EGFR mutation analysis in small samples by high-resolution melting analysis (HRMA), which is a rapid method using PCR amplification with a dye to analyze the melting curves in NSCLC. EXPERIMENTAL DESIGN: We designed a prospective study to compare the sensitivity and specificity of HRMA and DNA sequencing with laser capture microdissection. Eligible patients with lung lesions were screened by bronchoscopy or percutaneous needle biopsy to histologically confirm the diagnosis, followed by surgical resection of the NSCLC. Small diagnostic specimens were analyzed for EGFR mutations by HRMA, and the surgically resected specimens were examined for mutations by HRMA and DNA sequencing. RESULTS: The analyses for EGFR mutations were conducted in 52 eligible cases of the 92 enrolled patients. EGFR mutations were detected in 18 (34.6%) patients. The results of HRMA from surgically resected specimens as well as DNA sequencing revealed 100% sensitivity and specificity. On the other hand, the sensitivity and specificity of HRMA from the small diagnostic specimens were 83.3% and 100%, respectively. CONCLUSIONS: In this study, we showed that HRMA is a highly accurate method for detecting DEL and L858R mutations in patients with NSCLC, although it is necessary to consider the identification of patients with a false-negative result when the analysis is conducted using small samples. PMID- 18676745 TI - Imaging of urokinase-type plasminogen activator receptor expression using a 64Cu labeled linear peptide antagonist by microPET. AB - PURPOSE: Malignant tumors are capable of degrading the surrounding extracellular matrix, resulting in local invasion or metastasis. Urokinase-type plasminogen activator (uPA) and its cell surface receptor (uPAR) are central molecules in one of the major protease systems involved in extracellular matrix degradation. Noninvasive imaging of this receptor in vivo with radiolabeled peptides that specifically target uPAR may therefore be useful to decipher the potential invasiveness of malignant lesions. EXPERIMENTAL DESIGN: In this study, we developed a (64)Cu-labeled uPAR-binding peptide for positron emission tomography (PET) imaging. A linear, high-affinity uPAR-binding peptide antagonist AE105 was conjugated with 1,4,7,10-tetraazadodecane-N,N',N'',N'''-tetraacetic acid (DOTA) and labeled with (64)Cu for microPET imaging of mice bearing U87MG human glioblastoma (uPAR positive) and MDA-MB-435 human breast cancer (uPAR negative). RESULTS: Surface plasmon resonance measurements show that AE105 with DOTA conjugated at the alpha-amino group (DOTA-AE105) has high affinity toward uPAR. microPET imaging reveals a rapid and high accumulation of (64)Cu-DOTA-AE105 in uPAR-positive U87MG tumors (10.8 +/- 1.5%ID/g at 4.5 hours, n = 3) but not in uPAR-negative MDA-MB-435 tumors (1.2 +/- 0.6%ID/g at 4.5 hours, n = 3). Specificity of this peptide-based imaging of uPAR was validated by further control experiments. First, a nonbinding variant of AE105 carrying a single amino acid replacement (Trp-->Glu) does not target U87MG tumors in vivo. Second, targeting of U87MG tumors by (64)Cu-DOTA-AE105 is specifically inhibited by a nonlabeled antagonist. CONCLUSION: The successful demonstration of the ability of a (64)Cu labeled uPAR-specific probe to visualize uPAR expression in vivo may allow clinical translation of this class of radiopharmaceuticals for uPAR positive cancer detection and patient stratification for uPA/uPAR system-based cancer therapy. PMID- 18676746 TI - Pattern of serum autoantibodies allows accurate distinction between a tumor and pathologies of the same organ. AB - PURPOSE: Recent studies impressively showed the diagnostic potential of seroreactivity patterns for different tumor types, offering the prospect for low cost screening of numerous tumor types simultaneously. One of the major challenges toward this goal is to prove that seroreactivity profiles do not only allow for identifying a tumor but also allow for distinguishing tumors from other pathologies of the same organ. EXPERIMENTAL DESIGN: We chose glioma as a model system and tested 325 sera (88 glioma, 95 intracranial tumors, 60 other brain pathologies, and 82 healthy controls) for seroreactivity on a panel of 35 antigens. RESULTS: We were able to discriminate between glioma and all other sera with cross-validated specificity of 86.1%, sensitivity of 85.2%, and accuracy of 85.8%. We obtained comparably good results for the separation of glioma versus nontumor brain pathologies and glioma versus other intracranial tumors. CONCLUSION: Our study provides first evidence that seroreactivity patterns allow for an accurate discrimination between a tumor and pathologies of the same organ even between different tumor types of the same organ. PMID- 18676747 TI - The retinoblastoma family member pRb2/p130 is an independent predictor of survival in human soft tissue sarcomas. AB - PURPOSE: pRb2/p130, a member of the Retinoblastoma gene family, has been shown to be a powerful prognostic factor in several malignancies. We sought to evaluate pRb2/p130 protein expression and its clinical effect in patients affected with soft tissue sarcomas (STS). EXPERIMENTAL DESIGN: Expression of pRb2/p130 was evaluated by immunohistochemistry on formalin-fixed, paraffin-embedded sections in 41 STSs. Results obtained were correlated with clinicopathologic variables and disease-free and overall survival (OS) in univariate and multivariate analysis. RESULTS: Expression of pRb2/p130 was diminished in 25 (61%) tumors, whereas the remaining ones (39%) were classified as high expressors. No correlation between pRb2/p130 expression and clinicopathologic variables was observed. However, a direct relationship between pRb2/p130 expression and clinical outcome of the patients was found in the subgroup of nonmetastatic tumors (n = 31). In univariate analysis, reduced pRb2/p130 expression was a negative prognostic factor and correlated with shorter disease-free survival (P = 0.021) and OS (P = 0.017) survival. In multivariate analysis, reduced pRb2/p130 expression was confirmed to be an independent predictor of shorter OS when considered together with tumor stage and grading (risk ratio, 7.893; confidence interval, 1.618 38.509; P = 0.011). CONCLUSIONS: This study shows for the first time the potential prognostic value of pRb2/130 expression evaluated on formalin-fixed, paraffin-embedded sections in STSs patients. pRb2/p130 immunoreactivity can be used to predict OS in patients with nonmetastatic STSs and, therefore, may represent a new prognostic marker. PMID- 18676748 TI - Prognostic significance of CD55 expression in breast cancer. AB - PURPOSE: Our recent study revealed that CD55-high population in breast cancer cell line was resistant to apoptosis and formed colonies in vitro more efficiently than CD55-low population. The present study was conducted to examine whether CD55-high population in breast cancer cell line possesses higher tumorigenic potential in vivo and presence of CD55-high cells in breast cancer affects clinicopathologic behavior of patients. EXPERIMENTAL DESIGN: CD55-high and CD55-low population was sorted from breast cancer cell line, injected into immunodeficient mice, and the resultant tumor volume was measured. CD55 expression was immunohistochemically examined in clinical samples from 74 cases with breast cancers, and cases with >1% of tumor cells showing high level of CD55 expression were categorized as CD55 high. RESULTS: The xenotransplanted tumor volume derived from CD55-high population was significantly larger than that from CD55-low population. Fifty (67.6%) of 74 cases of breast cancer were CD55-high. A significant correlation was observed between CD55-high character and relapse rate (P < 0.001). Univariate analysis showed that tumor size (P = 0.005) and CD55 expression (P = 0.005) were unfavorable prognostic factors. Multivariate analysis revealed that the tumor size (P = 0.013) and CD55 expression (P = 0.011) were independent prognostic factors. CONCLUSIONS: CD55 play an important role in tumorigenesis of breast cancer, and presence of small population of cells with strong CD55 expression would be sufficient to predict poor prognosis of patients. PMID- 18676749 TI - Genetic polymorphisms of metastasis suppressor gene NME1 and breast cancer survival. AB - PURPOSE: Ample evidence supports an important role of tumor metastasis suppressor genes in cancer metastatic processes. We evaluated the association of genetic polymorphisms of metastasis suppressor gene NME1 with breast cancer prognosis in a follow-up study of patients with primary breast cancer and further investigated the functions of these polymorphisms. EXPERIMENTAL DESIGN: NME1 genotypes were analyzed in a cohort of 1,134 breast cancer patients recruited as part of the Shanghai Breast Cancer Study who were followed for a median of 7.1 years. In vitro biochemical analyses were carried out to examine the function of NME1 gene polymorphisms. RESULTS: Single nucleotide polymorphisms (SNP) in the promoter region of the NME1 gene were found to be associated with breast cancer prognosis. Patients carrying the C allele in rs16949649 were associated with higher breast cancer-specific mortality [hazard ratio (HR), 1.4; 95% confidence interval (95% CI), 1.1-1.9] compared with those carrying the wild-type allele, and the association was more evident in patients with an early-stage cancer (HR, 1.7; 95% CI, 1.2-2.5). SNP rs2302254 was also associated with breast cancer prognosis, and the association was statistically significant for the risk of breast cancer relapse, metastasis, and death (HR, 1.3; 95% CI, 1.0-1.6). In vitro biochemical analyses showed that minor alleles in rs2302254 and rs3760468, which is in strong linkage disequilibrium with rs16949646, altered nuclear proteins binding capacity and reduced NME1 promoter activity, supporting the results from an association study of these SNPs with breast cancer survival. CONCLUSION: Promoter polymorphisms in the NME1 gene may alter its expression and influence breast cancer survival. PMID- 18676750 TI - Three-gene expression signature predicts survival in early-stage squamous cell carcinoma of the lung. AB - PURPOSE: Adjuvant treatment may improve survival in early-stage squamous cell carcinoma (SCC) of the lung; however, the absolute gain is modest and mainly limited to stage II-IIIA. Current staging methods are imprecise indications of prognosis, but high-risk patients can be identified by gene expression profiling and considered for adjuvant therapy. EXPERIMENTAL DESIGN: The expression of 29 genes was assessed by reverse transcriptase quantitative PCR in frozen primary tumor specimens obtained from 66 SCC patients who had undergone surgical resection. Expression values were dichotomized using the median as a cutoff value. We used a risk score to develop a gene expression model for the prediction of survival. RESULTS: The univariate analysis of gene expression in the training cohort identified 10 genes with significant prognostic value: CSF1, EGFR, CA IX, PH4, KIAA0974, ANLN, VEGFC, NTRK1, FN1, and INR1. In the multivariate Cox model, CSF1 (hazard ratio, 3.5; P = 0.005), EGFR (hazard ratio, 2.7; P = 0.02), CA IX (hazard ratio, 0.2; P < 0.0001), and tumor size >4 cm (hazard ratio, 2.7; P = 0.02) emerged as significant markers for survival. The high prognostic value of a risk score based on the expression of the three genes (CSF1, EGFR, and CA IX) was positively validated in a separate cohort of 26 patients in an independent laboratory (P = 0.05). CONCLUSIONS: The three-gene signature is strongly associated with prognosis in early-stage SCC. Positive independent validation suggests its suitability for selecting SCC patients with an increased risk of death who might benefit from adjuvant treatment. PMID- 18676751 TI - T-cell coregulatory molecule expression in urothelial cell carcinoma: clinicopathologic correlations and association with survival. AB - PURPOSE: Aberrant expression of T-cell coregulatory molecules has been investigated as a mechanism by which certain cancers may evade host immune surveillance. We evaluated expression of the T-cell coregulators B7-H1, B7-H3, and PD-1 in urothelial cell carcinoma (UCC) of the bladder. EXPERIMENTAL DESIGN: Immunohistochemistry for B7-H1, B7-H3, and PD-1 was done on paraffin-embedded sections from 318 consecutive patients with UCC who underwent radical cystectomy. Expression was correlated with clinicopathologic outcomes and postoperative survival. RESULTS: B7-H3 was widely expressed in UCC, as 222 of 314 (70.7%) tumors showed positive staining. Expression of B7-H3 in UCC was significantly increased compared with adjacent, nontumor urothelium, as a median of 70% of tumor cells expressed B7-H3, compared with 20% of cells in nontumor specimens (P < 0.001). The increase in B7-H3 expression was independent of tumor stage (P = 0.13). Expression of B7-H1 by UCC tumors (P < 0.001) and PD-1 by tumor infiltrating lymphocytes (P = 0.012) were significantly associated with increased pathologic stage. Patients who had received intravesical bacillus Calmette-Guerin before cystectomy tended to show increased expression of B7-H3 (P = 0.023) and PD 1 (P = 0.071) but were less likely to express B7-H1 (P = 0.027). Moreover, for the subset of patients with organ-confined disease (n = 167), B7-H1 expression independently predicted all-cause mortality after cystectomy (hazard ratio, 3.18; 95% confidence interval, 1.74-5.79; P < 0.001). CONCLUSIONS: B7-H3 is highly expressed in UCC across tumor stages, whereas B7-H1 and PD-1 expression are associated with advanced disease. B7-H1 expression predicts mortality after cystectomy for patients with organ-confined tumors. These molecules may represent novel diagnostic or prognostic markers, as well as therapeutic targets, for patients with UCC. PMID- 18676752 TI - Quantitative analysis of the transrenal excretion of circulating EBV DNA in nasopharyngeal carcinoma patients. AB - PURPOSE: The existence of transrenal clearance of circulating cell-free DNA is controversial. In this study, we used NPC as a model to investigate if circulating EBV DNA can be excreted into urine and to quantify the contribution of renal excretion to the clearance of plasma EBV DNA. EXPERIMENTAL DESIGN: Quantitative analysis of urine EBV DNA was done for 74 NPC patients using real time PCR with two different amplicon sizes. The urine concentration of EBV DNA was expressed as copies per millimole of creatinine (copies/mmol Cr) to minimize the effects of interindividual variations in hydration status. RESULTS: EBV DNA was detectable in the urine of 56% NPC patients using a 59-bp real-time PCR assay. The median urine EBV DNA concentrations measured by the 59- and 76-bp assays were 7,040 and 290 copies/mmol Cr, respectively. Patients with detectable urine EBV DNA had significantly higher plasma concentrations, with a positive correlation between the plasma and urine concentrations of EBV DNA. The fraction of plasma EBV DNA excreted into the urine was 0.0026% of that for creatinine. CONCLUSIONS: We have shown that circulating EBV DNA can be excreted transrenally into urine in NPC patient and the fraction of excretion is negatively associated with the size of the DNA molecules. Because there is a positive correlation between plasma and urine EBV DNA concentration, urine EBV DNA analysis may potentially be applicable as an ultra-noninvasive test for the monitoring and prognostication of NPC patients. PMID- 18676753 TI - ASPM is a novel marker for vascular invasion, early recurrence, and poor prognosis of hepatocellular carcinoma. AB - PURPOSE: Abnormal spindle-like microcephaly associated (ASPM) plays an important role in neurogenesis and cell proliferation. This study is to elucidate its role in hepatocellular carcinoma (HCC), particularly early tumor recurrence (ETR) and prognosis. EXPERIMENTAL DESIGN: We used reverse transcription-PCR assays to measure the ASPM mRNA levels in 247 HCC and correlated with clinicopathologic and molecular features. RESULTS: ASPM mRNA levels were high in fetal tissues but very low in most adult tissues. ASPM mRNA was overexpressed in 162 HCC (66%) but not in benign liver tumors. ASPM overexpression correlated with high alpha fetoprotein (P = 1 x 10(-8)), high-grade (grade II-IV) HCC (P = 2 x 10(-6)), high stage (stage IIIA-IV) HCC (P = 1 x 10(-8)), and importantly ETR (P = 1 x 10(-8)). ETR is the most critical unfavorable clinical prognostic factor. Among the various independent histopathologic (tumor size, tumor grade and tumor stage) and molecular factors (p53 mutation, high alpha-fetoprotein, and ASPM overexpression), tumor stage was the most crucial histologic factor (odds ratio, 14.7; 95% confidence interval, 6.65-33.0; P = 1 x 10(-8)), whereas ASPM overexpression (odds ratio, 6.49; P = 1 x 10(-8)) is the most important molecular factor associated with ETR. ASPM overexpression was associated with vascular invasion and ETR in both p53-mutated (all P values = 1 x 10(-8)) and non-p53 mutated HCC (P = 1 x 10(-8) and 0.00088, respectively). Hence, patients with APSM overexpressing HCC had lower 5-year survival (P = 0.000001) in both p53-mutated (P = 0.00008) and non-p53-mutated HCC (P = 0.0027). In low-stage (stage II) HCC, ASPM overexpression also correlated with higher ETR (P = 0.008). CONCLUSION: ASPM overexpression is a molecular marker predicting enhanced invasive/metastatic potential of HCC, higher risk of ETR regardless of p53 mutation status and tumor stage, and hence poor prognosis. PMID- 18676754 TI - Tumor cell gene expression changes following short-term in vivo exposure to single agent chemotherapeutics are related to survival in multiple myeloma. AB - Changes in global gene expression patterns in tumor cells following in vivo therapy may vary by treatment and provide added or synergistic prognostic power over pretherapy gene expression profiles (GEP). This molecular readout of drug cell interaction may also point to mechanisms of action/resistance. In newly diagnosed patients with multiple myeloma (MM), microarray data were obtained on tumor cells prior to and 48 hours after in vivo treatment using dexamethasone (n = 45) or thalidomide (n = 42); in the case of relapsed MM, microarray data were obtained prior to (n = 36) and after (n = 19) lenalidomide administration. Dexamethasone and thalidomide induced both common and unique GEP changes in tumor cells. Combined baseline and 48-hour changes in GEP in a subset of genes, many related to oxidative stress and cytoskeletal dynamics, were predictive of outcome in newly diagnosed MM patients receiving tandem transplants. Thalidomide-altered genes also changed following lenalidomide exposure and predicted event-free and overall survival in relapsed patients receiving lenalidomide as a single agent. Combined with baseline molecular features, changes in GEP following short-term single-agent exposure may help guide treatment decisions for patients with MM. Genes whose drug-altered expression were found to be related to survival may point to molecular switches related to response and/or resistance to different classes of drugs. PMID- 18676755 TI - K-Ras mutations and treatment outcome in colorectal cancer patients receiving exclusive fluoropyrimidine therapy. AB - PURPOSE: K-Ras mutations predict resistance to anti-epidermal growth factor receptor (EGFR) monoclonal antibodies. Because combinations of anti-EGFR with 5 fluorouracil (5-FU)-based chemotherapy are promising treatments, we analyzed the effect of K-Ras mutations in patients having received exclusive 5-FU therapy. EXPERIMENTAL DESIGN: This study was conducted on 93 stage IV colorectal cancer patients with unresectable measurable liver metastasis receiving 5-FU-leucovorin (56 men and 37 women; 77 cancer deaths). Liver metastases (n = 93) along with primary tumors (n = 48) were analyzed for K-Ras mutations (codons 12 and 13), p53 mutations (exons 4-9), p53 polymorphism (codon 72), thymidylate synthase (TS) polymorphism (28-bp repeats including G>C mutation), methylenetetrahydrofolate reductase polymorphism (677C>T, 1298A>C), thymidylate synthase (TS) activity, dihydropyrimidine dehydrogenase activity, folylpolyglutamate synthase activity, and p53 protein expression. RESULTS: Thirty-six of 93 (38.7%) metastases were K Ras mutated (30 at codon 12 and 6 at codon 13). Mutated primary tumors (16 of 48) matched perfectly with mutated metastases. The additional analyzed tumor markers were not different between K-Ras mutated and wild-type tumors. The objective response rate was 37%: 44.4% in K-Ras mutated versus 32.1% in wild-type K-Ras metastasis (P = 0.27). Low TS activity in metastasis was the only significant predictor of tumor response (P = 0.047). K-Ras status did not influence specific survival. CONCLUSIONS: The present data indicate a perfect concordance of K-Ras mutations between primary and liver metastasis and suggest that any predictive and/or prognostic value of K-Ras mutations in treatments combining anti-EGFR monoclonal antibodies with 5-FU should be exclusively linked to the anti-EGFR agent. PMID- 18676756 TI - A phase I trial of the oral, multikinase inhibitor sorafenib in combination with carboplatin and paclitaxel. AB - PURPOSE: This study evaluated the safety, maximum tolerated dose, pharmacokinetics, and antitumor activity of sorafenib, a multikinase inhibitor, combined with paclitaxel and carboplatin in patients with solid tumors. PATIENTS AND METHODS: Thirty-nine patients with advanced cancer (24 with melanoma) received oral sorafenib 100, 200, or 400 mg twice daily on days 2 to 19 of a 21 day cycle. All patients received carboplatin corresponding to AUC6 and 225 mg/m(2) paclitaxel on day 1. Pharmacokinetic analyses were done for sorafenib on days 2 and 19 of cycle 1 and for paclitaxel on day 1 of cycles 1 and 2. Pretreatment tumor samples from 17 melanoma patients were analyzed for BRAF mutations. RESULTS: Sorafenib was well tolerated at the doses evaluated. The most frequent severe adverse events were hematologic toxicities (grade 3 or 4 in 33 patients, 85%). Twenty-seven (69%) patients had sorafenib-related adverse events, the most frequent of which were dermatologic events (26 patients, 67%). Exposure to paclitaxel was not altered by intervening treatment with sorafenib. Treatment with sorafenib, paclitaxel, and carboplatin resulted in one complete response and nine partial responses, all among patients with melanoma. There was no correlation between BRAF mutational status and treatment responses in patients with melanoma. CONCLUSIONS: The recommended phase II doses are oral 400 mg twice daily sorafenib, carboplatin at an AUC6 dose, and 225 mg/m(2) paclitaxel. The tumor responses observed with this combined regimen in patients with melanoma warrant further investigation. PMID- 18676757 TI - Combination chemotherapy and ALVAC-CEA/B7.1 vaccine in patients with metastatic colorectal cancer. AB - PURPOSE: The combination of vaccines and chemotherapy holds promise for cancer therapy, but the effect of cytotoxic chemotherapy on vaccine-induced antitumor immunity is unknown. This study was conducted to assess the effects of systemic chemotherapy on ALVAC-CEA/B7.1-induced T-cell immunity in patients with metastatic colorectal cancer. EXPERIMENTAL DESIGN: Patients with metastatic colorectal cancer were treated with fluorouracil, leucovorin, and irinotecan and were also given ALVAC-CEA/B7.1 vaccine with or without tetanus toxoid adjuvant. Eligible patients were randomized to ALVAC followed by chemotherapy and booster vaccination (group 1), ALVAC and tetanus toxoid followed by chemotherapy (group 2), or chemotherapy alone followed by ALVAC in patients without disease progression (group 3). Humoral immune responses were measured by standard ELISA assay, and carcinoembryonic antigen (CEA)-specific T-cell responses were measured by IFN-gamma enzyme-linked immunospot assay. RESULTS: One hundred eighteen patients were randomized to receive either ALVAC before and concomitantly with chemotherapy (n = 39), ALVAC with tetanus adjuvant before and concomitantly with chemotherapy (n = 40), or chemotherapy followed by ALVAC (n = 39). Serious adverse events were largely gastrointestinal (n = 30) and hematologic (n = 24). Overall, 42 patients (40.4%) showed objective clinical responses. All patients developed antibody responses against ALVAC, but increased anti-CEA antibody titers were detected in only three patients. Increases in CEA-specific T cells were detected in 50%, 37%, and 30% of patients in groups 1, 2, and 3, respectively. There were no differences in clinical or immune responses between the treatment groups. CONCLUSION: The combination of ALVAC-CEA/B7.1 vaccine and systemic chemotherapy has an acceptable safety profile in patients with metastatic colorectal cancer. Systemic chemotherapy did not affect the generation of CEA-specific T-cell responses following vaccination. PMID- 18676758 TI - A pilot study of consolidative immunotherapy in patients with high-risk pediatric sarcomas. AB - PURPOSE: Patients with metastatic or recurrent Ewing's sarcoma family of tumors and alveolar rhabdomyosarcoma have <25% 5-year survival in most studies. This study administered a novel immunotherapy regimen aimed at consolidating remission in these patients. EXPERIMENTAL DESIGN: Fifty-two patients with translocation positive, recurrent, or metastatic Ewing's sarcoma family of tumors or alveolar rhabdomyosarcoma underwent prechemotherapy cell harvest via apheresis for potential receipt of immunotherapy. Following completion of standard multimodal therapy, 30 patients ultimately initiated immunotherapy and were sequentially assigned to three cohorts. All cohorts received autologous T cells, influenza vaccinations, and dendritic cells pulsed with peptides derived from tumor specific translocation breakpoints and E7, a peptide known to bind HLA-A2. Cohort 1 received moderate-dose recombinant human interleukin-2 (rhIL-2), cohort 2 received low-dose rhIL-2, and cohort 3 did not receive rhIL-2. RESULTS: All immunotherapy recipients generated influenza-specific immune responses, whereas immune responses to the translocation breakpoint peptides occurred in 39%, and only 25% of HLA-A2(+) patients developed E7-specific responses. Toxicity was minimal. Intention-to-treat analysis revealed a 31% 5-year overall survival for all patients apheresed (median potential follow-up 7.3 years) with a 43% 5-year overall survival for patients initiating immunotherapy. CONCLUSIONS: Consolidative immunotherapy is a scientifically based and clinically practical approach for integrating immunotherapy into a multimodal regimen for chemoresponsive cancer. Patients receiving immunotherapy experienced minimal toxicity and favorable survival. The robust influenza immune responses observed suggest that postchemotherapy immune incompetence will not fundamentally limit this approach. Future studies will seek to increase efficacy by using more immunogenic antigens and more potent dendritic cells. PMID- 18676759 TI - Mismatch repair deficiency does not mediate clinical resistance to temozolomide in malignant glioma. AB - PURPOSE: A major mechanism of resistance to methylating agents, including temozolomide, is the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT). Preclinical data indicates that defective DNA mismatch repair (MMR) results in tolerance to temozolomide regardless of AGT activity. The purpose of this study was to determine the role of MMR deficiency in mediating resistance in samples from patients with both newly diagnosed malignant gliomas and those who have failed temozolomide therapy. EXPERIMENTAL DESIGN: The roles of AGT and MMR deficiency in mediating resistance in glioblastoma multiforme were assessed by immunohistochemistry and microsatellite instability (MSI), respectively. The mutation status of the MSH6 gene, a proposed correlate of temozolomide resistance, was determined by direct sequencing and compared with data from immunofluorescent detection of MSH6 protein and reverse transcription-PCR amplification of MSH6 RNA. RESULTS: Seventy percent of newly diagnosed and 78% of failed-therapy glioblastoma multiforme samples expressed nuclear AGT protein in > or = 20% of cells analyzed, suggesting alternate means of resistance in 20% to 30% of cases. Single loci MSI was observed in 3% of patient samples; no sample showed the presence of high MSI. MSI was not shown to correlate with MSH6 mutation or loss of MSH6 protein expression. CONCLUSIONS: Although high AGT levels may mediate resistance in a portion of these samples, MMR deficiency does not seem to be responsible for mediating temozolomide resistance in adult malignant glioma. Accordingly, the presence of a fraction of samples exhibiting both low AGT expression and MMR proficiency suggests that additional mechanisms of temozolomide resistance are operational in the clinic. PMID- 18676760 TI - Motexafin lutetium-photodynamic therapy of prostate cancer: short- and long-term effects on prostate-specific antigen. AB - PURPOSE: The time course of serum prostate-specific antigen (PSA) response to photodynamic therapy (PDT) of prostate cancer was measured. EXPERIMENTAL DESIGN: Seventeen patients were treated in a phase I trial of motexafin lutetium-PDT. PDT dose was calculated in each patient as the product of the ex vivo measured pre PDT photosensitizer level and the in situ measured light dose. Serum PSA level was measured within 2 months before PDT (baseline), and at day 1; weeks 1 to 3; months 1, 2, and 3; months 4 to 6; and months 7 to 11 after PDT. RESULTS: At 24 hours after PDT, serum PSA increased by 98% +/- 36% (mean +/- SE) relative to baseline levels (P = 0.007). When patients were dichotomized based on median PDT dose, those who received high PDT dose showed a 119% +/- 52% increase in PSA compared with a 54% +/- 27% increase in patients treated at low PDT dose. Patients treated with high versus low PDT dose showed a median biochemical delay of 82 versus 43 days (P = 0.024), with biochemical delay defined as the length of time between PDT and a nonreversible increase in PSA to a value greater than or equal to baseline. CONCLUSIONS: Results show PDT to induce large, transient increases in serum PSA levels. Patients who experienced high PDT dose showed greater short-term increase in PSA and a significantly more durable PSA response (biochemical delay). These data strongly promote the need for individualized delivery of PDT dose and assessment of treatment effect in PDT of prostate cancer. Information gained from such patient-specific measurements could facilitate the introduction of multiple PDT sessions in patients who would benefit. PMID- 18676761 TI - Lung cancer with epidermal growth factor receptor exon 20 mutations is associated with poor gefitinib treatment response. AB - PURPOSE: Clinical reports about responsiveness to gefitinib treatment in patients of non-small cell lung cancer (NSCLC) with mutations in exon 20 of epidermal growth factor receptor (EGFR) are limited. To increase understanding of the influence of exon 20 mutations on NSCLC treatment with gefitinib, we investigated the clinical features of lung cancer in patients with exon 20 mutations and analyzed the gefitinib treatment response. EXPERIMENTAL DESIGN: We surveyed the clinical data and mutational studies of NSCLC patients with EGFR exon 20 mutations in the National Taiwan University Hospital and reviewed the literature reports about EGFR exon 20 mutations and the gefitinib treatment response. RESULTS: Twenty-three patients with mutations in exon 20 were identified. Nine (39%) had coexisting mutations in EGFR exons other than exon 20. Sixteen patients received gefitinib treatment, and a response was noted in 4 patients. The gefitinib response rate of NSCLC with exon 20 mutations was 25%, far lower than those with deletions in exon 19 and L858R mutations. Interestingly, different exon 20 mutations and coexisting mutations seemed to have a different influence on gefitinib response. CONCLUSIONS: EGFR exon 20 mutations of NSCLC patients result in poorer responsiveness to gefitinib treatment, but variability exists between different individuals. PMID- 18676763 TI - Efficacy of systemically administered mutant vesicular stomatitis virus (VSVDelta51) combined with radiation for nasopharyngeal carcinoma. AB - PURPOSE: Nasopharyngeal carcinoma (NPC) is a malignancy of the head and neck region that is associated with EBV latency. Curative treatments for NPC achieve modest survival rates, underscoring a need to develop novel therapies. We evaluated the therapeutic potential of a mutant vesicular stomatitis virus (VSVDelta51) as single treatment modality or in combination with ionizing radiation (RT) in NPC. EXPERIMENTAL DESIGN: MTS assay was used to assess cell viability in vitro; apoptosis was measured using propidium iodide staining and caspase activation. In vivo experiments were conducted using tumor-bearing nude mice with or without local RT (4 Gy). Apoptosis was assessed in excised tumor sections with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. RESULTS: Our data showed that NPC cells are exquisitely sensitive to VSVDelta51 oncolysis, which correlated with the presence of EBV. Efficacy of VSVDelta51 against NPC cells was further augmented when combined with RT. A single systemic injection of VSVDelta51 achieved 50% survival in treated mice, which increased to 83% when combined with local tumor RT. In addition to induction of apoptosis, an antiangiogenic effect of VSVDelta51 was observed in vivo, suggesting a novel tumoricidal mechanism for VSVDelta51. This virus also prevented growth of NPC sphere-forming cells in vitro, showing potential utility in targeting NPC-initiating cells. CONCLUSIONS: Our data represent the first report showing that EBV-positive NPC cells are exquisitely sensitive to VSVDelta51 oncolysis and documenting the successful utilization of this combinatorial regimen as a novel curative therapeutic strategy for NPC. PMID- 18676764 TI - Oxidation of ovarian epithelial cancer cells by hypochlorous acid enhances immunogenicity and stimulates T cells that recognize autologous primary tumor. AB - PURPOSE: Hypochlorous acid, a product of neutrophil myeloperoxidase, is a powerful enhancer of antigen processing and presentation. In this study, we examine whether ovarian epithelial cells (SK-OV-3) exposed to hypochlorous acid can stimulate T cells from patients with ovarian epithelial cancer that recognize common tumor antigens as well as autologous tumor. EXPERIMENTAL DESIGN: T cells from human leukocyte antigen (HLA)-A2(+) and HLA-A2(-) patients or healthy controls were stimulated with autologous dendritic cells cocultured with the generic ovarian tumor line SK-OV-3, previously exposed to hypochlorous acid. RESULTS: Hypochlorous acid-treated SK-OV-3 cells drove expansion of CD8(+) T cells from HLA-A2(+) individuals, which recognized the HLA-A2-restricted tumor antigen epitopes of HER-2/neu (E75 and GP2) and MUC1 (M1.1 and M1.2). Up to 4.1% of the T cells were positive for the HER-2/neu KIFGSLAFL epitope using pentamer staining. Dendritic cells loaded with oxidized SK-OV-3 cells and further matured with CD40 agonistic antibody or monophosphoryl lipid A additionally induced CD4(+) class II-restricted responses. Critically, T cells stimulated with mature oxidized SK-OV-3 (but not a control oxidized melanoma cell line) directly recognized autologous tumor cells isolated from patient ascites. CONCLUSIONS: Immunization with mature dendritic cells loaded with a generic oxidized tumor cell line stimulates a polyclonal antitumor response that recognizes autologous tumor. These findings suggest a new immunotherapeutic strategy to extend remission in ovarian cancer. PMID- 18676762 TI - T-cell responses to survivin in cancer patients undergoing radiation therapy. AB - PURPOSE: The goal of this study was to determine if radiation therapy (RT) of human cancer enhances or diminishes tumor-specific T-cell reactivity. This is important if immunotherapy is to be harnessed to improve the outcome of cancer radiotherapy. EXPERIMENTAL DESIGN: Lymphocytes were isolated from colorectal cancer (CRC) patients before, during, and after presurgical chemoradiotherapy. Similar samples were taken from prostate cancer patients receiving standard RT. The level of CD8(+) T cells capable of binding tetramers for the tumor-associated antigen survivin, which is overexpressed in both cancer types, was enumerated in HLA-A*0201 patient samples. CD4(+), CD25(high), Foxp3(+) cells were also enumerated to evaluate therapy-induced changes in T(regulatory) cells. For CRC patients, most of whom were enrolled in a clinical trial, pathologic response data were available, as well as biopsy and resection specimens, which were stained for cytoplasmic and intranuclear survivin. RESULTS: Survivin-specific CD8(+) T lymphocytes were detected in the peripheral blood of CRC and prostate cancer patients and increased after therapy in some, but not all, patients. Increases were more common in CRC patients whose tumor was downstaged after chemoradiotherapy. Biopsy specimens from this cohort generally had higher nuclear to cytoplasmic survivin expression. T(regulatory) cells generally increased in the circulation following therapy but only in CRC patients. CONCLUSION: This study indicates that RT may increase the likelihood of some cancer patients responding to immunotherapy and lays a basis for future investigations aimed at combining radiation and immunotherapy. PMID- 18676765 TI - Sorafenib potently inhibits papillary thyroid carcinomas harboring RET/PTC1 rearrangement. AB - PURPOSE: Papillary thyroid carcinomas (PTC) are the most common type of thyroid malignancy with one of the two mutations, RET/PTC rearrangement or BRAF mutation. Both mutations are able to activate the MEK/ERK signaling transduction pathway and result in the activation of transcription factors that regulate cellular proliferation, differentiation, and apoptosis. Sorafenib (Nexavar, BAY 43-9006) is a multikinase inhibitor, and in this study, we tested its effects on PTC cells carrying either mutation. EXPERIMENTAL DESIGN: The effects of sorafenib on cell proliferation and signaling were evaluated in vitro on PTC cells using growth curves, cell cycle analysis, and immunoblotting. Using an orthotopic mouse model, we determined the antitumor effects of sorafenib in vivo. RESULTS: The concentration needed for 50% growth inhibition (GI(50)) by sorafenib was 0.14 mumol/L for the PTC cells with the RET/PTC1 rearrangement, and 2.5 mumol/L for PTC cells with a BRAF mutation, both readily achievable serum concentrations. After 3 weeks of oral administration of sorafenib (80 mg/kg/d) in mice, small (94% reduction compared with controls) or no tumor growth was detected in mice inoculated with PTC cells bearing the RET/PTC1 rearrangement, whereas the tumor volume of the orthotopic tumor implants of PTC cells with a BRAF mutation was reduced 53% to 54% (as compared with controls). CONCLUSIONS: PTC cells carrying the RET/PTC1 rearrangement were more sensitive to sorafenib than PTC cells carrying a BRAF mutation. Because RET/PTC rearrangements are unique to thyroid carcinomas, our findings support the clinical evaluation of sorafenib for patients with PTC and the identification of patients most likely to respond to sorafenib treatment. PMID- 18676766 TI - Radiation-mediated up-regulation of gene expression from replication-defective adenoviral vectors: implications for sodium iodide symporter gene therapy. AB - PURPOSE: To assess the effects of external beam radiotherapy (EBRT) on adenoviral mediated transgene expression in vitro and in vivo and to define an optimal strategy for combining sodium iodide symporter (NIS)-mediated (131)I therapy with EBRT. EXPERIMENTAL DESIGN: Expression of reporter genes [NIS, green fluorescent protein (GFP), beta-galactosidase (lacZ), and luciferase (Luc)] from replication deficient adenoviruses was assessed in tumor cell lines under basal conditions and following irradiation. The effects of viral multiplicity of infection (MOI) and EBRT dose on the magnitude and duration of gene expression were determined. In vivo studies were done with Ad-CMV-GFP and Ad-RSV-Luc. RESULTS: EBRT increased NIS, GFP, and beta-galactosidase expression in colorectal, head and neck, and lung cancer cells. Radiation dose and MOI were important determinants of response to EBRT, with greatest effects at higher EBRT doses and lower MOIs. Radiation exerted both transductional (through increased coxsackie-adenoviral receptor and integrin alpha(v)) and nontransductional effects, irrespective of promoter sequence (CMV, RSV, hTR, or hTERT). Analysis of the schedule of EBRT followed by viral infection revealed maximal transduction at 24 hours. Radiation maintained increasing radioiodide uptake from Ad-hTR-NIS over 6 days, in direct contrast to reducing levels in unirradiated cells. The effects of EBRT in increasing and maintaining adenovirus-mediated transgene expression were also seen in vivo using GFP- and luciferase-expressing adenoviral vectors. CONCLUSIONS: Radiation increased the magnitude and duration of NIS gene expression from replication deficient adenoviruses. The transductional effect is maximal at 24 hours, but radioiodide uptake is maintained at an elevated level over 6 days after infection. PMID- 18676767 TI - Radiation therapy with tositumomab (B1) anti-CD20 monoclonal antibody initiates extracellular signal-regulated kinase/mitogen-activated protein kinase-dependent cell death that overcomes resistance to apoptosis. AB - PURPOSE: The use of targeted radiation therapy (RT) in conjunction with anti-CD20 monoclonal antibodies (mAb) delivers high clinical response rates in B-cell lymphomas as part of radioimmunotherapy. The mechanisms underlying these impressive responses, particularly in patients whose lymphomas have become refractory to chemotherapy, are poorly understood. EXPERIMENTAL DESIGN: In this study, we have investigated the signaling pathways and mode of cell death induced in B-cell lymphoma cells after the combination of RT and either type I (rituximab) or type II (tositumomab/B1) anti-CD20 mAb. RESULTS: Increased tumor cell death was observed when RT was combined with tositumomab, but not rituximab. This additive cell death was found to be mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK)-dependent and could be reversed with mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) inhibitors, as well as small interfering RNA targeting MEK1/2. Furthermore, we found that this increased death was associated with ERK1/2 nuclear accumulation after tositumomab treatment, which was enhanced in combination with RT. Importantly, although Bcl-2 overexpression resulted in resistance to RT-induced apoptosis, it had no effect on the tumor cell death induced by tositumomab plus RT, indicating a nonapoptotic form of cell death. CONCLUSIONS: These findings indicate that RT and type II anti-CD20 mAb combine to stimulate a prodeath function of the MEK-ERK1/2 pathway, which is able to overcome apoptotic resistance potentially explaining the efficacy of this modality in treating patients with chemoresistant disease. PMID- 18676768 TI - Suppression of tumor formation by a cyclooxygenase-2 inhibitor and a peroxisome proliferator-activated receptor gamma agonist in an in vivo mouse model of spontaneous breast cancer. AB - PURPOSE: Activation of COX-2 and inhibition of PPARgamma have been observed in human and animal models of breast cancer. Both inhibition of COX-2 and activation of PPARgamma can inhibit proliferation of breast cancer cells in vitro. Here, we examine the effects of the COX-2 inhibitor celecoxib and the PPARgamma agonist N (9-fluorenyl-methyloxycarbonyl)-l-leucine (F-L-Leu) on mouse breast tumor cells in vitro and in vivo. EXPERIMENTAL DESIGN: We created and characterized a mouse mammary adenocarcinoma cell (MMAC-1) line from C3 (1)-SV40 tumor antigen mice to study COX-2 and PPARgamma expression and response to celecoxib and F-L-Leu in vitro. To study the in vivo effects, C3 (1)-SV40 tumor antigen mice were given either control diet or diets containing three different concentrations of celecoxib and F-L-Leu as well as a combination of both agents. Mice were then followed for tumor formation up to 1 year. RESULTS: MMAC-1 cells express both COX 2 and PPARgamma mRNA and exhibited cooperative growth inhibition with a combination of celecoxib and F-L-Leu. In mice, the median age of death due to mammary tumors was significantly delayed in celecoxib-treated animals at all three concentrations but was not significantly affected by F-L-Leu treatment alone. A combination of celecoxib and F-L-Leu was significantly more effective than celecoxib alone. CONCLUSIONS: Our findings suggest that a combination of a COX-2 inhibitor and PPARgamma agonist can delay breast cancer in a mouse model and suggest that these agents should be studied in the context of human populations with high breast cancer risk. PMID- 18676770 TI - The targeted immunocytokine L19-IL2 efficiently inhibits the growth of orthotopic pancreatic cancer. AB - PURPOSE: Effective control of pancreatic cancer has been hampered primarily by the lack of tumor specificity of current treatment modalities. The highly specific antibody-mediated delivery of therapeutic agents to the tumor microenvironment might overcome this problem. We therefore investigated the therapeutic efficacy of the targeted immunocytokine L19-Interleukin-2 (L19-IL2), consisting of the human single-chain Fv antibody L19, which is highly specific for the extradomain B (ED-B) of fibronectin, and the human cytokine IL-2, in pancreatic cancer. EXPERIMENTAL DESIGN: Therapeutic effects of L19-IL-2, IL-2, and gemcitabine on tumor growth and metastasis were evaluated in orthotopic mouse models for pancreatic cancer. Immunohistochemistry was done to define ED-B expression, tumor necrosis, apoptosis, proliferation, and invasion of macrophages and natural killer (NK) cells. NK cells were depleted by i.v. injection of an anti-asialo-GM-1 antibody. RESULTS: ED-B is selectively expressed in human pancreatic cancer and in primary tumors and metastases of the mouse models. L19 IL-2 therapy was clearly superior to untargeted IL-2 or gemcitabine and inhibited tumor growth and metastasis with remarkable long-term tumor control. Therapeutic effects were associated with the induction of extensive tumor necrosis and inhibition of tumor cell proliferation. Immunohistochemistry revealed an increase of macrophages and NK cells in the tumor tissue, suggesting immune-mediated mechanisms. The functional relevance of NK cells for the therapeutic effect of the targeted immunocytokine L19-IL-2 was confirmed by NK cell depletion, which completely abolished its antitumor efficacy. CONCLUSIONS: These preclinical results strongly encourage the initiation of clinical studies using L19-IL-2 in pancreatic cancer. PMID- 18676769 TI - Human Mena+11a isoform serves as a marker of epithelial phenotype and sensitivity to epidermal growth factor receptor inhibition in human pancreatic cancer cell lines. AB - PURPOSE: hMena, member of the enabled/vasodilator-stimulated phosphoprotein family, is a cytoskeletal protein that is involved in the regulation of cell motility and adhesion. The aim of this study was to determine whether or not the expression of hMena isoforms correlated with sensitivity to EGFR tyrosine kinase inhibitors and could serve as markers with potential clinical use. EXPERIMENTAL DESIGN: Human pancreatic ductal adenocarcinoma cell lines were characterized for in vitro sensitivity to erlotinib, expression of HER family receptors, markers of epithelial to mesenchymal transition, and expression of hMena and its isoform hMena(+11a). The effects of epidermal growth factor (EGF) and erlotinib on hMena expression as well as the effect of hMena knockdown on cell proliferation were also evaluated. RESULTS: hMena was detected in all of the pancreatic tumor cell lines tested as well as in the majority of the human tumor samples [primary (92%) and metastatic (86%)]. Intriguingly, in vitro hMena(+11a) isoform was specifically associated with an epithelial phenotype, EGFR dependency, and sensitivity to erlotinib. In epithelial BxPC3 cells, epidermal growth factor up regulated hMena/hMena(+11a) and erlotinib down-regulated expression. hMena knockdown reduced cell proliferation and mitogen-activated protein kinase and AKT activation in BxPC3 cells, and promoted the growth inhibitory effects of erlotinib. CONCLUSIONS: Collectively, our data indicate that the hMena(+11a) isoform is associated with an epithelial phenotype and identifies EGFR-dependent cell lines that are sensitive to the EGFR inhibitor erlotinib. The availability of anti-hMena(+11a)-specific probes may offer a new tool in pancreatic cancer management if these results can be verified prospectively in cancer patients. PMID- 18676771 TI - Silencing of transforming growth factor-beta1 in situ by RNA interference for breast cancer: implications for proliferation and migration in vitro and metastasis in vivo. AB - PURPOSE: Overexpression of transforming growth factor (TGF)-beta has been implicated in promoting immune suppression, tumor angiogenesis, tumor cell migration, and invasion in many cancers, including carcinoma of the breast. Thus, targeted down-regulation of TGF-beta1 expression in breast cancer in situ and determination of its implications would provide new treatment approaches for disease management. EXPERIMENTAL DESIGN: Small interfering RNA constructs targeting TGF-beta1 were validated and used to develop clonal derivatives of the metastatic breast cancer cell line MDA-MB-435. The cells were used in several in vitro analyses, including migration, invasion, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide, apoptosis, and signaling assays. A wound-healing assay was used to determine migration of the cells in culture and a Boyden chamber transwell assay was used for invasion. Further, the clones were used in an in vivo mouse model for the kinetics of tumor growth and gene expression in the primary site and in lungs upon metastasis. RESULTS: Inhibition of TGF-beta1 expression in MDA-MB-435 cells showed a 35% decrease in migration and a 55% decrease in invasion in vitro, with a 50% increase in proliferation and no effect on apoptosis. In vivo analysis indicated a 90% decrease in the number of mice bearing macroscopic lung metastases; however, the primary tumors did not show any difference in the growth kinetics when compared with the parental MDA-MB-435 cells. Analysis of TGF-beta signaling pathways in the clonal derivatives showed a decrease in Smad2 activation and an increase in AKT and extracellular signal regulated kinase activation. Interestingly, analysis of TGF-beta receptor expression showed a decrease in both receptor I and II expression in TGF-beta1 silenced cells. These results suggest that inhibition of TGF-beta1 ligand may act as a negative feedback loop to disrupt the function of all TGF-beta isoforms. CONCLUSIONS: Therapies targeting the TGF-beta signaling pathway may be more effective in late-stage disease to prevent organ metastasis but not primary tumor formation and may be combined with other tumor-targeted therapies normally limited by increased circulating TGF-beta levels. PMID- 18676773 TI - Synergistic inhibition of lung tumorigenesis by a combination of green tea polyphenols and atorvastatin. AB - PURPOSE: The present study investigated the possible synergistic inhibitory effect of a novel combination of polyphenon E (PPE, a standardized green tea polyphenol preparation) and atorvastatin (trade name Lipitor) in a mouse tumorigenesis model and in human lung cancer H1299 and H460 cell lines. EXPERIMENTAL DESIGN: Female A/J mice were given two weekly i.p. injections of 4 (methylnitrosaminao)-1-(3-pyridyl)-1-butanone (150 mg/kg total dose); 1 week later, mice were treated with PPE (0.25% or 0.5% in drinking fluid), atorvastatin (200 or 400 ppm in diet), or PPE (0.25%) plus atorvastatin (200 ppm) for 16 weeks. The interaction of these two agents was also studied in human lung cancer H1299 and H460 cells. RESULTS: The individual agents, PPE or atorvastatin, were not effective in inhibiting lung tumorigenesis. The low-dose combination of PPE and atorvastatin, however, significantly reduced both the tumor multiplicity and tumor burden (by 56% and 55%, respectively, P < 0.05). Isobologram analysis of the interaction of the two agents indicated that the combination synergistically decreased tumor multiplicity (P = 0.0006) and tumor burden (P = 0.0009). The inhibition was associated with enhanced apoptosis and suppressed myeloid cell leukemia 1 (Mcl-1) level in adenoma as determined by immunohistochemistry and Western blots. Treatment with combinations of PPE and atorvastatin also synergistically decreased the number of viable H1299 and H460 cells as determined by isobologram analysis. This synergistic effect was associated with increased apoptosis as determined by the terminal deoxyribonucleotide transferase-mediated nick-end labeling assay. The combination of PPE and atorvastatin was more efficient in reducing the antiapoptotic protein Mcl-1 level and increasing the cleaved caspase-3 and cleaved poly(ADP)-ribose polymerase level than the single agent treatment. CONCLUSIONS: The present work showed that PPE and atorvastatin synergistically inhibited 4-(methylnitrosaminao)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis in mice and the growth of lung cancer H1299 and H460 cells, possibly through enhanced apoptosis. The results provide leads for future research on the application of this combination for the prevention and treatment of lung cancer. PMID- 18676772 TI - Telomere maintenance in laser capture microdissection-purified Barrett's adenocarcinoma cells and effect of telomerase inhibition in vivo. AB - PURPOSE: The aims of this study were to investigate telomere function in normal and Barrett's esophageal adenocarcinoma (BEAC) cells purified by laser capture microdissection and to evaluate the effect of telomerase inhibition in cancer cells in vitro and in vivo. EXPERIMENTAL DESIGN: Epithelial cells were purified from surgically resected esophagi. Telomerase activity was measured by modified telomeric repeat amplification protocol and telomere length was determined by real-time PCR assay. To evaluate the effect of telomerase inhibition, adenocarcinoma cell lines were continuously treated with a specific telomerase inhibitor (GRN163L) and live cell number was determined weekly. Apoptosis was evaluated by Annexin labeling and senescence by beta-galactosidase staining. For in vivo studies, severe combined immunodeficient mice were s.c. inoculated with adenocarcinoma cells and following appearance of palpable tumors, injected i.p. with saline or GRN163L. RESULTS: Telomerase activity was significantly elevated whereas telomeres were shorter in BEAC cells relative to normal esophageal epithelial cells. The treatment of adenocarcinoma cells with telomerase inhibitor, GRN163L, led to loss of telomerase activity, reduction in telomere length, and growth arrest through induction of both the senescence and apoptosis. GRN163L-induced cell death could also be expedited by addition of the chemotherapeutic agents doxorubicin and ritonavir. Finally, the treatment with GRN163L led to a significant reduction in tumor volume in a subcutaneous tumor model. CONCLUSIONS: We show that telomerase activity is significantly elevated whereas telomeres are shorter in BEAC and suppression of telomerase inhibits proliferation of adenocarcinoma cells both in vitro and in vivo. PMID- 18676774 TI - CHEK2 1100delC is a susceptibility allele for HNPCC-related colorectal cancer. AB - PURPOSE: The pathogenic CHEK2 1100delC variant is firmly established as a breast cancer susceptibility allele. Dutch CHEK2 1100delC breast cancer families frequently also include colorectal cancer cases, and the variant is particularly prevalent among breast cancer families with hereditary breast and colorectal cancer. Yet, it is still unclear whether CHEK2 1100delC also confers a colorectal cancer risk independent of its breast cancer risk. EXPERIMENTAL DESIGN: CHEK2 1100delC was genotyped in the index cases of 369 Dutch colorectal cancer families that had been excluded for familial breast cancer. The cohort included 132 cases with familial adenomatous polyposis (FAP) and FAP-related disease, and 237 cases with hereditary nonpolyposis colorectal cancer (HNPCC) and HNPCC-related disease. RESULTS: None of the FAP/FAP-related cases carried the CHEK2 1100delC variant. In contrast, CHEK2 1100delC was present in 10 of 237 (4.2%) HNPCC/HNPCC-related cases that was significantly more prevalent than the 1.0% Dutch population frequency (odds ratio, 4.3; 95% confidence interval, 1.7-10.7; P = 0.002). Nine of the 10 CHEK2 1100delC colorectal cancer cases met the revised Amsterdam and/or Bethesda criteria. The 10 CHEK2 1100delC colorectal cancer families had a high risk cancer inheritance pattern, including 35 colorectal cancer cases, 9 cases with polyps, and 21 cases with other tumor types. CONCLUSION: Our analysis provides strong evidence that the 1100delC variant of CHEK2 confers a colorectal cancer risk in HNPCC/HNPCC-related families, supporting the hypothesis that CHEK2 is a multiorgan cancer susceptibility gene. PMID- 18676775 TI - Thy1 up-regulates FasL expression in lung myofibroblasts via Src family kinases. AB - We have previously demonstrated that myofibroblasts from lungs with bleomycin induced fibrosis overexpress FasL molecules. Two subpopulations of fibroblasts, distinguished by their expression of Thy1 molecules, have been shown in the lungs of both mice and humans. Thy1-mediated FasL induction has been reported in T cells through the use of anti-Thy1 (G7). We therefore sought to determine whether FasL expression in lung myofibroblasts is associated with and/or dependent on Thy1 expression, and to examine the underlying mechanism of regulation. We show that myofibroblast Thy1 expression is associated with increased FasL expression. Moreover, we directly show that Thy1 activation induces FasL up-regulation. Initially, Thy1 activation causes translocation of FasL to the membrane surface, and later induces de novo synthesis of FasL at the mRNA and protein levels. In contrast to Thy1 activation, TNF-alpha and IFN-gamma do not induce FasL myofibroblast up-regulation. Using Src family kinase (SFKs) inhibitor (PP2), we show the general involvement of SFKs in Thy1 signal transduction leading to FasL up-regulation; and, using specific siRNA, we show the particular involvement of Fyn, one protein in the SFK family. These results demonstrate that Thy1 in myofibroblasts is not just a marker, but is a functional protein that transmits signals into the cell, up-regulating its FasL expression. PMID- 18676777 TI - Cost-effectiveness analyses for injury prevention initiatives in low- and middle income countries. PMID- 18676776 TI - Nicotine induces resistance to chemotherapy by modulating mitochondrial signaling in lung cancer. AB - Continued smoking causes tumor progression and resistance to therapy in lung cancer. Carcinogens possess the ability to block apoptosis, and thus may induce development of cancers and resistance to therapy. Tobacco carcinogens have been studied widely; however, little is known about the agents that inhibit apoptosis, such as nicotine. We determine whether mitochondrial signaling mediates antiapoptotic effects of nicotine in lung cancer. A549 cells were exposed to nicotine (1 muM) followed by cisplatin (35 muM) plus etoposide (20 muM) for 24 hours. We found that nicotine prevented chemotherapy-induced apoptosis, improved cell survival, and caused modest increases in DNA synthesis. Inhibition of mitogen-activated protein kinase (MAPK) and Akt prevented the antiapoptotic effects of nicotine and decreased chemotherapy-induced apoptosis. Small interfering RNA MAPK kinase-1 blocked antiapoptotic effects of nicotine, whereas small interfering RNA MAPK kinase-2 blocked chemotherapy-induced apoptosis. Nicotine prevented chemotherapy-induced reduction in mitochondrial membrane potential and caspase-9 activation. Antiapoptotic effects of nicotine were blocked by mitochondrial anion channel inhibitor, 4,4'diisothiocyanatostilbene 2,2'disulfonic acid. Chemotherapy enhanced translocation of proapoptotic Bax to the mitochondria, whereas nicotine blocked these effects. Nicotine up-regulated Akt-mediated antiapoptotic X-linked inhibitor of apoptosis protein and phosphorylated proapoptotic Bcl2-antagonist of cell death. The A549-rho0 cells, which lack mitochondrial DNA, demonstrated partial resistance to chemotherapy induced apoptosis, but blocked the antiapoptotic effects of nicotine. Accordingly, we provide evidence that nicotine modulates mitochondrial signaling and inhibits chemotherapy-induced apoptosis in lung cancer. The mitochondrial regulation of nicotine imposes an important mechanism that can critically impair the treatment of lung cancer, because many cancer-therapeutic agents induce apoptosis via the mitochondrial death pathway. Strategies aimed at understanding nicotine-mediated signaling may facilitate the development of improved therapies in lung cancer. PMID- 18676778 TI - Surveillance alone is not the answer. PMID- 18676779 TI - Cost-effectiveness of traffic enforcement: case study from Uganda. AB - BACKGROUND: In October 2004, the Ugandan Police department deployed enhanced traffic safety patrols on the four major roads to the capital Kampala. OBJECTIVE: To assess the costs and potential effectiveness of increasing traffic enforcement in Uganda. METHODS: Record review and key informant interviews were conducted at 10 police stations along the highways that were patrolled. Monthly data on traffic citations and casualties were reviewed for January 2001 to December 2005; time series (ARIMA) regression was used to assess for a statistically significant change in traffic deaths. Costs were computed from the perspective of the police department in $US 2005. Cost offsets from savings to the health sector were not included. RESULTS: The annual cost of deploying the four squads of traffic patrols (20 officers, four vehicles, equipment, administration) is estimated at $72,000. Since deployment, the number of citations has increased substantially with a value of $327 311 annually. Monthly crash data pre- and post-intervention show a statistically significant 17% drop in road deaths after the intervention. The average cost-effectiveness of better road safety enforcement in Uganda is $603 per death averted or $27 per life year saved discounted at 3% (equivalent to 9% of Uganda's $300 GDP per capita). CONCLUSION: The costs of traffic safety enforcement are low in comparison to the potential number of lives saved and revenue generated. Increasing enforcement of existing traffic safety norms can prove to be an extremely cost-effective public health intervention in low-income countries, even from a government perspective. PMID- 18676780 TI - Ecological level analysis of the relationship between smoking and residential fire mortality. AB - OBJECTIVES: To examine the association between tobacco smoking and residential fire mortality and to investigate whether this association is explained by the confounding effects of selected socioeconomic factors (ie, educational attainment and median household income). DESIGN: An ecological analysis relating state-level residential-fire mortality to state-level percentages of adults who smoke was conducted. Negative binomial rate regression was used to model this relationship, simultaneously controlling for the selected socioeconomic factors. RESULTS: After educational attainment and median household income had been controlled for, smoking percentages among adults correlated significantly with state-level, population-based residential-fire mortality (estimated relative rate for a 1% decrease in smoking = 0.93; 95% CI 0.89 to 0.97). CONCLUSIONS: Mortality from residential fires is high in states with high smoking rates. This relationship cannot be explained solely by the socioeconomic factors examined in this study. PMID- 18676781 TI - The burden of fatal and non-fatal injury in rural India. AB - BACKGROUND: Little is known about the burden or causes of injury in rural villages in India. OBJECTIVE: To examine injury-related mortality and morbidity in villages in the state of Andhra Pradesh, India. METHODS: A verbal-autopsy based mortality surveillance study was used to collect mortality data on all ages from residents in 45 villages in 2003-2004. In early 2005, a morbidity survey in adults was carried out using stratified random sampling in 20 villages. Participants were asked about injuries sustained in the preceding 12 months. Both fatal and non-fatal injuries were coded using classification methods derived from ICD-10. RESULTS: Response rates for the mortality surveillance and morbidity survey were 98% and 81%, respectively. Injury was the second leading cause of death for all ages, responsible for 13% (95% CI 11% to 15%) of all deaths. The leading causes of fatal injury were self-harm (36%), falls (20%), and road traffic crashes (13%). Non-fatal injury was reported by 6.7% of survey participants, with the leading causes of injury being falls (38%), road traffic crashes (25%), and mechanical forces (16.1%). Falls were more common in women, with most (72.3%) attributable to slipping and tripping. Road traffic injuries were sustained mainly by men and were primarily the result of motorcycle crashes (48.8%). DISCUSSION: Injury is an important contributor to disease burden in rural India. The leading causes of injury-falls, road traffic crashes, and suicides-are all preventable. It is important that effective interventions are developed and implemented to minimize the impact of injury in this region. PMID- 18676782 TI - Helmet use and the risk of neck or cervical spine injury among users of motorized two-wheel vehicles. AB - OBJECTIVE: To quantify the effects of wearing a helmet on head and facial injury among users of motorized two-wheel vehicles and to determine if helmet use increases the risk of neck and cervical spine injury. DESIGN/METHOD: A population based study involving injured riders from the Rhone Road Trauma Registry from 1996 to 2005. Victims were only included if they had an injury to a body region other than (or in addition to) the head, face, neck, or cervical spine. Thus, inclusion was not affected by helmet use by the rider. The risk of head, face, neck, and cervical spine injury was assessed, with helmet use as the exposure of interest using logistic regression analyses. Adjusted odds ratios and corresponding confidence intervals were calculated. RESULTS: Helmet use significantly decreased the risk of head and facial injuries. The adjusted odds ratios for non-helmeted riders were 2.43 (95% CI 2.05 to 2.87) and 3.02 (95% CI 2.48 to 3.67), respectively. There was no association between helmet use and the occurrence of neck or cervical spine injuries. The adjusted odds ratios for non helmeted riders were 0.86 (95% CI 0.60 to 1.23) and 1.04 (95% CI 0.78 to 1.39), respectively. CONCLUSION: Helmets protect users of motorized two-wheel vehicles against head and facial injury without increasing the risk of neck or cervical spine injury. PMID- 18676783 TI - The Yorkhill CHIRPP story: a qualitative evaluation of 10 years of injury surveillance at a Scottish children's hospital. AB - BACKGROUND: The Canadian Hospitals Injury Reporting and Prevention Program (CHIRPP) is an emergency department-based injury surveillance system that was devised in Canada and has been in operation since 1990. CHIRPP was imported to Glasgow's Royal Hospital for Sick Children at Yorkhill in 1996 and ran for 10 years. OBJECTIVE: To critically review CHIRPP at Yorkhill (Y-CHIRPP). The following two key questions were posed. (1) Did Y-CHIRPP fail, and, if so, why? (2) What generalisable lessons can be learned about injury surveillance? METHODS: A retrospective qualitative review of Y-CHIRPP was carried out. In gathering information, the aims were to: (a) describe the processes involved in running Y CHIRPP; (b) identify changes made to that process over the 10 years; (c) determine the strengths and weaknesses of Y-CHIRPP. RESULTS: Taken together, and with reference to the WHO attributes of a good surveillance system, the findings suggest that Y-CHIRPP largely met the criteria of simplicity, flexibility, and acceptability. Criteria that were not, or only intermittently, met were reliability, utility, sustainability, and timeliness. CONCLUSIONS: Y-CHIRPP was, at best, a partial success. To maintain the viability of an injury surveillance system and to secure the long-term support of hospital staff, it is important that the system is perceived as an injury prevention service tool and not a research method. Experience with Y-CHIRPP suggests that injury surveillance requires three supporting posts: an emergency department staff member, a data analyst, and someone with responsibility for developing and/or lobbying for the implementation of preventive measures. PMID- 18676784 TI - Improving the predictive ability of the ICD-based Injury Severity Score. AB - OBJECTIVE: To assess whether the use of integrated hospitalization and mortality data sources and/or the inclusion of comorbidity improve the predictive ability of the International Classification of Disease (ICD)-based Injury Severity Score (ICISS). DESIGN: Models using either the ICISS based solely on hospital discharge data or one of nine modified ICISSs as the predictor variable were assessed on their ability to predict survival using logistic regression modeling. SETTING: New Zealand. Patients or SUBJECTS: Inpatients, with an S00-T89 ICD-10-AM principal diagnosis, and fatalities, with any S00-T89 ICD-10-AM diagnosis, occurring in 2000-2003. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Models were compared in terms of their discrimination (concordance), calibration, and goodness-of-fit. RESULTS: 186 835 cases including 9968 deaths met the inclusion criterion. The modified ICISS that included both mortality data and Charlson comorbid conditions at the ICD-10-AM level had the best concordance and high calibration. Calibration curves indicated that scores using hospital discharge data only to calculate survival risk ratios underestimated mortality, whereas scores using hospital discharge and mortality data overestimated mortality. CONCLUSIONS: Valid measurement of injury severity is important for both meaningful research and surveillance and to assist in classifying information to meet specific injury policy, prevention, and control needs. This study suggests that the predictive ability of ICISS would be improved if both mortality and comorbidity data were included in its calculation. PMID- 18676785 TI - An empirical approach for defining acceptable levels of risk: a case study in team sports. AB - OBJECTIVES: To determine acceptable levels of risk in sport and to compare these with values used in occupational settings. DESIGN: Cross-sectional, questionnaire based study. SETTINGS: Seven soccer and 11 rugby union teams. SUBJECTS: 140 male athletes and 108 male and 100 female spectators associated with soccer and rugby union teams. MAIN OUTCOMES: Views on acceptable frequencies with which athletes sustain acute injuries of various levels of severity. RESULTS: The responses of athletes and spectators were similar, although spectators consistently indicated a higher acceptable frequency of injury than athletes. There were no significant differences in responses as a function of respondents' gender and age. The results confirmed an inverse relationship between the acceptable frequency of occurrence and the severity of injury, although the relationships identified by the risk-averse and risk-taking minorities within the sample population were widely different. CONCLUSION: The mean frequency-severity risk relationship identified by athletes and spectators in soccer and rugby was similar to the relationship routinely used for risk assessments in industry and commerce. PMID- 18676786 TI - Examining the sensitivity of an injury surveillance program using population based estimates. AB - This study uses population-based estimates to assess the sensitivity and representativeness of an injury surveillance system using a 1-year population based approach. Data from the Ottawa Canadian Hospitals Injury Reporting and Prevention Program (CHIRPP) site (Children's Hospital of Eastern Ontario) were compared with those from six expansion sites. The overall sensitivity of CHIRPP was 43% of all treated injuries and 57% of injuries treated at emergency departments. CHIRPP was less likely to be representative for older children and more likely to capture children with more severe injuries. The limitations related to using CHIRPP for representing population-based injury remain fairly stable over time. A one-time population-based sample can provide useful information to add to routinely collected injury surveillance. PMID- 18676787 TI - It takes a village to prevent falls: reconceptualizing fall prevention and management for older adults. AB - Systematic evidence reviews support the efficacy of physical activity programs and multifactorial strategies for fall prevention. However, community settings in which fall prevention programs occur often differ substantially from the research settings in which efficacy was first demonstrated. Because of these differences, alternative approaches are needed to judge the adequacy of fall prevention activities occurring as part of standard medical care or community efforts. This paper uses the World Health Organization Innovative Care for Chronic Conditions (ICCC) framework to rethink how fall prevention programs might be implemented routinely in both medical and community settings. Examples of innovative programs and policies that provide fall prevention strategies consistent with the ICCC framework are highlighted, and evidence where available is provided on the effects of these strategies on processes and outcomes of care. Finally, a "no wrong door" approach to fall prevention and management is proposed, in which older adults who are found to be at risk of falls in either a medical or community setting are linked to a standard fall risk evaluation across three domains (physical activity, medical risks, and home safety). PMID- 18676788 TI - Improving infrastructure for injury control: a call for policy action. PMID- 18676789 TI - Systematic reviews on preventing fall-related injuries in older people. PMID- 18676790 TI - Securing the future of injury prevention: people and institutions. PMID- 18676791 TI - Injury research advocacy a high priority. PMID- 18676792 TI - Preventing violence and reducing its impact: how development agencies can help. PMID- 18676795 TI - The Sixth Washington Dental Service Foundation Distinguished Professor Symposium. November 2-3, 2007, Seattle. Introduction to the symposium proceedings. PMID- 18676796 TI - Combining normative and psychosocial perceptions for assessing orthodontic treatment needs. AB - Whereas perceptions of malocclusion by the public are mainly subjective, currently orthodontic treatment needs are predominantly determined using normative need. There are considerable differences between normative and subjective perceptions of orthodontic need. Yet clinical measures determine current provision of orthodontic services, whereas subjective oral health-related quality of life (OHRQoL) measures are seldom used and play a small part in need assessment despite predicting perceived need. The sociodental approach to assessing orthodontic treatment needs overcomes deficiencies of the sole use of normative need. It is a gradual integration process, estimating orthodontic needs by combining normative and psychosocial perspectives, as well as considering behavioral factors affecting outcomes of orthodontic treatment and scientific evidence about the effectiveness of interventions. To be appropriately used in needs assessment, an OHRQoL measure should provide condition-specific estimates of the impact of malocclusion on daily life. The sociodental approach attempts to replicate the characteristics of good clinical practice using a structured, rational, and coherent system for assessing orthodontic needs and, therefore, is a useful tool in planning oral health services. PMID- 18676797 TI - Does orthodontic treatment affect patients' quality of life? AB - The oral-facial region is usually an area of significant concern for the individual because it draws the most attention from other people in interpersonal interactions and is the primary source of vocal, physical, and emotional communication. As a result, patients who seek orthodontic treatment are concerned with improving their appearance and social acceptance, often more than they are with improving their oral function or health. Enhancing these aspects of quality of life is an important motive for undergoing orthodontic treatment. Regardless of age, patients' and their parents' or caregivers' expectations about improvements in oral function, esthetics, social acceptance, and body image are important for both general dentists and orthodontists to consider when advising patients about these procedures and during the treatment process. This review of research on the impact of conventional and surgical orthodontics on quality of life examines the association between oral health-related quality of life and severity and type of malocclusion, as well as the impact of treatment and patient characteristics on quality of life. The article will emphasize the importance of clinicians' having a clear understanding, before initiating treatment, of their patients' quality of life and their expectations about improvements in specific domains of quality of life. PMID- 18676798 TI - Orthodontic and nonorthodontic root resorption: their impact on clinical dental practice. AB - Occasionally, general dentists are challenged with providing restorative treatment for a postorthodontic patient who has had moderate to severe root resorption. When this happens, a number of questions about the cause of such resorption and the appropriate treatment arise in the dentist's mind. This article will describe the orthodontic and restorative management of three patients with severe maxillary incisor root resorption, provide a thorough discussion of the currently available literature on the topic of root resorption, and answer clinical questions regarding this relatively infrequent but devastating sequel to orthodontic treatment. PMID- 18676799 TI - Whole-tooth regeneration: it takes a village of scientists, clinicians, and patients. AB - A team of senior scientists was formed in 2006 to create a blueprint for the regeneration of whole human teeth along with all of the supporting structure of the dentition. The team included experts from diverse fields, each with a reputation for stellar accomplishment. Participants attacked the scientific issues of tooth regeneration but, more importantly, each agreed to work collaboratively with experts from other disciplines to form a learning organization. A commitment to learn from one another produced a unique interdisciplinary and multidisciplinary team. Inspired by the Kennedy space program to send a man to the moon, with its myriad of problems and solutions that no one discipline could solve, this tooth regeneration team devised an ambitious plan that sought to use stem cell biology, engineering, and computational biology to replicate the developmental program for odontogenesis. In this manner, team members envisioned a solution that consisted of known or knowable fundamentals. They proposed a laboratory-grown tooth rudiment that would be capable of executing the complete program for odontogenesis when transplanted to a suitable host, recreating all of the dental tissues, periodontal ligament, cementum, and alveolar bone associated with the canonical tooth. This plan was designed to bring regenerative medicine fully into the dental surgery suite, although a lack of funding has so far prevented the plan from being carried out. PMID- 18676800 TI - Effects of malocclusions and orthodontics on periodontal health: evidence from a systematic review. AB - Many patients seek orthodontic treatment for esthetic improvement. These patients mostly present with mal-alignment of the anterior teeth. The positive effects of orthodontic treatment on their appearance and self-esteem are easy to envision. However, does orthodontic treatment provide dental health benefits in addition to the esthetic benefits? Do malocclusions harm the periodontium? Is correcting malocclusions with orthodontic treatment beneficial for periodontal health? The purpose of this study is to present evidence available on this topic. Two systematic reviews were conducted to address these questions: does a malocclusion affect periodontal health, and does orthodontic treatment affect periodontal health? Inclusion and exclusion criteria were established for both reviews, and an electronic search and a hand search were conducted. Several papers were included in both reviews, but the overall quality of the studies was weak. The first review found a correlation between the presence of a malocclusion and periodontal disease. Subjects with greater malocclusion have more severe periodontal disease. This may be dependent on oral health status. One should keep in mind that an association does not necessarily mean causation. The second review identified an absence of reliable evidence on the effects of orthodontic treatment on periodontal health. The existing low-quality evidence suggests that orthodontic therapy results in small detrimental effects to the periodontium. The results of both reviews do not warrant recommendation for orthodontic treatment to prevent future periodontal problems, except for specific unusual malocclusions. PMID- 18676801 TI - Bisphosphonates and osteonecrosis of the jaws. AB - Bisphosphonates are important drugs that are increasingly prescribed to reduce the morbidity associated with osteoclast-mediated bone diseases. Shortly after the turn of the century, a variety of case reports described a necrosis of the jaw bone in patients using bisphosphonates. Currently, an exposed area of necrotic jaw bone present for at least eight weeks in patients using bisphosphonates has been defined as a bisphosphonate-associated osteonecrosis (BON) by the American Dental Association. BON may occur spontaneously but is more frequently associated with local trauma to the jaw. At this time, a causal relationship between BON and bisphosphonates has not been demonstrated. This review will evaluate current data related to the occurrence, risk, prevention, treatment, and management of BON. PMID- 18676802 TI - TMJ disorders: future innovations in diagnostics and therapeutics. AB - Because their etiologies and pathogenesis are poorly understood, temporomandibular joint (TMJ) diseases are difficult to diagnose and manage. All current approaches to treatments of TMJ diseases are largely palliative. Definitive and rational diagnoses or treatments can only be achieved through a comprehensive understanding of the etiologies, predisposing factors, and pathogenesis of TMJ diseases. While much work remains to be done in this field, novel findings in biomedicine and developments in imaging and computer technologies are beginning to provide us with a vision of future innovations in the diagnostics and therapeutics of TMJ disorders. These advances include the identification and use of local or systemic biomarkers to diagnose disease or monitor improvements in therapy; the use of imaging technologies for earlier and more sensitive diagnostics; and the use of biomedicine, biomimetics, and imaging to design and manufacture bioengineered joints. Such advances are likely to help to customize and enhance the quality of care we provide to patients with TMJ disorders. PMID- 18676803 TI - Esthetic orthodontic treatment using the invisalign appliance for moderate to complex malocclusions. AB - In this report, three patients were treated with a new treatment protocol for Invisalign to demonstrate that a variety of complex malocclusions can be successfully treated using this protocol, including correction of moderate crowding, correction of moderate Class II division 1, and deep overbite. Previous studies of Invisalign showed significant limitations for more complex orthodontic treatment, although a few recent case reports have shown successfully completed moderate to difficult orthodontic malocclusions. One reason for the discrepancy is that the earlier studies were done during the first four years of the appliance development (now ten years of clinical use), when significant problems existed with accomplishing bodily movement, torquing of roots, extrusions, and rotations of premolars and canines. The new protocol included new methods for anterior/posterior corrections, showing on the computer the effect of elastics for Class II treatment simulated as a one-stage anterior/posterior movement at the end of treatment. Staging for interproximal reduction (IPR) is now automatically staged when there is better access to interproximal contacts to avoid IPR where significant overlap between teeth is present to avoid performing IPR on surfaces that may be damaged by instruments such as burs, strips, and disks when cut on a sharp angle. Staging for tooth movements is now also done to enable combination movements to occur simultaneously for each tooth with the tooth that needs to move the most (the lead tooth) determining the minimum number of stages required. All other teeth move at a slower rate than the lead tooth throughout the duration of treatment. Attachments are now placed in the middle of the crown automatically for rotation and automatically sized in proportion to the clinical crown. Use of 1 mm thick (buccal-lingual dimension) horizontal beveled rectangular attachments is standard on premolars for retention of aligners during intrusive movements, such as leveling the lower curve of Spee in deep overbite for extrusions and for control of the tooth long axis during torquing movements. Staging of tooth movements now track linear and rotational velocities of teeth separately with the number of treatment stages determined by the lead tooth based on its rotational or linear maximum velocities at no more than two degrees of rotation per stage. Simultaneous movements are done for all teeth providing visible space (approximately 0.05 mm) between teeth during movements past other teeth using expansion instead of IPR as a primary way to increase space available for correction of crowding. PMID- 18676804 TI - The terminal 760 kb region on 4p16 is unlikely to be the critical interval for growth delay in Wolf-Hirschhorn syndrome. PMID- 18676805 TI - Pluripotent stem cell lines. AB - The derivation of human embryonic stem cells 10 years ago ignited an explosion of public interest in stem cells, yet this achievement depended on prior decades of research on mouse embryonic carcinoma cells and embryonic stem cells. In turn, the recent derivation of mouse and human induced pluripotent stem cells depended on the prior studies on mouse and human embryonic stem cells. Both human embryonic stem cells and induced pluripotent stem cells can self-renew indefinitely in vitro while maintaining the ability to differentiate into advanced derivatives of all three germ layers, features very useful for understanding the differentiation and function of human tissues, for drug screen and toxicity testing, and for cellular transplantation therapies. Here we review the family of pluripotent cell lines derived from early embryos and from germ cells, and compare them with the more recently described induced pluripotent stem cells. PMID- 18676807 TI - Transcription in the nucleus and mRNA decay in the cytoplasm are coupled processes. AB - Maintaining appropriate mRNAs levels is vital for any living cell. mRNA synthesis in the nucleus by RNA polymerase II core enzyme (Pol II) and mRNA decay by cytoplasmic machineries determine these levels. Yet, little is known about possible cross-talk between these processes. The yeast Rpb4/7 is a nucleo cytoplasmic shuttling heterodimer that interacts with Pol II and with mRNAs and is required for mRNA decay in the cytoplasm. Here we show that interaction of Rpb4/7 with mRNAs and eventual decay of these mRNAs in the cytoplasm depends on association of Rpb4/7 with Pol II in the nucleus. We propose that, following its interaction with Pol II, Rpb4/7 functions in transcription, interacts with the transcript cotranscriptionally and travels with it to the cytoplasm to stimulate mRNA decay. Hence, by recruiting Rpb4/7, Pol II governs not only transcription but also mRNA decay. PMID- 18676808 TI - Inhibitor of differentiation 4 drives brain tumor-initiating cell genesis through cyclin E and notch signaling. AB - Cellular origins and genetic factors governing the genesis and maintenance of glioblastomas (GBM) are not well understood. Here, we report a pathogenetic role of the developmental regulator Id4 (inhibitor of differentiation 4) in GBM. In primary murine Ink4a/Arf(-/-) astrocytes, and human glioma cells, we provide evidence that enforced Id4 can drive malignant transformation by stimulating increased cyclin E to produce a hyperproliferative profile and by increased Jagged1 expression with Notch1 activation to drive astrocytes into a neural stem like cell state. Thus, Id4 plays an integral role in the transformation of astrocytes via its combined actions on two-key cell cycle and differentiation regulatory molecules. PMID- 18676806 TI - On the origin of the beta cell. AB - The major forms of diabetes are characterized by pancreatic islet beta-cell dysfunction and decreased beta-cell numbers, raising hope for cell replacement therapy. Although human islet transplantation is a cell-based therapy under clinical investigation for the treatment of type 1 diabetes, the limited availability of human cadaveric islets for transplantation will preclude its widespread therapeutic application. The result has been an intense focus on the development of alternate sources of beta cells, such as through the guided differentiation of stem or precursor cell populations or the transdifferentiation of more plentiful mature cell populations. Realizing the potential for cell-based therapies, however, requires a thorough understanding of pancreas development and beta-cell formation. Pancreas development is coordinated by a complex interplay of signaling pathways and transcription factors that determine early pancreatic specification as well as the later differentiation of exocrine and endocrine lineages. This review describes the current knowledge of these factors as they relate specifically to the emergence of endocrine beta cells from pancreatic endoderm. Current therapeutic efforts to generate insulin-producing beta-like cells from embryonic stem cells have already capitalized on recent advances in our understanding of the embryonic signals and transcription factors that dictate lineage specification and will most certainly be further enhanced by a continuing emphasis on the identification of novel factors and regulatory relationships. PMID- 18676809 TI - Structural and functional analysis of the Crb2-BRCT2 domain reveals distinct roles in checkpoint signaling and DNA damage repair. AB - Schizosaccharomyces pombe Crb2 is a checkpoint mediator required for the cellular response to DNA damage. Like human 53BP1 and Saccharomyces cerevisiae Rad9 it contains Tudor(2) and BRCT(2) domains. Crb2-Tudor(2) domain interacts with methylated H4K20 and is required for recruitment to DNA dsDNA breaks. The BRCT(2) domain is required for dimerization, but its precise role in DNA damage repair and checkpoint signaling is unclear. The crystal structure of the Crb2-BRCT(2) domain, alone and in complex with a phosphorylated H2A.1 peptide, reveals the structural basis for dimerization and direct interaction with gamma-H2A.1 in ionizing radiation-induced foci (IRIF). Mutational analysis in vitro confirms the functional role of key residues and allows the generation of mutants in which dimerization and phosphopeptide binding are separately disrupted. Phenotypic analysis of these in vivo reveals distinct roles in the DNA damage response. Dimerization mutants are genotoxin sensitive and defective in checkpoint signaling, Chk1 phosphorylation, and Crb2 IRIF formation, while phosphopeptide binding mutants are only slightly sensitive to IR, have extended checkpoint delays, phosphorylate Chk1, and form Crb2 IRIF. However, disrupting phosphopeptide binding slows formation of ssDNA-binding protein (Rpa1/Rad11) foci and reduces levels of Rad22(Rad52) recombination foci, indicating a DNA repair defect. PMID- 18676810 TI - A chromatin-wide transition to H4K20 monomethylation impairs genome integrity and programmed DNA rearrangements in the mouse. AB - H4K20 methylation is a broad chromatin modification that has been linked with diverse epigenetic functions. Several enzymes target H4K20 methylation, consistent with distinct mono-, di-, and trimethylation states controlling different biological outputs. To analyze the roles of H4K20 methylation states, we generated conditional null alleles for the two Suv4-20h histone methyltransferase (HMTase) genes in the mouse. Suv4-20h-double-null (dn) mice are perinatally lethal and have lost nearly all H4K20me3 and H4K20me2 states. The genome-wide transition to an H4K20me1 state results in increased sensitivity to damaging stress, since Suv4-20h-dn chromatin is less efficient for DNA double strand break (DSB) repair and prone to chromosomal aberrations. Notably, Suv4-20h dn B cells are defective in immunoglobulin class-switch recombination, and Suv4 20h-dn deficiency impairs the stem cell pool of lymphoid progenitors. Thus, conversion to an H4K20me1 state results in compromised chromatin that is insufficient to protect genome integrity and to process a DNA-rearranging differentiation program in the mouse. PMID- 18676811 TI - A comprehensive synthetic genetic interaction network governing yeast histone acetylation and deacetylation. AB - Histone acetylation and deacetylation are among the principal mechanisms by which chromatin is regulated during transcription, DNA silencing, and DNA repair. We analyzed patterns of genetic interactions uncovered during comprehensive genome wide analyses in yeast to probe how histone acetyltransferase (HAT) and histone deacetylase (HDAC) protein complexes interact. The genetic interaction data unveil an underappreciated role of HDACs in maintaining cellular viability, and led us to show that deacetylation of the histone variant Htz1p at Lys 14 is mediated by Hda1p. Studies of the essential nucleosome acetyltransferase of H4 (NuA4) revealed acetylation-dependent protein stabilization of Yng2p, a potential nonhistone substrate of NuA4 and Rpd3C, and led to a new functional organization model for this critical complex. We also found that DNA double-stranded breaks (DSBs) result in local recruitment of the NuA4 complex, followed by an elaborate NuA4 remodeling process concomitant with Rpd3p recruitment and histone deacetylation. These new characterizations of the HDA and NuA4 complexes demonstrate how systematic analyses of genetic interactions may help illuminate the mechanisms of intricate cellular processes. PMID- 18676812 TI - Vezf1 regulates genomic DNA methylation through its effects on expression of DNA methyltransferase Dnmt3b. AB - The zinc finger protein vascular endothelial zinc finger 1 (Vezf1) has been implicated in the development of the blood vascular and lymphatic system in mice, and has been characterized as a transcriptional activator in some systems. The chicken homolog, BGP1, has binding sites in the beta-globin locus, including the upstream insulator element. We report that in a mouse embryonic stem cell line deletion of both copies of Vezf1 results in loss of DNA methylation at widespread sites in the genome, including Line1 elements and minor satellite repeats, some imprinted genes, and several CpG islands. Loss of methylation appears to arise from a substantial decrease in the abundance of the de novo DNA methyltransferase, Dnmt3b. These results suggest that naturally occurring mutations in Vezf1/BGP1 might have widespread effects on DNA methylation patterns and therefore on epigenetic regulation of gene expression. PMID- 18676813 TI - PU.1 expression is modulated by the balance of functional sense and antisense RNAs regulated by a shared cis-regulatory element. AB - The transcription factor PU.1 is an important regulator of hematopoiesis; precise expression levels are critical for normal hematopoietic development and suppression of leukemia. We show here that noncoding antisense RNAs are important modulators of proper dosages of PU.1. Antisense and sense RNAs are regulated by shared evolutionarily conserved cis-regulatory elements, and we can show that antisense RNAs inhibit PU.1 expression by modulating mRNA translation. We propose that such antisense RNAs will likely be important in the regulation of many genes and may be the reason for the large number of overlapping complementary transcripts with so far unknown function. PMID- 18676814 TI - Encoding NF-kappaB temporal control in response to TNF: distinct roles for the negative regulators IkappaBalpha and A20. AB - TNF-induced NF-kappaB activity shows complex temporal regulation whose different phases lead to distinct gene expression programs. Combining experimental studies and mathematical modeling, we identify two temporal amplification steps-one determined by the obligate negative feedback regulator IkappaBalpha-that define the duration of the first phase of NF-kappaB activity. The second phase is defined by A20, whose inducible expression provides for a rheostat function by which other inflammatory stimuli can regulate TNF responses. Our results delineate the nonredundant functions implied by the knockout phenotypes of ikappabalpha and a20, and identify the latter as a signaling cross-talk mediator controlling inflammatory and developmental responses. PMID- 18676815 TI - A branched-chain fatty acid is involved in post-embryonic growth control in parallel to the insulin receptor pathway and its biosynthesis is feedback regulated in C. elegans. AB - Growth and development of multicellular organisms are controlled by signaling systems that sense nutrition availability and metabolic status. We report a novel and surprising factor in Caenorhabditis elegans development, the monomethyl branched-chain fatty acid C17ISO, a product of leucine catabolism. We show here that C17ISO is an essential constituent in a novel mechanism that acts in parallel with the food-sensing DAF-2 (insulin receptor)/DAF-16 (FOXO) signaling pathway to promote post-embryonic development, and that the two pathways converge on a common target repressing cell cycle. We show that C17ISO homeostasis is regulated by a SREBP-1c-mediated feedback mechanism that is different from the SREBP-1c-mediated regulation of common fatty acid biosynthesis, as well as by peptide uptake and transport. Our data suggest that C17ISO may act as a chemical/nutritional factor in a mechanism that regulates post-embryonic development in response to the metabolic state of the organism. PMID- 18676816 TI - Laminin-511 is an epithelial message promoting dermal papilla development and function during early hair morphogenesis. AB - Hair morphogenesis takes place through reciprocal epithelial and mesenchymal signaling; however, the mechanisms controlling signal exchange are poorly understood. Laminins are extracellular proteins that play critical roles in adhesion and signaling. Here we demonstrate the mechanism of how laminin-511 controls hair morphogenesis. Dermal papilla (DP) from laminin-511 mutants showed developmental defects by E16.5, including a failure to maintain expression of the key morphogen noggin. This maintenance was critical as exogenous introduction of noggin or sonic hedgehog (Shh) produced downstream from noggin was sufficient to restore hair follicle development in lama5(-/-) (laminin-511-null) skin. Hair development required the beta1 integrin binding but not the heparin binding domain of laminin-511. Previous studies demonstrated that Shh signaling requires primary cilia, microtubule-based signaling organelles. Laminin-511 mutant DP showed decreased length and structure of primary cilia in vitro and in vivo. Laminin-511, but not laminin-111, restored primary cilia formation in lama5(-/-) mesenchyme and triggered noggin expression in an Shh- and PDGF-dependent manner. Inhibition of laminin-511 receptor beta1 integrin disrupted DP primary cilia formation as well as hair development. These studies show that epithelial-derived laminin-511 is a critical early signal that directs ciliary function and DP maintenance as a requirement for hair follicle downgrowth. PMID- 18676817 TI - A highly conserved molecular switch binds MSY-3 to regulate myogenin repression in postnatal muscle. AB - Myogenin is the dominant transcriptional regulator of embryonic and fetal muscle differentiation and during maturation is profoundly down-regulated. We show that a highly conserved 17-bp DNA cis-acting sequence element located upstream of the myogenin promoter (myogHCE) is essential for postnatal repression of myogenin in transgenic animals. We present multiple lines of evidence supporting the idea that repression is mediated by the Y-box protein MSY-3. Electroporation in vivo shows that myogHCE and MSY-3 are required for postnatal repression. We further show that, in the C2C12 cell culture system, ectopic MSY-3 can repress differentiation, while reduced MSY-3 promotes premature differentiation. MSY-3 binds myogHCE simultaneously with the homeodomain protein Pbx in postnatal innervated muscle. We therefore propose a model in which the myogHCE motif operates as a switch by specifying opposing functions; one that was shown previously is regulated by MyoD and Pbx and it specifies a chromatin opening, gene-activating function at the time myoblasts begin to differentiate; the other includes MYS-3 and Pbx, and it specifies a repression function that operates during and after postnatal muscle maturation in vivo and in myoblasts before they begin to differentiate. PMID- 18676818 TI - Genomic medicine in Mexico: initial steps and the road ahead. AB - Mexico faces important demographic and epidemiological transitions with significant implications to patterns of disease, disability, and death. On the one hand, there are problems of underdevelopment and, on the other, the emerging challenges of the chronic and degenerative diseases of the industrialized world. For these diseases, prevention becomes a key strategy for alleviating a major burden to the economy and health of the Mexican population. Genomic medicine has become a priority to the Mexican government as a means of finding new strategies to tackle common diseases. In 2000, strategic planning for genomic medicine began, from a feasibility study and a multi-institutional consortium effort, to the creation of a National Institute of Genomic Medicine by the Mexican congress in 2004. Current research programs in genomic medicine in Mexico include the construction of a haplotype map of the Mexican population, several genome-wide association studies for common diseases, such as diabetes, obesity, cardiovascular disease, and cancer, as well as translational medicine projects that include biomarkers discovery for several kinds of cancer, pharmacogenomics, and nutrigenomics. Although this strategy has been successful, there are challenges that still need to be addressed, including increased investment in science and technology to stimulate a more vigorous and competitive research environment, development of more effective basic and clinical research synergies, recruitment and training of more human resources in genomic medicine, developing mechanisms to stimulate translational research, and developing a more modern regulatory framework to ensure that genomic medicine will successfully contribute to improve healthcare in the Mexican population. PMID- 18676820 TI - An MCMC algorithm for haplotype assembly from whole-genome sequence data. AB - In comparison to genotypes, knowledge about haplotypes (the combination of alleles present on a single chromosome) is much more useful for whole-genome association studies and for making inferences about human evolutionary history. Haplotypes are typically inferred from population genotype data using computational methods. Whole-genome sequence data represent a promising resource for constructing haplotypes spanning hundreds of kilobases for an individual. In this article, we propose a Markov chain Monte Carlo (MCMC) algorithm, HASH (haplotype assembly for single human), for assembling haplotypes from sequenced DNA fragments that have been mapped to a reference genome assembly. The transitions of the Markov chain are generated using min-cut computations on graphs derived from the sequenced fragments. We have applied our method to infer haplotypes using whole-genome shotgun sequence data from a recently sequenced human individual. The high sequence coverage and presence of mate pairs result in fairly long haplotypes (N50 length ~ 350 kb). Based on comparison of the sequenced fragments against the individual haplotypes, we demonstrate that the haplotypes for this individual inferred using HASH are significantly more accurate than the haplotypes estimated using a previously proposed greedy heuristic and a simple MCMC method. Using haplotypes from the HapMap project, we estimate the switch error rate of the haplotypes inferred using HASH to be quite low, ~1.1%. Our Markov chain Monte Carlo algorithm represents a general framework for haplotype assembly that can be applied to sequence data generated by other sequencing technologies. The code implementing the methods and the phased individual haplotypes can be downloaded from (http://www.cse.ucsd.edu/users/vibansal/HASH/). PMID- 18676821 TI - The Arf/p53 pathway in cancer and aging. AB - Arf and p53 are regarded among the most relevant tumor suppressors based on their ubiquitous and frequent inactivation in human cancer. The Arf/p53 pathway protects cells against several types of damage and this is the basis of its tumor suppressor activity. Interestingly, aging is a process associated with the accumulation of damage derived from chronic stresses of small magnitude. In agreement with its damage protection role, it has been recently described that the Arf/p53 pathway not only protects mammalian organisms from cancer but also from aging. However, there is also evidence that p53, under certain circumstances, such as when constitutively active, can induce aging. We discuss here the current evidence linking the Arf/p53 pathway to the process of aging and present a unified model. PMID- 18676819 TI - The opossum genome: insights and opportunities from an alternative mammal. AB - The strategic importance of the genome sequence of the gray, short-tailed opossum, Monodelphis domestica, accrues from both the unique phylogenetic position of metatherian (marsupial) mammals and the fundamental biologic characteristics of metatherians that distinguish them from other mammalian species. Metatherian and eutherian (placental) mammals are more closely related to one another than to other vertebrate groups, and owing to this close relationship they share fundamentally similar genetic structures and molecular processes. However, during their long evolutionary separation these alternative mammals have developed distinctive anatomical, physiologic, and genetic features that hold tremendous potential for examining relationships between the molecular structures of mammalian genomes and the functional attributes of their components. Comparative analyses using the opossum genome have already provided a wealth of new evidence regarding the importance of noncoding elements in the evolution of mammalian genomes, the role of transposable elements in driving genomic innovation, and the relationships between recombination rate, nucleotide composition, and the genomic distributions of repetitive elements. The genome sequence is also beginning to enlarge our understanding of the evolution and function of the vertebrate immune system, and it provides an alternative model for investigating mechanisms of genomic imprinting. Equally important, availability of the genome sequence is fostering the development of new research tools for physical and functional genomic analyses of M. domestica that are expanding its versatility as an experimental system for a broad range of research applications in basic biology and biomedically oriented research. PMID- 18676822 TI - Metastasis suppressor function of tumor necrosis factor-related apoptosis inducing ligand-R in mice: implications for TRAIL-based therapy in humans? AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising candidate for cancer therapy, as it can induce apoptosis specifically in tumor cells but not in normal cells. Although earlier mouse tumor studies revealed a strong tissue dependency of TRAIL and its death receptor in suppressing primary tumorigenesis or experimental metastases, we recently found that TRAIL-R inhibits lymph node metastases without affecting primary tumor formation in a mouse model of multistage skin tumorigenesis. This finding uncouples the role of TRAIL in primary tumorigenesis from metastasis formation, likely by sensitization of previously TRAIL-resistant tumor cells upon detachment, an early step required for metastasis formation. Therefore, TRAIL-R is a novel metastasis suppressor, suggesting that TRAIL-related tumor therapy might be most effective in primary tumors and early metastatic cancers, before selection for TRAIL resistance occurs. PMID- 18676823 TI - 17Beta-estradiol mobilizes bone marrow-derived endothelial progenitor cells to tumors. AB - Neovascularization is critical for tumor growth and development. The cellular mediators for this process are yet to be defined. We discovered that bone marrow derived endothelial progenitor cells (BM-EPC), having the phenotype (CD133+, CD34+, VEGFR-2+), initiate neovascularization in response to TG1-1 mammary cells implanted in the inguinal mammary gland of Tie-2 GFP transgenic mice. The fluorescence tag allowed for tracing the migration of green fluorescent protein tagged endothelial progenitor cells to tumor tissues. We discovered that 17-beta estradiol supplementation of ovariectomized mice significantly enhanced BM-EPC induced neovascularization and secretion of angiogenic factors within the tumor microenvironment. Cell-based system analyses showed that estrogen-stimulated BM EPCs secreted paracrine factors which enhanced TG1-1 cell proliferation and migration. Furthermore, TG1-1 cell medium supplemented with estrogen-induced BM EPC mediated tubulogenesis, which was an experimental in vivo representation of the neovasculature. Our data provide evidence of BM-EPC mammary tumor cell interactions and identify a novel cellular mediator of tumor progression that can be exploited clinically. PMID- 18676824 TI - Targeting cancer stem cells through L1CAM suppresses glioma growth. AB - Malignant gliomas are lethal cancers that display striking cellular heterogeneity. A highly tumorigenic glioma tumor subpopulation, termed cancer stem cells or tumor-initiating cells, promotes therapeutic resistance and tumor angiogenesis. Therefore, targeting cancer stem cells may improve patient survival. We interrogated the role of a neuronal cell adhesion molecule, L1CAM, in glioma stem cells as L1CAM regulates brain development and is expressed in gliomas. L1CAM(+) and CD133(+) cells cosegregated in gliomas, and levels of L1CAM were higher in CD133(+) glioma cells than normal neural progenitors. Targeting L1CAM using lentiviral-mediated short hairpin RNA (shRNA) interference in CD133(+) glioma cells potently disrupted neurosphere formation, induced apoptosis, and inhibited growth specifically in glioma stem cells. We identified a novel mechanism for L1CAM regulation of cell survival as L1CAM knockdown decreased expression of the basic helix-loop-helix transcription factor Olig2 and up-regulated the p21(WAF1/CIP1) tumor suppressor in CD133(+) glioma cells. To determine if targeting L1CAM was sufficient to reduce glioma stem cell tumor growth in vivo, we targeted L1CAM in glioma cells before injection into immunocompromised mice or directly in established tumors. In each glioma xenograft model, shRNA targeting of L1CAM expression in vivo suppressed tumor growth and increased the survival of tumor-bearing animals. Together, these data show that L1CAM is required for maintaining the growth and survival of CD133(+) glioma cells both in vitro and in vivo, and L1CAM may represent a cancer stem cell-specific therapeutic target for improving the treatment of malignant gliomas and other brain tumors. PMID- 18676825 TI - COOH-terminal binding protein regulates expression of the p16INK4A tumor suppressor and senescence in primary human cells. AB - The p16/pocket-protein pathway sets a balance between tumor suppression and capacity for tissue regeneration. Understanding the upstream signaling pathway that turns on the expression of p16 is required both for knowing the tumorigenic stresses from which this pathway provides protection and for appreciating the selective pressure that leads to the loss of this pathway in most human tumors. We report that COOH-terminal binding protein (CtBP), a physiologically regulated transcriptional corepressor that dimerizes to hold together repressive complexes, regulates p16 expression in primary human fibroblasts and keratinocytes. Interfering with CtBP-mediated repression increased p16 expression and accelerated senescence. CtBP had little influence on the expression of the alternate product of the CDKN2A tumor-suppressor gene, p14(ARF). Loss of CtBP mediated repression diminished the Polycomb-based epigenetic histone mark that is reported to favor silencing of p16 via DNA methylation. Enhancing CtBP-mediated repression by growing cells in low oxygen increased the association of CtBP with the p16 promoter, as assessed by chromatin immunoprecipitation, and reduced p16 expression. Stresses and stimuli that reduce CtBP-mediated repression are associated with increased p16 expression; therefore, CtBP may provide a common final target for regulating the balance among tumor suppression, regenerative capacity, and senescence. PMID- 18676826 TI - Serum-soluble B7x is elevated in renal cell carcinoma patients and is associated with advanced stage. AB - B7x is the newest member of the B7-CD28 family and is thought to dampen immune responses via negative costimulation. Tumor expression of B7x was recently described in renal cell carcinoma (RCC) and was associated with poor outcome. We developed an assay to detect serum-soluble B7x (sB7x) and investigated 101 patients with clear cell RCC who underwent nephrectomy between 2003 and 2007. For controls, we obtained serum from 101 sex-matched blood donors within the same age range. Following an ELISA for sB7x, detectable levels (>0.1 ng/mL) of sB7x were observed in 53 RCC patients compared with 18 controls (P < 0.001). Median (range) concentrations of sB7x for RCC patients and controls were 14.4 ng/mL (0.1-56.9) and 2.7 ng/mL (0.2-37.1), respectively. For RCC patients with detectable sB7x, median levels were significantly higher for patients with a tumor thrombus (19.2 versus 6.6 ng/mL; P = 0.007), positive lymph nodes (41.3 versus 10.3 ng/mL; P = 0.018), and distant metastases at nephrectomy (43.3 versus 8.5 ng/mL; P = 0.002) and tended to be higher in patients with high-grade tumors (18.8 versus 8.5; P = 0.090). Additionally, median sB7x levels for tumor-node-metastasis stage I to IV RCC were 6.6, 10.3, 14.5, and 43.3 ng/mL, respectively (P = 0.012). In this first evaluation of sB7x in RCC, we show that RCC patients are more likely to have detectable sB7x compared with controls and higher sB7x levels correlate with advanced tumor stage. These early results merit further investigation of this serum marker for potential diagnostic and prognostic purposes. PMID- 18676827 TI - Methadone, commonly used as maintenance medication for outpatient treatment of opioid dependence, kills leukemia cells and overcomes chemoresistance. AB - The therapeutic opioid drug methadone (d,l-methadone hydrochloride) is the most commonly used maintenance medication for outpatient treatment of opioid dependence. In our study, we found that methadone is also a potent inducer of cell death in leukemia cells and we clarified the unknown mechanism of methadone induced cell killing in leukemia cells. Methadone inhibited proliferation in leukemia cells and induced cell death through apoptosis induction and activated apoptosis pathways through the activation of caspase-9 and caspase-3, down regulation of Bcl-x(L) and X chromosome-linked inhibitor of apoptosis, and cleavage of poly(ADP-ribose) polymerase. In addition, methadone induced cell death not only in anticancer drug-sensitive and apoptosis-sensitive leukemia cells but also in doxorubicin-resistant, multidrug-resistant, and apoptosis resistant leukemia cells, which anticancer drugs commonly used in conventional therapies of leukemias failed to kill. Depending on caspase activation, methadone overcomes doxorubicin resistance, multidrug resistance, and apoptosis resistance in leukemia cells through activation of mitochondria. In contrast to leukemia cells, nonleukemic peripheral blood lymphocytes survived after methadone treatment. These findings show that methadone kills leukemia cells and breaks chemoresistance and apoptosis resistance. Our results suggest that methadone is a promising therapeutic approach not only for patients with opioid dependence but also for patients with leukemias and provide the foundation for new strategies using methadone as an additional anticancer drug in leukemia therapy, especially when conventional therapies are less effective. PMID- 18676828 TI - Cyclic AMP response element-binding protein overexpression: a feature associated with negative prognosis in never smokers with non-small cell lung cancer. AB - Lung cancer is the leading cause of cancer deaths worldwide. Recent advances in targeted therapies hold promise for the development of new treatments for certain subsets of cancer patients by targeting specific signaling molecule. Based on the identification of the transcription factor cyclic AMP response element-binding protein (CREB) as an important regulator of growth of several types of cancers and our recent findings of its importance in normal differentiation of bronchial epithelial cells, we hypothesized that CREB plays an important pathobiologic role in lung carcinogenesis. We conducted this initial study to determine whether the expression and activation status of CREB are altered in non-small cell lung cancer (NSCLC) and of any prognostic importance in NSCLC patients. We found that the expression levels of mRNA and protein of CREB and phosphorylated CREB (p CREB) were significantly higher in most of the NSCLC cell lines and tumor specimens than in the normal human tracheobronchial epithelial cells and adjacent normal lung tissue, respectively. Analysis of CREB mRNA expression and the CREB gene copy number showed that CREB overexpression occurred mainly at the transcriptional level. Immunohistochemical analysis of tissue microarray slides containing sections of NSCLC specimens obtained from 310 patients showed that a decreased survival duration was significantly associated with overexpression of CREB or p-CREB in never smokers but not in current or former smokers with NSCLC. These are the first reported results illustrating the potential of CREB as a molecular target for the prevention and treatment of NSCLC, especially in never smokers. PMID- 18676829 TI - Defective transcription/repair factor IIH recruitment to specific UV lesions in trichothiodystrophy syndrome. AB - Most trichothiodystrophy (TTD) patients present mutations in the xeroderma pigmentosum D (XPD) gene, coding for a subunit of the transcription/repair factor IIH (TFIIH) complex involved in nucleotide excision repair (NER) and transcription. After UV irradiation, most TTD/XPD patients are more severely affected in the NER of cyclobutane pyrimidine dimers (CPD) than of 6-4 photoproducts (6-4PP). The reasons for this differential DNA repair defect are unknown. Here we report the first study of NER in response to CPDs or 6-4PPs separately analyzed in primary fibroblasts. This was done by using heterologous photorepair; recombinant adenovirus vectors carrying photolyases enzymes that repair CPD or 6-4PP specifically by using the energy of light were introduced in different cell lines. The data presented here reveal that some TTD/XPD mutations affect the recruitment of TFIIH specifically to CPDs, but not to 6-4PPs. This deficiency is further confirmed by the inability of TTD/XPD cells to recruit, specifically for CPDs, NER factors that arrive in a TFIIH-dependent manner later in the NER pathway. For 6-4PPs, we show that TFIIH complexes carrying an NH(2) terminal XPD mutated protein are also deficient in recruitment of NER proteins downstream of TFIIH. Treatment with the histone deacetylase inhibitor trichostatin A allows the recovery of TFIIH recruitment to CPDs in the studied TTD cells and, for COOH-terminal XPD mutations, increases the repair synthesis and survival after UV, suggesting that this defect can be partially related with accessibility of DNA damage in closed chromatin regions. PMID- 18676831 TI - A six-gene signature predicting breast cancer lung metastasis. AB - The lungs are a frequent target of metastatic breast cancer cells, but the underlying molecular mechanisms are unclear. All existing data were obtained either using statistical association between gene expression measurements found in primary tumors and clinical outcome, or using experimentally derived signatures from mouse tumor models. Here, we describe a distinct approach that consists of using tissue surgically resected from lung metastatic lesions and comparing their gene expression profiles with those from nonpulmonary sites, all coming from breast cancer patients. We show that the gene expression profiles of organ-specific metastatic lesions can be used to predict lung metastasis in breast cancer. We identified a set of 21 lung metastasis-associated genes. Using a cohort of 72 lymph node-negative breast cancer patients, we developed a 6-gene prognostic classifier that discriminated breast primary cancers with a significantly higher risk of lung metastasis. We then validated the predictive ability of the 6-gene signature in 3 independent cohorts of breast cancers consisting of a total of 721 patients. Finally, we show that the signature improves risk stratification independently of known standard clinical variables and a previously established lung metastasis signature based on an experimental breast cancer metastasis model. PMID- 18676830 TI - An integrative genomic and proteomic analysis of PIK3CA, PTEN, and AKT mutations in breast cancer. AB - Phosphatidylinositol 3-kinase (PI3K)/AKT pathway aberrations are common in cancer. By applying mass spectroscopy-based sequencing and reverse-phase protein arrays to 547 human breast cancers and 41 cell lines, we determined the subtype specificity and signaling effects of PIK3CA, AKT, and PTEN mutations and the effects of PIK3CA mutations on responsiveness to PI3K inhibition in vitro and on outcome after adjuvant tamoxifen. PIK3CA mutations were more common in hormone receptor-positive (34.5%) and HER2-positive (22.7%) than in basal-like tumors (8.3%). AKT1 (1.4%) and PTEN (2.3%) mutations were restricted to hormone receptor positive cancers. Unlike AKT1 mutations that were absent from cell lines, PIK3CA (39%) and PTEN (20%) mutations were more common in cell lines than tumors, suggesting a selection for these but not AKT1 mutations during adaptation to culture. PIK3CA mutations did not have a significant effect on outcome after adjuvant tamoxifen therapy in 157 hormone receptor-positive breast cancer patients. PIK3CA mutations, in comparison with PTEN loss and AKT1 mutations, were associated with significantly less and inconsistent activation of AKT and of downstream PI3K/AKT signaling in tumors and cell lines. PTEN loss and PIK3CA mutation were frequently concordant, suggesting different contributions to pathophysiology. PTEN loss rendered cells significantly more sensitive to growth inhibition by the PI3K inhibitor LY294002 than did PIK3CA mutations. Thus, PI3K pathway aberrations likely play a distinct role in the pathogenesis of different breast cancer subtypes. The specific aberration present may have implications for the selection of PI3K-targeted therapies in hormone receptor-positive breast cancer. PMID- 18676832 TI - Neutrophil gelatinase-associated lipocalin: a novel suppressor of invasion and angiogenesis in pancreatic cancer. AB - Neutrophil gelatinase-associated lipocalin (NGAL) is a 25-kDa secreted acute phase protein, which is also up-regulated in multiple cancers, including breast, lung, and pancreas. Recently, NGAL has been proposed as an early biomarker in pancreatic cancer (PaCa). However, its biological role in PaCa is unknown. In this study, we examined in vitro and in vivo the functional role of NGAL in PaCa. Well- to moderately differentiated PaCa cells (AsPC-1, BxPC-3, and Capan-2) expressed high levels of NGAL but moderately to poorly differentiated PaCa cells (PANC-1 and MIAPaCa-2) expressed undetectable NGAL levels. Immunohistochemistry of untreated tissue microarray showed specific NGAL staining in resected PaCa specimens (P = 0.0167). Stable NGAL overexpression (MIAPaCa-2 and PANC-1) significantly blocked PaCa cell adhesion and invasion in vitro and vice versa with stable PaCa clones (BxPC-3 and AsPC-1). Moreover, NGAL overexpression reduced focal adhesion kinase (FAK) tyrosine-397 phosphorylation in PaCa cells. Furthermore, NGAL overexpression potently decreased angiogenesis in vitro partly through reduced vascular endothelial growth factor (VEGF) production and vice versa. Stable NGAL overexpression or underexpression had no effect on PaCa cell survival, viability, and response to chemotherapeutic drugs. Finally, MIAPaCa-2 cells overexpressing NGAL reduced tumor volume (P = 0.012), local and distant metastasis (P = 0.002), and angiogenesis (P = 0.05) with no effect on K-67 proliferation index (P > 0.1) in an orthotopic nude mouse PaCa model. Collectively, our results suggest that NGAL reduces adhesion/invasion partly by suppressing FAK activation and inhibits angiogenesis partly by blocking VEGF production in PaCa cells. Thus, NGAL is a potential suppressor of invasion and angiogenesis in advanced PaCa. PMID- 18676833 TI - Down-regulation of myeloid cell leukemia-1 through inhibiting Erk/Pin 1 pathway by sorafenib facilitates chemosensitization in breast cancer. AB - Myeloid cell leukemia-1 (Mcl-1), a Bcl-2-like antiapoptotic protein, plays a role in cell immortalization and chemoresistance in a number of human malignancies. A peptidyl-prolyl cis/trans isomerase, Pin1 is involved in many cellular events, such as cell cycle progression, cell proliferation, and differentiation through isomerizing prophosphorylated substrates. It has been reported that down regulation of Pin1 induces apoptosis, and that Erk phosphorylates and up regulates Mcl-1; however, the underlying mechanisms for the two phenomena are not clear yet. Here, we showed that Pin 1 stabilizes Mcl-1, which is required for Mcl 1 posphorylation by Erk. First, we found expression of Mcl-1 and Pin1 were positively correlated and associated with poor survival in human breast cancer. We then showed that Erk could phosphorylate Mcl-1 at two consensus residues, Thr 92 and 163, which is required for the association of Mcl-1 and Pin1, resulting in stabilization of Mcl-1. Moreover, Pin1 is also required for the up-regulation of Mcl-1 by Erk activation. Based on this newly identified mechanism of Mcl-1 stabilization, two strategies were used to overcome Mcl-1-mediated chemoresistance: inhibiting Erk by Sorafenib, an approved clinical anticancer drug, or knocking down Pin1 by using a SiRNA technique. In conclusion, the current report not only unravels a novel mechanism to link Erk/Pin1 pathway and Mcl-1-mediated chemoresistance but also provides a plausible combination therapy, Taxol (Paclitaxel) plus Sorafenib, which was shown to be effective in killing breast cancer cells. PMID- 18676834 TI - Ugene, a newly identified protein that is commonly overexpressed in cancer and binds uracil DNA glycosylase. AB - Expression microarrays identified a novel transcript, designated as Ugene, whose expression is absent in normal colon and colon adenomas, but that is commonly induced in malignant colon cancers. These findings were validated by real-time PCR and Northern blot analysis in an independent panel of colon cancer cases. In addition, Ugene expression was found to be elevated in many other common cancer types, including breast, lung, uterus, and ovary. Immunofluorescence of V5-tagged Ugene revealed it to have a nuclear localization. In a pull-down assay, uracil DNA glycosylase 2 (UNG2), an important enzyme in the base excision repair (BER) pathway, was identified as a partner protein that binds to Ugene. Coimmunoprecipitation and Western blot analysis confirmed the binding between the endogenous Ugene and UNG2 proteins. Using deletion constructs, we find that Ugene binds to the first 25 amino acids of the UNG2 NH(2) terminus. We suggest that Ugene induction in cancer may contribute to the cancer phenotype by interacting with the BER pathway. PMID- 18676835 TI - Loss of integrin alpha1beta1 ameliorates Kras-induced lung cancer. AB - The collagen IV binding receptor integrin alpha1beta1 has been shown to regulate lung cancer due to its proangiogenic properties; however, it is unclear whether this receptor also plays a direct role in promoting primary lung tumors. To investigate this possibility, integrin alpha1-null mice were crossed with KrasLA2 mice that carry an oncogenic mutation of the Kras gene (G12D) and develop spontaneous primary tumors with features of non-small cell lung cancer. We provide evidence that KrasLA2/alpha1-null mice have a decreased incidence of primary lung tumors and longer survival compared with KrasLA2/alpha1 wild-type controls. Tumors from KrasLA2/alpha1-null mice were also smaller, less vascularized, and exhibited reduced cell proliferation and increased apoptosis, as determined by proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase-mediated dUTP nick-end staining, respectively. Moreover, tumors from the KrasLA2/alpha1-null mice showed diminished extracellular signal-regulated kinase (ERK) but enhanced p38 mitogen-activated protein kinase activation. Primary lung tumor epithelial cells isolated from KrasLA2/alpha1-null mice showed a significant decrease in anchorage-independent colony formation, collagen mediated cell proliferation, ERK activation, and, most importantly, tumorigenicity when injected into nude mice compared with KrasLA2/alpha1 wild type tumor cells. These results indicate that loss of the integrin alpha1 subunit decreases the incidence and growth of lung epithelial tumors initiated by oncogenic Kras, suggesting that both Kras and integrin alpha1beta1 cooperate to drive the growth of non-small cell lung cancer in vivo. PMID- 18676836 TI - MUC1 oncoprotein blocks death receptor-mediated apoptosis by inhibiting recruitment of caspase-8. AB - Stimulation of the death receptor superfamily induces the activation of caspase-8 and thereby the apoptotic response. The MUC1 oncoprotein is aberrantly overexpressed by diverse human malignancies and inhibits stress-induced apoptosis. The present results show that MUC1 blocks activation of caspase-8 and apoptosis in the response of malignant cells to tumor necrosis factor alpha, tumor necrosis factor-related apoptosis-inducing ligand, and Fas ligand. The results show that MUC1 associates constitutively with caspase-8. The MUC1 cytoplasmic domain (MUC1-CD) binds directly to the caspase-8 p18 fragment upstream to the catalytic Cys(360) site. The results also show that MUC1-CD binds to Fas-associated death domain (FADD) at the death effector domain. In nonmalignant epithelial cells, MUC1 interacts with caspase-8 and FADD as an induced response to death receptor stimulation. The functional significance of these interactions is supported by the demonstration that MUC1 competes with caspase-8 for binding to FADD and blocks recruitment of caspase-8 to the death inducing signaling complex. These findings indicate that MUC1 is of importance to the physiologic regulation of caspase-8 activity and that overexpression of MUC1, as found in human malignancies, could contribute to constitutive inhibition of death receptor signaling pathways. PMID- 18676837 TI - BRAF V600E disrupts AZD6244-induced abrogation of negative feedback pathways between extracellular signal-regulated kinase and Raf proteins. AB - AZD6244 (ARRY 142886) is a potent and selective mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor currently in early clinical trials. We examined the activity of AZD6244 in a panel of non-small cell lung cancer and a panel of cell lines representing many cancer types using in vitro growth assays. AZD6244 induced G(0)-G(1) cell cycle arrest in sensitive cell lines that primarily included cells containing the BRAF V600E mutation. In these cells, G(0)-G(1) arrest is accompanied by the up regulation of the cell cycle inhibitors p21(WAF1) and p27(Kip1) and down regulation of cyclin D1. In the majority of cell lines tested, including those with K-ras or non-V600E BRAF mutations, AZD6244 induced the accumulation of phospho-MEK, an effect not observed in the most sensitive BRAF V600E-containing cells. Accumulation of phospho-MEK in non-V600E-containing cell lines is due to abrogation of negative feedback pathways. BRAF V600E disrupts negative feedback signaling, which results in enhanced baseline phospho-MEK expression. Exogenous expression of BRAF V600E disrupts feedback inhibition but does not sensitize cells to AZD6244. Specific suppression of endogenous BRAF V600E does not confer resistance to AZD6244 but enhances sensitivity to AZD6244. Thus, our findings show that BRAF V600E marks cells with an in vitro requirement for MAPK signaling to support proliferation. These cells are exquisitely sensitive to AZD6244 (IC(50), <100 nmol/L), have high baseline levels of phospho-MEK, and lack feedback inhibition between ERK and Raf. These data suggest an approach to identifying cells that may be sensitive to AZD6244 and other MEK inhibitors. PMID- 18676838 TI - RhoA-dependent regulation of cell migration by the tumor suppressor hSNF5/INI1. AB - Malignant rhabdoid tumors (MRT) are extremely aggressive pediatric tumors caused by the inactivation of the hSNF5/INI1 tumor suppressor gene, which encodes a core member of the SWI/SNF chromatin remodeling complex. Roles for hSNF5/INI1 in cell cycle and differentiation have been documented. Based on the observation that MRTs are highly invasive, we investigated a role for hSNF5/INI1 in cell migration. MRT cell lines exhibit high migration properties that are dramatically reduced upon hSNF5/INI1 expression. This effect is associated with the disorganization of the actin stress fiber network and is mediated by the inhibition of the activity of the small GTPase RhoA, through a nuclear, SWI/SNF dependent transcriptional mechanism. We further show that the knockdown of hSNF5/INI1 in epithelial 293T or MCF7 cells results in increased cell size, loss of cell-cell adhesions, and enhanced migration, associated with an increased RhoA activity. Finally, we show that the SNF5 homology domain is required for hSNF5/INI1-mediated inhibition of migration, and that a missense mutation (S284L) associated with cancer is sufficient to impair hSNF5/INI1 function in migration. We conclude that the inhibition of migration is another crucial tumor suppressor function of hSNF5/INI1, in addition to its previously described functions in proliferation and differentiation, and that its loss-of-function in MRTs may account for the high invasiveness and metastatic potential of these tumors. PMID- 18676839 TI - Genomic profiling of microRNA and messenger RNA reveals deregulated microRNA expression in prostate cancer. AB - MicroRNAs are small noncoding RNAs that regulate the expression of protein-coding genes. To evaluate the involvement of microRNAs in prostate cancer, we determined genome-wide expression of microRNAs and mRNAs in 60 primary prostate tumors and 16 nontumor prostate tissues. The mRNA analysis revealed that key components of microRNA processing and several microRNA host genes, e.g., MCM7 and C9orf5, were significantly up-regulated in prostate tumors. Consistent with these findings, tumors expressed the miR-106b-25 cluster, which maps to intron 13 of MCM7, and miR-32, which maps to intron 14 of C9orf5, at significantly higher levels than nontumor prostate. The expression levels of other microRNAs, including a number of miR-106b-25 cluster homologues, were also altered in prostate tumors. Additional differences in microRNA abundance were found between organ-confined tumors and those with extraprostatic disease extension. Lastly, we found evidence that some microRNAs are androgen-regulated and that tumor microRNAs influence transcript abundance of protein-coding target genes in the cancerous prostate. In cell culture, E2F1 and p21/WAF1 were identified as targets of miR-106b, Bim of miR-32, and exportin-6 and protein tyrosine kinase 9 of miR-1. In summary, microRNA expression becomes altered with the development and progression of prostate cancer. Some of these microRNAs regulate the expression of cancer related genes in prostate cancer cells. PMID- 18676840 TI - Reciprocal relationship between O6-methylguanine-DNA methyltransferase P140K expression level and chemoprotection of hematopoietic stem cells. AB - Retroviral-mediated delivery of the P140K mutant O(6)-methylguanine-DNA methyltransferase (MGMT(P140K)) into hematopoietic stem cells (HSC) has been proposed as a means to protect against dose-limiting myelosuppressive toxicity ensuing from chemotherapy combining O(6)-alkylating agents (e.g., temozolomide) with pseudosubstrate inhibitors (such as O(6)-benzylguanine) of endogenous MGMT. Because detoxification of O(6)-alkylguanine adducts by MGMT is stoichiometric, it has been suggested that higher levels of MGMT will afford better protection to gene-modified HSC. However, accomplishing this goal would potentially be in conflict with current efforts in the gene therapy field, which aim to incorporate weaker enhancer elements to avoid insertional mutagenesis. Using a panel of self inactivating gamma-retroviral vectors that express a range of MGMT(P140K) activity, we show that MGMT(P140K) expression by weaker cellular promoter/enhancers is sufficient for in vivo protection/selection following treatment with O(6)-benzylguanine/temozolomide. Conversely, the highest level of MGMT(P140K) activity did not promote efficient in vivo protection despite mediating detoxification of O(6)-alkylguanine adducts. Moreover, very high expression of MGMT(P140K) was associated with a competitive repopulation defect in HSC. Mechanistically, we show a defect in cellular proliferation associated with elevated expression of MGMT(P140K), but not wild-type MGMT. This proliferation defect correlated with increased localization of MGMT(P140K) to the nucleus/chromatin. These data show that very high expression of MGMT(P140K) has a deleterious effect on cellular proliferation, engraftment, and chemoprotection. These studies have direct translational relevance to ongoing clinical gene therapy studies using MGMT(P140K), whereas the novel mechanistic findings are relevant to the basic understanding of DNA repair by MGMT. PMID- 18676841 TI - Kinetic stabilization of microtubule dynamics by estramustine is associated with tubulin acetylation, spindle abnormalities, and mitotic arrest. AB - Estramustine (EM) alone or in combination with other anticancer agents is clinically used for the treatment of hormone refractory prostate cancer. Furthermore, EM has been shown to potently inhibit the proliferation of different types of cancer cells in culture apparently by targeting microtubules; however, the antiproliferative mechanism of action of EM is not clear. In this work, we have shown that EM strongly suppressed the dynamic instability of individual microtubules in MCF-7 cells by reducing the rates of growing and shortening excursions and increasing the time microtubule spent in the pause state. At its half maximal proliferation inhibitory concentration (IC(50)), EM exerted strong suppressive effects on the dynamics of microtubules in MCF-7 cells without detectably affecting either the organization or the polymerized mass of microtubules. At relatively high concentrations (5 x IC(50)), EM significantly depolymerized microtubules in the cells. Furthermore, the microtubules were found highly acetylated, supporting the conclusion that they were stabilized by the drug. EM treatment induced spindle abnormalities in MCF-7 cells, and a major population of the arrested mitotic cells was multipolar. EM also perturbed the microtubule-kinetochore interaction, thereby activating the spindle assembly checkpoint and leading to apoptotic cell death. PMID- 18676842 TI - Par-4 binds to topoisomerase 1 and attenuates its DNA relaxation activity. AB - The regulation of DNA relaxation by topoisomerase 1 (TOP1) is essential for DNA replication, transcription, and recombination events. TOP1 activity is elevated in cancer cells, yet the regulatory mechanism restraining its activity is not understood. We present evidence that the tumor suppressor protein prostate apoptosis response-4 (Par-4) directly binds to TOP1 and attenuates its DNA relaxation activity. Unlike camptothecin, which binds at the TOP1-DNA interface to form cleavage complexes, Par-4 interacts with TOP1 via its leucine zipper domain and sequesters TOP1 from the DNA. Par-4 knockdown by RNA interference enhances DNA relaxation and gene transcription activities and promotes cellular transformation in a TOP1-dependent manner. Conversely, attenuation of TOP1 activity either by RNA interference or Par-4 overexpression impedes DNA relaxation, cell cycle progression, and gene transcription activities and inhibits transformation. Collectively, our findings suggest that Par-4 serves as an intracellular repressor of TOP1 catalytic activity and regulates DNA topology to suppress cellular transformation. PMID- 18676843 TI - Loss of MLL PHD finger 3 is necessary for MLL-ENL-induced hematopoietic stem cell immortalization. AB - Reciprocal chromosomal translocations at the MLL gene locus result in expression of novel fusion proteins, such as MLL-ENL, associated with leukemia. The three PHD finger cassette, one of the highly conserved domains in MLL, is absent in all fusion proteins. This domain has been shown to interact with Cyp33, a cyclophilin which enhances the recruitment of histone deacetylases (HDAC) to the MLL repression domain and mediates HOX gene repression. Insertion of the third PHD finger of MLL into MLL-ENL allows the recruitment of Cyp33 and, subsequently, HDAC1 to the fusion protein. Furthermore, expression of the fusion protein with the PHD finger insertion mediates the down-regulation of the HOXC8 gene expression in a Cyp33-dependent manner. Finally, the addition of the PHD finger domain or the third PHD finger alone into MLL-ENL blocks the hematopoietic stem cell immortalization potential of the fusion protein in serial plating colony assays. Insertion of only the first and second PHD fingers has no such effect. These data support the hypothesis that the binding of Cyp33 to the MLL third PHD finger switches the MLL function from transactivation to repression. In the immortalizing MLL fusion protein, the loss of the PHD fingers, in combination with the gain of the activation domain of ENL or of other partner proteins, makes the fusion protein a constitutive transactivator. This leads to constitutive overexpression of MLL target genes that block stem cell commitment and promote stem cell renewal, probably the first step in MLL-related leukemogenesis. PMID- 18676844 TI - RUNX3 methylation reveals that bladder tumors are older in patients with a history of smoking. AB - Exposure to tobacco smoke is associated with increased DNA methylation at certain genes in both lung and bladder tumors. We sought to identify interactions in bladder cancer between DNA methylation and a history of smoking, along with any possible effect of aging. We measured DNA methylation in 342 transitional cell carcinoma tumors at BCL2, PTGS2 (COX2), DAPK, CDH1 (ECAD), EDNRB, RASSF1A, RUNX3, TERT, and TIMP3. The prevalence of methylation at RUNX3, a polycomb target gene, increased as a function of age at diagnosis (P = 0.031) and a history of smoking (P = 0.015). RUNX3 methylation also preceded methylation at the other eight genes (P < 0.001). It has been proposed that DNA methylation patterns constitute a "molecular clock" and can be used to determine the "age" of normal tissues (i.e., the number of times the cells have divided). Because RUNX3 methylation increases with age, is not present in normal urothelium, and occurs early in tumorigenesis, it can be used for the first time as a molecular clock to determine the age of a bladder tumor. Doing so reveals that tumors from smokers are "older" than tumors from nonsmokers (P = 0.009) due to tumors in smokers either initiating earlier or undergoing more rapid cell divisions. Because RUNX3 methylation is acquired early on in tumorigenesis, then its detection in biopsy or urine specimens could provide a marker to screen cigarette smokers long before any symptoms of bladder cancer are present. PMID- 18676845 TI - Fibroblast growth factor 2 restrains Ras-driven proliferation of malignant cells by triggering RhoA-mediated senescence. AB - Fibroblast growth factor 2 (FGF2) is considered to be a bona fide oncogenic factor, although results from our group and others call this into question. Here, we report that exogenous recombinant FGF2 irreversibly inhibits proliferation by inducing senescence in Ras-dependent malignant mouse cells, but not in immortalized nontumorigenic cell lines. We report the following findings in K-Ras dependent malignant Y1 adrenocortical cells and H-Ras V12-transformed BALB-3T3 fibroblasts: (a) FGF2 inhibits clonal growth and tumor onset in nude and immunocompetent BALB/c mice, (b) FGF2 irreversibly blocks the cell cycle, and (c) FGF2 induces the senescence-associated beta-galactosidase with no accompanying signs of apoptosis or necrosis. The tyrosine kinase inhibitor PD173074 completely protected malignant cells from FGF2. In Y1 adrenal cells, reducing the constitutively high levels of K-Ras-GTP using the dominant-negative RasN17 mutant made cells resistant to FGF2 cytotoxicity. In addition, transfection of the dominant-negative RhoA-N19 into either Y1 or 3T3-B61 malignant cell lines yielded stable clonal transfectants that were unable to activate RhoA and were resistant to the FGF2 stress response. We conclude that in Ras-dependent malignant cells, FGF2 interacts with its cognate receptors to trigger a senescence-like process involving RhoA-GTP. Surprisingly, attempts to select FGF2-resistant cells from the Y1 and 3T3-B61 cell lines yielded only rare clones that (a) had lost the overexpressed ras oncogene, (b) were dependent on FGF2 for proliferation, and (c) were poorly tumorigenic. Thus, FGF2 exerted a strong negative selection that Ras dependent malignant cells could rarely overcome. PMID- 18676846 TI - Transforming growth factor-beta (TGF-beta) and TGF-beta-associated kinase 1 are required for R-Ras-mediated transformation of mammary epithelial cells. AB - Transforming growth factor-beta (TGF-beta) cooperates with oncogenic members of the Ras superfamily to promote cellular transformation and tumor progression. Apart from the classic (H-, K-, and N-) Ras GTPases, only the R-Ras subfamily (R Ras, R-Ras2/TC21, and R-Ras3/M-Ras) has significant oncogenic potential. In this study, we show that oncogenic R-Ras transformation of EpH4 cells requires TGF beta signaling. When murine EpH4 cells were stably transfected with a constitutively active R-Ras(G38V) mutant, they were no longer sensitive to TGF beta-mediated growth inhibition and showed increased proliferation and transformation in response to exogenous TGF-beta. R-Ras/EpH4 cells require TGF beta signaling for transformation to occur and they produce significantly elevated levels of endogenous TGF-beta, which signals in an autocrine fashion. The effects of TGF-beta are independent of Smad2/3 activity and require activation of TGF-beta-associated kinase 1 (TAK1) and its downstream effectors c Jun NH(2)-terminal kinase and p38 mitogen-activated protein kinase as well as the phosphoinositide 3-kinase/Akt and mammalian target of rapamycin pathways. Thus, TAK1 is a novel link between TGF-beta signaling and oncogenic R-Ras in the promotion of tumorigenesis. PMID- 18676847 TI - Vascular endothelial growth factor reduces tamoxifen efficacy and promotes metastatic colonization and desmoplasia in breast tumors. AB - Clinical studies have shown that decreased tamoxifen effectiveness correlates with elevated levels of vascular endothelial growth factor (VEGF)-A(165) in biopsy samples of breast cancers. To investigate the mechanisms underlying tamoxifen resistance and metastasis, we engineered the estrogen receptor (ER) positive MCF-7 human breast cancer cell line to express VEGF to clinically relevant levels in a doxycycline-regulated manner. Induction of VEGF expression in orthotopically implanted xenografts that were initially tamoxifen responsive and noninvasive resulted in tamoxifen-resistant tumor growth and metastasis to the lungs. Lung metastases were also observed in a VEGF-dependent manner following tail vein injection of tumor cells. At both primary and metastatic sites, VEGF-overexpressing tumors exhibited extensive fibroblastic stromal content, a clinical feature called desmoplasia. VEGF-induced metastatic colonies were surrounded by densely packed stromal cells before detectable angiogenesis, suggesting that VEGF is involved in the initiation of desmoplasia. Because expression of VEGF receptors R1 and R2 was undetectable in these tumor cells, the observed VEGF effects on reduction of tamoxifen efficacy and metastatic colonization are most likely mediated by paracrine signaling that enhances tumor/stromal cell interactions and increases the level of desmoplasia. This study reveals new roles for VEGF in breast cancer progression and suggests that combination of antiestrogens and VEGF inhibitors may prolong tamoxifen sensitivity and prevent metastasis in patients with ER-positive tumors. PMID- 18676848 TI - Inhibition of metastatic outgrowth from single dormant tumor cells by targeting the cytoskeleton. AB - Metastatic breast cancer may emerge from latent tumor cells that remain dormant at disseminated sites for many years. Identifying mechanisms regulating the switch from dormancy to proliferative metastatic growth has been elusive due to the lack of experimental models of tumor cell dormancy. We characterized the in vitro growth characteristics of cells that exhibit either dormant (D2.0R, MCF-7, and K7M2AS1.46) or proliferative (D2A1, MDA-MB-231, and K7M2) metastatic behavior in vivo. Although these cells proliferate readily in two-dimensional culture, we show that when grown in three-dimensional matrix, distinct growth properties of the cells were revealed that correlate to their dormant or proliferative behavior at metastatic sites in vivo. In three-dimensional culture, cells with dormant behavior in vivo remained cell cycle arrested with elevated nuclear expression of p16 and p27. The transition from quiescence to proliferation of D2A1 cells was dependent on fibronectin production and signaling through integrin beta1, leading to cytoskeletal reorganization with filamentous actin (F-actin) stress fiber formation. We show that phosphorylation of myosin light chain (MLC) by MLC kinase (MLCK) through integrin beta1 is required for actin stress fiber formation and proliferative growth. Inhibition of integrin beta1 or MLCK prevents transition from a quiescent to proliferative state in vitro. Inhibition of MLCK significantly reduces metastatic outgrowth in vivo. These studies show that the switch from dormancy to metastatic growth may be regulated, in part, through epigenetic signaling from the microenvironment, leading to changes in the cytoskeletal architecture of dormant cells. Targeting this process may provide therapeutic strategies for inhibition of the dormant-to-proliferative metastatic switch. PMID- 18676849 TI - Effect of ablation or inhibition of stromal matrix metalloproteinase-9 on lung metastasis in a breast cancer model is dependent on genetic background. AB - Matrix metalloproteinases (MMP) are a family of enzymes with a myriad of functions. Lately, we have come to realize that broad-spectrum inhibition of these enzymes, as was tried unsuccessfully in multiple phase III trials in cancer patients, is likely unwise given the protumorigenic and antitumorigenic functions of various family members. Here, we used the multistage mammary tumor model MMTV PyVT to investigate roles for either MMP7 or MMP9 in tumor progression. We found no effect of genetic ablation of MMP7 or MMP9 on the multifocal tumors that developed in the mammary glands. Lack of MMP7 also had no effect on the development of lung metastases, suggesting that MMP7 is irrelevant in this model. In contrast, MMP9 deficiency was associated with an 80% decrease in lung tumor burden. The predominant cellular source of MMP9 was myeloid cells, with neutrophils being the largest contributor in tumor-bearing lungs. Experimental metastasis assays corroborated the role of host-derived MMP9 in lung metastasis and also facilitated determination of a time frame most relevant for the MMP9 mediated effect. The lung tumors from MMP9-deficient mice showed decreased angiogenesis. Surprisingly, the antimetastatic outcome of MMP9 ablation seemed to be dependent on strain. Only mice that had genetic background derived from C57BL/6 showed reduced metastasis, whereas mice fully of the FVB/N background showed no significant effect. These strain-specific responses were also observed in a study using a highly selective pharmacologic inhibitor of MMP9 and thus suggest that responses to MMP inhibition are controlled by genetic differences. PMID- 18676850 TI - A pivotal role for heat shock protein 90 in Ewing sarcoma resistance to anti insulin-like growth factor 1 receptor treatment: in vitro and in vivo study. AB - Ewing Sarcoma (ES) shows several deregulated autocrine loops mediating cell survival and proliferation. Therefore, their blockade is a promising therapeutic approach. We previously reported the in vitro effect of insulin-like growth factor 1 receptor (IGF1R)/KIT pathway blockade on ES cell lines, and we now extend our observations to changes induced by this treatment in interacting proteins/networks. A proteomic analysis revealed that Heat Shock Protein (HSP)90 was differentially expressed between ES cell lines sensitive and resistant to specific IGF1R/KIT inhibitors. We therefore inhibited HSP90 with 17-allylamino-17 demethoxygeldanamycin (17-AAG) and siRNA, and observed that ES cell line growth and survival were reduced, especially in the resistant cell lines. Conversely, HSP90 induced-expression conferred resistance to anti-IGF1R/KIT treatment in the sensitive cell lines. 17-AAG treatment induced HSP90 client protein degradation, including AKT, KIT, or IGF1R, by inhibiting their physical interaction with HSP90. Xenograft models developed with A673 ES cell line confirmed that HSP90 inhibition, alone or combined with IGF1R inhibition, significantly reduced tumor growth and expression of client proteins. Remarkably, using two independent clinical sample sets, we have found that nearly half of IGF1R-positive tumors also show HSP90 overexpression. This delineates a subset of patients that could benefit from combination of anti-HSP90 agents when considering IGF1R-targeting therapies. Importantly, sensitivity to drugs such as ADW/IMA depends not only on the levels of expression and basal activation of IGF1R/KIT, but also, and for the first time reported in ES, on the development of the stress response mechanism. Accordingly, HSP90 expression could be a predictive factor of response to IGF1R targeting therapies. PMID- 18676851 TI - Phosphatidylinositol 3-kinase inhibition broadly sensitizes glioblastoma cells to death receptor- and drug-induced apoptosis. AB - The aberrant activity of the phosphatidylinositol 3-kinase (PI3K) pathway has been reported to correlate with adverse clinical outcome in human glioblastoma in vivo. However, the question of how this survival network can be successfully targeted to restore the sensitivity of glioblastoma to apoptosis induction has not yet been answered. Here, we report that inhibition of PI3K by LY294002 broadly sensitizes wild-type and mutant PTEN glioblastoma cells to both death receptor- and chemotherapy-induced apoptosis, whereas mammalian target of rapamycin (mTOR) inhibition is not sufficient to restore apoptosis sensitivity. LY294002 significantly enhances apoptosis triggered by tumor necrosis factor related apoptosis-inducing ligand (TRAIL), agonistic anti-CD95 antibodies, or several anticancer drugs (i.e., doxorubicin, etoposide, and vincristine) in a highly synergistic manner. In addition, LY294002 cooperates with TRAIL or doxorubicin to suppress colony formation, thus also showing a strong effect on long-term survival. Similarly, genetic knockdown of PI3K subunits p110alpha and/or p110beta by RNA interference (RNAi) primes glioblastoma cells for TRAIL- or doxorubicin-mediated apoptosis. In contrast to PI3K inhibition, pharmacologic or genetic blockade of mTOR by RAD001 (everolimus), rapamycin, or RNAi fails to enhance TRAIL- or doxorubicin-induced apoptosis. Analysis of apoptosis pathways reveals that PI3K inhibition acts in concert with TRAIL or doxorubicin to trigger mitochondrial membrane permeabilization, caspase activation, and caspase dependent apoptosis, which are abolished by the caspase inhibitor N benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone. Most importantly, PI3K inhibition by LY294002 sensitizes primary cultured glioblastoma cells obtained from surgical specimens to TRAIL- or chemotherapy-induced cell death. By showing that PI3K inhibition broadly primes glioblastoma cells for apoptosis, our findings provide the rationale for using PI3K inhibitors in combination regimens to enhance TRAIL- or chemotherapy-induced apoptosis in glioblastoma. PMID- 18676852 TI - Oct-3/4 expression reflects tumor progression and regulates motility of bladder cancer cells. AB - Cancer and embryonic stem cells exhibit similar behavior, including immortal, undifferentiated, and invasive activities. Here, we show that in clinical samples bladder tumors with intense expression of stem cell marker Oct-3/4 (also known as POU5F1) are associated with further disease progression, greater metastasis, and shorter cancer-related survival compared with those with moderate and low expressions. Expression of Oct-3/4 is detected in human bladder transitional cell carcinoma samples and cell lines. Overexpression of Oct-3/4 enhances, whereas knockdown of Oct-3/4 expression by RNA interference reduces, migration and invasion of bladder cancer cells. Oct-3/4 can up-regulate fibroblast growth factor-4 and matrix metalloproteinase-2 (MMP-2), MMP-9, and MMP-13 production, which may contribute to tumor metastasis. Finally, we show that Ad5WS4, an E1B-55 kD-deleted adenovirus driven by the Oct-3/4 promoter, exerts potent antitumor activity against bladder cancer in a syngeneic murine tumor model. Therefore, our results implicate that Oct-3/4 may be useful as a novel tumor biological and prognostic marker and probably as a potential therapeutic target for bladder cancer. PMID- 18676853 TI - A small-molecule E2F inhibitor blocks growth in a melanoma culture model. AB - HLM006474 was identified using a computer-based virtual screen and the known crystal structure of the DNA-bound E2F4/DP2 heterodimer. Treatment of multiple cell lines with HLM006474 resulted in the loss of intracellular E2F4 DNA-binding activity as measured by electrophoretic mobility shift assay within hours. Overnight exposure to HLM006474 resulted in down-regulation of total E2F4 protein as well as known E2F targets. The effects of HLM006474 treatment on different cell lines varied but included a reduction in cell proliferation and an increase in apoptosis. HLM006474 induced apoptosis in a manner distinct from cisplatin and doxorubicin. E2F4-null mouse embryonic fibroblasts were less sensitive than wild type counterparts to the apoptosis-inducing activity of the compound, revealing its biological specificity. A375 cells were extremely sensitive to the apoptosis inducing activity of the compound in two-dimensional culture, and HLM006474 was a potent inhibitor of melanocytes proliferation and subsequent invasion in a three dimensional tissue culture model system. Together, these results suggest that interference with E2F activity using small molecules may have clinical application in cancer therapy. PMID- 18676854 TI - Differential cellular internalization of anti-CD19 and -CD22 immunotoxins results in different cytotoxic activity. AB - B-cell malignancies routinely express surface antigens CD19 and CD22. Immunotoxins against both antigens have been evaluated, and the immunotoxins targeting CD22 are more active. To understand this disparity in cytotoxicity and guide the screening of therapeutic targets, we compared two immunotoxins, FMC63(Fv)-PE38-targeting CD19 and RFB4(Fv)-PE38 (BL22)-targeting CD22. Six lymphoma cell lines have 4- to 9-fold more binding sites per cell for CD19 than for CD22, but BL22 is 4- to 140-fold more active than FMC63(Fv)-PE38, although they have a similar cell binding affinity (Kd, approximately 7 nmol/L). In 1 hour, large amounts of BL22 are internalized (2- to 3-fold more than the number of CD22 molecules on the cell surface), whereas only 5.2% to 16.6% of surface bound FMC63(Fv)-PE38 is internalized. The intracellular reservoir of CD22 decreases greatly after immunotoxin internalization, indicating that it contributes to the uptake of BL22. Treatment of cells with cycloheximide does not reduce the internalization of BL22. Both internalized immunotoxins are located in the same vesicles. Our results show that the rapid internalization of large amounts of BL22 bound to CD22 makes CD22 a better therapeutic target than CD19 for immunotoxins and probably for other immunoconjugates that act inside cells. PMID- 18676855 TI - Topology of NGEP, a prostate-specific cell:cell junction protein widely expressed in many cancers of different grade level. AB - New gene expressed in prostate (NGEP) is a prostate-specific polytopic membrane protein found at high concentrations at cell:cell contact regions. To determine if NGEP is a useful target for antibody-based therapy of prostate cancer, we performed an immunohistochemical analysis of 126 human prostate carcinoma samples using polyclonal anti-NGEP sera and found that 91% of the cancers express NGEP protein. To elucidate the topology of NGEP and guide the development of monoclonal antibodies (mAb) reacting with the extracellular regions of NGEP, a hemagglutinin epitope tag was inserted at several positions within the NGEP sequence. The tagged proteins were expressed in 293T cells and locations of the tags were determined by immunofluorescence in intact or permeabilized cells. The results indicate that NGEP contains eight transmembrane domains with both the NH(2) and COOH termini of NGEP located inside the cell. We produced mAb to three regions that are predicted to be intracellular based on the epitope tag data (amino acids 1-352, 441-501, and 868-933), and as predicted, the mAb only detected the protein in permeabilized cells. NGEP is a glycoprotein with predicted glycosylation sites at N809 and N824. When these residues were converted to glutamine, glycosylation was abolished, confirming that the residues are extracellular. Our findings on the expression and the orientation of the NGEP protein serve as an important framework for the development of mAb targeting the extracellular regions of NGEP that could be used for prostate cancer immunotherapy. PMID- 18676856 TI - Bobel-24 and derivatives induce caspase-independent death in pancreatic cancer regardless of apoptotic resistance. AB - The poor prognosis of pancreatic cancer and poor sensitivity to current therapeutics, associated with resistance to apoptosis, urge the search for new drugs. We previously described the induction of caspase-independent mithochondrial death in leukemia cells by Bobel-24 (AM-24) and derivatives. Here, we explored whether these compounds induce a similar cytotoxicity in human pancreatic carcinoma cell lines (NP18, NP9, NP31, and NP29). Bobel-24 or Bobel-16 induced cytotoxicity and DNA synthesis inhibition in all cell lines and apoptosis in all lines, except for NP9. Caspase and/or poly(ADP-ribose) polymerase-1 (PARP 1) activity inhibition experiments showed that cytotoxicity was mainly induced through apoptosis in NP18 and through a caspase-independent process in NP9. Moreover, in NP29 or NP31 cell lines, both caspase-dependent and caspase independent cell death mechanisms coexisted. Cell death was associated with reactive oxygen species (ROS) production, mitochondrial depolarization, cytochrome c and apoptosis-inducing factor (AIF) release, AIF nuclear translocation, and lysosomal cathepsin release. Inhibition of ROS production, mitochondrial pore permeability, PARP-1, or phospholipase A2 partially prevented cell death. Moreover, cathepsin B inhibition or down-regulation by small interfering RNA partially blocked cell death. In conclusion, Bobel-24 and derivatives trigger caspase-independent lysosomal and mitochondrial death in all tested human pancreatic cancer lines, irrespective of their degree of apoptotic sensitivity, becoming the only active cytotoxic mechanism in the apoptosis resistant NP9 line. This mechanism may overcome the resistance to apoptosis observed in pancreatic carcinoma when treated with current genotoxic drugs. PMID- 18676858 TI - Gonadotropin-releasing hormone receptor levels and cell context affect tumor cell responses to agonist in vitro and in vivo. AB - Activation of gonadotropin-releasing hormone (GnRH) receptors inhibits proliferation of transformed cells derived from reproductive tissues and in transfected cell lines. Hence, GnRH receptors represent a therapeutic target for direct action of GnRH analogues on certain proliferating cells. However, more cell biological data are required to develop this particular application of GnRH analogues. Therefore, we compared the effects of GnRH receptor activation in transfected HEK293 cells (HEK293([SCL60])) with transfected human ovarian cancer cell lines SKOV3 and EFO21, human hepatoblastoma HepG2 cells, and rat neuroblastoma B35 cells. Marked differences in receptor levels, magnitude of inositol phosphate generation, and dynamics of inositol phosphate turnover occurred in the different cells. Activation of GnRH receptors, expressed at high or moderate levels, inhibited the growth of HEK293([SCL60]) and B35 cells, respectively. Western blotting detected markers of apoptosis [cleaved poly(ADP ribose) polymerase, caspase-9] in HEK293([SCL60]) and B35 following treatment with 100 nmol/L d-Trp(6)-GnRH-I. Cell growth inhibition was partially or completely rescued with inhibitor Q-VD-OPh or Ro32-0432. Low levels of GnRH receptor expression in transfected SKOV3, EFO21, or HepG2 activated intracellular signaling but did not induce apoptosis or significantly affect cell proliferation. Tumor xenografts prepared from HEK293([SCL60]) regressed during treatment with d-Trp(6)-GnRH-I and growth of xenografts derived from transfected B35 was slowed. SKOV3 xenografts were not growth inhibited. Therefore, differences in levels of GnRH receptor and signaling differentially affect the apoptotic machinery within cell lines and contribute to the cell type-specific effects of GnRH on growth. Further studies should exploit the growth-inhibitory potential of GnRH receptor activation in abnormal cells in diseased human tissues. PMID- 18676857 TI - Oxidative stress plays a critical role in inactivating mutant BRAF by geldanamycin derivatives. AB - The geldanamycin derivatives 17-allylamino-17-demethoxygeldanamycin (17-AAG) and 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG) are promising chemotherapeutic drugs that inhibit heat shock protein 90 (HSP90) function. Previous studies have shown that 17-AAG/DMAG treatment induces the degradation of mutant BRAF (V600E) and inhibits the activation of mitogen-activated protein/extracellular signal-regulated kinase 1/2 (MEK1/2). We have found, however, that HSP90 inhibition alone is not sufficient for efficient BRAF(V600E) degradation in some cells. HSP90 inhibitors structurally unrelated to geldanamycin, radicicol and novobiocin, while inducing the degradation of the HSP90 client protein RAF-1 fail to induce BRAF(V600E) degradation or inhibit MEK1/2 activation in HT29 human colon cancer cells. Moreover, after treatment with 17-DMAG, the kinase activity of residual, undegraded BRAF(V600E) was also lost. Incubation of cells with a reactive oxygen species (ROS) scavenger, N acetyl cysteine, partially restored kinase activity and also partially prevented BRAF(V600E) degradation due to 17-DMAG treatment. Conversely, treatment with the ROS producing drug menadione clearly inhibited MEK1/2 and reduced BRAF(V600E). These results suggest that in addition to direct inhibition of HSP90, the antitumor effect of geldanamycin and its derivatives is also mediated though the production of ROS, which may directly inactivate tumorigenic mutant BRAF(V600E). PMID- 18676859 TI - Impaired dendritic cell function in aging leads to defective antitumor immunity. AB - We recently reported that bone marrow-derived dendritic cells (DC) from aged miced are less effective than their young counterparts in inducing the regression of B16-ovalbumin (OVA) melanomas. To examine the underlying mechanisms, we investigated the effect of aging on DC tumor antigen presentation and migration. Although aging does not affect the ability of DCs to present OVA peptide((257 264)), DCs from aged mice are less efficient than those from young mice in stimulating OVA-specific T cells in vitro. Phenotypic analysis revealed a selective decrease in DC-specific/intracellular adhesion molecule type-3-grabbing nonintegrin (DC-SIGN) level in aged DCs. Adoptive transfer experiments showed defective in vivo DC trafficking in aging. This correlates with impaired in vitro migration and defective CCR7 signaling in response to CCL21 in aged DCs. Interestingly, vaccination of young mice using old OVA peptide((257-264))-pulsed DCs (OVA PP-DC) resulted in impaired activation of OVA-specific CD8(+) T cells in vivo. Effector functions of these T cells, as determined by IFN-gamma production and cytotoxic activity, were similar to those obtained from mice vaccinated with young OVA PP-DCs. A decreased influx of intratumor CD8(+) T cells was also observed. Importantly, although defective in vivo migration could be restored by increasing the number of old DCs injected, the aging defect in DC tumor surveillance and OVA-specific CD8(+) T-cell induction remained. Taken together, our findings suggest that defective T-cell stimulation contributes to the observed impaired DC tumor immunotherapeutic response in aging. PMID- 18676860 TI - The Wilms' tumor antigen is a novel target for human CD4+ regulatory T cells: implications for immunotherapy. AB - Compelling evidences indicate a key role for regulatory T cells (T(reg)) on the host response to cancer. The Wilms' tumor antigen (WT1) is overexpressed in several human leukemias and thus considered as promising target for development of leukemia vaccine. However, recent studies indicated that the generation of effective WT1-specific cytotoxic T cells can be largely affected by the presence of T(regs). We have generated T-cell lines and clones that specifically recognized a WT1-84 (RYFKLSHLQMHSRKH) peptide in an HLA-DRB1*0402-restricted manner. Importantly, they recognized HLA-DRB1*04-matched fresh leukemic cells expressing the WT1 antigen. These clones exerted a T helper 2 cytokine profile, had a CD4(+)CD25(+)Foxp3(+)GITR(+)CD127(-) T(reg) phenotype, and significantly inhibited the proliferative activity of allogeneic T cells independently of cell contact. Priming of alloreactive T cells in the presence of T(regs) strongly inhibited the expansion of natural killer (NK), NK T, and CD8(+) T cells and had an inhibitory effect on NK/NK T cytotoxic activity but not on CD8(+) T cells. Furthermore, priming of T cells with the WT1-126 HLA-A0201-restricted peptide in the presence of T(regs) strongly inhibited the induction of anti-WT1-126 CD8(+) CTL responses as evidenced by both very low cytotoxic activity and IFN-gamma production. Moreover, these T(reg) clones specifically produced granzyme B and selectively induced apoptosis in WT1-84-pulsed autologous antigen-presenting cells but not in apoptotic-resistant DR4-matched leukemic cells. Importantly, we have also detected anti-WT1-84 interleukin-5(+)/granzyme B(+)/Foxp3(+) CD4(+) T(regs) in five of eight HLA-DR4(+) acute myeloid leukemia patients. Collectively, our in vitro and in vivo findings strongly suggest important implications for the clinical manipulation of T(regs) in cancer patients. PMID- 18676862 TI - Tumor-associated MICA is shed by ADAM proteases. AB - The immunoreceptor NKG2D promotes immunosurveillance of malignant cells and protects the host from tumor initiation by activating natural killer cells and costimulating CD8 T cells. NKG2D-mediated recognition of malignant cells by cytotoxic lymphocytes is enabled through the tumor-associated expression of NKG2D ligands (NKG2DL) resulting from cellular or genotoxic stress. Shedding of NKG2DL is thought to constitute a major countermechanism of tumor cells to subvert NKG2D mediated immunosurveillance. Here, we report that the prototypical NKG2DL MICA is released by proteolytic cleavage in the stalk of the MICA ectodomain, where deletions, but not alanine substitutions, impede MICA shedding. Small compound mediated stimulation and inhibition of MICA shedding adduced characteristics that indicated an involvement of members of the "a disintegrin and metalloproteinase" (ADAM) family. Accordingly, MICA shedding by tumor cells was inhibited by silencing of the related ADAM10 and ADAM17 proteases, which are known to promote tumor growth by releasing epidermal growth factor receptor ligands. Collectively, our data show that ADAM10 and ADAM17 are critically involved in the tumor associated proteolytic release of soluble MICA facilitating tumor immune escape. Hence, therapeutic blockade of ADAM10 and ADAM17 seems promising for cancer treatment by targeting both growth and immune escape of tumors. PMID- 18676861 TI - Antitumor activity of immunotoxins with T-cell receptor-like specificity against human melanoma xenografts. AB - In this study, we have explored the use of Fab-toxin proteins (immunotoxin) to target antigen-specific MHC-peptide complexes of in vitro and in vivo cancer cells. A human phage display library was used to screen for T-cell receptor (TCR) like antibodies that are highly specific for the peptide melanoma-associated antigen MART-1(26-35) presented by HLA-A201. We also used previously selected TCR like antibodies specific for the peptide melanoma-associated antigen gp100(280 288) presented by HLA-A201. The recombinant immunotoxin constructs were generated by fusing the targeting Fab fragment to a truncated form of Pseudomonas exotoxin, PE38KDEL. These immunotoxins bound with high affinity to the EBV-transformed JY cell line pulsed with the aforementioned peptides and internalized within 30 min. A significant inhibition of protein synthesis, which resulted in cell death, was detected at 24 h. MART-1-specific and gp100-specific immunotoxins bound and killed HLA-A201 melanoma MART-1(+) and gp100(+) cell lines that were presented at natural levels but do not bind to HLA-A201(-) or to HLA-A201(+) MART-1(-) and gp100(-) cell lines. In severe combined immunodeficient mice, MART-1 and gp100 immunotoxins significantly and discriminately inhibited human melanoma growth. These results show that MHC class I/peptide complexes can serve as a specific target for passive immunotherapy of cancer. PMID- 18676863 TI - Rat prolactinoma cell growth regulation by epidermal growth factor receptor ligands. AB - Epidermal growth factor (EGF) regulates pituitary development, hormone synthesis, and cell proliferation. Although ErbB receptor family members are expressed in pituitary tumors, the effects of EGF signaling on pituitary tumors are not known. Immunoprecipitation and Western blot confirmed EGF receptor (EGFR) and p185(c neu) protein expression in GH3 lacto-somatotroph but not in adrenocorticotropic hormone-secreting AtT20 pituitary tumor cells. EGF (5 nmol/L) selectively enhanced baseline ( approximately 4-fold) and serum-induced (>6-fold) prolactin (PRL) mRNA levels, whereas gefitinib, an EGFR antagonist, suppressed serum induced cell proliferation and Pttg1 expression, blocked PRL gene expression, and reversed EGF-mediated somatotroph-lactotroph phenotype switching. Downstream EGFR signaling by ERK, but not phosphoinositide-3-kinase or protein kinase C, mediated the gefitinib response. Tumors in athymic mice implanted s.c. with GH3 cells resulted in weight gain accompanied by increased serum PRL, growth hormone, and insulin growth factor 1. Gefitinib decreased tumor volumes and peripheral hormone levels by approximately 30% and restored normal mouse body weight patterns. Mice treated with gefitinib exhibited decreased tumor tissue ERK1/2 phosphorylation and down-regulated tumor PRL and Pttg1 mRNA abundance. These results show that EGFR inhibition controls tumor growth and PRL secretion in experimental lacto somatotroph tumors. EGFR inhibitors could therefore be useful for the control of PRL secretion and tumor load in prolactinomas resistant to dopaminergic treatment, or for those prolactinomas undergoing rare malignant transformation. PMID- 18676864 TI - The HER4/4ICD estrogen receptor coactivator and BH3-only protein is an effector of tamoxifen-induced apoptosis. AB - Greater than 40% of breast cancer patients treated with tamoxifen exhibit de novo or acquired tumor resistance. Recent clinical evidence indicates that loss of expression of HER4 is an independent marker for tamoxifen resistance. In direct corroboration with clinical observations, suppression of HER4 expression in the tamoxifen-sensitive MCF-7 and T47D breast tumor cell lines resulted in resistance to tamoxifen-induced apoptosis. Furthermore, HER4 expression was lost in three independent MCF-7 models of acquired tamoxifen resistance. The HER4 intracellular domain (4ICD) is an independently signaling nuclear protein that functions as a potent ERalpha coactivator. In addition, mitochondrial 4ICD functions as a proapoptotic BH3-only protein. Tamoxifen disrupts an estrogen-driven interaction between ERalpha and 4ICD while promoting mitochondrial accumulation of the 4ICD BH3-only protein. BCL-2 inhibition of tamoxifen-induced apoptosis and tamoxifen activation of BAK, independent of BAX, further supports a role for 4ICD during tamoxifen-induced apoptosis. Finally, reintroduction of HER4, but not HER4 with a mutated BH3 domain, restores tamoxifen sensitivity to tamoxifen-resistant TamR cells in a xenograft model. Clinically, breast cancer patients with tumor expression of nuclear 4ICD responded to tamoxifen therapy with no clinical failures reported after 14 years of follow-up, whereas 20% of patients lacking nuclear 4ICD expression succumbed to their disease within 10 years of diagnosis. Our identification of the HER4/4ICD BH3-only protein as a critical mediator of tamoxifen action provides a clinically important role for 4ICD in human cancer and reveals a potential tumor marker to predict patient response to tamoxifen therapy. PMID- 18676865 TI - Targeted overexpression of vav3 oncogene in prostatic epithelium induces nonbacterial prostatitis and prostate cancer. AB - Our previous study revealed that Vav3 oncogene is overexpressed in human prostate cancer, activates androgen receptor (AR), and stimulates growth in prostate cancer cells. The purpose of this study is to further determine the potential role of Vav3 in prostate cancer development in genetically engineered mouse model. We generated Vav3 transgenic mice by targeted overexpression of a constitutive active Vav3 in the prostatic epithelium. We found that overexpression of Vav3 led to development of mouse prostatic intraepithelial neoplasia and prostate cancer at the age of as early as 3 months. The AR signaling axis and phosphatidylinositol 3-kinase-Akt signaling were elevated in the prostate glands of Vav3 transgenic mice. In addition to prostate cancer, Vav3 transgenic mice developed significant nonbacterial chronic prostatitis in the prostate gland with notable infiltration of lymphomononuclear cells (monocytes, lymphocytes, and plasma cells), which was associated with elevated incidence of prostate cancer. DNA microarray and signaling pathway analysis revealed that the top diseases and disorders were inflammatory diseases and cancer of the prostate gland in Vav3 transgenic mice. In vitro analysis showed that overexpression of Vav3 in prostate cancer cells enhanced nuclear factor-kappaB (NF-kappaB) activity, implicating an underlying mechanism of innate inflammatory response induced by elevated Vav3 activity. These data showed that Vav3 overexpression in the prostate epithelium enhanced both the AR signaling axis and NF-kappaB mediated pathway, which potentially contributed to the development of nonbacterial prostatitis and prostate cancer. PMID- 18676866 TI - Androgen levels increase by intratumoral de novo steroidogenesis during progression of castration-resistant prostate cancer. AB - Although systemic androgen deprivation prolongs life in advanced prostate cancer, remissions are temporary because patients almost uniformly progress to a state of a castration-resistant prostate cancer (CRPC) as indicated by recurring PSA. This complex process of progression does not seem to be stochastic as the timing and phenotype are highly predictable, including the observation that most androgen regulated genes are reactivated despite castrate levels of serum androgens. Recent evidence indicates that intraprostatic levels of androgens remain moderately high following systemic androgen deprivation therapy, whereas the androgen receptor (AR) remains functional, and silencing the AR expression following castration suppresses tumor growth and blocks the expression of genes known to be regulated by androgens. From these observations, we hypothesized that CRPC progression is not independent of androgen-driven activity and that androgens may be synthesized de novo in CRPC tumors leading to AR activation. Using the LNCaP xenograft model, we showed that tumor androgens increase during CRPC progression in correlation to PSA up-regulation. We show here that all enzymes necessary for androgen synthesis are expressed in prostate cancer tumors and some seem to be up-regulated during CRPC progression. Using an ex vivo radiotracing assays coupled to high-performance liquid chromatography radiometric/mass spectrometry detection, we show that tumor explants isolated from CRPC progression are capable of de novo conversion of [(14)C]acetic acid to dihydrotestosterone and uptake of [(3)H]progesterone allows detection of the production of six other steroids upstream of dihydrotestosterone. This evidence suggests that de novo androgen synthesis may be a driving mechanism leading to CRPC progression following castration. PMID- 18676867 TI - Diagnostic and prognostic microRNAs in stage II colon cancer. AB - MicroRNAs (miRNA) are a class of small noncoding RNAs with important posttranscriptional regulatory functions. Recent data suggest that miRNAs are aberrantly expressed in many human cancers and that they may play significant roles in carcinogenesis. Here, we used microarrays to profile the expression of 315 human miRNAs in 10 normal mucosa samples and 49 stage II colon cancers differing with regard to microsatellite status and recurrence of disease. Several miRNAs were differentially expressed between normal tissue and tumor microsatellite subtypes, with miR-145 showing the lowest expression in cancer relative to normal tissue. Microsatellite status for the majority of cancers could be correctly predicted based on miRNA expression profiles. Furthermore, a biomarker based on miRNA expression profiles could predict recurrence of disease with an overall performance accuracy of 81%, indicating a potential role of miRNAs in determining tumor aggressiveness. The expression levels of miR-320 and miR-498, both included in the predictive biomarker, correlated with the probability of recurrence-free survival by multivariate analysis. We successfully verified the expression of selected miRNAs using real-time reverse transcription PCR assays for mature miRNAs, whereas in situ hybridization was used to detect the accumulation of miR-145 and miR-320 in normal epithelial cells and adenocarcinoma cells. Functional studies showed that miR-145 potently suppressed growth of three different colon carcinoma cell lines. In conclusion, our results suggest that perturbed expression of numerous miRNAs in colon cancer may have a functional effect on tumor cell behavior, and, furthermore, that some miRNAs with prognostic potential could be of clinical importance. PMID- 18676868 TI - Implications of apurinic/apyrimidinic endonuclease in reactive oxygen signaling response after cisplatin treatment of dorsal root ganglion neurons. AB - Peripheral neuropathy is one of the major side effects of the anticancer drug cisplatin. Although previous work suggests that this neuropathy correlates with formation of DNA adducts in sensory neurons, growing evidence suggests that cisplatin also increases the generation of reactive oxygen species (ROS), which could cause DNA damage. Apurinic/apyrimidinic endonuclease/redox factor-1 (Ape1/Ref-1) is a multifunctional protein involved in DNA base excision repair of oxidative DNA damage and in redox regulation of a number of transcription factors. Therefore, we asked whether altering Ape1 functions would influence cisplatin-induced neurotoxicity. Sensory neurons in culture were exposed to cisplatin for 24 hours and several end points of toxicity were measured, including production of ROS, cell death, apoptosis, and release of the immunoreactive calcitonin gene-related peptide (iCGRP). Reducing expression of Ape1 in neuronal cultures using small interfering RNA (siRNA) enhances cisplatin induced cell killing, apoptosis, ROS generation, and cisplatin-induced reduction in iCGRP release. Overexpressing wild-type Ape1 attenuates all the toxic effects of cisplatin in cells containing normal endogenous levels of Ape1 and in cells with reduced Ape1 levels after Ape1siRNA treatment. Overexpressing the redox deficient/repair competent C65-Ape1 provides partial rescue, whereas the repair deficient Ape1 (N226A + R177A) does not protect neurons from cisplatin toxicity. We also observe an increase in phosphorylation of p53 after a decrease in Ape1 levels in sensory neuronal cultures. These results strongly support the notion that Ape1 is a potential translational target such that protecting Ape1 levels and particularly its DNA repair function could reduce peripheral neuropathy in patients undergoing cisplatin treatment. PMID- 18676869 TI - Long-term dietary cadmium intake and postmenopausal endometrial cancer incidence: a population-based prospective cohort study. AB - Environmental pollutants mimicking the effects of estrogen are suggested to contribute to the high incidence of hormone-related cancers, but supporting data are sparse. A potent estrogen-like activity of the pollutant cadmium, mediated via the estrogen receptor-alpha, has been shown in vivo. We prospectively examined the association between cadmium exposure and incidence of postmenopausal endometrial cancer. The Swedish Mammography Cohort is a population-based prospective cohort of 30,210 postmenopausal women free of cancer diagnose at baseline (1987) and who completed a food frequency questionnaire at baseline and in 1997. We estimated the dietary cadmium intake based on the questionnaire data and the cadmium content in all foods. During 16.0 years (484,274 person-years) of follow-up between the baseline and mid-2006, we ascertained 378 incident cases of endometrioid adenocarcinoma. The average estimated dietary cadmium intake was 15 mug/day (80% from cereals and vegetables). Cadmium intake was statistically significantly associated with increased risk of endometrial cancer in all women; the multivariate relative risk (RR) was 1.39 [95% confidence interval (CI), 1.04 1.86; P(trend) = 0.019], comparing highest tertile versus lowest. Among never smoking women with body mass index (BMI) of <27 kg/m(2), the RR was 1.86 (95% CI, 1.13-3.08; P(trend) = 0.009). We observed a 2.9-fold increased risk (95% CI, 1.05 7.79) associated with long-term cadmium intake consistently above the median at both baseline 1987 and in 1997 in never-smoking women with low bioavailable estrogen (BMI of <27 kg/m(2) and nonusers of postmenopausal hormones). Our results support the hypothesis that cadmium may exert estrogenic effects and thereby increase the risk of hormone-related cancers. PMID- 18676870 TI - Variants in inflammation genes and the risk of biliary tract cancers and stones: a population-based study in China. AB - To evaluate the role of chronic inflammation in the development of gallstones and biliary tract cancer, we examined the risk associated with 62 single nucleotide polymorphisms (SNPs), in 22 inflammation-related genes, in a population-based case-control study conducted in Shanghai, China, where the incidence of biliary tract cancer has been increasing in recent decades. The study included 411 cases with biliary tract cancer (237 gallbladder, 127 extrahepatic bile duct, and 47 ampulla of Vater), 895 with biliary stones, and 786 controls randomly selected from the population. Unconditional logistic regression was used to calculate odds ratios and 95% confidence intervals for the association of individual SNPs and haplotypes with biliary stones and biliary tract cancer. Of the 62 SNPs examined, 14 were related to the risk of biliary cancer and stones. Specifically, variants in the IL8, IL8RB, RNASEL, and NOS2 genes were associated with biliary stones, whereas VEGF variants were associated with gallbladder cancer. Of the 10 genes with multiple SNPs from which we inferred haplotypes, only one IL8RB haplotype, consisting of 3 SNPs (rs2230054, rs1126579, and rs1126580), was associated with the risk of bile duct cancer (P = 0.003) and biliary stones (P = 0.02), relative to the most frequent haplotype. In summary, common variants in genes that influence inflammatory responses may predispose to gallstones and biliary tract cancer, suggesting the need for future studies into the immunologic and inflammatory pathways that contribute to biliary diseases, including cancer. PMID- 18676871 TI - Carcinogen-altered genes in rat esophagus positively modulated to normal levels of expression by both black raspberries and phenylethyl isothiocyanate. AB - Our recent study identified 2,261 dysregulated genes in the esophagi of rats that received a 1-week exposure to the carcinogen N-nitrosomethylbenzylamine (NMBA). We further reported that 1,323 of these genes were positively modulated to near normal levels of expression in NMBA-treated animals that consumed dietary phenylethyl isothiocyanate (PEITC), a constituent of cruciferous vegetables. Herein, we report our results with companion animals that were fed a diet containing 5% freeze-dried black raspberries (BRB) instead of PEITC. We found that 462 of the 2,261 NMBA-dysregulated genes in rat esophagus were restored to near-normal levels of expression by BRB. Further, we have identified 53 NMBA dysregulated genes that are positively modulated by both PEITC and BRB. These 53 common genes include genes involved in phase I and II metabolism, oxidative damage, and oncogenes and tumor suppressor genes that regulate apoptosis, cell cycling, and angiogenesis. Because both PEITC and BRB maintain near-normal levels of expression of these 53 genes, their dysregulation during the early phase of NMBA-induced esophageal cancer may be especially important in the genesis of the disease. PMID- 18676872 TI - Loss of cannabinoid receptor 1 accelerates intestinal tumor growth. AB - Although endocannabinoid signaling is important for certain aspects of gastrointestinal homeostasis, the role of the cannabinoid receptors (CB) in colorectal cancer has not been defined. Here we show that CB1 expression was silenced in human colorectal cancer due to methylation of the CB1 promoter. Our genetic and pharmacologic studies reveal that loss or inhibition of CB1 accelerated intestinal adenoma growth in Apc(Min/+) mice whereas activation of CB1 attenuated intestinal tumor growth by inducing cell death via down-regulation of the antiapoptotic factor survivin. This down-regulation of survivin by CB1 is mediated by a cyclic AMP-dependent protein kinase A signaling pathway. These results indicate that the endogenous cannabinoid system may represent a potential therapeutic target for prevention or treatment of colorectal cancer. PMID- 18676873 TI - Aberrant catalytic cycle and impaired lipid transport into intracellular vesicles in ABCA3 mutants associated with nonfatal pediatric interstitial lung disease. AB - The ATP-binding cassette transporter ABCA3 mediates uptake of choline phospholipids into intracellular vesicles and is essential for surfactant metabolism in lung alveolar type II cells. We have shown previously that ABCA3 mutations in fatal surfactant deficiency impair intracellular localization or ATP hydrolysis of ABCA3 protein. However, the mechanisms underlying the less severe phenotype of patients with ABCA3 mutation are unclear. In this study, we characterized ABCA3 mutant proteins identified in pediatric interstitial lung disease (pILD). E292V (intracellular loop 1), E690K (adjacent to Walker B motif in nucleotide binding domain 1), and T1114M (8th putative transmembrane segment) mutant proteins are localized mainly in intracellular vesicle membranes as wild type protein. Lipid analysis and sucrose gradient fractionation revealed that the transport function of E292V mutant protein is moderately preserved, whereas those of E690K and T1114M mutant proteins are severely impaired. Vanadate-induced nucleotide trapping and photoaffinity labeling of wild-type and mutant proteins using 8-azido-[(32)P]ATP revealed an aberrant catalytic cycle in these mutant proteins. These results demonstrate the importance of a functional catalytic cycle in lipid transport of ABCA3 and suggest a pathophysiological mechanism of pILD due to ABCA3 mutation. PMID- 18676874 TI - Cross talk between paxillin and Rac is critical for mediation of barrier protective effects by oxidized phospholipids. AB - We previously reported that the barrier-protective effects of oxidized 1 palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (OxPAPC) on pulmonary endothelial cells (ECs) delineate the role of Rac- and Cdc42-dependent mechanisms and described the involvement of the focal adhesion (FA) protein paxillin in enhancement of the EC barrier upon OxPAPC challenge. This study examined a potential role of paxillin in the feedback mechanism of Rac regulation by FAs in OxPAPC-stimulated ECs. Our results demonstrate that OxPAPC induced Rac-dependent, Rho-independent peripheral accumulation of paxillin-containing FAs and time dependent paxillin phosphorylation. Molecular inhibition of Rac decreased association of paxillin with the Rac-specific guanine nucleotide exchange factor beta-PIX. Molecular inhibition of paxillin also attenuated OxPAPC-induced enhancement of adherens junctions critical for the EC barrier-protective response, accumulation of vascular endothelial cadherin in the membrane fractions, and decreased activation of Rac and its effector p21-activated kinase (PAK1). Expression of paxillin with a mutated PAK1-dependent phosphorylation site (S273A) attenuated OxPAPC-induced PAK1 activation and the EC barrier-protective response. These results suggest that PAK1-specific paxillin phosphorylation at Ser(273) is critically involved in the positive-feedback regulation of the Rac PAK1 pathway and may contribute to sustained enhancement of the EC barrier caused by oxidized phospholipids. PMID- 18676875 TI - Proteomic analysis of CTGF-activated lung fibroblasts: identification of IQGAP1 as a key player in lung fibroblast migration. AB - Connective tissue growth factor (CTGF, CCN2) is overexpressed in lung fibroblasts isolated from patients with interstitial lung disease (ILD) and systemic sclerosis (SSc, scleroderma) and is considered to be a molecular marker of fibrosis. To understand the significance of elevated CTGF, we investigated the changes in lung fibroblast proteome in response to CTGF overexpression. Using 2 dimensional gel electrophoresis followed by in-gel proteolytic digestion and mass spectrometric analysis, we identified 13 proteins affected by CTGF. Several of the CTGF-induced proteins, such as pro-alpha (I) collagen and cytoskeletal proteins vinculin, moesin, and ezrin, are known to be elevated in pulmonary fibrosis, whereas 9 of 13 proteins have not been studied in pulmonary fibrosis and are, therefore, novel CTGF-responsive molecules that may have important roles in ILD. Our study demonstrates that 1 of the novel CTGF-induced proteins, IQ motif containing GTPase activating protein (IQGAP) 1, is elevated in lung fibroblasts isolated from scleroderma patients with ILD. IQGAP1 is a scaffold protein that plays a pivotal role in regulating migration of endothelial and epithelial cells. Scleroderma lung fibroblasts and normal lung fibroblasts treated with CTGF demonstrated increased rate of migration in a wound healing assay. Depletion of IQGAP1 expression by small interfering RNA inhibited CTGF induced migration and MAPK ERK1/2 phosphorylation in lung fibroblasts. MAPK inhibitor U0126 decreased CTGF-induced cell migration and did not interfere with CTGF-induced IQGAP1 expression, suggesting that MAPK pathway is downstream of IQGAP1. These findings further implicate the importance of CTGF in lung tissue repair and fibrosis and propose that CTGF-induced migration of lung fibroblasts to the damaged tissue is mediated via IQGAP1 and MAPK signaling pathways. PMID- 18676877 TI - Arabidopsis protein disulfide isomerase-5 inhibits cysteine proteases during trafficking to vacuoles before programmed cell death of the endothelium in developing seeds. AB - Protein disulfide isomerase (PDI) oxidizes, reduces, and isomerizes disulfide bonds, modulates redox responses, and chaperones proteins. The Arabidopsis thaliana genome contains 12 PDI genes, but little is known about their subcellular locations and functions. We demonstrate that PDI5 is expressed in endothelial cells about to undergo programmed cell death (PCD) in developing seeds. PDI5 interacts with three different Cys proteases in yeast two-hybrid screens. One of these traffics together with PDI5 from the endoplasmic reticulum through the Golgi to vacuoles, and its recombinant form is functionally inhibited by recombinant PDI5 in vitro. Peak PDI5 expression in endothelial cells precedes PCD, whereas decreasing PDI5 levels coincide with the onset of PCD-related cellular changes, such as enlargement and subsequent collapse of protein storage vacuoles, lytic vacuole shrinkage and degradation, and nuclear condensation and fragmentation. Loss of PDI5 function leads to premature initiation of PCD during embryogenesis and to fewer, often nonviable, seeds. We propose that PDI5 is required for proper seed development and regulates the timing of PCD by chaperoning and inhibiting Cys proteases during their trafficking to vacuoles before PCD of the endothelial cells. During this transitional phase of endothelial cell development, the protein storage vacuoles become the de facto lytic vacuoles that mediate PCD. PMID- 18676878 TI - Functional diversification of CLAVATA3-related CLE proteins in meristem maintenance in rice. AB - Postembryonic development in plants depends on the activity of the shoot apical meristem (SAM) and root apical meristem (RAM). In Arabidopsis thaliana, CLAVATA signaling negatively regulates the size of the stem cell population in the SAM by repressing WUSCHEL. In other plants, however, studies of factors involved in stem cell maintenance are insufficient. Here, we report that two proteins closely related to CLAVATA3, FLORAL ORGAN NUMBER2 (FON2) and FON2-LIKE CLE PROTEIN1 (FCP1/Os CLE402), have functionally diversified to regulate the different types of meristem in rice (Oryza sativa). Unlike FON2, which regulates the maintenance of flower and inflorescence meristems, FCP1 appears to regulate the maintenance of the vegetative SAM and RAM. Constitutive expression of FCP1 results in consumption of the SAM in the vegetative phase, and application of an FCP1 CLE peptide in vitro disturbs root development by misspecification of cell fates in the RAM. FON1, a putative receptor of FON2, is likely to be unnecessary for these FCP1 functions. Furthermore, we identify a key amino acid residue that discriminates between the actions of FCP1 and FON2. Our results suggest that, although the basic framework of meristem maintenance is conserved in the angiosperms, the functions of the individual factors have diversified during evolution. PMID- 18676879 TI - Clinical features and treatment outcome of nasal-type NK/T-cell lymphoma of Waldeyer ring. AB - The clinical characteristics and prognosis remain unclear for nasal-type NK/T cell lymphoma of Waldeyer ring (WR-NKTL). The aim of this study is to determine the clinical features and outcome. Ninety-one patients with WR-NKTL were reviewed. According to the Ann Arbor system, 15, 56, 12, and 8 patients had stage I, II, III, and IV. Of patients with stage I and II, 54 received combined chemotherapy and radiotherapy (CMT), 13 received radiotherapy alone, and 4 patients received chemotherapy alone. All 20 patients with stage III/IV received primary chemotherapy. The disease is characterized by predominance in young males, good performance, a propensity for nodal involvement, frequent stage II through IV diseases, low frequency of elevated LDH, low-risk international prognostic index (IPI), high sensitivity to radiotherapy, and intermediate sensitivity to chemotherapy. The 5-year overall survival and progression-free survival for all patients were 65% and 51%, respectively. The age, B symptoms, stage, and IPI were important prognostic factors. CMT tended to improve the survival compared with radiotherapy alone for patients with stage I and II diseases. Both nodal involvement and distant extranodal dissemination were the primary failure patterns. WR-NKTL appears to have distinct clinical characteristics and favorable outcomes. PMID- 18676880 TI - Molecular mechanisms of thrombus formation in ischemic stroke: novel insights and targets for treatment. AB - In ischemic stroke, treatment options are limited. Therapeutic thrombolysis is restricted to the first few hours after stroke, and the utility of current platelet aggregation inhibitors, including GPIIb/IIIa receptor antagonists, and anticoagulants is counterbalanced by the risk of intracerebral bleeding complications. Numerous attempts to establish neuroprotection in ischemic stroke have been unfruitful. Thus, there is strong demand for novel treatment strategies. Major advances have been made in understanding the molecular functions of platelet receptors such as glycoprotein Ib (GPIb) and GPVI and their downstream signaling pathways that allow interference with their function. Inhibition of these receptors in the mouse stroke model of transient middle cerebral artery occlusion prevented infarctions without increasing the risk of intracerebral bleeding. Similarly, it is now clear that the intrinsic coagulation factor XII (FXII) and FXI play a functional role in thrombus formation and stabilization during stroke: their deficiency or blockade protects from cerebral ischemia without overtly affecting hemostasis. Based on the accumulating evidence that thrombus formation and hemostasis are not inevitably linked, new concepts for prevention and treatment of ischemic stroke may eventually emerge without the hazard of severe bleeding complications. This review discusses recent advances related to antithrombotic strategies in experimental stroke research. PMID- 18676882 TI - Penetration of chlorhexidine into human skin. AB - This study evaluated a model of skin permeation to determine the depth of delivery of chlorhexidine into full-thickness excised human skin following topical application of 2% (wt/vol) aqueous chlorhexidine digluconate. Skin permeation studies were performed on full-thickness human skin using Franz diffusion cells with exposure to chlorhexidine for 2 min, 30 min, and 24 h. The concentration of chlorhexidine extracted from skin sections was determined to a depth of 1,500 microm following serial sectioning of the skin using a microtome and analysis by high-performance liquid chromatography. Poor penetration of chlorhexidine into skin following 2-min and 30-min exposures to chlorhexidine was observed (0.157 +/- 0.047 and 0.077 +/- 0.015 microg/mg tissue within the top 100 microm), and levels of chlorhexidine were minimal at deeper skin depths (less than 0.002 microg/mg tissue below 300 microm). After 24 h of exposure, there was more chlorhexidine within the upper 100-microm sections (7.88 +/- 1.37 microg/mg tissue); however, the levels remained low (less than 1 microg/mg tissue) at depths below 300 microm. There was no detectable penetration through the full thickness skin. The model presented in this study can be used to assess the permeation of antiseptic agents through various layers of skin in vitro. Aqueous chlorhexidine demonstrated poor permeation into the deeper layers of the skin, which may restrict the efficacy of skin antisepsis with this agent. This study lays the foundation for further research in adopting alternative strategies for enhanced skin antisepsis in clinical practice. PMID- 18676881 TI - Antiviral effects of lamivudine, emtricitabine, adefovir dipivoxil, and tenofovir disoproxil fumarate administered orally alone and in combination to woodchucks with chronic woodchuck hepatitis virus infection. AB - Adefovir dipivoxil (ADV) and tenofovir disoproxil fumarate (TDF) are nucleotide analogs that inhibit the replication of wild-type hepatitis B virus (HBV) and lamivudine (3TC)-resistant virus in HBV-infected patients, including those who are coinfected with human immunodeficiency virus. The combination of ADV or TDF with other nucleoside analogs is a proposed strategy for managing antiviral drug resistance during the treatment of chronic HBV infection. The antiviral effect of oral ADV or TDF, alone or in combination with 3TC or emtricitabine (FTC), against chronic woodchuck hepatitis virus (WHV) infection was evaluated in a placebo controlled study in the woodchuck, an established and predictive model for antiviral therapy. Once-daily treatment for 48 weeks with ADV plus 3TC or TDF plus FTC significantly reduced serum WHV viremia levels from the pretreatment level by 6.2 log(10) and 6.1 log(10) genome equivalents/ml serum, respectively, followed by TDF plus 3TC (5.6 log(10) genome equivalents/ml), ADV alone (4.8 log(10) genome equivalents/ml), ADV plus FTC (one survivor) (4.4 log(10) genome equivalents/ml), TDF alone (2.9 log(10) genome equivalents/ml), 3TC alone (2.7 log(10) genome equivalents/ml), and FTC alone (2.0 log(10) genome equivalents/ml). Individual woodchucks across all treatment groups also demonstrated pronounced declines in serum WHV surface antigen, characteristically accompanied by declines in hepatic WHV replication and the hepatic expression of WHV antigens. Most woodchucks had prompt recrudescence of WHV replication after drug withdrawal, but individual woodchucks across treatment groups had sustained effects. No signs of toxicity were observed for any of the drugs or drug combinations administered. In conclusion, the oral administration of 3TC, FTC, ADV, and TDF alone and in combination was safe and effective in the woodchuck model of HBV infection. PMID- 18676883 TI - Novel type of staphylococcal cassette chromosome mec in a methicillin-resistant Staphylococcus aureus strain isolated in Sweden. AB - We identified a novel type of staphylococcal cassette chromosome mec (SCCmec) element carried by methicillin-resistant Staphylococcus aureus (MRSA) strain JCSC6082 isolated in Sweden. The SCCmec element was demarcated by characteristic nucleotide sequences at both ends and was integrated at the 3' end of orfX. The element carried a novel combination of a type 5 ccr gene complex and class C1 mec gene complex. The J regions of the element were homologous to those of the SCCmercury element of S. aureus strain 85/2082, with nucleotide identity greater than 99%. However, the novel SCCmec element from JCSC6082 did not carry the mer operon nor Tn554, suggesting that evolution to SCCmec could have been from a common ancestor by acquisition of the class C1 mec gene complex. The novel SCCmec element from JCSC6082 was flanked by a novel SCC-like chromosome cassette (CC6082), which was demarcated by two direct repeats and could be excised from the chromosome independently of the SCCmec element. Our data suggest that novel SCCmec elements can be generated on the staphylococcal chromosome through the recombination between extant SCC elements and mec gene complexes. PMID- 18676884 TI - Macrolide antibiotic-mediated downregulation of MexAB-OprM efflux pump expression in Pseudomonas aeruginosa. AB - Macrolide antibiotics modulate the quorum-sensing system of Pseudomonas aeruginosa. We tested the effect of macrolide antibiotics on the cell density dependent expression of the MexAB-OprM efflux pump and found that 1.0 mug/ml (MIC/6.25) of azithromycin suppressed the expression of MexAB-OprM by about 70%, with the result that the cells became two- to fourfold more susceptible to antibiotics such as aztreonam, tetracycline, carbenicillin, chloramphenicol, and novobiocin. PMID- 18676885 TI - Development of caspofungin resistance following prolonged therapy for invasive candidiasis secondary to Candida glabrata infection. AB - We report a case of Candida glabrata invasive candidiasis that developed reduced susceptibility to caspofungin during prolonged therapy. Pre- and posttreatment isolates were confirmed to be isogenic, and sequencing of hot spots known to confer echinocandin resistance revealed an F659V substitution within the FKS2 region of the glucan synthase complex. PMID- 18676886 TI - Low penetration of oseltamivir and its carboxylate into cerebrospinal fluid in healthy Japanese and Caucasian volunteers. AB - Oseltamivir is a potent, well-tolerated antiviral for the treatment and prophylaxis of influenza. Although no relationship with treatment could be demonstrated, recent reports of abnormal behavior in young individuals with influenza who were receiving oseltamivir have generated renewed interest in the central nervous system (CNS) tolerability of oseltamivir. This single-center, open-label study explored the pharmacokinetics of oseltamivir and oseltamivir carboxylate (OC) in the plasma and cerebrospinal fluid (CSF) of healthy adult volunteers over a 24-hour interval to determine the CNS penetration of both these compounds. Four Japanese and four Caucasian males were enrolled in the study. Oseltamivir and OC concentrations in CSF were low (mean of observed maximum concentrations [C(max)], 2.4 ng/ml [oseltamivir] and 19.0 ng/ml [OC]) versus those in plasma (mean C(max), 115 ng/ml [oseltamivir] and 544 ng/ml [OC]), with corresponding C(max) CSF/plasma ratios of 2.1% (oseltamivir) and 3.5% (OC). Overall exposure to oseltamivir and OC in CSF was also comparatively low versus that in plasma (mean area under the concentration-time curve CSF/plasma ratio, 2.4% [oseltamivir] and 2.9% [OC]). No gross differences in the pharmacokinetics of oseltamivir or OC were observed between the Japanese and Caucasian subjects. Oseltamivir was well tolerated. This demonstrates that the CNS penetration of oseltamivir and OC is low in Japanese and Caucasian adults. Emerging data support the idea that oseltamivir and OC have limited potential to induce or exacerbate CNS adverse events in individuals with influenza. A disease- rather than drug related effect appears likely. PMID- 18676887 TI - In vivo pharmacodynamics of ceftobiprole against multiple bacterial pathogens in murine thigh and lung infection models. AB - Ceftobiprole medocaril is the parenteral prodrug of ceftobiprole, a novel pyrrolidinone broad-spectrum cephalosporin with in vitro and in vivo bactericidal activities against methicillin-resistant Staphylococcus aureus (MRSA) and penicillin-resistant Streptococcus pneumoniae (PRSP). We have used murine thigh and lung infection models in neutropenic and normal mice to characterize the in vivo pharmacokinetic (PK)-pharmacodynamic (PD) activities of ceftobiprole against multiple strains of S. aureus (including MRSA), S. pneumoniae (including PRSP), and gram-negative bacilli. Serum levels of ceftobiprole following the administration of multiple doses were determined by a microbiological assay. In vivo bactericidal activities and postantibiotic effects (PAEs) of ceftobiprole against MRSA and PRSP strains were determined from serial CFU/thigh values following single doses of ceftobiprole (40 and 160 mg/kg of body weight). Dose fractionation studies were used to determine which PK-PD index correlated best with activity. Magnitudes of the PK-PD indices were calculated from MICs and PK parameters. A sigmoid dose-response model was used to estimate the dose (mg/kg/24 h) required to achieve a static and 2-log(10) kill effects over 24 h. PK results showed area under the concentration-time curve/dose values of 1.8 to 2.8 and half lives of 0.29 to 0.51 h. MICs ranged from 0.015 to 2 microg/ml. Ceftobiprole demonstrated time-dependent killing; its in vivo PAEs varied from 3.8 h to 4.8 h for MRSA and from 0 to 0.8 h for PRSP. The time above MIC (T > MIC) correlated best with efficacy for both MRSA and PRSP. The T > MIC values required for the static doses were significantly longer (P < 0.001) for Enterobacteriaceae (36 to 45%) than for S. aureus (14 to 28%) and S. pneumoniae (15 to 22%). The drug showed activities in the lung model similar to those in the thigh model. The presence of neutrophils significantly enhanced the activity of ceftobiprole against S. pneumoniae but only slightly against Klebsiella pneumoniae. Based on its PD profile, ceftobiprole is a promising new beta-lactam agent with activity against gram-negative and gram-positive organisms including MRSA and PRSP. PMID- 18676888 TI - Antagonistic effect of rifampin on the efficacy of high-dose levofloxacin in staphylococcal experimental foreign-body infection. AB - Since levofloxacin at high doses was more active than levofloxacin at conventional doses and was the best therapy alone in a rat model of staphylococcal foreign-body infection, in this study we tested how these differences affect the activities of their respective combinations with rifampin in vitro and in vivo. In vitro studies were performed in the log and stationary phases. By using this model, rifampin at 25 mg/kg of body weight/12 h, levofloxacin at 100 mg/kg/day, levofloxacin at 100 mg/kg/day plus rifampin, levofloxacin at 50 mg/kg/day, levofloxacin at 50 mg/kg/day plus rifampin, or a control treatment was administered for 7 days; and therapy with for levofloxacin at 100 mg/kg/day alone and rifampin alone was prolonged to 14 days. We screened for the appearance of resistant strains. Killing curves in the log phase showed a clear antagonism with levofloxacin at concentrations >or=2x MIC and rifampin and tended to occur in the stationary phase. At the end of 7 days of therapy, levofloxacin at 100 mg/kg/day was the best treatment and decreased the bacterial counts from tissue cage fluid (P < 0.05 compared with the results for groups except those receiving rifampin alone). At the end of 14 days of therapy with levofloxacin at 100 mg/kg/day, levofloxacin at 100 mg/kg/day plus rifampin, and the control treatment, the bacterial counts on the coverslips were 2.24 (P < 0.05 compared with the results with the combined therapy), 3.36, and 5.4 log CFU/ml, respectively. No rifampin or levofloxacin resistance was detected in any group except that receiving rifampin alone. In conclusion, high-dose levofloxacin was the best treatment and no resistant strains appeared; the addition of rifampin showed an antagonistic effect. The efficacy of the rifampin-levofloxacin combination is not significantly improved by the dosage of levofloxacin. PMID- 18676889 TI - Novel insertion sequence- and transposon-mediated genetic rearrangements in genomic island SGI1 of Salmonella enterica serovar Kentucky. AB - Salmonella genomic island 1 (SGI1) is an integrative mobilizable element that harbors a multidrug resistance (MDR) gene cluster. Since its identification in epidemic Salmonella enterica serovar Typhimurium DT104 strains, variant SGI1 MDR gene clusters conferring different MDR phenotypes have been identified in several S. enterica serovars and classified as SGI1-A to -O. A study was undertaken to characterize SGI1 from serovar Kentucky strains isolated from travelers returning from Africa. Several strains tested were found to contain the partially characterized variant SGI1-K, recently described in a serovar Kentucky strain isolated in Australia. This variant contained only one cassette array, aac(3)-Id aadA7, and an adjacent mercury resistance module. Here, the uncharacterized part of SGI1-K was sequenced. Downstream of the mer module similar to that found in Tn21, a mosaic genetic structure was found, comprising (i) part of Tn1721 containing the tetracycline resistance genes tetR and tet(A); (ii) part of Tn5393 containing the streptomycin resistance genes strAB, IS1133, and a truncated tnpR gene; and (iii) a Tn3-like region containing the tnpR gene and the beta-lactamase bla(TEM-1) gene flanked by two IS26 elements in opposite orientations. The rightmost IS26 element was shown to be inserted into the S044 open reading frame of the SGI1 backbone. This variant MDR region was named SGI1-K1 according to the previously described variant SGI1-K. Other SGI1-K MDR regions due to different IS26 locations, inversion, and partial deletions were characterized and named SGI1-K2 to -K5. Two new SGI1 variants named SGI1-P1 and -P2 contained only the Tn3-like region comprising the beta-lactamase bla(TEM-1) gene flanked by the two IS26 elements inserted into the SGI1 backbone. Three other new variants harbored only one IS26 element inserted in place of the MDR region of SGI1 and were named SGI1-Q1 to -Q3. Thus, in serovar Kentucky, the SGI1 MDR region undergoes recombinational and insertional events of transposon and insertion sequences, resulting in a higher diversity of MDR gene clusters than previously reported and consequently a higher diversity of MDR phenotypes. PMID- 18676890 TI - A clinically relevant assessment of posterior cruciate ligament and posterolateral corner injuries. Evaluation of isolated and combined deficiency. AB - BACKGROUND: The treatment of symptomatic posterior cruciate ligament injuries of the knee is controversial. Identification of concomitant injuries of the posterolateral corner is important to optimize surgical and clinical outcomes, although this diagnosis is sometimes difficult. The purpose of this study was to determine the physical examination and stress radiography parameters of isolated injuries of the posterior cruciate ligament and combined injuries of the posterior cruciate ligament and posterolateral corner. Our hypothesis was that Grade-3 posterior drawer laxity is an indicator of a concomitant injury of the posterolateral corner. METHODS: Ten pairs of cadaver knees (a total of twenty knees) were evaluated with posterior drawer testing, dial testing, and stress radiography. Stress radiography was performed by applying a 200-N posterior drawer at 90 degrees of knee flexion. The knees were tested while intact and then retested following the sequential resection of the posterior cruciate ligament followed by the posterolateral corner structures. RESULTS: All intact specimens were rated as Grade 0 on posterior drawer testing. Sectioning of the posterior cruciate ligament resulted in Grade-2 posterior drawer in all specimens. The additional resection of the posterolateral corner resulted in Grade-3 posterior drawer in all specimens. Dial testing of the intact knees resulted in a mean (and standard error) of 10.5 degrees +/- 1.0 degrees and 10.5 degrees +/- 0.80 degrees of external rotation at 30 degrees and 90 degrees, respectively. This increased significantly to 15.1 degrees +/- 1.1 degrees and 16.2 degrees +/- 0.89 degrees, respectively, following sectioning of the posterior cruciate ligament (p < 0.05). After resection of the posterolateral corner, rotation was further increased to a mean of 21.6 degrees +/- 1.5 degrees at 30 degrees and 27.5 degrees +/- 1.6 degrees at 90 degrees (p < 0.05). On stress radiography, the average posterior displacements measured 2.9 +/- 0.5 mm in the intact specimens, 12.7 +/- 1.0 mm after resection of the posterior cruciate ligament, and 22.3 +/- 1.6 mm after the additional resection of the posterolateral corner (p < 0.05). The corrected posterior displacement, calculated by subtracting the displacement in the intact knees, was 9.8 mm after resection of the posterior cruciate ligament and 19.4 mm after the additional resection of the posterolateral corner. CONCLUSIONS: A grade of 3 on posterior drawer testing and >10 mm of posterior tibial translation on stress radiography correlate with the presence of a posterolateral corner injury in addition to a complete disruption of the posterior cruciate ligament. PMID- 18676891 TI - Computer-assisted surgery for subtalar arthrodesis. A study in cadavers. AB - BACKGROUND: Despite considerable recent interest in computer navigation for orthopaedic surgery, few investigations of computer-assisted surgery for foot and ankle operations have been reported. The purpose of the present study was to compare subtalar arthrodesis with and without computer navigation in a cadaver model. METHODS: Subtalar arthrodesis was performed on thirty-six matched-pair cadaver lower extremities with intact soft tissues, with an attempt being made to orient two screws in the optimal configuration based on unpublished data from a preceding biomechanical study. Each matched pair was randomly assigned either to a group of surgeons who were experienced in subtalar arthrodesis or to a group of inexperienced operators. Neither surgical group was experienced in computer assisted surgery. We compared optimal first-pass guidewire placement, fluoroscopic time, total operative time, screw placement accuracy, and adverse screw placement events between conventional (fluoroscopically guided) and computer-assisted subtalar arthrodesis. RESULTS: The number of passes needed to achieve optimal guidewire placement decreased with the use of computer assistance for both experienced surgeons and inexperienced operators (p < 0.001), with ideal placement occurring on the first attempt in 95% of the procedures performed with use of computer assistance. While the experienced surgeons required less time and fewer guidewire passes during conventional subtalar arthrodesis than the inexperienced operators did (p < 0.001), both groups used less fluoroscopy with computer assistance (p < 0.001). There was no significant difference in operative time between the two techniques when performed by the inexperienced operators, yet the total procedure time doubled for the experienced surgeons when the procedure was performed with use of computer assistance (p < 0.001). There was no significant difference between experienced surgeons and inexperienced operators or between conventional and computer-assisted subtalar arthrodesis with respect to adverse screw placement events or the ability to accurately place both screws. CONCLUSIONS: Computer-assisted subtalar arthrodesis resulted in screw placement accuracy that was equivalent to that of conventional (fluoroscopically guided) subtalar arthrodesis while decreasing the number of suboptimal guidewire passes and fluoroscopic time. The computer-assisted surgery technique increased the operative time for surgeons who were more experienced in conventional subtalar arthrodesis, but there was no difference in operative time for the group of operators who were inexperienced in subtalar arthrodesis. PMID- 18676892 TI - Cell count and differential of aspirated fluid in the diagnosis of infection at the site of total knee arthroplasty. AB - BACKGROUND: Although there is no absolute diagnostic test for periprosthetic infection, the synovial fluid leukocyte count and neutrophil percentage have been reported to have high sensitivity and specificity. However, the cutoff values for these tests are not agreed upon. We sought to identify definite cutoff values for both the fluid leukocyte count and the neutrophil percentage that may help to diagnose infection at the site of a prosthetic joint. METHODS: We analyzed synovial fluid that had been aspirated preoperatively from 429 knees that had undergone revision arthroplasty at three different academic institutions; 161 knees were found to be infected, and 268 knees were not. Using receiver operating characteristic curves, we determined cutoff values for the fluid leukocyte count and neutrophil differential with an optimal balance of sensitivity and specificity for the diagnosis of periprosthetic infection. The sensitivity, specificity, and predictive values were calculated for those cutoff values. The erythrocyte sedimentation rate and C-reactive protein level cutoff values of 30 mm/hr and 10 mg/L, respectively, were combined with the cutoff values for the fluid leukocyte count and neutrophil percentage. RESULTS: The cutoff values for optimal accuracy in the diagnosis of infection were >1100 cells/10(-3)cm(3) for the fluid leukocyte count and >64% for the neutrophil differential. When both tests yielded results below their cutoff values, the negative predictive value of the combination increased to 98.2% (95% confidence interval, 95.5% to 99.5%), whereas when both tests yielded results greater than their cutoff values, infection was confirmed in 98.6% (95% confidence interval, 94.9% to 99.8%) of the cases in our cohort. Similarly, when both the neutrophil percentage and the C reactive protein level were less than the cutoff values of 64% and 10 mg/L, respectively, the presence of periprosthetic infection was very unlikely. CONCLUSIONS: The synovial fluid leukocyte count and differential are useful adjuncts to the erythrocyte sedimentation rate and the C-reactive protein level in the preoperative workup of infection at the site of a total knee arthroplasty. The present study identified cutoff values for the leukocyte count (>1100 cells/10(-3)cm(3)) and neutrophil percentage (>64%) that can be used to diagnose infection. Combining the peripheral blood tests with the synovial fluid cell count and differential can improve their diagnostic value. PMID- 18676893 TI - Erythropoietin accelerates functional recovery after peripheral nerve injury. AB - BACKGROUND: Erythropoietin is a naturally occurring hormone with multiple effects on a number of different cell types. Recent data have suggested neuroprotective and perhaps even neurotrophic roles for erythropoietin. We hypothesized that these functional effects could be demonstrable in standard models of peripheral nerve injury. METHODS: Experiments were undertaken to evaluate the effect of erythropoietin on the previously reported standard course of healing of sciatic injuries in mice. The injury groups included mice that were subjected to (1) sham surgery, (2) a calibrated sciatic crush injury, (3) transection of the sciatic nerve followed by epineural repair, or (4) a transection followed by burial of the proximal stump in the adjacent muscle tissue (neurectomy). Either erythropoietin or saline solution was administered to the mice in each of these experimental groups twenty-four hours preinjury, immediately after surgical creation of the injury, twenty-four hours postinjury, or one week postinjury. All mice were evaluated on the basis of the published model for recovery of sciatic nerve motor function by measuring footprint parameters at specific times after the injury. Immunohistochemistry was also performed to assess the erythropoietin receptor expression profile at the site of injury. RESULTS: In general, the mice treated with erythropoietin recovered sciatic nerve motor function significantly faster than did the untreated controls. This conclusion was based on a sciatic function index that was 60% better in the erythropoietin-treated mice at seven days postinjury (p < 0.05). Although the group that had been given the erythropoietin immediately postinjury showed the best enhancement of recovery, the timing of the administration of the drug was not critical. Histological analysis demonstrated enhanced erythropoietin-receptor positivity in the nerves that recovered fastest, suggesting that accelerated healing correlates with expression of the receptor in nerve tissue. CONCLUSIONS: Erythropoietin treatment of an acute sciatic nerve crush injury leads to an effect consistent with functional neuroprotection. This protective effect may have clinical relevance, especially since it was detectable even when erythropoietin had been administered up to one week after injury. PMID- 18676894 TI - Results of internal fixation of Pauwels type-3 vertical femoral neck fractures. AB - BACKGROUND: It has been postulated that femoral neck fractures with a more vertical fracture line (i.e., a high Pauwels angle) may experience more shear forces and therefore may be predisposed to nonunion or loss of fixation. Although there is controversy regarding which fixation method is ideal, we are aware of no large clinical series in which the treatment outcomes of these fractures were evaluated. The purpose of this multicenter study was to evaluate a large consecutive series of high shear angle (>70 degrees) femoral neck fractures to learn more about the outcomes, complications, and performance of various internal fixation strategies. METHODS: Between January 1993 and January 2005, seventy-six Pauwels type-3 (Orthopaedic Trauma Association [OTA] type-31B2.3) femoral neck fractures were treated in seventy-five patients with a mean age of forty-two years. Fourteen patients were lost to follow-up. Sixty-two fractures in sixty-one patients were followed to union or revision surgery, with a mean duration of follow-up of twenty-four months. Thirty-seven fractures were treated with cannulated screws and twenty-five, with a fixed-angle device. The reduction quality, accuracy of implant placement, time to surgery, influence of capsular decompression, and rates of nonunion and osteonecrosis were evaluated. RESULTS: Fifty-nine (95%) of the fractures had good-to-excellent reduction, and three had a fair reduction. There was a nonunion of eight (14%) of the fifty-nine fractures with a good-to-excellent reduction and two of the three with a fair reduction. There was a septic nonunion of one fracture treated with a dynamic hip screw. There was an aseptic nonunion of seven (19%) of the thirty-seven fractures treated with screw fixation alone as compared with two (8%) of the twenty-five fractures treated with a fixed-angle device. Osteonecrosis occurred after treatment of seven (11%) of the sixty-two fractures. CONCLUSIONS: Despite timely, excellent reduction and accurate implant placement in the vast majority of cases, the nonunion rate was 19% for fractures treated with cannulated screws alone and 8% for those treated with a fixed-angle device. Although these failure rates are not significantly different, we believe that this study documents the challenging nature of this fracture pattern and the ideal fixation device remains undefined. PMID- 18676895 TI - Emergency brake response time after first metatarsal osteotomy. AB - BACKGROUND: There is sparse information upon which to recommend a return to activity after foot surgery. The purpose of this study was to determine whether six weeks is sufficient time for the emergency brake response time to return to preoperative levels after a first metatarsal osteotomy for symptomatic hallux valgus. METHODS: We conducted a prospective, observational study to assess the effect of surgery on emergency brake response time in a group of twenty-eight patients undergoing a unilateral first metatarsal osteotomy on the right side for symptomatic hallux valgus. A custom-made driving simulator was used to assess total brake response time, reaction time, and brake time. Patients were assessed preoperatively and at two and six weeks postoperatively. A control group of twenty-eight individuals matched for age, driving status, and sex was included for baseline comparison. RESULTS: Total brake response time, reaction time, and brake time were significantly lower for the control cohort compared with the preoperative values recorded in the study cohort (p < 0.05). Only seven of the twenty-eight study patients were able to complete the assessment at two weeks; the remainder were not able to complete it because of postoperative pain. All patients were able to comfortably complete the study at six weeks. In the study cohort, the total brake response time, reaction time, and brake time had significantly improved compared with preoperative recordings (p < 0.05). CONCLUSIONS: By six weeks after surgery, emergency braking time in patients undergoing a first metatarsal osteotomy is similar to that of healthy individuals. PMID- 18676896 TI - Trigger finger: prognostic indicators of recurrence following corticosteroid injection. AB - BACKGROUND: Corticosteroid injections are commonly used in the treatment of flexor tenosynovitis in adults. The present study was performed in an attempt to identify prognostic indicators of symptom recurrence one year after corticosteroid injection for the treatment of trigger digits. METHODS: One hundred and thirty consecutive patients with trigger digits treated with corticosteroid injection were prospectively enrolled. Exclusion criteria were prior treatment and inflammatory arthritis. Demographic data and information on existing comorbidities were identified with a questionnaire. Patients were contacted at one year after treatment to determine symptom recurrence. Kaplan Meier analysis and the Cox regression model were used to estimate recurrence rates and identify predictors. RESULTS: One hundred and twenty-four trigger digits in 119 patients (average age, 62.3 years) were included. The most commonly involved digits were the thumb (35% of the digits), ring finger (31%), and long finger (23%). Seventy digits (56%) had a recurrence of symptoms at a median of 5.6 months (range, 0.5 to 13.1 months) after the injection. Twenty-two digits (18%) underwent surgical release at a median of 7.4 months after the injection. According to the Kaplan-Meier analysis, the estimated rate of freedom from symptom recurrence was 70% (95% confidence interval, 63% to 77%) at six months and 45% (95% confidence interval, 36% to 54%) at twelve months and the estimated rate of freedom from surgical release was 95% (95% confidence interval, 92% to 98%) at six months and 83% (95% confidence interval, 77% to 89%) at twelve months. Insulin-dependent diabetes mellitus was identified as a strong predictor of symptom recurrence (p < 0.01). Younger age (p < 0.01), involvement of other digits prior to presentation (p < 0.01), and a history of other tendinopathies of the upper extremity (p = 0.02) were all independent predictors of a surgical release. The duration and severity of symptoms were not predictive of poor outcomes following injections. CONCLUSIONS: At one year following injection, 56% of the digits had a recurrence of symptoms. Younger age, insulin-dependent diabetes mellitus, involvement of multiple digits, and a history of other tendinopathies of the upper extremity were associated with a higher rate of treatment failure. Symptoms often recurred several months after the injection. PMID- 18676898 TI - Simulated pivot-shift testing with single and double-bundle anterior cruciate ligament reconstructions. AB - BACKGROUND: One of the principal rationales for performing a double-bundle reconstruction of the anterior cruciate ligament is the suggestion that it may be superior to a single-bundle reconstruction in restoring a normal pivot-shift sign. The purpose of this study was to measure the abilities of single-bundle and anatomic double-bundle reconstructions to restore normal knee kinematics and graft forces during a simulated pivot-shift test. METHODS: Graft force and knee kinematics were recorded during a simulated pivot-shift event with and without the anterior cruciate ligament and after graft reconstructions. With a single bundle, the graft was tensioned to restore anterior-posterior laxity at 30 degrees of flexion. With double-bundle reconstructions, the anteromedial graft was first tensioned as above and then the posterolateral graft tension was set with use of one of four protocols: posterolateral tension = anteromedial tension at 10 degrees of flexion (DB1); posterolateral tension = anteromedial tension at 30 degrees (DB2); posterolateral tension = (anteromedial tension + 30 N) at 10 degrees (DB3); and posterolateral tension = (anteromedial tension + 30 N) at 30 degrees (DB4). RESULTS: A single-bundle reconstruction restored all displacements and rotations during the pivot shift to the intact knee levels. The mean tibial rotations and lateral plateau displacements during the pivot shift with DB2, DB3, and DB4 reconstructions were less than those in the intact knee and also less than those in a single-bundle reconstruction. Before the pivot shift, the mean graft forces with all reconstructions were greater than that of the intact knee; the mean graft forces with the DB3 and DB4 reconstructions were also greater than that of a single-bundle reconstruction. After the pivot shift, the mean graft forces for all reconstructions were less than the levels before the pivot shift with single-bundle forces lower than intact knee levels and DB4 forces higher than intact knee levels. CONCLUSIONS: Reduction or elimination of the pivot-shift sign is an important goal for anterior cruciate ligament reconstruction. In our model, the results show that a single-bundle reconstruction was sufficient to restore intact knee kinematics during a simulated pivot-shift event. The higher graft forces with some double-bundle graft-tensioning protocols reduced the coupled rotations and displacements from an applied valgus moment to less than the intact levels. This overcorrection should theoretically make the knee less likely to pivot but could have unknown clinical consequences. PMID- 18676897 TI - Inpatient compared with home-based rehabilitation following primary unilateral total hip or knee replacement: a randomized controlled trial. AB - BACKGROUND: Home-based rehabilitation is increasingly utilized to reduce health care costs; however, with a shorter hospital stay, the possibility arises for an increase in adverse clinical outcomes. We evaluated the effectiveness and cost of care of home-based compared with inpatient rehabilitation following primary total hip or knee joint replacement. METHODS: We randomized 234 patients, using block randomization techniques, to either home-based or inpatient rehabilitation following total joint replacement. All patients followed standardized care pathways and were evaluated, with use of validated outcome measures (Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC], Short Form-36, and patient satisfaction), prior to surgery and at three and twelve months following surgery. The primary outcome was the WOMAC function score at three months after surgery. RESULTS: The mean length of stay (and standard deviation) in the acute care hospital was 6.3 +/- 2.5 days for the group designated for inpatient rehabilitation prior to transfer to that facility compared with 7.0 +/- 3.0 days for the home-based rehabilitation group prior to discharge home (p = 0.06). The mean length of stay in inpatient rehabilitation was 17.7 +/- 8.6 days. The mean number of postoperative home-based rehabilitation visits was eight. The prevalence of postoperative complications up to twelve months postoperatively was similar in both groups, which each had a 2% rate of dislocation and a 3% rate of clinically important deep venous thrombosis. The prevalence of infection was 0% in the home-based group and 2% in the inpatient group. None of these differences was clinically important. Both groups showed substantial improvements at three and twelve months, with no significant differences between the groups with respect to WOMAC, Short Form-36, or patient satisfaction scores (p > 0.05). The total episode-of-care costs (in Canadian dollars) for the inpatient rehabilitation and home-based rehabilitation arms were $14,532 and $11,082, respectively (p < 0.01). CONCLUSIONS: Despite concerns about early hospital discharge, there was no difference in pain, functional outcomes, or patient satisfaction between the group that received home-based rehabilitation and the group that had inpatient rehabilitation. On the basis of our findings, we recommend the use of a home-based rehabilitation protocol following elective primary total hip or knee replacement as it is the more cost-effective strategy. PMID- 18676899 TI - Lateral external fixation--a new surgical technique for displaced unreducible supracondylar humeral fractures in children. AB - BACKGROUND: Percutaneous Kirschner wire fixation represents the classic treatment for displaced supracondylar humeral fractures in childhood. This type of treatment first requires satisfactory reduction of the fracture. Failure to achieve a satisfactory reduction or inadequate stabilization can result in instability of the fracture fragments, which can result in either an unsatisfactory cosmetic or functional outcome. In our experience, these problems can be overcome with the use of a small lateral external fixator. METHODS: Between 1999 and 2005, thirty-one of 170 Gartland type-III supracondylar humeral fractures were treated with a lateral external fixator. The outcome of treatment was analyzed with regard to limb alignment, elbow movement, cosmetic appearance, and patient satisfaction. RESULTS: In twenty-eight of the thirty-one patients, a satisfactory reduction was achieved with closed methods. All children except one had a normal or good range of movement. The cosmetic result was excellent in all cases. All of the children and their parents stated that they would choose this treatment again. CONCLUSIONS: The use of a small lateral external fixator seems to be a safe alternative for the treatment of displaced supracondylar fractures of the humerus when a closed reduction appears to be unattainable by means of manipulation alone or when sufficient stability is not achieved with standard methods of Kirschner wire fixation. PMID- 18676900 TI - Comparison of thoracolumbar motion produced by manual and Jackson-table-turning methods. Study of a cadaveric instability model. AB - BACKGROUND: Patients who have sustained a spinal cord injury remain at risk for further neurologic deterioration until the spine is adequately stabilized. To our knowledge, no study has previously addressed the effects of different bed-to operating room table transfer techniques on thoracolumbar spinal motion in an instability model. We hypothesized that the conventional logroll technique used to transfer patients from a supine position to a prone position on the operating room table has the potential to confer significantly more motion to the unstable thoracolumbar spine than the Jackson technique. METHODS: Three-column instability was surgically created at the L1 level in seven cadavers. Two protocols were tested. The manual technique entailed performing a standard logroll of a supine cadaver to a prone position on an operating room Jackson table. The Jackson technique involved sliding the supine cadaver to the Jackson table, securing it to the table, and then rotating it into a prone position. An electromagnetic tracking device measured motion--i.e., angular motion (flexion-extension, lateral bending, and axial rotation) and linear translation (axial, medial-lateral, and anterior-posterior) between T12 and L2. RESULTS: The logroll technique created significantly more motion than the Jackson technique as measured with all six parameters. Manual logroll transfers produced an average of 13.8 degrees to 18.1 degrees of maximum angular displacement and 16.6 to 28.3 mm of maximum linear translation. The Jackson technique resulted in an average of 3.1 degrees to 5.8 degrees of maximum angular displacement (p < 0.001) and 4.0 to 10.0 mm of maximum linear translation (p < 0.05). CONCLUSIONS: Compared with the logroll, the Jackson-table transfer method provides superior immobilization of an unstable thoracolumbar spine during transfer of supine cadavers to a prone position on the operating room table. CLINICAL RELEVANCE: This study addresses in-hospital patient safety. Performing the Jackson turn requires approximately half as many people as required for a manual logroll. This study suggests that the Jackson technique should be considered for supine-to-prone transfer of patients with known or suspected instability of the thoracolumbar spine. PMID- 18676901 TI - Long-term follow-up of radial shortening osteotomy for Kienbock disease. AB - BACKGROUND: Three previous studies have investigated the long-term outcome of radial osteotomy in the treatment of Kienbock disease. However, none used patient based assessment. The purpose of this study was to investigate the long-term clinical and radiographic outcomes of this osteotomy, including the subjective evaluation of the patient with use of the Disabilities of the Arm, Shoulder and Hand (DASH) Questionnaire. METHODS: A DASH questionnaire was sent to nineteen patients with Kienbock disease who had undergone a radial shortening osteotomy, and thirteen replied. The mean age at the time of surgery was thirty-nine years. On the basis of the Lichtman classification, six patients had stage-II, four had stage-IIIA, and three had stage-IIIB disease. Prior to surgery, ulnar variance was positive in six patients, neutral in four, and negative in three. The mean duration of follow-up was twenty-one years. Clinical evaluation, including calculation of the modified Mayo wrist score, and radiographic evaluation were also performed on twelve of the thirteen patients. RESULTS: The mean DASH score was 8 points (range, 0 to 23 points), and patient satisfaction was high. Compared with the findings in the contralateral wrist, the mean range of motion was 81% in flexion and 82% in extension and mean grip strength was 88%. The mean modified Mayo wrist score was 83 points, and the clinical results were excellent in six patients, good in five, and moderate in one. The DASH scores tended to be worse in patients with Lichtman stage-IIIB disease. Follow-up radiographs revealed that the Lichtman stage had progressed in six of the twelve patients. CONCLUSIONS: Although most patients had mild wrist pain, patient satisfaction and the clinical results were satisfactory following a radial shortening osteotomy. This procedure is a reliable long-term treatment for Lichtman stage-II and IIIA disease and may be a reasonable option for patients with stage-IIIB disease. PMID- 18676902 TI - Changes in dynamic foot pressure after surgical treatment of valgus deformity of the hindfoot in cerebral palsy. AB - BACKGROUND: Calcaneal lengthening osteotomy and extra-articular arthrodesis of the subtalar joint are two methods used for the correction of valgus deformity of the heel and forefoot abduction. The purpose of this study was to compare the operative results of these procedures in patients with cerebral palsy who were able to walk. We focused primarily on changes in radiographic parameters and how altered mobility of the subtalar joint by the two operative methods would modify pressure distribution over the plantar surface of the foot. METHODS: A total of eighty-one feet in forty-seven patients were included in the study. The mean age at the time of surgery was eight years and one month, and the mean follow-up period was thirty-nine months. The subjects were divided into two groups; Group I consisted of thirty-seven feet in twenty-two patients who underwent a calcaneal lengthening osteotomy, and Group II comprised forty-four feet in twenty-five patients who underwent an extra-articular subtalar arthrodesis. Preoperative and final follow-up radiographs and dynamic pedobarographs were used to evaluate the results. RESULTS: The feet in both groups were found to be similarly deformed before surgery, by radiographic measurements and dynamic foot-pressure analysis. Both operative procedures led to improved radiographic indices; however, calcaneal pitch failed to improve after the subtalar arthrodesis. After surgery, the relative vertical impulse was decreased for the hallux, first metatarsal head, and medial aspect of the midfoot in both groups, while it was increased for the lateral aspect of the midfoot and calcaneus. On the other hand, postoperatively, the relative vertical impulse of the medial aspect of the midfoot was higher and the relative vertical impulse of the first through fourth metatarsal heads was lower in the group that had subtalar arthrodesis compared with the group that had a calcaneal lengthening osteotomy and the normal control subjects. CONCLUSIONS: Extra-articular subtalar arthrodesis appears to be an effective means to achieve predictable correction of severe valgus deformity of the heel in patients with cerebral palsy who are able to walk; however, supination deformity of the forefoot remains and calcaneal equinus is not corrected. On the other hand, we believe that the calcaneal lengthening osteotomy is the treatment of choice because postoperative foot-pressure distribution more closely approximates the normal foot-pressure distribution. PMID- 18676903 TI - Percutaneous spine biopsy: a meta-analysis. AB - BACKGROUND: Percutaneous spine biopsy has widely replaced open biopsy. We conducted a meta-analysis to evaluate the effect of the inner diameter of the biopsy needle and the method of imaging guidance on the adequacy and accuracy of tissue samples and to evaluate the complication rates associated with the different needle diameters and imaging guidance methods. METHODS: We searched MEDLINE for studies that evaluated either the adequacy (whether or not a diagnosis could be made on the basis of pathologic examination) or the accuracy (whether or not the primary diagnosis was correct) of samples obtained by means of percutaneous spine biopsy. These articles and their relevant references subsequently were reviewed twice and were evaluated against the inclusion criteria, yielding twenty-five studies. The inclusion criterion was the use of a biopsy instrument (a fine needle or trephine with an identifiable inner diameter) under the guidance of imaging (fluoroscopy or computed tomography) for the evaluation of an identified spine lesion, with the report of either adequacy or accuracy. Meta-analysis with use of the random-effects model was used to analyze the data. RESULTS: The adequacy, accuracy, and complication rates increased with the inner diameter of the needles, but, with the numbers available, only the complication rate increased significantly (p = 0.01). Although the use of a computed tomography scan slightly increased the adequacy and accuracy of the samples, these increases were not significant. The complication rate associated with the use of computed tomography was 3.3%, compared with 5.3% for fluoroscopy. CONCLUSIONS: As the outcomes associated with computed tomography were not significantly different from those associated with fluoroscopy, the decision to use one or the other requires the consideration of other factors, such as the type, level, and vertebral location of the lesion as well as the expertise of the physician. In situations in which the use of a needle with a small inner diameter is highly effective (for example, in cases of metastatic lesions), the clinician should first consider using a needle with a smaller inner diameter to obtain the biopsy specimen because of the higher complication rate associated with large bore needles. However, in cases of sclerotic lesions, in which obtaining an adequate sample can be difficult, the use of a needle with a larger inner diameter is desirable. PMID- 18676904 TI - Infantile thoracolumbar kyphosis secondary to lumbar hypoplasia. AB - BACKGROUND: There is a normal transition from the kyphotic alignment that is present in the spine of the newborn to the normal sagittal contours that are present in the adult spine. Although abnormal kyphosis at the thoracolumbar junction in infants is rare, it can result from congenital anomalies and bone dysplasias. We report the cases of seven otherwise normal infants who had thoracolumbar kyphosis due to lumbar hypoplasia, with total spontaneous resolution over time. METHODS: The medical records of seven patients who met the inclusion criteria were reviewed. Clinical data and radiographs were analyzed to rule out the presence of congenital anomalies or bone dysplasias. Progression of kyphosis was measured on serial radiographs made with the patient sitting and standing. RESULTS: All patients were managed conservatively with observation alone. No patient had a neurologic deficit. The average age at the time of the initial diagnosis was 5.3 months, and the average duration of follow-up was 5.7 years. Three cases of kyphosis were secondary to L1 hypoplasia, and four were secondary to L2 hypoplasia. The average initial kyphosis was 34.2 degrees, which progressed to 0.4 degrees of lordosis at the time of the latest follow-up. CONCLUSIONS: Thoracolumbar kyphosis in normal infants secondary to lumbar hypoplasia may resolve spontaneously. After congenital anomalies and bone dysplasias are ruled out, a period of observation is advised. PMID- 18676905 TI - Geographic variation in epidural steroid injection use in medicare patients. AB - BACKGROUND: The rates of epidural steroid injections have increased dramatically over time, with conflicting evidence regarding the efficacy of epidural steroid injections for the treatment of various low-back pain disorders. Given the uncertainty about their role, we sought to evaluate the geographic variation in the use of epidural steroid injections for low back pain within the United States. We also sought to determine whether greater rates of epidural steroid injections are associated with lower rates of lumbar surgery. METHODS: We used the 2001 Medicare Physician Part-B claims to examine the geographic variation in the use of epidural steroid injections. Current Procedural Technology codes were used to identify the number of procedures performed as well as the percentage of injections that were fluoroscopically guided. Procedure rates were analyzed with use of several geographic indicators, including state, United States Census Bureau regions (Northeast, South, Midwest, and West), and health referral regions as defined by the Dartmouth Atlas of Health Care. RESULTS: In 2001, there was a 7.7-fold difference between the state with the lowest rate (Hawaii at 5.2 per 1000) and the state with the highest rate (Alabama at 39.9 per 1000). The variation among health referral regions, which are smaller in size, was even greater, with an 18.4-fold difference from 5.6 per 1000 in Honolulu, Hawaii, to 103.6 per 1000 in Palm Springs, California. Higher statewide rates of epidural steroid injections were associated with significantly higher rates of lumbar surgery (p = 0.001). In areas with high injection rates, a significantly higher percentage of patients who sought care for low back pain received injections (p < 0.001). In addition, in areas with high injection rates, a significantly higher percentage of patients who presented with low back pain received both injections and lumbar surgery within the same year (p < 0.001). CONCLUSIONS: There is substantial geographic variation in the rates of epidural steroid injections within the United States. Southern states tend to have the highest procedure rates, whereas northeastern states have the lowest. Injection rates are positively correlated with lumbar surgery rates; therefore, epidural steroid injections do not appear to be substituting for lumbar surgeries or reducing overall rates of lumbar surgery. PMID- 18676907 TI - Endoscopic surgery for chronic osteomyelitis extending across the physis. A report of two cases. PMID- 18676906 TI - Ability of lower-extremity injury severity scores to predict functional outcome after limb salvage. AB - BACKGROUND: Lower-extremity injury severity scoring systems were developed to assist surgeons in decision-making regarding whether to amputate or perform limb salvage after high-energy trauma to the lower extremity. These scoring systems have been shown to not be good predictors of limb amputation or salvage. This study was performed to evaluate the clinical utility of the five commonly used lower-extremity injury severity scoring systems as predictors of final functional outcome. METHODS: We analyzed data from a cohort of patients who participated in a multicenter prospective study of clinical and functional outcomes after high energy lower-extremity trauma. Injury severity was assessed with use of the Mangled Extremity Severity Score; the Limb Salvage Index; the Predictive Salvage Index; the Nerve Injury, Ischemia, Soft-Tissue Injury, Skeletal Injury, Shock, and Age of Patient Score; and the Hannover Fracture Scale-98. Functional outcomes were measured with use of the physical and psychosocial domains of the Sickness Impact Profile at both six months and two years following hospital discharge. Four hundred and seven subjects for whom the reconstruction regimen was considered successful at six months were included in the analysis. We used partial correlation statistics and multiple linear regression models to quantify the association between injury severity scores and Sickness Impact Profile outcomes with the subjects' ages held constant. RESULTS: The mean age of the patients was thirty-six years (interquartile range, twenty-six to forty-four years); 75.2% were male and 24.8% were female. The median Sickness Impact Profile scores were 15.2 and 6.0 points at six and twenty-four months, respectively. The analysis showed that none of the scoring systems were predictive of the Sickness Impact Profile outcomes at six or twenty-four months to any reasonable degree. Likewise, none were predictive of patient recovery between six and twenty-four months postoperatively as measured by a change in the scores in either the physical or the psychosocial domain of the Sickness Impact Profile. CONCLUSIONS: Currently available injury severity scores are not predictive of the functional recovery of patients who undergo successful limb reconstruction. PMID- 18676909 TI - Avoiding complications in the care of fractures of the tibia. PMID- 18676908 TI - Direct demonstration of viable Staphylococcus aureus biofilms in an infected total joint arthroplasty. A case report. PMID- 18676910 TI - AOA Symposium. Barriers (threats) to clinical research. PMID- 18676911 TI - Topics in medical economics: medical malpractice. PMID- 18676913 TI - Patients can provide a valid assessment of quality of life, functional status, and general health on the day they undergo knee surgery. PMID- 18676915 TI - Comparison of external and percutaneous pin fixation with plate fixation for intra-articular distal radial fractures. PMID- 18676917 TI - In vivo efficacy of antimicrobial-coated devices. PMID- 18676919 TI - Primary arthroscopic stabilization for a first-time anterior dislocation of the shoulder. PMID- 18676920 TI - Femoral nerve block for diaphyseal and distal femoral fractures in the emergency department. PMID- 18676922 TI - The economic impact of reprocessing external fixation components. PMID- 18676924 TI - Music playing in a cast room reduced heart rate in children. PMID- 18676925 TI - Manipulation under anesthesia did not enhance the benefit conferred by home exercises for frozen shoulder. PMID- 18676926 TI - Percutaneous screw fixation led to faster recovery and return to work than immobilization for fractures of the waist of the scaphoid. PMID- 18676927 TI - The future of the orthopaedic clinician-scientist. Part I: The potential role of MD-PhD students considering orthopaedic surgery. AB - BACKGROUND: There is currently a severe shortage of clinician-scientists, who fill a vital role in musculoskeletal care. One way to address this shortage is to recruit more MD-PhD students into orthopaedics. We analyzed data from a national survey of MD-PhD students to assess this potential. METHODS: A total of 868 students from thirteen MD-PhD training programs were requested to fill out a multiple-choice online survey concerning their education and future goals. RESULTS: We achieved a response rate of 56.7% (492 of 868). Seven (1.4%) of the 492 respondents listed orthopaedics as their primary clinical interest, and thirty (6.1%) listed it as one of their three strongest clinical interests. Among the thirty respondents, seven (23%) were senior students, five (17%) were women, and none were minorities. In comparison, 33% of the 462 respondents in the nonorthopaedic cohort were women and 12.1% were a member of a minority group (p < 0.05). Among twenty-three students who had a secondary orthopaedic interest, only one-third had a primary surgical interest. Both the thirty with a strong clinical interest in orthopaedics and the others without a strong interest in orthopaedics showed similar intent on becoming physician-scientists (a score of 2.73 and 3.30, respectively) and an interest in an academic career (90.0% and 90.3%, respectively) (p > 0.05 for both). The orthopaedic group showed significantly greater interest in clinical care as a primary activity than did the nonorthopaedic group (63.3% compared with 30.7%; p < 0.0005). Eighty-seven percent of those in the orthopaedic group reported research as their most likely primary or secondary activity. CONCLUSIONS: This study suggests that there is a relatively strong interest in orthopaedic surgery (patient care and research) among MD-PhD students nationally, creating the potential to recruit approximately 100 new orthopaedic clinician-scientists every eight years (the average MD-PhD training period). Extrapolation indicates that there is the ability to double the number of orthopaedic clinician-scientists in the United States over the next fifty years. Therefore, efforts should be made to attract these students (especially women and those in underrepresented minority groups) to orthopaedic surgery. The study further suggests recruiting broadly-we should not be biased toward students late in training and just those with surgical interests. PMID- 18676928 TI - What's new in orthopaedic research. PMID- 18676930 TI - Modern metal-on-metal hip resurfacing: important observations from the first ten years. PMID- 18676931 TI - Improved survivorship of hybrid metal-on-metal hip resurfacing with second generation techniques for Crowe-I and II developmental dysplasia of the hip. AB - BACKGROUND: The purpose of this study was to determine the influence of improved femoral fixation techniques on the survivorship of metal-on-metal total hip resurfacing prostheses in patients with developmental dysplasia of the hip and to report the long-term results of our patients managed earlier with first generation fixation techniques. METHODS: One hundred and three hips (ninety patients) were resurfaced for osteoarthritis secondary to developmental dysplasia. The mean age of the patients was forty-seven years, and 77% were women. Most hips (94%) were Crowe class I, but 43% had femoral head defects of >1 cm in size. The clinical results of these hips were compared with those of a group of patients with other etiologies, largely dominated by idiopathic osteoarthritis (78%). RESULTS: All clinical scores improved significantly (p < 0.0001) and were comparable with those of patients with other etiologies except for the postoperative activity scores, which were lower (7.0 compared with 7.5). Range of motion was greater for the patients with dysplasia than for the patients with other etiologies. Seven hips that were resurfaced with the first-generation femoral fixation techniques and one hip that was resurfaced with the second and third-generation techniques had conversion to total hip arthroplasty. This difference was found to be significant (p = 0.032) in a multivariate, time dependent analysis after adjustment for other covariates known to affect prosthetic survival. There was no loosening of the acetabular component in this series. CONCLUSIONS: The current improvements in the short-term to midterm results after resurfacing in patients with developmental dysplasia of the hip in whom more current techniques were used are encouraging and allow for greater expectations regarding the elimination of short-term failures and improved long term durability of resurfacing in this population. PMID- 18676932 TI - Results and lessons learned from a United States hip resurfacing investigational device exemption trial. AB - BACKGROUND: Improvements in metal-on-metal bearings have made hybrid hip surface replacement a potential alternative for the young active patient with end-stage hip disease. Possible advantages include greater hip joint stability, bone preservation, and decreased osteolysis. In this study, we compared the clinical and radiographic results of a new resurfacing device with those in a historical group of standard total hip arthroplasties. METHODS: In 2001, the Cormet 2000 Hip Resurfacing Investigational Device Exemption study was initiated at twelve centers. A total of 337 patients treated with unilateral hip surface replacement with the Cormet device were enrolled in that study. These patients were compared with 266 patients in a previous study who had undergone unilateral total hip arthroplasty with ceramic bearing surfaces. Clinical and radiographic results were compared at similar time intervals. A newly recommended performance standard, the composite clinical success score, was used to assess non inferiority of the hip resurfacing compared with the total hip arthroplasty used in the historical comparison population. RESULTS: At the time of follow-up, at a minimum of two years, the Harris hip scores were comparable between the resurfacing and total hip arthroplasty groups. Statistical evaluation of the composite clinical success scores confirmed the non-inferiority hypothesis. Revision was required in twenty-four patients in the resurfacing group and five patients in the total hip arthroplasty group. The most common cause of revision following resurfacing was failure of the femoral component (fracture of the femoral neck or loosening of the femoral component). CONCLUSIONS: Careful review of this study population revealed several important criteria for successful introduction of this resurfacing device into the United States. These include careful patient selection based on clinical and radiographic parameters and attention to various surgical details of implantation. These findings can be used to focus the training process for surgeons who wish to add implantation of this device to their surgical armamentarium. Such efforts should help to ensure safe and effective introduction of this new technology. PMID- 18676933 TI - Total hip resurfacing in patients who are sixty years of age or older. AB - BACKGROUND: Metal-on-metal total hip resurfacing arthroplasty has been associated with excellent early results in patients who are younger than sixty years of age, but it remains controversial whether this procedure is appropriate in older patients. The purpose of the present study was to compare the clinical and radiographic outcomes after hip resurfacing in two cohorts of patients: those who were sixty years of age or older, and those who were younger than sixty years of age. METHODS: Between November 2002 and August 2005, thirty-five patients (forty hips) who were sixty years of age or older were managed with metal-on-metal total hip resurfacing arthroplasty. The outcomes of these patients were compared with those of 130 patients (153 hips) who were younger than sixty years of age but otherwise had similar preoperative parameters and who had resurfacing arthroplasty performed during the same time interval and by the same surgeon. We evaluated Harris hip scores, Short Form-12 scores, and complications as well as radiographic alignment and radiolucencies. RESULTS: At a mean follow-up time of thirty-six months, the mean Harris hip scores improved from 52 points to 94 points in the older patient cohort and from 53 points to 92 points in the younger patient cohort. The final Short Form-12 scores of the two groups were also similar. Two patients who were sixty years or older and five of the younger patients required conversion to a conventional total hip arthroplasty. Femoral neck fracture was the reason for one conversion in each group. There were no impending radiographic failures in either cohort. CONCLUSIONS: Although national registries indicate that the risk of femoral neck fracture is higher in older patients, the present study found that these patients had excellent clinical outcomes that were similar to those of patients who were younger. We await longer follow-up results to determine further outcomes in these patients. PMID- 18676934 TI - Metal-on-metal hip resurfacing with an uncemented femoral component. A seven-year follow-up study. AB - BACKGROUND: Metal-on-metal hip resurfacing with hybrid fixation has been introduced as an alternative to standard total hip arthroplasty, especially for young and active patients. There are few studies in the literature on the midterm results of cementless femoral side resurfacing. The purpose of this study was to present our seven-year clinical results of a series of twenty cementless metal-on metal hip resurfacing procedures. METHODS: Between 1999 and 2000, eighteen patients (twenty hips) underwent primary metal-on-metal hip resurfacing with uncemented femoral and acetabular components. One patient was lost to follow-up. This left eleven men and six women, who had a mean age of forty-five years at the time of surgery. Clinical and radiographic examinations were performed prospectively, and the results were analyzed. RESULTS: The mean duration of follow-up was 7.4 years. There were four revisions, none of which was due to aseptic failure of the femoral component. Two were due to loosening of the acetabular component, one was due to a late hematogenous infection, and one was due to persistent pain despite normal radiographic findings. The mean preoperative Harris hip score was 54 points, and it increased to 94 points at the time of the last follow-up. Radiographic examination of the hips for which the procedure was successful revealed no femoral or acetabular radiolucencies, no migration of any implant, and no osteolysis. The radiographs of one patient (two hips) showed substantial narrowing of the femoral necks (a mean of 12%), which stabilized at three years postoperatively. This patient had a Harris hip score of 100 points for both hips at six years. CONCLUSIONS: This study suggests that cementless femoral fixation may be a viable alternative to fixation with cement in metal-on-metal hip resurfacing. Further study of this concept in larger numbers of patients is warranted. PMID- 18676935 TI - Hip resurfacing through an anterolateral approach. Surgical description and early review. AB - BACKGROUND: Recently, metal-on-metal hip resurfacing has enjoyed a resurgence as an alternative to hip arthroplasty in properly selected patients. The purpose of the present study was to report the early results of hip resurfacing through an anterolateral approach and to describe the technique with modifications that have been made as experience with the procedure has increased. METHODS: A total of fifty-seven hip resurfacing procedures in fifty-three patients were performed by a single surgeon with use of the Cormet Hip Resurfacing system through an anterolateral approach. The results were assessed on the basis of Harris hip scores, a radiographic analysis, and an analysis of failures. RESULTS: After a mean duration of follow-up of thirty-eight months, the mean Harris hip score was 99 points. Fifty-five hips were in patients with normal function scores, and two were in patients who reported mild dysfunction. Fifty hips had no pain, five had slight pain, one had mild pain, and one had moderate pain. Three hips had a failure of resurfacing and required revision. Two failures were secondary to femoral neck fractures, and one was the result of debonding of the surface coating of the acetabular component. CONCLUSIONS: The early results associated with a new generation of hip resurfacing devices are promising. The anterolateral approach affords the surgeon excellent exposure with the advantage of increased hip stability and potentially improved vascular supply of the femoral head as compared with the posterior approach. PMID- 18676936 TI - Metal-on-metal total hip resurfacing arthroplasty in the presence of extra articular deformities or implants. AB - BACKGROUND: Hip resurfacing has been proposed as an alternative to total hip replacement in patients who have proximal femoral deformities or retained hardware in the proximal aspect of the femur. In these situations, placement of a conventional stemmed hip prosthesis would be difficult or impossible, possibly necessitating a complex osteotomy or a custom prosthesis. The purpose of this study was to evaluate a series of patients who had extra-articular deformities of the proximal aspect of the femur and/or retained hardware and who were managed with a resurfacing hip prosthesis. METHODS: Fifteen patients (seventeen hips) who underwent metal-on-metal resurfacing hip replacements were studied. Ten patients (twelve hips) had bowing or other deformities of the femur secondary to trauma, multiple epiphyseal dysplasia, renal osteodystrophy, or proximal femoral focal deficiency. Five patients (five hips) had retained hardware. Twelve of the patients (thirteen hips) had previously been told by orthopaedic surgeons that, due to the deformity or retained hardware, they could not undergo conventional total hip arthroplasty without also undergoing ancillary surgical procedures. We evaluated perioperative factors (operative time and estimated blood loss), Harris hip scores, complications, and failure rates. RESULTS: At a mean follow-up time of three years (range, two to five years), fourteen patients (sixteen hips) were doing well clinically and radiographically. Assessment of the intraoperative records revealed minimal difficulty, with a mean operative time of 104 minutes and a mean blood loss of 621 mL. The mean Harris hip score was 92 points. One patient, a fifty-nine-year-old woman, underwent two subsequent revisions-one for the treatment of a femoral neck fracture, and one for the treatment of acetabular component loosening. CONCLUSIONS: Resurfacing hip arthroplasty offers an option for patients when placement of a conventional total hip prosthesis is difficult or impossible because of the presence of proximal femoral deformities or retained hardware in or on the proximal aspect of the femur. PMID- 18676937 TI - Anatomic diameter femoral heads in total hip arthroplasty: a preliminary report. AB - BACKGROUND: The stability of total hip replacements has been directly related to the diameter of the femoral head in several studies; however, durability has necessitated the use of femoral heads with a relatively small diameter. Recent developments in metal-on-metal technology have allowed for the use of femoral head bearings that are anatomic in diameter. In this case series, we report on the early results of patients who were at greater risk for dislocation because of anatomic deficiencies or increased range-of-motion activities and underwent hip arthroplasty with implants that had articulating surfaces approaching anatomic dimensions. METHODS: Thirty-four patients underwent forty total hip arthroplasties with use of a modular metal-on-metal articulation with an anatomic diameter femoral head and a press-fit stem. Thirty patients were active, and four patients were profoundly disabled and had bone or soft-tissue deficiencies that would increase the risk for dislocation. Dislocation precautions were maintained for six weeks, and patients were allowed extreme ranges of motion at three months. RESULTS: There were no dislocations. Active patients continued in extreme range-of-motion activities. Disabled patients improved but were limited by their comorbidities. CONCLUSIONS: Anatomic diameter femoral heads offer distinct theoretical advantages in total hip arthroplasty. These short-term results are encouraging, and further study of this new technology in a larger series with a longer follow-up period is warranted. PMID- 18676938 TI - Learning how to resurface cam-type femoral heads with acceptable accuracy and precision: the role of computed tomography-based navigation. AB - BACKGROUND: Resurfacing arthroplasty for cam-type deformities, which are a common cause of early osteoarthritis, is a technically demanding operation. Like any other arthroplasty, it requires both accuracy and precision. On the basis of the results of series reported by expert surgeons, we considered it desirable that this operation should be performed within +/-10 degrees of the desired angular orientation and +/-6 mm of entry-point translation in 95% of hips. Technological aids are now available to help surgeons achieve that level of accuracy. Three models of cam-type hips of increasing severity were used to assess the efficacy of three systems of instrumentation at delivering the required level of accuracy and precision. METHODS: Thirty-two students of surgical technology were instructed in hip resurfacing and shown detailed plans of the desired operative outcome for the three hips with cam-type deformity. They then used conventional instruments, imageless navigation, and computed tomography-based navigation to perform the operation as accurately as possible. RESULTS: Conventional instrumentation produced an unacceptably wide range of entry-point errors. Imageless navigation was able to deliver adequate accuracy and precision in varus valgus angulation and superoinferior translation, but was less satisfactory in version and anteroposterior translation. Computed tomography-based navigation enabled novice surgeons to navigate hips that had difficult cam-type deformity with acceptable precision in all four degrees of freedom measured. CONCLUSIONS: Only computed tomography-based navigation appears to be appropriate for delivering both the accuracy and the precision needed by surgeons on the steep part of their learning curve. Neither conventional neck-based instrumentation nor imageless navigation provided enough help for novice surgeons learning to perform this technically challenging operation. PMID- 18676939 TI - Imageless navigation in hip resurfacing: avoiding component malposition during the surgeon learning curve. AB - BACKGROUND: Studies suggest that hip arthroplasty procedures performed in specialty hospitals or by physicians in practices with a high surgical volume are associated with a decreased rate of adverse outcomes related to component malpositioning. Little is known, however, about the influence of imageless computer navigation systems on the procedural experience of the surgeon and the subsequent alignment of implants in the setting of hip resurfacing arthroplasty. METHODS: Seventy-one consecutive hip resurfacing arthroplasties in which the components were placed with use of computer-assisted navigation were reviewed retrospectively. Intraoperative femoral and acetabular component parameters were compared with postoperative radiographic alignment values. Within this single surgeon series, operative time, intraoperative cup inclination and femoral stem shaft angles, and postoperative cup inclination and femoral stem-shaft angles were measured and compared over the course of three discrete, sequential operative time periods. Patient demographic data and surgical parameters, including blood loss, surgical approach, and anesthesia time, were recorded. RESULTS: No significant difference was seen between the intraoperative and postoperative cup inclination angles. A significant difference was noted between the intraoperative and postoperative femoral stem-shaft angles; however, the mean angles in all groups had a valgus orientation when compared with the mean native neck angles. Over three sequential operative time periods, computer-assisted navigation produced consistent values with regard to intraoperative cup inclination (43 degrees , 44 degrees , and 40 degrees ) and postoperative radiographic alignment of the cup (46 degrees , 44 degrees , and 43 degrees ) and femoral stem (148 degrees , 147 degrees , and 144 degrees ), despite different levels of surgeon experience. Operative times significantly decreased with surgeon experience, showing the largest decrease after the first sequence interval (110, ninety-eight, and ninety-five minutes, respectively). There was a significant difference with evolving surgeon experience concerning intraoperative stem placement (144 degrees , 142 degrees , and 138 degrees , respectively) despite the mean values remaining well-clustered. No femoral notching occurred throughout the series. CONCLUSIONS: Computer-assisted navigation is a dependable and accurate method of positioning hip resurfacing components during arthroplasty, as measured by cup inclination, and a reliable technique for valgus stem placement and avoidance of notching. Furthermore, computer navigation allows for consistency of component alignment independent of procedural experience. PMID- 18676940 TI - Does computer-assisted surgery improve accuracy and decrease the learning curve in hip resurfacing? A radiographic analysis. AB - BACKGROUND: Hip resurfacing is a technically demanding procedure in which accurate positioning of the femoral component is critical to the avoidance of early implant failures. The purpose of this study was to assess the accuracy of computer-assisted placement of the femoral component and to evaluate the impact of computer-assisted surgery on the learning curve associated with this procedure. METHODS: The accuracy of positioning the femoral component was analyzed radiographically in hips undergoing resurfacing procedures performed by surgeons assigned to four different study groups: Group 1, in which the operations were performed with use of computer-assisted surgery by a fellowship trained surgeon who was experienced in performing resurfacing arthroplasty (surgical experience, more than 250 hip resurfacings); Group 2, in which the operations were performed with use of computer-assisted surgery by senior residents who were inexperienced in performing resurfacing arthroplasty and who were closely supervised by faculty; Group 3, in which the operations were performed with use of conventional instruments by fellowship-trained faculty members; and Group 4, in which the operations were performed with use of computer assisted surgery by a lesser experienced fellowship-trained faculty member (surgical experience, more than forty but less than seventy-five hip resurfacings) from Group 3. RESULTS: The range of error in varus or valgus angulation that was observed for navigated procedures was 6 degrees in Group 1, 7 degrees in Group 2, and 5 degrees in Group 4. Compared with the preoperative neck shaft angle value, the mean postoperative stem-shaft angle value increased by a mean of 4.7 degrees in Group 1, 7.2 degrees in Group 2, 6.5 degrees in Group 3, and 11.6 degrees in Group 4. When compared with the use of standard instrumentation, the use of computer-assisted surgery reduced the number of outliers and facilitated valgus insertion. CONCLUSIONS: In the present study, computer-assisted surgery resulted in improved accuracy and precision in positioning the femoral component. In addition, computer-assisted surgery led to a reduction in the length of the learning curve for beginners in hip resurfacing and improved the surgeon's ability to perform this procedure safely. PMID- 18676941 TI - Have contemporary hip resurfacing designs reached maturity? A review. AB - BACKGROUND: The increasing prevalence of hip replacements in young patients is expected to fuel the demand for hip resurfacing arthroplasty. Patient selection, surgical technique, and implant design can influence the clinical outcomes for these patients. In this review, we discuss whether contemporary hip resurfacing designs have reached maturity and suggest design considerations for future generation implants. These design-related factors include the amount and extent of cement fixation, adoption of cementless femoral fixation, optimization of implant position, minimization of stress-shielding, improvement in modularity or sizing and geometry options, metallurgy, development of alternative bearing options, and examination of in vivo cup deformation. In addition, this review is based on an understanding of the causes of failure of revision hip resurfacing arthroplasty, which is essential to help guide research, implant design, and clinical decision-making. PMID- 18676942 TI - Modes of implant failure after hip resurfacing: morphological and wear analysis of 267 retrieval specimens. AB - BACKGROUND: Resurfacing of the hip joint is experiencing a revival due to improvements in materials, design, and manufacturing techniques. Despite good midterm outcomes, the high early rate of failure and concerns about metal debris require a detailed morphological and wear analysis of retrieved resurfacing implants in order to understand failure mechanisms. METHODS: A worldwide collection of hip resurfacing revision devices was initiated, and 267 components were received. Devices were analyzed by patient demographics, radiographic positioning, and wear, as well as morphologically and histologically. Specimens were grouped into four different failure types. They were also stratified into rim-loaded or non-rim-loaded groups. Failures were also assessed by surgeon learning-curve effects. RESULTS: Time to failure was significantly different between the four revision-type groups: Specimens with fractures involving the implant rim were most common (46%) and failed earliest after surgery (mean of ninety-nine days), followed by fractures inside the femoral head (20%, 262 days) and loose cups (9%, 423 days). Revisions not due to fractures or cup loosening (25%) occurred at a mean of 722 days after surgery. Rim-loaded implants exhibited an average twenty-one to twenty-sevenfold higher wear rate than implants without rim-loading. Rim-loaded implants also showed a steeper mean cup inclination than their non-rim-loaded counterparts (59 degrees compared with 50 degrees ). Most failures occurred during the learning curve of the surgeon (the first fifty to 100 implantations). CONCLUSIONS: Failures on the femoral side usually occur within the first nine months after surgery and appear to be most directly related to the implantation technique or patient selection. Later failures are observed mainly due to acetabular problems, either due to dramatically increased wear or poor cup anchorage. Improper cup anteversion may be similar to or more important than cup inclination in producing excessive wear. PMID- 18676943 TI - Risk of periprosthetic femoral neck fracture after hip resurfacing arthroplasty: valgus compared with anatomic alignment. A biomechanical and clinical analysis. AB - BACKGROUND: Early clinical results of hip resurfacing arthroplasty have led to the recommendation to achieve a neck-shaft angle of 140 degrees when inserting the femoral component. In addition, the idea of adhering to an absolute angle when inserting instrumentation in hips with excessive anatomic varus or valgus neck-shaft angles has raised concern. A biomechanical analysis was completed in order to determine if the achieved valgus orientation of the femoral component reduced the risk of periprosthetic fracture. METHODS: Twenty fresh-frozen cadaveric femora were blindly assigned to be implanted with a neutral or valgus oriented hip-resurfacing femoral component. Bone mineral density scans were acquired for all femora. All specimens were loaded axially to failure at a rate of 0.21 mm per second. Radiographs of the specimens were measured in order to determine the relative valgus orientation of the femoral components and the change in offset. RESULTS: There was a significant increase in the ultimate failure load for the valgus-oriented components. While the bone density scans revealed that the bone mineral densities measured in the neutral and valgus oriented femoral components were almost identical, the ultimate failure load was found to be significantly increased for the valgus-oriented components (6955 N) compared with the neutral-oriented components (5254 N). For the valgus-oriented femoral components, two had failure at the subcapital level, seven had vertical shear fractures, and one had an anterior shear fracture. For the neutral-oriented components, five subcapital fractures and five vertical shear failures were observed. CONCLUSIONS: The study suggests that a valgus orientation decreases the risk of periprosthetic femoral neck fracture following hip resurfacing. It also brings into question the use of an absolute angle for all patients. Obtaining the maximum possible valgus angle, while avoiding notching, may in fact provide the optimum protection from periprosthetic femoral neck fractures. PMID- 18676944 TI - Cementing techniques for hip resurfacing arthroplasty: development of a laboratory model. AB - BACKGROUND: Hip resurfacing is extremely technique-sensitive, yet scientific investigations into cementing techniques are lacking. In this study, we tested open-cell, reticulated, carbon-foam materials in comparison with paired human femoral heads to validate a laboratory cementing model for resurfacing arthroplasty. METHODS: Paired human femoral heads prepared for resurfacing were compared with thirty and sixty-pore-per-inch fat-filled foam specimens. Two different cementing techniques were analyzed: manual application of high viscosity cement, and half component-filling with low-viscosity cement. Real-time measurements were made of cement pressure and temperature. Cement penetration areas and depths were quantified. RESULTS: We found no significant differences between the human femoral heads and the fat-filled thirty-pore-per-inch foam models in all measured variables (pressures at the top, chamfer and outer wall, temperature at the 5-mm and 15-mm subsurface). There was no significant difference in the cement penetration of the human femoral heads and the fat filled thirty-pore-per-inch foam models. There were a number of significant differences between the human femoral heads and the sixty-pore-per-inch foam models with use of the low-viscosity cement technique. The differences between the cementing techniques were greater than those between the three materials for most of the measurements. CONCLUSIONS: Fat-filling (to emulate bone marrow) of lower-density carbon foam more closely simulates the cement penetration resistance and thermal properties of human femoral heads than does the denser (unfilled) material. This model is sensitive to differences in cementing technique. PMID- 18676945 TI - Tribology and wear of metal-on-metal hip prostheses: influence of cup angle and head position. AB - BACKGROUND: Clinical studies have indicated that the angular position of the acetabular cup may influence wear in metal-on-metal total hip bearings. A high cup angle in comparison to the anatomical position may lead to the head being constrained by the superior lateral surface and rim of the cup, thus potentially changing the location of the contact zone between the head and the cup. The aim of this study was to test the hypothesis that both a steep cup angle and a lateralized position of the head can increase head contact on the superior rim of the cup, with the consequence of increased wear. METHODS: Hip-joint simulator studies of metal-on-metal bearings were undertaken with cup angles of 45 degrees and 55 degrees . The femoral head was either aligned to the center of the cup or placed in a position of microlateralization. Wear was measured gravimetrically over 5 million cycles. RESULTS: A steep cup angle of 55 degrees showed significantly higher long-term steady-state wear than a standard cup angle of 45 degrees (p < 0.01). The difference was fivefold. Microlateralization of the head resulted in a fivefold increase in steady-state wear compared with a centralized head. The combination of a steep cup angle and a microlateralized head increased the steady-state wear rate by tenfold compared with a standard cup angle with a centralized head. CONCLUSIONS: These studies support the hypothesis that both an increased cup angle and a lateral head position increase wear in metal-on-metal hip prostheses. PMID- 18676946 TI - Tribological considerations in primary and revision metal-on-metal arthroplasty. AB - BACKGROUND: Metal-on-metal hip bearings undergo biphasic wear, starting with a short period of high wear (bedding-in) and followed by low steady-state wear. Bedding-in is the process by which the cup wears locally to conform to the geometry of the head. This process reduces the maximum contact stress and allows for appropriate lubrication. A critical area of conformance and wear is required for the bearing to reach a low steady-wear state. Cups were analyzed in this study after primary and revision wear scenarios to determine this critical area for this specific bearing. METHODS: Forty and 56-mm cobalt-chromium resurfacing bearings with 150 and 400-microm clearances were wear tested in a hip simulator for 5 million cycles. The cups underwent an additional 5 million cycles of testing against new heads, simulating a revision scenario. The revision heads were manufactured to cause the highest mismatch with the pre-worn cups, resulting in polar or local annular contact. Cup wear area was determined from weight-loss measurements after each phase of testing. RESULTS: All bearings experienced a biphasic wear performance with a short period of high wear followed by low steady state wear. A consistent critical area of conformance was reached by all bearings after primary and revision testing conditions, regardless of bearing size, bearing clearance, or contact mode. CONCLUSIONS: An area of conformity (wear) reduces contact pressures, is beneficial for lubrication, and is critical to reach a low steady-state wear rate. This study shows that this critical area is consistent regardless of bearing size, clearance, or contact mode. Bearing designs that allow the proper formation of this conformance area should bed-in and reach a low steady-state wear rate. PMID- 18676947 TI - Characterization of the running-in period in total hip resurfacing arthroplasty: an in vivo and in vitro metal ion analysis. AB - BACKGROUND: Metal-on-metal total hip resurfacing arthroplasty is increasingly being performed in young and active patients. Preclinical in vitro testing of implants is usually performed with use of hip simulators, and the serum metal ion concentration is determined for the purpose of monitoring the patients. The goal of this study was to characterize the early running-in period in vivo and in vitro by characterizing metal ion levels. METHODS: A well-functioning total hip resurfacing prosthesis was implanted in fifteen consecutive patients, and the serum metal ion concentrations in these patients were then determined preoperatively and at intervals during the first postoperative year (at one, six, twelve, twenty-four, and fifty-two weeks). The number of walking cycles was measured with use of a computerized accelerometer in order to compare walking cycles to hip simulator cycles. In vitro, five similar components were investigated for 3 million cycles with use of a hip simulator. Serum samples were obtained at different time points, and wear was measured by quantifying wear particles and ions in the samples. All patient and simulation serum samples were analyzed with use of inductively coupled plasma-mass spectrometry. One simulator implant was investigated with use of scanning electron microscopy. RESULTS: The serum chromium and cobalt levels of the patients continuously increased during the first six months and showed an insignificant decrease thereafter. The molybdenum concentration was unchanged compared with preoperative values. In contrast, the simulator measurements showed a different wear pattern with a high wear running-in period and a low-wear steady-state phase. The running-in period was delayed by 300,000 cycles and lasted up to 1 million cycles. Scanning electron microscopic analysis showed a carbon-rich protein film predominantly in the early phases of simulation. Scratches were detected originating from pits filled with aluminum oxide and silicon oxide and from pulled-out carbides that were causing third-body wear. CONCLUSIONS: The simulator study allowed an exact characterization of the running-in period and showed a delayed onset of running in wear. In contrast, the clinical data showed a slow increase in measured ion concentrations. These different wear patterns are probably due to the effects of distribution, accumulation, and excretion of particles and ions in vivo. PMID- 18676949 TI - Metal ion levels in the blood of patients after hip resurfacing: a comparison between twenty-eight and thirty-six-millimeter-head metal-on-metal prostheses. AB - BACKGROUND: Metal ion toxicity, metal hypersensitivity, and metal carcinogenicity are causes for concern for patients with metal-on-metal hip replacements. Thus, understanding the biological fate of metal ions, and consequently their long-term systemic effects, is of great interest to patients and surgeons alike. METHODS: Inductively coupled plasma mass spectrometry was used to measure the levels of cobalt, chromium, and molybdenum ions in the blood of control patients (preoperative control pre-resurfacing patients), patients with a metal-on polyethylene total hip prosthesis, patients with a metal-on-metal total hip prosthesis with either a 28 or 36-mm femoral head, and patients with a hip resurfacing prosthesis. Since cobalt and chromium ions have the potential to induce oxidative stress through irreversible biochemical damage to macromolecules, the levels of ions were correlated to the concentration of three oxidative stress markers in the plasma of these patients. RESULTS: The median cobalt level was significantly lower (p < 0.001) in the 36-mm metal-on-metal total hip arthroplasty group (1.8 parts per billion [1.8 microg/L]) compared with the 28-mm metal-on-metal total hip arthroplasty group (2.5 parts per billion [2.5 microg/L]) and the hip resurfacing group (2.3 parts per billion [2.3 microg/L]) at six months postoperatively. The median chromium level was also significantly lower (p < 0.01) in the 36-mm metal-on-metal total hip arthroplasty group (0.25 parts per billion [0.25 microg/L]) compared with the 28-mm metal-on-metal total hip arthroplasty group (0.35 parts per billion [0.35 microg/L]) and the hip resurfacing group (0.50 parts per billion [0.50 microg/L]) at six months postoperatively. However, neither the median cobalt levels nor the median chromium levels were significantly different among the three metal-on-metal groups at one year. The median levels of molybdenum were not significantly different among the three groups at either six months or one year. In addition, there was no significant difference in the plasma concentration of oxidative stress markers in patients with metal-on-metal bearings compared with that in control patients. CONCLUSIONS: The blood metal ion levels in the hip resurfacing group were similar to those in the 28 and 36-mm-head metal-on-metal total hip arthroplasty groups. This study suggests that the increased metal ion levels had no effect on oxidative stress markers in the blood of these patients. PMID- 18676948 TI - Design considerations and life prediction of metal-on-metal bearings: the effect of clearance. AB - BACKGROUND: Clinical observations suggest that metal-on-metal arthroplasties that have been implanted for more than twenty years do fail. It is proposed that there are not two, but three distinct phases of wear life for any metal-on-metal implant system: bedding-in, steady state, and end point. In this study, we asked two questions: can we explain late failure due to wear, and will there be a late failure mechanism due to a change in the frictional torque? METHODS: In order to characterize wear failure, an analysis was made of five retrieved metal-on-metal couples that were mapped with use of a roundness machine. A geometrical model was developed on the basis of these observations, and wear at the end point was calculated. The literature on first-generation metal-on-metal implants retrieved for aseptic loosening was reviewed to assess the agreement with the retrieval findings as well as the wear model. RESULTS: A wear patch of an appreciable and constant size could be measured in all five retrieved couples. The end point of revision was observed to occur when the wear progression reached a contact area corresponding to approximately 75% of the projected diameter of the ball. The wear volume was calculated from the geometry. The available literature describing the wear characteristics of retrieved bearings after successful clinical use showed good agreement with the calculated wear model. CONCLUSIONS: During the implant life of long-term successful metal-on-metal devices, a wear patch develops, as evident from retrieved failed devices. Failure often occurs through loosening, and the observed wear patch is similar in size for devices measured by us and for those described in the literature. We hypothesized that failure by loosening occurs through the accumulation of wear, which eventually leads to high friction within the bearing and increased torsional forces across the joint and its fixation. PMID- 18676950 TI - New biosynthetic step in the melanin pathway of Wangiella (Exophiala) dermatitidis: evidence for 2-acetyl-1,3,6,8-Tetrahydroxynaphthalene as a novel precursor. AB - The predominant cell wall melanin of Wangiella dermatitidis, a black fungal pathogen of humans, is synthesized from 1,8-dihydroxynaphthalene (D2HN). An early precursor, 1,3,6,8-tetrahydroxynaphthalene (T4HN), in the pathway leading to D2HN is reportedly produced directly as a pentaketide by an iterative type I polyketide synthase (PKS). In contrast, the bluish-green pigment in Aspergillus fumigatus is produced after the enzyme Ayg1p converts the PKS product, the heptaketide YWA1, to T4HN. Previously, we created a new melanin-deficient mutant of W. dermatitidis, WdBrm1, by random molecular insertion. From this strain, the altered gene WdYG1 was cloned by a marker rescue strategy and found to encode WdYg1p, an ortholog of Ayg1p. In the present study, two gene replacement mutants devoid of the complete WdYG1 gene were derived to eliminate the possibility that the phenotype of WdBrm1 was due to other mutations. Characterization of the new mutants showed that they were phenotypically identical to WdBrm1. Chemical analyses of mutant cultures demonstrated that melanin biosynthesis was blocked, resulting in the accumulation of 2-acetyl-1,3,6,8-tetrahydroxynaphthalene (AT4HN) and its oxidative product 3-acetylflaviolin in the culture media. When given to an albino W. dermatitidis strain with an inactivated WdPKS1 gene, AT4HN was mostly oxidized to 3-acetylflaviolin and deacetylated to flaviolin. Under reduced oxygen conditions, cell-free homogenates of the albino converted AT4HN to D2HN. This is the first report of evidence that the hexaketide AT4HN is a melanin precursor for T4HN in W. dermatitidis. PMID- 18676951 TI - Sorting signals within the Saccharomyces cerevisiae sporulation-specific dityrosine transporter, Dtr1p, C terminus promote Golgi-to-prospore membrane transport. AB - During sporulation in Saccharomyces cerevisiae, the dityrosine transporter Dtr1p, which is required for formation of the outermost layer of the spore wall, is specifically expressed and transported to the prospore membrane, a novel double lipid-bilayer membrane. Dtr1p consists of 572 amino acids with predicted N- and C terminal cytoplasmic extensions and 12 transmembrane domains. Dtr1p missing the largest internal cytoplasmic loop was trapped in the endoplasmic reticulum in both mitotically dividing cells and cells induced to sporulate. Deletion of the carboxyl 15 amino acids, but not the N-terminal extension of Dtr1p, resulted in a protein that failed to localize to the prospore membrane and was instead observed in cytoplasmic puncta. The puncta colocalized with a cis-Golgi marker, suggesting that Dtr1p missing the last 15 amino acids was trapped in an early Golgi compartment. Deletion of the C-terminal 10 amino acids resulted in a protein that localized to the prospore membrane with a delay and accumulated in cytoplasmic puncta that partially colocalized with a trans-Golgi marker. Both full-length Dtr1p and Dtr1p missing the last 10 amino acids expressed in vegetative cells localized to the plasma membrane and vacuoles, while Dtr1p deleted for the carboxyl-terminal 15 amino acids was observed only at vacuoles, suggesting that transport to the prospore membrane is mediated by distinct signals from those that specify plasma membrane localization. Transfer-of-function experiments revealed that both the carboxyl transmembrane domain and the C-terminal tail are important for Golgi complex-to-prospore membrane transport. PMID- 18676952 TI - Beta1,2-xylosyltransferase Cxt1p is solely responsible for xylose incorporation into Cryptococcus neoformans glycosphingolipids. AB - The Manalpha1,3(Xylbeta1,2)Manalpha structural motif is common to both capsular polysaccharides of Cryptococcus neoformans and to cryptococcal glycosphingolipids. Comparative analysis of glycosphingolipid structural profiles in wild-type and mutant strains showed that the Xylbeta1,2-transferase (Cxt1p) that participates in capsular polysaccharide biosynthesis is also the sole transferase responsible for adding xylose to C. neoformans glycosphingolipids. PMID- 18676953 TI - A protein with similarity to PTEN regulates aggregation territory size by decreasing cyclic AMP pulse size during Dictyostelium discoideum development. AB - An interesting but largely unanswered biological question is how eukaryotic organisms regulate the size of multicellular tissues. During development, a lawn of Dictyostelium cells breaks up into territories, and within the territories the cells aggregate in dendritic streams to form groups of approximately 20,000 cells. Using random insertional mutagenesis to search for genes involved in group size regulation, we found that an insertion in the cnrN gene affects group size. Cells lacking CnrN (cnrN(-)) form abnormally small groups, which can be rescued by the expression of exogenous CnrN. Relayed pulses of extracellular cyclic AMP (cAMP) direct cells to aggregate by chemotaxis to form aggregation territories and streams. cnrN(-) cells overaccumulate cAMP during development and form small territories. Decreasing the cAMP pulse size by treating cnrN(-) cells with cAMP phosphodiesterase or starving cnrN(-) cells at a low density rescues the small territory phenotype. The predicted CnrN sequence has similarity to phosphatase and tensin homolog (PTEN), which in Dictyostelium inhibits cAMP-stimulated phosphatidylinositol 3-kinase signaling pathways. CnrN inhibits cAMP-stimulated phosphatidylinositol 3,4,5-trisphosphate accumulation, Akt activation, actin polymerization, and cAMP production. Our results suggest that CnrN is a protein with some similarities to PTEN and that it regulates cAMP signal transduction to regulate territory size. PMID- 18676954 TI - Dissecting colony development of Neurospora crassa using mRNA profiling and comparative genomics approaches. AB - Colony development, which includes hyphal extension, branching, anastomosis, and asexual sporulation, is a fundamental aspect of the life cycle of filamentous fungi; genetic mechanisms underlying these phenomena are poorly understood. We conducted transcriptional profiling during colony development of the model filamentous fungus Neurospora crassa, using 70-mer oligonucleotide microarrays. Relative mRNA expression levels were determined for six sections of defined age excised from a 27-h-old N. crassa colony. Functional category analysis showed that the expression of genes involved in cell membrane biosynthesis, polar growth, and cellular signaling was enriched at the periphery of the colony. The relative expression of genes involved in protein synthesis and energy production was enriched in the middle section of the colony, while sections of the colony undergoing asexual development (conidiogenesis) were enriched in expression of genes involved in protein/peptide degradation and unclassified proteins. A cross examination of the N. crassa data set with a published data set of Aspergillus niger revealed shared patterns in the spatiotemporal regulation of gene orthologs during colony development. At present, less than 50% of genes in N. crassa have functional annotation, which imposes the chief limitation on data analysis. Using an evolutionary approach, we observed that the expression of phylogenetically conserved groups of genes was enriched in the middle section of an N. crassa colony whereas expression of genes unique to euascomycete species and of N. crassa orphan genes was enriched at the colony periphery and in the older, conidiating sections of a fungal colony. PMID- 18676955 TI - Nucleus-specific importin alpha proteins and nucleoporins regulate protein import and nuclear division in the binucleate Tetrahymena thermophila. AB - The ciliate Tetrahymena thermophila, having both germ line micronuclei and somatic macronuclei, must possess a specialized nucleocytoplasmic transport system to import proteins into the correct nucleus. To understand how Tetrahymena can target proteins to distinct nuclei, we first characterized FG repeat containing nucleoporins and found that micro- and macronuclei utilize unique subsets of these proteins. This finding implicates these proteins in the differential permeability of the two nuclei and implies that nuclear pores with discrete specificities are assembled within a single cell. To identify the import machineries that interact with these different pores, we characterized the large families of karyopherin homologs encoded within the genome. Localization studies of 13 putative importin (imp) alpha- and 11 imp beta-like proteins revealed that imp alpha-like proteins are nucleus specific--nine localized to the germ line micronucleus--but that most imp beta-like proteins localized to both types of nuclei. These data suggest that micronucleus-specific proteins are transported by specific imp alpha adapters. The different imp alpha proteins exhibit substantial sequence divergence and do not appear to be simply redundant in function. Disruption of the IMA10 gene encoding an imp alpha-like protein that accumulates in dividing micronuclei results in nuclear division defects and lethality. Thus, nucleus-specific protein import and nuclear function in Tetrahymena are regulated by diverse, specialized karyopherins. PMID- 18676956 TI - Protein import into hydrogenosomes of Trichomonas vaginalis involves both N terminal and internal targeting signals: a case study of thioredoxin reductases. AB - The parabasalian flagellate Trichomonas vaginalis harbors mitochondrion-related and H(2)-producing organelles of anaerobic ATP synthesis, called hydrogenosomes, which harbor oxygen-sensitive enzymes essential to its pyruvate metabolism. In the human urogenital tract, however, T. vaginalis is regularly exposed to low oxygen concentrations and therefore must possess antioxidant systems protecting the organellar environment against the detrimental effects of molecular oxygen and reactive oxygen species. We have identified two closely related hydrogenosomal thioredoxin reductases (TrxRs), the hitherto-missing component of a thioredoxin-linked hydrogenosomal antioxidant system. One of the two hydrogenosomal TrxR isoforms, TrxRh1, carried an N-terminal extension resembling known hydrogenosomal targeting signals. Expression of hemagglutinin-tagged TrxRh1 in transfected T. vaginalis cells revealed that its N-terminal extension was necessary to import the protein into the organelles. The second hydrogenosomal TrxR isoform, TrxRh2, had no N-terminal targeting signal but was nonetheless efficiently targeted to hydrogenosomes. N-terminal presequences from hydrogenosomal proteins with known processing sites, i.e., the alpha subunit of succinyl coenzyme A synthetase (SCSalpha) and pyruvate:ferredoxin oxidoreductase A, were investigated for their ability to direct mature TrxRh1 to hydrogenosomes. Neither presequence directed TrxRh1 to hydrogenosomes, indicating that neither extension is, by itself, sufficient for hydrogenosomal targeting. Moreover, SCSalpha lacking its N-terminal extension was efficiently imported into hydrogenosomes, indicating that this extension is not required for import of this major hydrogenosomal protein. The finding that some hydrogenosomal enzymes require N-terminal signals for import but that in others the N-terminal extension is not necessary for targeting indicates the presence of additional targeting signals within the mature subunits of several hydrogenosome-localized proteins. PMID- 18676957 TI - Involvement of Sib proteins in the regulation of cellular adhesion in Dictyostelium discoideum. AB - Molecular mechanisms ensuring cellular adhesion have been studied in detail in Dictyostelium amoebae, but little is known about the regulation of cellular adhesion in these cells. Here, we show that cellular adhesion is regulated in Dictyostelium, notably by the concentration of a cellular secreted factor accumulating in the medium. This constitutes a quorum-sensing mechanism allowing coordinated regulation of cellular adhesion in a Dictyostelium population. In order to understand the mechanism underlying this regulation, we analyzed the expression of recently identified Dictyostelium adhesion molecules (Sib proteins) that present features also found in mammalian integrins. sibA and sibC are both expressed in vegetative Dictyostelium cells, but the expression of sibC is repressed strongly in conditions where cellular adhesion decreases. Analysis of sibA and sibC mutant cells further suggests that variations in the expression levels of sibC account largely for changes in cellular adhesion in response to environmental cues. PMID- 18676958 TI - Characterization of a serine proteinase mediating encystation of Acanthamoeba. AB - Members of the genus Acanthamoeba, amphizoic protozoan parasites, are causative agents of granulomatous amoebic encephalitis and amoebic keratitis. Proteinases play a role in various biologic actions in Acanthamoeba, including host tissue destruction, pathogenesis, and digestion of phagocytosed food. Interestingly, we found that encystation of Acanthamoeba was inhibited by the serine proteinase inhibitor phenylmethanesulfonyl fluoride. In this study, we characterize a serine proteinase that is involved in mediating the encystation of Acanthamoeba. This encystation-mediating serine proteinase (EMSP) is shown to be highly expressed during encystation by real-time PCR and Western blot analysis. Chemically synthesized small interfering RNA against EMSP inhibited the expression of EMSP mRNA and significantly reduced the encystation efficiency of Acanthamoeba. An EMSP-enhanced green fluorescent protein fusion protein localized to vesicle-like structures within the amoeba. Using LysoTracker analysis, these vesicular structures were confirmed to be lysosomes. After incubation of the transfected amoeba in encystment media, small fluorescent vesicle-like structures gathered and formed ball-like structures, which were identified as colocalizing with the autophagosome. Taken together, these results indicate that EMSP plays an important role in the differentiation of Acanthamoeba by promoting autolysis. PMID- 18676959 TI - Comprehensive evaluation of apolipoprotein H gene (APOH) variation identifies novel associations with measures of lipid metabolism in GENOA. AB - Apolipoprotein H (apoH, also named beta-2 glycoprotein I) is found on several classes of lipoproteins, and is involved in the activation of lipoprotein lipase in lipid metabolism. We have comprehensively investigated the association of variation in the apoH gene (APOH) with lipid traits in hepatic cholesterol transport, dietary cholesterol transport (DCT), and reverse cholesterol transport (RCT). Our study population consisted of families from the Genetic Epidemiology Network of Arteriopathy multicenter study that include African Americans, Mexican Americans, and European Americans. We individually tested 36 single-nucleotide polymorphisms (SNPs) that span the APOH locus, including nonsynonymous variants that result in known apoH charge isoforms. In addition, we constructed haplotypes from SNPs in the 5' promoter region that comprise cis-acting regulatory elements, as well as haplotypes for multiple amino acid substitutions. We found point-wise significant associations of APOH variants with various lipid measures in the three racial groups. The strongest associations were found for DCT traits (triglyceride and apoE levels) in Mexican Americans with a nonsynonymous variant (SNP 14917, Cys306Gly) that may alter apoH protein folding in a region involved in phospholipid binding. In conclusion, family-based analyses of APOH variants have identified associations with measures of lipid metabolism in three American racial groups. PMID- 18676960 TI - Measuring depression at the end of life: is the Hamilton Depression Rating Scale a valid instrument? AB - Depression at the end of life is a common mental health issue with serious implications for quality of life and decision making. This study investigated the reliability and validity of one of the most frequently used measures of depression, the Hamilton Depression Rating Scale (HAM-D) in 422 patients with terminal cancer admitted to a palliative care hospital. The HAM-D demonstrated high reliability and concurrent validity with depression diagnosis, based on Structured Clinical Interview for DSM-IV Axis I Disorders, non-patient research version (SCID-I/NP Depression Module). Receiver Operating Characteristic (ROC) analyses generated an area under the curve of .91, indicating high sensitivity and specificity. The HAM-D strongly correlated with several measures of distress (e.g., hopelessness, desire for hastened death, and suicidal ideation). Factor analysis generated 4 factors (anxiety, depressed mood, insomnia, and somatic symptoms), accounting for 42% of the variance. Implications for assessing depression in this population are discussed. PMID- 18676961 TI - Medications to modify lung function decline in chronic obstructive pulmonary disease: some hopeful signs. PMID- 18676962 TI - Lung stress and strain in acute respiratory distress syndrome: good ideas for clinical management? PMID- 18676963 TI - A unique spirometric phenotype in COPD. PMID- 18676964 TI - The considerable, often neglected, impact of pulmonary tuberculosis on the prevalence of COPD. PMID- 18676965 TI - Rapid bed-side myocardial perfusion imaging: is a dream coming true? PMID- 18676966 TI - A novel approach for a novel combination: a trans-septal biopsy of left atrial mass in recurrent phyllodes tumour. AB - A 69-year-old female with a previous hemisternectomy for recurrent phyllodes tumour was referred for echocardiography because of progressive dyspnoea, orthopnea, and a diastolic mitral rumble on auscultation. Transthoracic echocardiography revealed a large left atrial mass. Although a diagnosis of myxoma was likely, there was concern that this was a cardiac metastasis. A tissue diagnosis was mandatory before further thoracic surgery could be considered. This was obtained via a trans-septal puncture using transoesophageal echocardiographic guidance. Histology confirmed a diagnosis of myxoma, and our patient underwent successful surgery. This case highlights the importance of obtaining accurate tissue diagnosis and of excluding metastatic disease in patients with a cardiac mass and a history of tumour, prior to deciding whether surgical excision is warranted. PMID- 18676967 TI - Syphilitic aortitis causing critical obstruction of coronary ostia. PMID- 18676968 TI - Giant calcified cardiomegaly. PMID- 18676969 TI - More good reasons for adherence to statin therapy during acute coronary syndromes. PMID- 18676970 TI - Concentration of apolipoprotein B is comparable with the apolipoprotein B/apolipoprotein A-I ratio and better than routine clinical lipid measurements in predicting coronary heart disease mortality: findings from a multi-ethnic US population. AB - AIMS: Prospective studies indicate that apolipoprotein measurements predict coronary heart disease (CHD) risk; however, evidence is conflicting, especially in the US. Our aim was to assess whether measurements of apolipoprotein B (apoB) and apolipoprotein A-I (apoA-I) can improve the ability to predict CHD death beyond what is possible based on traditional cardiovascular (CV) risk factors and clinical routine lipid measurements. METHODS AND RESULTS: We analysed prospectively associations of apolipoprotein measurements, traditional CV risk factors, and clinical routine lipid measurements with CHD mortality in a multi ethnic representative subset of 7594 US adults (mean age 45 years; 3881 men and 3713 women, median follow-up 124 person-months) from the Third National Health and Nutrition Examination Survey mortality study. Multiple Cox-proportional hazards regression was applied. There were 673 CV deaths of which 432 were from CHD. Concentrations of apoB [hazard ratio (HR) 1.98, 95% confidence interval (CI) 1.09-3.61], apoA-I (HR 0.48, 95% CI 0.27-0.85) and total cholesterol (TC) (HR 1.17, 95% CI 1.02-1.34) were significantly related to CHD death, whereas high density lipoprotein cholesterol (HDL-C) (HR 0.68, 95% CI 0.45-1.05) was borderline significant. Both the apoB/apoA-I ratio (HR 2.14, 95% CI 1.11-4.10) and the TC/HDL-C ratio (HR 1.10, 95% CI 1.04-1.16) were related to CHD death. Only apoB (HR 2.01, 95% CI 1.05-3.86) and the apoB/apoA-I ratio (HR 2.09, 95% CI 1.04-4.19) remained significantly associated with CHD death after adjusting for CV risk factors. CONCLUSION: In the US population, apolipoprotein measurements significantly predict CHD death, independently of conventional lipids and other CV risk factors (smoking, dyslipidaemia, hypertension, obesity, diabetes and C reactive protein). Furthermore, the predictive ability of apoB alone to detect CHD death was better than any of the routine clinical lipid measurements. Inclusion of apolipoprotein measurements in future clinical guidelines should not be discarded. PMID- 18676971 TI - Modelling germ cell development in vitro. AB - Germ cells have a critical role in mediating the generation of genetic diversity and transmitting this information across generations. Furthermore, gametogenesis is unique as a developmental process in that it generates highly-specialized haploid gametes from diploid precursor stem cells through meiosis. Despite the importance of this process, progress in elucidating the molecular mechanisms underpinning mammalian germ cell development has been retarded by the lack of an efficient and reproducible system of in vitro culture for the expansion and trans meiotic differentiation of germline cells. The dearth of such a culture system has rendered the study of germ cell biology refractory to the application of new high-throughput technologies such as RNA interference, leaving in vivo gene targeting approaches as the only option to determine the function of genes believed to be involved in gametogenesis. Recent reports detailing the derivation of gametes in vitro from stem cells may provide the first steps in developing new tools to solve this problem. This review considers the developments made in modelling germ cell development using stem cells, and some of the challenges that need to be overcome to make this a useful tool for studying gametogenesis and to realize any future clinical application. PMID- 18676972 TI - Predicting pathway membership via domain signatures. AB - MOTIVATION: Functional characterization of genes is of great importance for the understanding of complex cellular processes. Valuable information for this purpose can be obtained from pathway databases, like KEGG. However, only a small fraction of genes is annotated with pathway information up to now. In contrast, information on contained protein domains can be obtained for a significantly higher number of genes, e.g. from the InterPro database. RESULTS: We present a classification model, which for a specific gene of interest can predict the mapping to a KEGG pathway, based on its domain signature. The classifier makes explicit use of the hierarchical organization of pathways in the KEGG database. Furthermore, we take into account that a specific gene can be mapped to different pathways at the same time. The classification method produces a scoring of all possible mapping positions of the gene in the KEGG hierarchy. Evaluations of our model, which is a combination of a SVM and ranking perceptron approach, show a high prediction performance. Moreover, for signaling pathways we reveal that it is even possible to forecast accurately the membership to individual pathway components. AVAILABILITY: The R package gene2pathway is a supplement to this article. PMID- 18676973 TI - Optimizing amino acid groupings for GPCR classification. AB - MOTIVATION: There is much interest in reducing the complexity inherent in the representation of the 20 standard amino acids within bioinformatics algorithms by developing a so-called reduced alphabet. Although there is no universally applicable residue grouping, there are numerous physiochemical criteria upon which one can base groupings. Local descriptors are a form of alignment-free analysis, the efficiency of which is dependent upon the correct selection of amino acid groupings. RESULTS: Within the context of G-protein coupled receptor (GPCR) classification, an optimization algorithm was developed, which was able to identify the most efficient grouping when used to generate local descriptors. The algorithm was inspired by the relatively new computational intelligence paradigm of artificial immune systems. A number of amino acid groupings produced by this algorithm were evaluated with respect to their ability to generate local descriptors capable of providing an accurate classification algorithm for GPCRs. PMID- 18676974 TI - Novel dominant-negative prion protein mutants identified from a randomized library. AB - Prion diseases are untreatable neurodegenerative disorders characterized by accumulation of PrP(Sc), an aggregated isoform of the cellular prion protein (PrP(C)). We generated a library of PrP variants with random mutations in the helix-3 domain and screened for dominant-negative mutants (DNMs) that would inhibit replication of prions (the Rocky Mountain Laboratory strain) in infected N2a cells. Two of the identified PrP mutants, Q167R and Q218K, were already known to counteract prion replication, thereby validating the effectiveness of this approach. In addition, novel DNMs were found efficiently to antagonize PrP(Sc) propagation in cells. In contrast to Q167R and Q218K, the newly identified DNMs S221P and Y217C resided on the cell surface at a substantially lower level, suggesting that robust cell surface display of DNM might not be a general prerequisite for efficient prion antagonism. The newly identified DNMs point to useful target-selective therapeutic tools for the treatment of prion diseases. PMID- 18676975 TI - Engineering of a monomeric fluorescent protein AsGFP499 and its applications in a dual translocation and transcription assay. AB - The tetrameric green fluorescent protein AsGFP(499) from the sea anemone Anemonia sulcata was converted into a dimeric and monomeric protein by site-directed mutagenesis. The protein was engineered without prior knowledge of its crystal structure based on a sequence alignment of multiple proteins belonging to the GFP family. Crucial residues for oligomerisation of AsGFP(499) were predicted and selected for mutation. By introduction of a single site mutation (S103K) the A/B subunit was disrupted whereas two substitutions were necessary to separate the A/C subunit (T159K/F173E). This method can be applied as a general predictive method for designing monomeric proteins from multimeric fluorescent proteins. The maturation temperature was optimised to 37 degrees C by a combination of Site directed and random mutagenesis. In cell-based assays, the NFATc1A (nuclear factor of activated T-cells, subtype 1, isoform A)-AsGFP(499) chimera formed massive cytoplasmic aggregates in HeLa cells, which prevented the shuttling of NFATc1A into the nucleus and consequentially its transcriptional activity. In contrast, the cells expressing the NFATc1A in fusion with our engineered dimeric and monomeric fluorescent mutants were homogeneously distributed throughout the cytoplasm, making these stable cell lines functional in both translocation and transcriptonal assays. This new dual cellular assay will allow the screening and discovery of new drugs that target NFAT cellular processes. PMID- 18676976 TI - Applicability of an exposure model for the determination of emissions from mobile phone base stations. AB - Applicability of a model to estimate radiofrequency electromagnetic field (RF EMF) strength in households from mobile phone base stations was evaluated with technical data of mobile phone base stations available from the German Net Agency, and dosimetric measurements, performed in an epidemiological study. Estimated exposure and exposure measured with dosemeters in 1322 participating households were compared. For that purpose, the upper 10th percentiles of both outcomes were defined as the 'higher exposed' groups. To assess the agreement of the defined 'higher' exposed groups, kappa coefficient, sensitivity and specificity were calculated. The present results show only a weak agreement of calculations and measurements (kappa values between -0.03 and 0.28, sensitivity between 7.1 and 34.6). Only in some of the sub-analyses, a higher agreement was found, e.g. when measured instead of interpolated geo-coordinates were used to calculate the distance between households and base stations, which is one important parameter in modelling exposure. During the development of the exposure model, more precise input data were available for its internal validation, which yielded kappa values between 0.41 and 0.68 and sensitivity between 55 and 76 for different types of housing areas. Contrary to this, the calculation of exposure on the basis of the available imprecise data from the epidemiological study-is associated with a relatively high degree of uncertainty. Thus, the model can only be applied in epidemiological studies, when the uncertainty of the input data is considerably reduced. Otherwise, the use of dosemeters to determine the exposure from RF-EMF in epidemiological studies is recommended. PMID- 18676977 TI - The solution structure of the amino-terminal domain of human DNA polymerase epsilon subunit B is homologous to C-domains of AAA+ proteins. AB - DNA polymerases alpha, delta and epsilon are large multisubunit complexes that replicate the bulk of the DNA in the eukaryotic cell. In addition to the homologous catalytic subunits, these enzymes possess structurally related B subunits, characterized by a carboxyterminal calcineurin-like and an aminoproximal oligonucleotide/oligosaccharide binding-fold domain. The B subunits also share homology with the exonuclease subunit of archaeal DNA polymerases D. Here, we describe a novel domain specific to the N-terminus of the B subunit of eukaryotic DNA polymerases epsilon. The N-terminal domain of human DNA polymerases epsilon (Dpoe2NT) expressed in Escherichia coli was characterized. Circular dichroism studies demonstrated that Dpoe2NT forms a stable, predominantly alpha-helical structure. The solution structure of Dpoe2NT revealed a domain that consists of a left-handed superhelical bundle. Four helices are arranged in two hairpins and the connecting loops contain short beta-strand segments that form a short parallel sheet. DALI searches demonstrated a striking structural similarity of the Dpoe2NT with the alpha-helical subdomains of ATPase associated with various cellular activity (AAA+) proteins (the C-domain). Like C domains, Dpoe2NT is rich in charged amino acids. The biased distribution of the charged residues is reflected by a polarization and a considerable dipole moment across the Dpoe2NT. Dpoe2NT represents the first C-domain fold not associated with an AAA+ protein. PMID- 18676978 TI - A member of the Whirly family is a multifunctional RNA- and DNA-binding protein that is essential for chloroplast biogenesis. AB - 'Whirly' proteins comprise a plant-specific protein family whose members have been described as DNA-binding proteins that influence nuclear transcription and telomere maintenance, and that associate with nucleoids in chloroplasts and mitochondria. We identified the maize WHY1 ortholog among proteins that coimmunoprecipitate with CRS1, which promotes the splicing of the chloroplast atpF group II intron. ZmWHY1 localizes to the chloroplast stroma and to the thylakoid membrane, to which it is tethered by DNA. Genome-wide coimmunoprecipitation assays showed that ZmWHY1 in chloroplast extract is associated with DNA from throughout the plastid genome and with a subset of plastid RNAs that includes atpF transcripts. Furthermore, ZmWHY1 binds both RNA and DNA in vitro. A severe ZmWhy1 mutant allele conditions albino seedlings lacking plastid ribosomes; these exhibit the altered plastid RNA profile characteristic of ribosome-less plastids. Hypomorphic ZmWhy1 mutants exhibit reduced atpF intron splicing and a reduced content of plastid ribosomes; aberrant 23S rRNA metabolism in these mutants suggests that a defect in the biogenesis of the large ribosomal subunit underlies the ribosome deficiency. However, these mutants contain near normal levels of chloroplast DNA and RNAs, suggesting that ZmWHY1 is not directly required for either DNA replication or for global plastid transcription. PMID- 18676979 TI - Molecular interactions of ASPP1 and ASPP2 with the p53 protein family and the apoptotic promoters PUMA and Bax. AB - The apoptosis stimulating p53 proteins, ASPP1 and ASPP2, are the first two common activators of the p53 protein family that selectively enable the latter to regulate specific apoptotic target genes, which facilitates yes yet unknown mechanisms for discrimination between cell cycle arrest and apoptosis. To better understand the interplay between ASPP- and p53-family of proteins we investigated the molecular interactions between them using biochemical methods and structure based homology modelling. The data demonstrate that: (i) the binding of ASPP1 and ASPP2 to p53, p63 and p73 is direct; (ii) the C-termini of ASPP1 and ASPP2 interact with the DNA-binding domains of p53 protein family with dissociation constants, K(d), in the lower micro-molar range; (iii) the stoichiometry of binding is 1:1; (iv) the DNA-binding domains of p53 family members are sufficient for these protein-protein interactions; (v) EMSA titrations revealed that while tri-complex formation between ASPPs, p53 family of proteins and PUMA/Bax is mutually exclusive, ASPP2 (but not ASPP1) formed a complex with PUMA (but not Bax) and displaced p53 and p73. The structure-based homology modelling revealed subtle differences between ASPP2 and ASPP1 and together with the experimental data provide novel mechanistic insights. PMID- 18676980 TI - Characterization of the TaALMT1 protein as an Al3+-activated anion channel in transformed tobacco (Nicotiana tabacum L.) cells. AB - TaALMT1 encodes a putative transport protein associated with Al(3+)-activated efflux of malate from wheat root apices. We expressed TaALMT1 in Nicotiana tabacum L. suspension cells and conducted a detailed functional analysis. Protoplasts were isolated for patch-clamping from cells expressing TaALMT1 and from control cells (empty vector transformed). With malate(2-) as the permeant anion in the protoplast, an inward current (anion efflux) that reversed at positive potentials was observed in protoplasts expressing TaALMT1 in the absence of Al(3+). This current was sensitive to the anion channel antagonist niflumate, but insensitive to Gd(3+). External AlCl(3) (50 microM), but not La(3+) and Gd(3+), increased the inward current in TaALMT1-transformed protoplasts. The inward current was highly selective to malate over nitrate and chloride (P(mal) >> P(NO3) >or= P(Cl), P(mal)/P(Cl) >or=18, +/-Al(3+)), under conditions with higher anion concentration internally than externally. The anion currents displayed a voltage and time dependent deactivation at negative voltages. Voltage ramps revealed that inward rectification was caused by the imposed anion gradients. Single channels with conductances between 10 and 17 pS were associated with the deactivation of the current at negative voltages, agreeing with estimates from voltage ramps. This study of the electrophysiological function of the TaALMT1 protein in a plant heterologous expression system provides the first direct evidence that TaALMT1 functions as an Al(3+)-activated malate(2-) channel. We show that the Al(3+)-activated currents measured in TaALMT1-transformed tobacco cells are identical to the Al(3+)-activated currents observed in the root cells of wheat, indicating that TaALMT1 alone is likely to be responsible for those endogenous currents. PMID- 18676981 TI - Combating endometriosis by blocking proteasome and nuclear factor-kappaB pathways. AB - BACKGROUND: The objective of this study is to investigate the effect of pyrrolidine dithiocarbamate [PDTC; a nuclear factor-kappaB (NF-kappaB) inhibitor] and bortezomib (Velcade; a proteasome inhibitor) on the development of experimental endometriotic implants in rats. METHODS: Endometriosis was surgically induced in 30 rats using the method of Vernon and Wilson. Three weeks later the viability and volume of the implants were recorded and classified. Afterwards, rats were put into three groups with equal numbers. The groups were labelled as the control, the PDTC and the bortezomib groups. Seven days after treatment, a third laparotomy was done and the volume of implants was measured again. The animals were then sacrificed, and the implants were stained with Ki67, proliferating cell nuclear antigen (PCNA), CD34, CD31 and Masson's trichrome histochemical staining. RESULTS: In 80% of the implanted rats, vesicles at the suture region were observed, and the rats graded according to average vesicle diameter (D) as: Grade 1 (no vesicle, 20% of rats), Grade 2 (D < 2 mm, 33.3% of rats), Grade 3 (2 mm 4.5 mm, 26.7% of rats) and Grade 4 (D > 4.5 mm, 20% of rats). After treatment with PDTC or bortezomib, these percentages were decreased for Grades 3 and 4, and increased in Grade 1. The post-treatment implant volumes were decreased in the PDTC and bortezomib groups (P < 0.002 and P < 0.001), and slightly increased in the control group (P = 0.279). In the PDTC and bortezomib groups, CD34, CD31, PCNA and Ki67 expression levels were similar but were significantly reduced compared with the control group. CONCLUSIONS: PDTC and bortezomib may represent a novel therapeutic strategy for treatment of endometriosis. PMID- 18676982 TI - Maternal protein-energy supplementation does not affect adolescent blood pressure in The Gambia. AB - BACKGROUND: Birthweight, and by inference maternal nutrition during pregnancy, is thought to be an important determinant of offspring blood pressure but the evidence base for this in humans is lacking data from randomized controlled trials. METHODS: The offspring from a maternal prenatal protein-energy supplementation trial were enrolled into a follow-up study of chronic disease risk factors including blood pressure. Subjects were 11-17 years of age and blood pressure was measured in triplicate using an automated monitor (Omron 705IT). One thousand two-hundred sixty seven individuals (71% of potential participants) were included in the analysis. RESULTS: There was no difference in blood pressure between those whose mothers had consumed protein-energy biscuits during pregnancy and those whose mothers had consumed the same supplement post-partum. For systolic blood pressure the intention-to-treat regression coefficient was 0.46 (95% CI: -1.12, 2.04). Mean systolic blood pressure for control children was 110.2 (SD +/- 9.3) mmHg and for intervention children was 110.8 (SD +/- 8.8) mmHg. Mean diastolic blood pressure for control children was 64.7 (SD +/- 7.7) mmHg and for intervention children was 64.6 (SD +/- 7.6) mmHg. CONCLUSIONS: We have found no association between maternal prenatal protein-energy supplementation and offspring blood pressure in adolescence amongst rural Gambians. We found some evidence to suggest that offspring body composition may interact with the effect of maternal supplementation on blood pressure. PMID- 18676983 TI - Fish, human health and marine ecosystem health: policies in collision. AB - BACKGROUND: Health recommendations advocating increased fish consumption need to be placed in the context of the potential collapse of global marine capture fisheries. METHODS: Literature overview. RESULTS: In economically developed countries, official healthy eating advice is to eat more fish, particularly that rich in omega-3 oils. In many less economically developed countries, fish is a key human health asset, contributing >20% of animal protein intake for 2.6 billion people. Marine ecologists predict on current trends that fish stocks are set to collapse in 40 years, and propose increased restrictions on fishing, including no-take zones, in order to restore marine ecosystem health. Production of fishmeal for aquaculture and other non-food uses (22 MT in 2003) appears to be unsustainable. Differences in fish consumption probably contribute to within country and international health inequalities. Such inequalities are likely to increase if fish stocks continue to decline, while increasing demand for fish will accelerate declines in fish stocks and the health of marine ecosystems. CONCLUSIONS: Urgent national and international action is necessary to address the tensions arising from increasing human demand for fish and seafood, and rapidly declining marine ecosystem health. PMID- 18676984 TI - Meningioma and mobile phone use--a collaborative case-control study in five North European countries. AB - BACKGROUND: Use of mobile telephones has been suggested as a possible risk factor for intracranial tumours. To evaluate the effect of mobile phones on risk of meningioma, we carried out an international, collaborative case-control study of 1209 meningioma cases and 3299 population-based controls. METHODS: Population based cases were identified, mostly from hospitals, and controls from national population registers and general practitioners' patient lists. Detailed history of mobile phone use was obtained by personal interview. Regular mobile phone use (at least once a week for at least 6 months), duration of use, cumulative number and hours of use, and several other indicators of mobile phone use were assessed in relation to meningioma risk using conditional logistic regression with strata defined by age, sex, country and region. RESULTS: Risk of meningioma among regular users of mobile phones was apparently lower than among never or non regular users (odds ratio, OR = 0.76, 95% confidence interval, CI 0.65, 0.89). The risk was not increased in relation to years since first use, lifetime years of use, cumulative hours of use or cumulative number of calls. The findings were similar regardless of telephone network type (analogue/digital), age or sex. CONCLUSIONS: Our results do not provide support for an association between mobile phone use and risk of meningioma. PMID- 18676985 TI - Differential health reporting by education level and its impact on the measurement of health inequalities among older Europeans. AB - BACKGROUND: This study aims to establish whether health reporting differs by education level and, if so, to determine the extent to which this biases the measurement of health inequalities among older Europeans. METHODS: Data are from the Survey of Health, Ageing and Retirement in Europe (SHARE) covering eight countries. Differential reporting of health by education is identified from ratings of anchoring vignettes that describe fixed health states. Ratings of own health in six domains (mobility, pain, sleep, breathing, emotional health and cognition) are corrected for differences in reporting using an extended ordered probit model. For each country and health domain, we compare the corrected with the uncorrected age-sex standardized high-to-low education rate ratio for the absence of a health problem. RESULTS: Before correction for reporting differences across the 48 combinations of country by health domain, there was no inequality in health by education (P > 0.05) in 32 of 48 cases. However, there were reporting differences by education (P < 0.05) in 29 out of 48 cases. In general, higher educated older Europeans are more likely to rate a given health state negatively (except for Spain and Sweden). Correcting for these differences generally increases health inequalities (except for Spain and Sweden) and results in the emergence of inequalities in 18 cases (P < 0.05), which may be considered 'statistically significant'. The greatest impact is in Belgium, Germany and The Netherlands, where inequalities (P < 0.05) appear only after adjustment in four of the six health domains. CONCLUSIONS: These results emphasize the need to account for differences in the reporting of health. Measured health inequalities by education are often underestimated, and even go undetected, if no account is taken of these reporting differences. PMID- 18676986 TI - Serum levels of tumour necrosis factor family members a proliferation-inducing ligand (APRIL) and B lymphocyte stimulator (BLyS) are inversely correlated in systemic lupus erythematosus. AB - OBJECTIVE: To determine whether serum levels of a proliferation-inducing ligand (APRIL) are altered in patients with systemic lupus erythematosus (SLE), and correlate with disease parameters. METHODS: Clinical and biological parameters were analysed for 43 patients that fulfilled American College of Rheumatology (ACR) criteria for SLE classification and were positive for anti-double-stranded DNA (dsDNA) antibodies at least once in their medical records. Tests included measurement of serum levels of the tumour necrosis factor (TNF) family members APRIL and B lymphocyte stimulator (BLyS; a cytokine shown to promote SLE disease). RESULTS: Median APRIL levels were elevated in patients with SLE compared to patients with osteoarthritis and healthy controls, but did not correlate with the SLE Disease Activity Index (SLEDAI). APRIL serum levels showed an inverse correlation with BLyS serum levels (r = -0.339; p = 0.03). For patients with SLE with positive anti-dsDNA titres (>40 arbitrary units (AU)/ml) at inclusion (n = 25), circulating APRIL was inversely correlated with BLyS levels (r = -0.465; p = 0.022) and anti-dsDNA antibody titres (r = -0.411; p = 0.046). In a follow-up study at their second visit, 27 patients showed an inverse correlation of APRIL serum levels with BLyS (r = -0.398; p = 0.03) as well as with anti-dsDNA (r = -0.408; p = 0.03) titres and SLEDAI (r = -0.408; p = 0.01). CONCLUSION: The inverse correlation observed between APRIL and BLyS suggests that APRIL acts as a protective factor. APRIL and BLyS may thus have opposite roles in SLE, which must be considered when defining therapeutic applications of these cytokines. PMID- 18676987 TI - Study of the impact of laws regulating tobacco consumption on the prevalence of passive smoking in Spain. AB - BACKGROUND: In 2005, the Spanish parliament passed the Spanish anti-smoking law. This legislation restricted tobacco smoking in public places, including recreation venues (bars and restaurants), but smoking was not completely prohibited in bars and restaurants. The law was enforced in January 2006. With the objective of analysing the impact that this law has had on the general Spanish population, the Spanish Society of Pulmonology and Thoracic Surgery (SEPAR) designed and implemented a survey of a representative sample of the general Spanish population on two separate occasions: in 2005 and in 2007 (12 months after the ban came into effect). METHODS: Two epidemiological, observational and cross-sectional surveys were performed among a random and representative sample of the general Spanish population, using the Computer Assisted Telephone Interview system. RESULTS: In the first survey, a total of 6533 subjects were interviewed, of whom 3907 (59.8%) were non-smokers and in the second, a total of 3289 subjects were interviewed, of whom 2174 (65.9%) were non smokers. The overall prevalence of exposure to environmental tobacco smoke (ETS) decreased from 49.5% in 2005 to 37.9% in 2007 (22% reduction). The greatest reduction in prevalence of ETS exposure was in workplaces (from 25.8% to 11%, a decrease of 58.8%). Smaller reductions occurred in the home (from 29.5% to 21.4%, a decrease of 27%) and in recreation venues (from 37.4% to 31.8%, a decrease of 8%). CONCLUSIONS: Implementation of the smoking ban resulted in a significant decrease in exposure to ETS. PMID- 18676989 TI - Mice deficient for the chromosome 21 ortholog Itsn1 exhibit vesicle-trafficking abnormalities. AB - Enlarged early endosomes in the neurons of young Down syndrome (DS) and pre Alzheimer's disease (AD) brains suggest that a disturbance in endocytosis is one of the earliest hallmarks of AD pathogenesis in both conditions. We identified a chromosome 21 gene, Intersectin-1 (ITSN1) that is up-regulated in DS brains and has a putative function in endocytosis and vesicle trafficking. To elucidate the function of ITSN1 and assess its contribution to endocytic defects associated with DS and AD, we generated Itsn1 null mice. In knockout mice we found alterations in a number of parameters associated with endocytic and vesicle trafficking events. We found a reduced number of exocytosis events in chromaffin cells and a slowing of endocytosis in neurons. Endosome size was increased in neurons and NGF levels were reduced in the septal region of the brain. Our data is the first indication that Itsn1 has a role in endocytosis in an in vivo mammalian model, and that a disruption in Itsn1 expression causes a disturbance in vesicle trafficking and endocytic function in the brain. These results imply a role for ITSN1 in the early endocytic anomalies reported in DS brains which may have ramifications for the onset of AD. PMID- 18676988 TI - A high-density association screen of 155 ion transport genes for involvement with common migraine. AB - The clinical overlap between monogenic Familial Hemiplegic Migraine (FHM) and common migraine subtypes, and the fact that all three FHM genes are involved in the transport of ions, suggest that ion transport genes may underlie susceptibility to common forms of migraine. To test this leading hypothesis, we examined common variation in 155 ion transport genes using 5257 single nucleotide polymorphisms (SNPs) in a Finnish sample of 841 unrelated migraine with aura cases and 884 unrelated non-migraine controls. The top signals were then tested for replication in four independent migraine case-control samples from the Netherlands, Germany and Australia, totalling 2835 unrelated migraine cases and 2740 unrelated controls. SNPs within 12 genes (KCNB2, KCNQ3, CLIC5, ATP2C2, CACNA1E, CACNB2, KCNE2, KCNK12, KCNK2, KCNS3, SCN5A and SCN9A) with promising nominal association (0.00041 < P < 0.005) in the Finnish sample were selected for replication. Although no variant remained significant after adjusting for multiple testing nor produced consistent evidence for association across all cohorts, a significant epistatic interaction between KCNB2 SNP rs1431656 (chromosome 8q13.3) and CACNB2 SNP rs7076100 (chromosome 10p12.33) (pointwise P = 0.00002; global P = 0.02) was observed in the Finnish case-control sample. We conclude that common variants of moderate effect size in ion transport genes do not play a major role in susceptibility to common migraine within these European populations, although there is some evidence for epistatic interaction between potassium and calcium channel genes, KCNB2 and CACNB2. Multiple rare variants or trans-regulatory elements of these genes are not ruled out. PMID- 18676991 TI - Reliability of different Doppler ultrasound quantification methods and devices in the assessment of therapeutic response in arthritis. AB - OBJECTIVE: To evaluate the metric qualities of power Doppler ultrasound (PDUS) for different quantification methods and devices in the assessment of arthritis during anti-inflammatory treatment. METHODS: Twenty-four patients with active arthritis and first-time treatment with the TNF-alpha inhibitor adalimumab underwent sequential clinical, laboratory and US examination at baseline, weeks 2, 6 and 12. 2D and 3D PDUS was performed by two independent investigators. The images and movies were scored from 0 to 3 and the amount of colour pixels and voxels was calculated. In addition, the resistance index of a synovial artery was measured. Thirteen patients were examined with a second US machine. RESULTS: Treatment response was already observed at week 2 with a significant reduction of 2D (P < 0.01) and 3D scores (P < 0.001). A moderate correlation to 28-joint disease activity score was found for 3D voxel count (r(s) = 0.35, P < 0.001). Interobserver agreement was kappa or ICC >or= 0.8 for all methods except the resistance index (ICC = 0.60). Intermachine agreement was kappa = 0.57 for 2D PDUS score. CONCLUSIONS: The study demonstrates good to excellent interobserver and moderate intermachine reliability of different PDUS assessment methods in a longitudinal open-label study. PMID- 18676992 TI - 4-Hydroxynonenal-modified Ro 60 autoantigen accelerates autoimmunity in experimental animals. PMID- 18676993 TI - Radiation pneumonitis caused by a migrated brachytherapy seed lodged in the lung. AB - We report a case of radiation pneumonitis caused by a migrated seed lodged in the lung after prostate brachytherapy. A 71-year-old man underwent transperineal interstitial permanent prostate brachytherapy for localized prostate cancer. On the day after brachytherapy, a routine postimplant chest X-ray revealed migration of one seed to the lower lobe of the left lung. After 1 month, pulmonary opacities were observed in the left lower lobe but not near the seed. He was diagnosed with bacterial pneumonia, and antibiotic therapy was commenced. Two months after brachytherapy, the patient's symptoms, laboratory data and pulmonary opacities improved; however, an abnormal shadow (consolidation) developed around the migrated seed. Lung consolidation disappeared almost completely 12 months after brachytherapy without any medical treatment. The abnormal shadow probably represented radiation pneumonitis. To the best of our knowledge, this is the first report of radiation pneumonitis caused by a migrated brachytherapy seed in the lung. PMID- 18676994 TI - Label-free quantitative analysis of one-dimensional PAGE LC/MS/MS proteome: application on angiotensin II-stimulated smooth muscle cells secretome. AB - A widely used method for protein identification couples prefractionation of protein samples by one-dimensional (1D) PAGE with LC/MS/MS. We developed a new label-free quantitative algorithm by combining measurements of spectral counting, ion intensity, and peak area on 1D PAGE-based proteomics. This algorithm has several improvements over other label-free quantitative algorithms: (i) Errors in peak detection are reduced because the retention time is based on each LC/MS/MS run and actual precursor m/z. (ii) Detection sensitivity is increased because protein quantification is based on the combination of peptide count, ion intensity, and peak area. (iii) Peak intensity and peak area are calculated in each LC/MS/MS run for all slices from 1D PAGE for every single identified protein and visualized as a Western blot image. The sensitivity and accuracy of this algorithm were demonstrated by using standard curves (17.4 fmol to 8.7 pmol), complex protein mixtures (30 fmol to 1.16 pmol) of known composition, and spiked protein (34.8 fmol to 17.4 pmol) in complex proteins. We studied the feasibility of this approach using the secretome of angiotensin II (Ang II)-stimulated vascular smooth muscle cells (VSMCs). From the VSMC-conditioned medium, 629 proteins were identified including 212 putative secreted proteins. 26 proteins were differently expressed in control and Ang II-stimulated VSMCs, including 18 proteins not previously reported. Proteins related to cell growth (CYR61, protein NOV, and clusterin) were increased, whereas growth arrest-specific 6 (GAS6) and growth/differentiation factor 6 were decreased by Ang II stimulation. Ang II stimulated changes of plasminogen activator inhibitor-1, GAS6, cathepsin B, and periostin were validated by Western blot. In conclusion, a novel label-free quantitative analysis of 1D PAGE-LC/MS/MS-based proteomics has been successfully applied to the identification of new potential mediators of Ang II action and may provide an alternative to traditional protein staining methods. PMID- 18676995 TI - Ligand binding to truncated hemoglobin N from Mycobacterium tuberculosis is strongly modulated by the interplay between the distal heme pocket residues and internal water. AB - The survival of Mycobacterium tuberculosis requires detoxification of host *NO. Oxygenated Mycobacterium tuberculosis truncated hemoglobin N catalyzes the rapid oxidation of nitric oxide to innocuous nitrate with a second-order rate constant (k'(NOD) approximately 745 x 10(6) m(-1) x s(-1)), which is approximately 15-fold faster than the reaction of horse heart myoglobin. We ask what aspects of structure and/or dynamics give rise to this enhanced reactivity. A first step is to expose what controls ligand/substrate binding to the heme. We present evidence that the main barrier to ligand binding to deoxy-truncated hemoglobin N (deoxy trHbN) is the displacement of a distal cavity water molecule, which is mainly stabilized by residue Tyr(B10) but not coordinated to the heme iron. As observed in the Tyr(B10)/Gln(E11) apolar mutants, once this kinetic barrier is lowered, CO and O(2) binding is very rapid with rates approaching 1-2 x 10(9) m(-1) x s(-1). These large values almost certainly represent the upper limit for ligand binding to a heme protein and also indicate that the iron atom in trHbN is highly reactive. Kinetic measurements on the photoproduct of the *NO derivative of met trHbN, where both the *NO and water can be directly followed, revealed that water rebinding is quite fast (approximately 1.49 x 10(8) s(-1)) and is responsible for the low geminate yield in trHbN. Molecular dynamics simulations, performed with trHbN and its distal mutants, indicated that in the absence of a distal water molecule, ligand access to the heme iron is not hindered. They also showed that a water molecule is stabilized next to the heme iron through hydrogen-bonding with Tyr(B10) and Gln(E11). PMID- 18676997 TI - Clinical and laboratory investigations of the outbreaks of foot-and-mouth disease in southern England in 2007. AB - A case of foot-and-mouth disease (fmd) on a cattle farm in Normandy, Surrey, was confirmed on Friday August 3, 2007, the first case in the uk since 2001. The infection was detected nearby on a second farm on August 6. On September 12, fmd was confirmed on a farm approximately 20 km from Normandy in Egham, and this was followed by cases on five more farms in that area in the next three weeks. The majority of the infected farms consisted of multiple beef cattle holdings in semi urban areas. In total, 1578 animals were culled on the infected farms, and fmd virus infection was confirmed in 278 of them by the detection of viral antigen, genome or antibodies to the virus, or by clinical signs. This paper describes the findings from animal inspections on the infected farms, including the estimated ages of the fmd lesions and the numbers of animals infected. It also summarises the test results from samples taken for investigation, including the detection of preclinically viraemic animals by using real-time reverse transcriptase-pcr. PMID- 18676998 TI - Evaluation of indirect biomarkers for detecting corticosteroids used as illegal growth promoters in beef cattle. AB - The histological status of the thymus, blood cortisol concentration and circulating neutrophil:lymphocyte ratio were evaluated in 349 slaughtered beef cattle, to assess the potential of these parameters as indirect biomarkers of the illegal use of corticosteroids in meat production. The livers of 20 of the animals were analysed chemically for residues of corticosteroids. The morphology of the thymus was examined for adipose tissue infiltration, cortical atrophy and 'starry sky' appearance, and on the basis of these characteristics, the animals were considered to be negative, suspected or positive for illegal corticosteroid treatment. The animals considered to be negative had a mean cortisol concentration that was significantly higher (29 ng/ml) than that of the animals suspected for corticosteroid treatment (22 ng/ml). Using the chemical analysis as the gold standard for identifying illegally treated animals, the histological examination of the thymus had a sensitivity of 100 per cent and a specificity of 85 per cent. The samples that were positive by chemical analysis had cortisol concentrations of less than 2.0 ng/ml, whereas the mean cortisol concentration of the negative samples was 10.3 ng/ml. PMID- 18676996 TI - An iron responsive element-like stem-loop regulates alpha-hemoglobin-stabilizing protein mRNA. AB - Hemoglobin production during erythropoiesis is mechanistically coupled to the acquisition and metabolism of iron. We discovered that iron regulates the expression of alpha-hemoglobin-stabilizing protein (AHSP), a molecular chaperone that binds and stabilizes free alpha-globin during hemoglobin synthesis. In primates, the 3'-untranslated region (UTR) of AHSP mRNA contains a nucleotide sequence resembling iron responsive elements (IREs), stem-loop structures that regulate gene expression post-transcriptionally by binding iron regulatory proteins (IRPs). The AHSP IRE-like stem-loop deviates from classical consensus sequences and binds IRPs poorly in electrophoretic mobility shift assays. However, in cytoplasmic extracts, AHSP mRNA co-immunoprecipitates with IRPs in a fashion that is dependent on the stem-loop structure and inhibited by iron. Moreover, this interaction enhances AHSP mRNA stability in erythroid and heterologous cells. Our findings demonstrate that IRPs can regulate mRNA expression through non-canonical IREs and extend the repertoire of known iron regulated genes. In addition, we illustrate a new mechanism through which hemoglobin may be modulated according to iron status. PMID- 18676999 TI - Computed tomography for the diagnosis and treatment monitoring of bacterial pneumonia in Indian pythons (Python molurus). AB - Eight Indian pythons (Python molurus) with clinical and microbiological evidence of pneumonia were examined by computed tomography (ct) before and after treatment. The results were assessed subjectively and measurements were taken following a standard protocol. Changes in the lung tissue of all the pythons were diagnosed, and the extent of the disease could be assessed. ct examinations after treatment showed an improvement in the six pythons whose clinical condition had improved, but in the other two pythons they demonstrated the severity of the disease. The subjective assessments were superior to the evaluation of measurements of attenuation in regions of interest. However, the average and the maximum attenuation provided additional information on the extent of the disease. Except for one python with only mild clinical signs, the attenuation after successful treatment was still higher than in healthy pythons. PMID- 18677000 TI - Epidemiological survey on farms with documented occurrence of equine proliferative enteropathy due to Lawsonia intracellularis. PMID- 18677002 TI - Local veterinary inspector/Official Veterinarian fee review. PMID- 18677001 TI - Bovine tuberculosis in a badger (Meles meles) in Spain. PMID- 18677003 TI - Macrocyclic lactone resistance in cyathostomins. PMID- 18677004 TI - Investigating suspect bluetongue disease incidents. PMID- 18677005 TI - Equine metabolic syndrome and Cushing's disease clinical trial. PMID- 18677007 TI - Representing the profession. PMID- 18677008 TI - Welfare standards in livery yards. PMID- 18677009 TI - Clinical subsequent fractures cluster in time after first fractures. AB - OBJECTIVES: The risk of subsequent fractures is double the risk of having a first fracture. We analysed whether this risk is constant or not over time. METHODS: A population-based study in 4140 postmenopausal women, aged between 50 and 90 years, on radiographic confirmed clinical fractures from menopause onwards analysed by Cox regression. RESULTS: A total of 924 (22%) women had a first fracture and 243 (26% of 924) a subsequent fracture. Of all first fractures, 4% occurred in each year from menopause onwards, while after a first fracture 23% of all subsequent fractures occurred within 1 year and 54% within 5 years. When calculated from time of first fracture, the relative risk (RR) of subsequent fracture was 2.1 (95% CI 1.7 to 2.6) and remained increased over 15 years. When calculated for specific time intervals after a first fracture, the RR was 5.3 (95% CI 4.0 to 6.6) within 1 year, 2.8 (95% CI 2.0 to 3.6) within 2-5 years, 1.4 (95% CI 1.0 to 1.8) within 6-10 years and 0.41 (95% CI 0.29 to 0.53) after >10 years. CONCLUSIONS: From menopause onwards, clinical fractures cluster in time, indicating the need for early action to prevent subsequent fractures. PMID- 18677010 TI - Associations of obesity with modifiable risk factors for the development of cardiovascular disease in patients with rheumatoid arthritis. AB - OBJECTIVES: To assess the association of body mass index (BMI) with modifiable cardiovascular disease (CVD) risk factors in patients with rheumatoid arthritis (RA). METHODS: BMI, disease activity, selected CVD risk factors and CVD medication were assessed in 378 (276 women) patients with RA. Patients exceeding accepted thresholds in >or=3 CVD risk factors were classified as having the metabolic syndrome (MetS). RESULTS: BMI independently associated with hypertension (OR = 1.28 (95% CI = 1.22 to 1.34); p = 0.001), high-density lipoprotein (OR = 1.10 (95% CI = 1.06 to 1.15); p = 0.025), insulin resistance (OR = 1.13 (95% CI = 1.08 to 1.18); p = 0.000) and MetS (OR = 1.15 (95% CI = 1.08 to 1.21); p = 0.000). In multivariable analyses, BMI had the strongest associations with CVD risk factors (F(1-354) = 8.663, p = 0.000), and this was followed by lipid-lowering treatment (F(1-354) = 7.651, p = 0.000), age (F(1-354) = 7.541, p = 0.000), antihypertensive treatment (F(1-354) = 4.997, p = 0.000) and gender (F(1-354) = 4.707, p = 0.000). Prevalence of hypertension (p = 0.004), insulin resistance (p = 0.005) and MetS (p = 0.000) was significantly different between patients with RA who were normal, overweight and obese, and BMI differed significantly according to the number of risk factors present (p = 0.000). CONCLUSIONS: Increasing BMI associates with increased CVD risk independently of many confounders. RA-specific BMI cut-off points better identify patients with RA at increased CVD risk. Weight-loss regimens should be developed and applied in order to reduce CVD in patients with RA. PMID- 18677011 TI - The natural history of bone marrow lesions in community-based adults with no clinical knee osteoarthritis. AB - OBJECTIVE: Although bone marrow lesions (BML) have been implicated in the pathogenesis of osteoarthritis, their natural history in a healthy population is unknown. This study in a healthy, pain-free population aimed to examine the natural history of BML; factors associated with incidence and progression of BML over 2 years and whether incident BML are associated with the development of pain. METHODS: 271 subjects with no clinical knee osteoarthritis, being pain free at baseline, underwent magnetic resonance imaging of their dominant knee at baseline and 2 years later. The presence of BML was assessed. RESULTS: In knees initially free of BML, incident BML developed in 14% of people over the study period. Increased body mass index (BMI; odds ratio (OR) 1.15, 95% CI 1.06 to 1.2, p = 0.001) was associated with incident BML. Those who developed a BML were more likely to develop knee pain compared with those in whom no BML developed (OR 4.2, 95% CI 1.2 to 15.1, p = 0.03). Among those in whom BML were present at baseline, 46% completely resolved. There was no association between age, gender and BMI and persistence of BML over 2 years. CONCLUSION: In this healthy population, the rate of incident BML is lower than previously described in a population with osteoarthritis. Incident BML are associated with increased BMI and the development of pain. Approximately half the BML present at baseline resolved. These data suggest that in pain-free people with no clinical knee osteoarthritis, BML are reversible and may provide a target for interventions aimed at the prevention of knee osteoarthritis. PMID- 18677012 TI - Anti-tumour necrosis factor therapy in patients with refractory Takayasu arteritis: long-term follow-up. AB - OBJECTIVE: To assess the efficacy of anti-tumour necrosis factor (TNF) therapy to induce remission in patients with Takayasu arteritis (TAK) refractory to other immunosuppressive therapies. METHODS: Retrospective single-centre study of 25 patients with refractory TAK. RESULTS: Patients were treated with infliximab (IFX) or etanercept (ETA) for up to 7 years; 21 with IFX (median 28 months (range 2-84)) and 9 with ETA (median 28 months (range 4-82)); 5 patients initially treated with ETA subsequently switched to IFX. Following anti-TNF therapy, remission was achieved and prednisone was discontinued in 15 patients (60%) and successfully tapered below 10 mg/day in an additional 7 patients (28%). Of 18 patients treated with other immunosuppressive agents concurrent with anti-TNF therapy, 9 (50%) could taper or discontinue the additional agent. Major relapses occurred in four patients that initially achieved stable remission. Four patients suffered adverse events, including one with opportunistic infections and one with breast cancer. CONCLUSIONS: In this group of patients with refractory TAK, anti TNF therapy was associated with remission in a majority of patients, facilitating dose reduction or discontinuation of prednisone and other immunosuppressive therapy. These findings strengthen the rationale for the conducting of a randomised controlled trial of anti-TNF therapy in TAK. PMID- 18677013 TI - A 39-year-old man with HIV-associated lipodystrophy. AB - Human immunodeficiency virus (HIV)-associated lipodystrophy refers to fat accumulation, also known as lipohypertrophy, and fat wasting, also known as lipoatrophy. Both conditions can be very disturbing to patients and have been associated with metabolic disturbances such as insulin resistance and hyperlipidemias. The prevalence of HIV-associated lipodystrophy ranges from 6% to 69% in the medical literature. Although no clear associations have been made between specific drugs and HIV lipohypertrophy, stavudine and zidovudine have been implicated in the development of HIV lipoatrophy. The case of Mr B, a 39 year-old man with HIV-associated lipodystrophy whose facial changes are a cause of significant distress, highlights the need for clinicians to be attuned to the psychological impact that lipodystrophy can have on patients, especially because it may serve as a disincentive to adherence to antiretroviral drug regimens, resulting in an increased risk of developing viral resistance. PMID- 18677015 TI - A person of status. PMID- 18677016 TI - Leaders warn of deadly HIV, TB collision. PMID- 18677017 TI - Innovative program offers HIV therapy to internally displaced persons in Uganda. PMID- 18677018 TI - Integrated care needed for patients with HIV, drug abuse, and mental illness. PMID- 18677019 TI - Universal screening for methicillin-resistant Staphylococcus aureus by hospitals. PMID- 18677020 TI - Universal screening for methicillin-resistant Staphylococcus aureus by hospitals. PMID- 18677021 TI - Universal screening for methicillin-resistant Staphylococcus aureus by hospitals. PMID- 18677022 TI - Sampling-based approach to determining outcomes of patients lost to follow-up in antiretroviral therapy scale-up programs in Africa. PMID- 18677023 TI - Low-dose physiological growth hormone in patients with HIV and abdominal fat accumulation: a randomized controlled trial. AB - CONTEXT: Antiretroviral therapy can be associated with visceral adiposity and metabolic complications, increasing cardiovascular risk, and reduced growth hormone (GH) secretion may be a contributing factor. OBJECTIVE: To investigate the effects of low-dose physiological GH administration on body composition, glucose, and cardiovascular parameters in patients with human immunodeficiency virus (HIV) having abdominal fat accumulation and relative GH deficiency. DESIGN, SETTING, AND PATIENTS: A randomized, double-blind, placebo-controlled trial of 56 patients with HIV, abdominal fat accumulation, and reduced GH secretion (peak GH <7.5 ng/mL) conducted at a US academic medical center between November 2003 and October 2007. INTERVENTION: Patients were randomly assigned to receive either subcutaneous GH or matching placebo titrated to the upper quartile of normal insulinlike growth factor 1 (IGF-1) range for 18 months. Starting dose was 2 microg/kg/d and increased to maximum dose of 6 microg/kg/d (average dose, 0.33 mg/d). MAIN OUTCOME MEASURES: Change in body composition assessed by computed tomographic scan and dual-energy x-ray absorptiometry. Secondary outcomes included glucose, IGF-1, blood pressure (BP), and lipids. Treatment effect was the difference in the change between GH and placebo groups, using all available data. RESULTS: Fifty-five patients (26 with GH and 29 with placebo) were included in the safety analyses and 52 patients (25 with GH and 27 with placebo) were included in the efficacy analyses. Visceral adipose tissue area (treatment effect [last-value-carried-forward analysis {n = 56}, -19 cm(2); 95% confidence interval {CI}, -37 to -0.3 cm(2)], -19 cm(2); 95% CI, -38 to -0.5 cm(2); P = .049); trunk fat (-0.8 kg; 95% CI, -1.5 to -0.04 kg; P = .04); diastolic BP (-7 mm Hg; 95% CI, -11 to -2 mm Hg; P = .006); and triglycerides (-7 mg/dL, P = .002) improved but 2 hour glucose levels on glucose tolerance testing increased in the GH group vs the placebo group (treatment effect, 22 mg/dL; 95% CI, 6-37 mg/dL; P = .009). The IGF 1 levels increased (treatment effect, 129 ng/mL; 95% CI, 95-164 ng/mL; P < .001). Adverse events were not increased for GH vs placebo (23%; 95% CI, 9%-44% vs 28%; 95% CI, 13%-47%; P = .70). CONCLUSIONS: In HIV-associated abdominal fat accumulation and relative GH deficiency, low-dose GH received for 18 months resulted in significantly reduced visceral fat and truncal obesity, triglycerides, and diastolic BP, but 2-hour glucose levels on glucose tolerance testing were increased. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00100698. PMID- 18677024 TI - Estimation of HIV incidence in the United States. AB - CONTEXT: Incidence of human immunodeficiency virus (HIV) in the United States has not been directly measured. New assays that differentiate recent vs long-standing HIV infections allow improved estimation of HIV incidence. OBJECTIVE: To estimate HIV incidence in the United States. DESIGN, SETTING, AND PATIENTS: Remnant diagnostic serum specimens from patients 13 years or older and newly diagnosed with HIV during 2006 in 22 states were tested with the BED HIV-1 capture enzyme immunoassay to classify infections as recent or long-standing. Information on HIV cases was reported to the Centers for Disease Control and Prevention through June 2007. Incidence of HIV in the 22 states during 2006 was estimated using a statistical approach with adjustment for testing frequency and extrapolated to the United States. Results were corroborated with back-calculation of HIV incidence for 1977-2006 based on HIV diagnoses from 40 states and AIDS incidence from 50 states and the District of Columbia. MAIN OUTCOME MEASURE: Estimated HIV incidence. RESULTS: An estimated 39,400 persons were diagnosed with HIV in 2006 in the 22 states. Of 6864 diagnostic specimens tested using the BED assay, 2133 (31%) were classified as recent infections. Based on extrapolations from these data, the estimated number of new infections for the United States in 2006 was 56,300 (95% confidence interval [CI], 48,200-64,500); the estimated incidence rate was 22.8 per 100,000 population (95% CI, 19.5-26.1). Forty-five percent of infections were among black individuals and 53% among men who have sex with men. The back-calculation (n = 1.230 million HIV/AIDS cases reported by the end of 2006) yielded an estimate of 55,400 (95% CI, 50,000-60,800) new infections per year for 2003-2006 and indicated that HIV incidence increased in the mid-1990s, then slightly declined after 1999 and has been stable thereafter. CONCLUSIONS: This study provides the first direct estimates of HIV incidence in the United States using laboratory technologies previously implemented only in clinic-based settings. New HIV infections in the United States remain concentrated among men who have sex with men and among black individuals. PMID- 18677025 TI - Outcomes of nevirapine- and efavirenz-based antiretroviral therapy when coadministered with rifampicin-based antitubercular therapy. AB - CONTEXT: Rifampicin-based antitubercular therapy reduces the plasma concentrations of nevirapine and efavirenz. The virological consequences of these interactions are not well described. OBJECTIVE: To assess the effectiveness and tolerability of concomitant efavirenz- or nevirapine-based combination antiretroviral therapy and rifampicin-based antitubercular therapy. DESIGN, SETTING, AND PARTICIPANTS: Cohort analysis of prospectively collected routine clinical data in a community-based South African antiretroviral treatment program. Antiretroviral treatment-naive adults enrolled between May 2001 and June 2006 were included in the analysis, and were followed up until the end of 2006. INTERVENTIONS: Patients starting antiretroviral therapy with or without concurrent antitubercular therapy received either efavirenz or nevirapine at standard doses. Patients developing tuberculosis while taking antiretroviral therapy that included nevirapine were either changed to efavirenz or continued taking nevirapine. MAIN OUTCOME MEASURES: Viral load of 400 copies/mL or more after 6, 12, and 18 months of antiretroviral therapy; time to the first viral load of 400 copies/mL or more; time to confirmed virological failure (2 consecutive values > or = 5000 copies/mL); time to death; and time to treatment limiting toxicity were assessed. RESULTS: The analysis included 2035 individuals who started antiretroviral therapy with efavirenz (1074 with concurrent tuberculosis) and 1935 with nevirapine (209 with concurrent tuberculosis). There were no differences in time to death or substitution of either antiretroviral drug for toxicity with and without concurrent tuberculosis. Patients starting nevirapine with concurrent tuberculosis were at a higher risk of elevated viral load most notably at 6 months (16.3%; 95% confidence interval [CI], 10.6%-23.5%) than those without tuberculosis (8.3%; 95% CI, 6.7%-10.0%; adjusted odds ratio [OR], 2.1; 95% CI, 1.2-3.4; and in the combined estimate, adjusted OR, 1.7; 95% CI, 1.2-2.6). In the time-to-event analysis of confirmed virological failure (2 consecutive values of > or = 5000 copies/mL), patients starting nevirapine with concurrent tuberculosis developed virological failure sooner (adjusted hazard ratio [HR] 2.2; 95% CI, 1.3-3.7). There were no differences between patients starting efavirenz with and without concurrent tuberculosis (adjusted OR, 1.1; 95% CI, 0.8-1.5 [combined estimate] and adjusted HR, 1.1; 95% CI, 0.6-2.0, respectively). There was no difference in time to virological rebound in patients free of tuberculosis and those developing tuberculosis during follow-up while taking nevirapine (adjusted HR, 1.0; 95% CI, 0.5-2.0) or efavirenz (adjusted HR, 0.8; 95% CI, 0.4-1.7). CONCLUSION: In this cohort study, virological outcomes were inferior when nevirapine-based antiretroviral therapy was commenced while taking antitubercular treatment (vs without concurrent tuberculosis) but comparable when starting efavirenz-based antiretroviral therapy (vs without concurrent tuberculosis) or when tuberculosis developed while taking established nevirapine- or efavirenz-based therapies. PMID- 18677026 TI - Risk factors for recent HIV infection in Uganda. AB - CONTEXT: Studies of factors associated with acquiring human immunodeficiency virus (HIV) are often based on prevalence data that might not reflect recent infections. OBJECTIVE: To determine demographic, biological, and behavioral factors for recent HIV infection in Uganda. DESIGN AND SETTING: Nationally representative household survey of cross-sectional design conducted in Uganda from August 2004 through January 2005; data were analyzed until November 2007. PARTICIPANTS: There were 11,454 women and 9905 men aged 15 to 59 years who were eligible. Questionnaires were completed for 10,826 women (95%) and 8830 men (89%); of those interviewed, blood specimens were collected for 10,227 women (94%) and 8298 men (94%). MAIN OUTCOME MEASURE: Specimens seropositive for HIV were tested with the BED IgG capture-based enzyme immunosorbent assay to identify recent seroconversions (median, 155 days) using normalized optical density of 0.8 and adjustments. RESULTS: Of the 1023 HIV infections with BED results, 172 (17%) tested as recent. In multivariate analysis, risk factors associated with recent HIV infection included female sex (adjusted odds ratio [aOR], 2.4; 95% confidence interval [CI], 1.1-5.2); current marital status (widowed vs never married, aOR, 6.1; 95% CI, 2.8-13.3; divorced vs never married, aOR, 3.0; 95% CI, 1.5-6.1); geographic region (north central Uganda vs central Uganda/Kampala, aOR, 2.6; 95% CI, 1.7-4.1); number of sex partners in past year (> or = 2 compared with none; aOR, 2.9; 95% CI, 1.6-5.5); herpes simplex virus type 2 infection (aOR, 3.9; 95% CI, 2.6-5.8); report of a sexually transmitted disease in the past year (aOR, 1.7; 95% CI, 1.2-2.4); and being an uncircumcised man (aOR, 2.5; 95% CI, 1.1 5.3). Among married participants, recent HIV infection was associated with never using condoms with partners outside of marriage (aOR, 3.2; 95% CI, 1.7-6.1) compared with individuals having no outside partners. The risk of incident HIV infection for married individuals who used condoms with at least 1 outside partner was similar to that of those who did not have any partners outside of marriage (aOR, 1.0; 95% CI, 0.3-2.7). CONCLUSION: A survey of individuals in Uganda who were tested with an HIV assay used to establish recent infection identified risk factors, which offers opportunities for prevention initiatives. PMID- 18677027 TI - Highly active antiretroviral therapy and survival in HIV-infected injection drug users. AB - CONTEXT: Highly active antiretroviral therapy (HAART) is often withheld from injection drug users (IDUs) infected with the human immunodeficiency virus (HIV) based on the belief that their unstable lifestyles may predetermine a markedly inferior outcome with HAART. However, long-term evaluations of HIV treatment outcomes among IDUs in comparison with other risk groups are not available. OBJECTIVE: To compare survival rates among HIV-infected patients initiating HAART with and without a history of injection drug use. DESIGN, SETTING, AND PATIENTS: Population-based, prospective cohort study (HAART Observational Medical Evaluation and Research [HOMER]) of 3116 antiretroviral-naive HIV-infected patients in a province-wide HIV/AIDS treatment program in British Columbia, Canada. Of the 3116 patients, 915 were IDUs (29.4%), 579 were female (18.6%), and the median age was 39.4 years (interquartile range, 33.3-46.4 years). Treatment with HAART was initiated between August 1, 1996, and June 30, 2006. The median duration of follow-up was 5.3 years (interquartile range, 2.8-8.3 years) for IDUs and 4.3 years (interquartile range, 2.0-7.6 years) for non-IDUs. Patients were followed up until June 30, 2007. Data were analyzed between November 1, 2007, and May 26, 2008. MAIN OUTCOME MEASURE: All-cause mortality. RESULTS: Overall, 622 individuals died (20.0%) during the study period (232 IDUs and 390 non-IDUs), for a crude mortality rate of 20.0% (95% confidence interval [CI], 18.4%-21.5%). At 84 months after the initiation of HAART, the product limit estimate of the cumulative all-cause mortality rate was similar between the 915 IDUs (26.5%; 95% CI, 23.2%-29.8%) and 2201 non-IDUs (21.6%; 95% CI, 16.9%-26.2%) (Wilcoxon P = .47). In multivariate time-updated Cox regression, the hazard ratio of mortality was similar between IDUs and non-IDUs (1.09; 95% CI, 0.92-1.29). CONCLUSION: In this study population, injection drug use was not associated with decreased survival among HIV-infected patients initiating HAART. PMID- 18677028 TI - Antiretroviral treatment of adult HIV infection: 2008 recommendations of the International AIDS Society-USA panel. AB - CONTEXT: The availability of new antiretroviral drugs and formulations, including drugs in new classes, and recent data on treatment choices for antiretroviral naive and -experienced patients warrant an update of the International AIDS Society-USA guidelines for the use of antiretroviral therapy in adult human immunodeficiency virus (HIV) infection. OBJECTIVES: To summarize new data in the field and to provide current recommendations for the antiretroviral management and laboratory monitoring of HIV infection. This report provides guidelines in key areas of antiretroviral management: when to initiate therapy, choice of initial regimens, patient monitoring, when to change therapy, and how best to approach treatment options, including optimal use of recently approved drugs (maraviroc, raltegravir, and etravirine) in treatment-experienced patients. DATA SOURCES AND STUDY SELECTION: A 14-member panel with expertise in HIV research and clinical care was appointed. Data published or presented at selected scientific conferences since the last panel report (August 2006) through June 2008 were identified. DATA EXTRACTION AND SYNTHESIS: Data that changed the previous guidelines were reviewed by the panel (according to section). Guidelines were drafted by section writing committees and were then reviewed and edited by the entire panel. Recommendations were made by panel consensus. CONCLUSIONS: New data and considerations support initiating therapy before CD4 cell count declines to less than 350/microL. In patients with 350 CD4 cells/microL or more, the decision to begin therapy should be individualized based on the presence of comorbidities, risk factors for progression to AIDS and non-AIDS diseases, and patient readiness for treatment. In addition to the prior recommendation that a high plasma viral load (eg, >100,000 copies/mL) and rapidly declining CD4 cell count (>100/microL per year) should prompt treatment initiation, active hepatitis B or C virus coinfection, cardiovascular disease risk, and HIV-associated nephropathy increasingly prompt earlier therapy. The initial regimen must be individualized, particularly in the presence of comorbid conditions, but usually will include efavirenz or a ritonavir-boosted protease inhibitor plus 2 nucleoside reverse transcriptase inhibitors (tenofovir/emtricitabine or abacavir/lamivudine). Treatment failure should be identified and managed promptly, with the goal of therapy, even in heavily pretreated patients, being an HIV-1 RNA level below assay detection limits. PMID- 18677029 TI - Mexico's evolving HIV epidemic. PMID- 18677030 TI - Universal access to HIV/AIDS treatment: promise and problems. PMID- 18677031 TI - Balancing disclosure of diagnosis and assent for research in children with HIV. PMID- 18677032 TI - The case against criminalization of HIV transmission. PMID- 18677033 TI - The past, present, and future of AIDS. PMID- 18677034 TI - JAMA patient page. HIV infection: the basics. PMID- 18677036 TI - Analysis of effects of posture on organ doses by internal photon emitters using voxel phantoms. AB - In order to clarify the effects of posture on absorbed doses in organs due to internal exposures, specific absorbed fractions (SAFs) for monoenergetic photons from 0.01 to 4 MeV were calculated using two Japanese adult male phantoms, JM2 (upright) and JM (supine), whose designs were based on CT images of two postures obtained from the same person. Differences in the SAFs of JM2 and JM are within a factor of 2 in most cases although significant differences in SAFs exist in several combinations of source and target organs for low energy photons. Organ doses due to photons from the intake of unit activity of eight radionuclides (126Ba, 128Ba, 133Ba, 130Cs, 131Cs, 178W, 179W and 190W) with different biokinetics and decay characteristics were calculated from the SAFs of JM2 and JM. It was found that the organ doses in the testes, bone marrow, lower large intestine wall, lungs and stomach wall, which are important for calculating effective dose, in the two postures roughly agree, the maximum difference being 14%. These results indicate that the impact of posture on the organ doses and effective doses from internal photon emitters is not significant. PMID- 18677035 TI - The epidermal growth factor homology domain of the LDL receptor drives lipoprotein release through an allosteric mechanism involving H190, H562, and H586. AB - The low density lipoprotein (LDL) receptor (LDLR) mediates efficient endocytosis of VLDL, VLDL remnants, and LDL. As part of the endocytic process, the LDLR releases lipoproteins in endosomes. The release process correlates with an acid dependent conformational change in the receptor from an extended, "open" state to a compact, "closed" state. The closed state has an intramolecular contact involving H190, H562, and H586. The current model for lipoprotein release holds that protonation of these histidines drives the conformational change that is associated with release. We tested the roles of H190, H562, and H586 on LDLR conformation and on lipoprotein binding, uptake, and release using variants in which the three histidines were replaced with alanine (AAA variant) or in which the histidines were replaced with charged residues that can form ionic contacts at neutral pH (DRK variant). Contrary to expectation, both the AAA and the DRK variants exhibited normal acid-dependent transitions from open to closed conformations. Despite this similarity, both the AAA and DRK mutations modulated lipoprotein release, indicating that H190, H562, and H586 act subsequent to the conformational transition. These observations also suggest that the intramolecular contact does not drive release through a competitive mechanism. In support of this possibility, mutagenesis experiments showed that beta-VLDL binding was inhibited by mutations at D203 and E208, which are exposed in the closed conformation of the LDLR. We propose that H190, H562, and H586 are part of an allosteric mechanism that drives lipoprotein release. PMID- 18677037 TI - Fractional-order viscoelasticity applied to describe uniaxial stress relaxation of human arteries. AB - Viscoelastic models can be used to better understand arterial wall mechanics in physiological and pathological conditions. The arterial wall reveals very slow time-dependent decays in uniaxial stress-relaxation experiments, coherent with weak power-law functions. Quasi-linear viscoelastic (QLV) theory was successfully applied to modeling such responses, but an accurate estimation of the reduced relaxation function parameters can be very difficult. In this work, an alternative relaxation function based on fractional calculus theory is proposed to describe stress relaxation experiments in strips cut from healthy human aortas. Stress relaxation (1 h) was registered at three incremental stress levels. The novel relaxation function with three parameters was integrated into the QLV theory to fit experimental data. It was based in a modified Voigt model, including a fractional element of order alpha, called spring-pot. The stress relaxation prediction was accurate and fast. Sensitivity plots for each parameter presented a minimum near their optimal values. Least-squares errors remained below 2%. Values of order alpha = 0.1-0.3 confirmed a predominant elastic behavior. The other two parameters of the model can be associated to elastic and viscous constants that explain the time course of the observed relaxation function. The fractional-order model integrated into the QLV theory proved to capture the essential features of the arterial wall mechanical response. PMID- 18677038 TI - Quantitative analysis of water proton spectral lineshape: a novel source of contrast in MRI. AB - Previous work in this laboratory has demonstrated improved anatomic and functional images produced from high spectral and spatial resolution (HiSS) MRI of the water proton signal. The present work tests the hypothesis that different Fourier components of the water resonance represent anatomically and/or physiologically distinct populations of water molecules within each small image voxel. HiSS datasets were acquired from tomatoes and rodent tumors at 4.7 T using echo-planar spectroscopic imaging (spatial and spectral resolutions were 117-150 microm and 1.5-3.1 Hz, respectively). Images of each Fourier component of the water resonance (referred to as Fourier component images, or FCIs) were produced. FCIs at frequencies offset from the peak of the water resonance ('off-peak' FCIs) were compared to images of the Fourier component with largest amplitude, i.e. the water peak-height image. Results demonstrate that off-peak FCIs differ significantly from the water peak-height image and that water resonances are often asymmetric. These results show that water signal at various frequency offsets from the peak of the water resonance come from water molecules in different anatomic/physiologic environments. Off-peak FCIs are a new source of structural and functional information and may have clinical utility. PMID- 18677039 TI - Efficient Monte Carlo modelling of individual tumour cell propagation for hypoxic head and neck cancer. AB - A Monte Carlo tumour model has been developed to simulate tumour cell propagation for head and neck squamous cell carcinoma. The model aims to eventually provide a radiobiological tool for radiation oncology clinicians to plan patient treatment schedules based on properties of the individual tumour. The inclusion of an oxygen distribution amongst the tumour cells enables the model to incorporate hypoxia and other associated parameters, which affect tumour growth. The object oriented program FORTRAN 95 has been used to create the model algorithm, with Monte Carlo methods being employed to randomly assign many of the cell parameters from probability distributions. Hypoxia has been implemented through random assignment of partial oxygen pressure values to individual cells during tumour growth, based on in vivo Eppendorf probe experimental data. The accumulation of up to 10 million virtual tumour cells in 15 min of computer running time has been achieved. The stem cell percentage and the degree of hypoxia are the parameters which most influence the final tumour growth rate. For a tumour with a doubling time of 40 days, the final stem cell percentage is approximately 1% of the total cell population. The effect of hypoxia on the tumour growth rate is significant. Using a hypoxia induced cell quiescence limit which affects 50% of cells with and oxygen levels less than 1 mm Hg, the tumour doubling time increases to over 200 days and the time of tumour growth for a clinically detectable tumour (10(9) cells) increases from 3 to 8 years. A biologically plausible Monte Carlo model of hypoxic head and neck squamous cell carcinoma tumour growth has been developed for real time assessment of the effects of multiple biological parameters which impact upon the response of the individual patient to fractionated radiotherapy. PMID- 18677040 TI - Sensitivity of different dose scoring methods on organ-specific neutron dose calculations in proton therapy. AB - Scattered doses, e.g. neutron doses in proton therapy, are of concern in radiation therapy. Although measured data are the gold standard, Monte Carlo simulations allow a more realistic consideration of patient anatomy via whole body phantoms. When calculating neutron doses with Monte Carlo techniques, the dose can be scored in different ways because neutrons deposit dose indirectly. The purpose of this study was to assess the differences in neutron dose predictions when using different dose scoring methods. Two methods were tested. In the first method, the organ dose was calculated by accumulating dose from each individual dose deposition event with a particle-specific radiation weighting factor applied. Alternatively, we applied a method where the calculation was done by averaging the dose over the total number of events irrespective of particle type and applying average neutron radiation weighting factors. In addition, we assessed the sensitivity of different neutron quality factor assignments based on two recommendations by the International Commission on Radiological Protection (ICRP). We found that the scoring procedure can lead to differences in the organ equivalent dose of about 25%. As to the ICRP definition of neutron quality factors, the most recent recommendation results in about 10% higher organ doses. PMID- 18677041 TI - Auto-propagation of contours for adaptive prostate radiation therapy. AB - The purpose of this work is to develop an effective technique to automatically propagate contours from planning CT to cone beam CT (CBCT) to facilitate CBCT guided prostate adaptive radiation therapy. Different from other disease sites, such as the lungs, the contour mapping here is complicated by two factors: (i) the physical one-to-one correspondence may not exist due to the insertion or removal of some image contents within the region of interest (ROI); and (ii) reduced contrast to noise ratio of the CBCT images due to increased scatter. To overcome these issues, we investigate a strategy of excluding the regions with variable contents by a careful design of a narrow shell signifying the contour of an ROI. For rectum, for example, a narrow shell with the delineated contours as its interior surface was constructed to avoid the adverse influence of the day-to day content change inside the rectum on the contour mapping. The corresponding contours in the CBCT were found by warping the narrow shell through the use of BSpline deformable model. Both digital phantom experiments and clinical case testing were carried out to validate the proposed ROI mapping method. It was found that the approach was able to reliably warp the constructed narrow band with an accuracy better than 1.3 mm. For all five clinical cases enrolled in this study, the method yielded satisfactory results even when there were significant rectal content changes between the planning CT and CBCT scans. The overlapped area of the auto-mapped contours over 90% to the manually drawn contours is readily achievable. The proposed approach permits us to take advantage of the regional calculation algorithm yet avoiding the nuisance of rectum/bladder filling and provide a useful tool for adaptive radiotherapy of prostate in the future. PMID- 18677043 TI - [Daily practice using guidelines for prevention and treatment of osteoporosis. Risk factors for fracture and guideline for prevention of fracture]. AB - In 2006, a set of guidelines was released in Japan regarding the initiation of medical treatment to prevent fragility fracture with risk factors considered. In February this year, a WHO working group announced the development of the Fracture Risk Assessment Tool (FRAX), which estimates fracture risk based on age, sex, bone density at the femoral neck (body mass index if bone density is not available), previous fragility fracture in adulthood, parental fracture history at the femoral neck, current smoking, steroid use, secondary osteoporosis/rheumatoid arthritis, and alcohol consumption. And then, the NOF released guidelines incorporating FRAX in the US. WHO recommends that the threshold of medical treatment should be set based on each country's medical circumstances and healthcare economic situation. PMID- 18677044 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Fracture risk assessment with Japanese guideline]. AB - The diagnostic criteria of osteoporosis in Japan is comprised of low bone mineral density and presence of fragile fractures as risk factors for further fractures. In addition, the importance of differential diagnosis is emphasized in this criteria. The risk factors for fragile fractures independent from low mineral density and the presence of fragile fractures were included in the criteria for pharmacotherapy in the 2006 edition of guideline for prevention and treatment of osteoporosis. These factors include smoking, excessive alcohol, and familial history of femoral neck fractures. It would be a research question whether the removal of lifestyle-related risk factors are related to the reduction of fracture risk. PMID- 18677042 TI - An imaging-based tumour growth and treatment response model: investigating the effect of tumour oxygenation on radiation therapy response. AB - A multiscale tumour simulation model employing cell-line-specific biological parameters and functional information derived from pre-therapy PET/CT imaging data was developed to investigate effects of different oxygenation levels on the response to radiation therapy. For each tumour voxel, stochastic simulations were performed to model cellular growth and therapeutic response. Model parameters were fitted to published preclinical experiments of head and neck squamous cell carcinoma (HNSCC). Using the obtained parameters, the model was applied to a human HNSCC case to investigate effects of different uniform and non-uniform oxygenation levels and results were compared for treatment efficacy. Simulations of the preclinical studies showed excellent agreement with published data and underlined the model's ability to quantitatively reproduce tumour behaviour within experimental uncertainties. When using a simplified transformation to derive non-uniform oxygenation levels from molecular imaging data, simulations of the clinical case showed heterogeneous tumour response and variability in radioresistance with decreasing oxygen levels. Once clinically validated, this model could be used to transform patient-specific data into voxel-based biological objectives for treatment planning and to investigate biologically optimized dose prescriptions. PMID- 18677045 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Bone mineral measurements and guideline]. AB - We reviewed a role of BMD measurements in Japanese Guidelines for the Prevention and Treatment of Osteoporosis. The BMD measurements are necessary in the diagnosis of osteoporosis, the determination of initiation of drug therapy, and the evaluation of fracture risk and the therapeutical response. It is important to understand not only the method and the site of the measurement but also the characteristics. On EBM, axial DXA, as the site and the method of BMD measurements, is recommended to all women >or= 65 y.o., and women <65 y.o., with risk factor. PMID- 18677046 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Osteoporosis prevention and use of the new management guideline for osteoporosis]. AB - Now that the goal of osteoporosis management has been clearly defined as prevention of osteoporosis-associated fractures, prevention strategy for these fractures becomes compellingly important. While the new management guideline for osteoporosis places an overriding emphasis on the prevention of osteoporosis, it goes on to stress the importance of pharmacological therapy even in patients at high risk of developing fractures, in whom pharmacological therapy would not be indicated in the earlier guideline, thus recommending that pharmacological therapy be extended to a larger proportion of patients who may benefit from such treatment. In this therapeutic strategy, not only pharmacological therapy in middle-aged and elderly individuals in perimenopause will be of vital importance, as in the earlier guideline, but also non-pharmacological intervention through modification of everyday lifestyle habits will become increasingly important in adolescents and young female adults in their developmental years. It is thus hoped that adherence to the new guideline will lead to a decrease in the frequency of reported fractures, including femoral neck fractures, in Japan. PMID- 18677047 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. How do we realize the bone quality in routine practice using Japanese guideline for osteoporosis prevention and treatment?]. AB - Bone quality is thought to encompass the structural and material properties of bone that are affected by turnover rate. The concept of bone quality is included in Japanese Guideline for Osteoporosis prevention and treatment. Evidence has accumulated that collagen cross-links play important roles in bone strength. We have demonstrated that the quantitative and qualitative deterioration of lysyl oxidase controlled and non-enzymatic cross-links (Advanced glycation end products, AGEs, Pentosidine) of collagen in patients with osteoporotic femoral neck fracture cases might be affected by hyperhomocysteinemia (Saito M, Calcif Tissue Int, 2006), oxidative stress, vitamin B status (Saito M, Osteoporos Int, 2006) . Recently, Shiraki et al. demonstrated that a functional polymorphism in methylenetetrahydrofolate reductase (MTHFR) polymorphism, T allele (C677T), may be a risk factor for future fracture in addition to the traditional risk factors (Shiraki M, Saito M, et al., J Bone Miner Metab, in press). In addition, we have reported that a higher urinary pentosidine was an independent risk factor, for vertebral fracture in a 5-year prospective study in Japanese women (Shiraki M, Saito M, et al., J Bone Miner Metab, 2008). If confirmed in large, prospective trials, measurement of serum homocysteine and serum or urinary excretion of pentosidine might be characterized as markers reflecting bone collagen deterioration. PMID- 18677048 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Proximal femoral BMD and measurement manual]. AB - The diagnosis of osteoporosis is made from the measurement of BMD. Since DXA at the hip best predicts hip fracture among the various methods of bone densitometry, hip DXA is currently recommended as the standard method for the diagnosis of osteoporosis in Europe and USA. In Japan, hip DXA has not been common because of difficulty in the measurement due to the short neck axis length compared with Caucasian. However, because of the recent development in DXA technology and the acknowledgment of the importance of hip BMD, hip DXA has recently garnered attention in Japan. In this paragraph, general guideline and points of attention in hip DXA have been summarized. PMID- 18677049 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Semi-quantitative method in the assessment of vertebral fracture]. AB - Assessment of vertebral fracture based on a thoraco-lumbar spine radiographs is a good tool to evaluate the risk of incident fracture. Presence of vertebral fracture indicates 4 to 7 fold increase of risk of future fracture. However, vertebral fracture are often underdiagnosed. Quantitative (QM) and semi quantitative (SQ) methods are designed for the assessment of prevalent and incident fracture. The advantages of SQ method are more convenient and reproducible as well as more excellent in the assessment of the risk of future fracture compared to QM. It should be recognized that untrained SQ reader produce a high false negative of grade 1 (mild fracture). PMID- 18677050 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Utilization of metabolic markers of bone in daily osteoporosis practice]. AB - The evaluation of metabolic state is thought to be useful for the risk evaluation of fractures and early evaluation of treatment effects. The evaluation may also be useful to maintain adherence to osteoporosis treatment, especially in patients without symptoms. The most markers are reimbursed, although number and timing of the evaluation are limited. The guidelines are published, and it is common practice to evaluate metabolic conditions of osteoporotic patients. They are especially useful to decision making for treatment not only in patients but also in physicians. The follow-up evaluation of the markers is useful in patients treated with bisphosphonate, raloxifene and/or vitamin K2 to maintain adherence. PMID- 18677051 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Use of clinical risk factors for osteoporotic fractures in the evaluation of risk of future fracture]. AB - A number of clinical risk factors that provide information on fracture risk over and above that given by BMD have been defined. WHO proposed FRAX (fracture risk assessment tool) in which the fracture probability could be calculated by the use of risk factors with or without BMD. The proposed independent risk factors are age, a prior fragility fracture, a parental history of hip fracture, smoking, use of systemic corticosteroids, excess alcohol intake, secondary osteoporosis, and rheumatoid arthritis. Management algorithms in postmenopausal women based on an health analysis have been proposed in several countries including UK, Sweden, Germany, USA and Canada. PMID- 18677052 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Recent evidence of estrogen therapy for osteoporosis]. AB - Results of WHI hormone trial have given the strong restriction on HRT. But recently the conception of adverse events of HRT is getting changed. Generally, it is considered that HRT under 60 years aged or early menopausal women make many benefits without elevation of risk of adverse events and selection of dose level or way to apply can also vary the risk of disease. Because any kind of estrogens has exact effects suppressing bone resorption and fracture, we can use them for osteopenia or osteoporotic women in climacteric state cost-effectively by taking age, dose level and application methods into consideration. PMID- 18677053 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Association between height loss and QOL in the elderly]. AB - Medical care of osteoporosis aims at preventing bone fractures and thereby maintaining and improving the quality of life (QOL). It was observed in some studies that height loss and vertebral fracture had an independent, significant impact on QOL in the elderly. It had been believed thus far that the phenomenon of height loss was attributable mainly to incurred spinal fracture and that it had no clinical value other than as a sign of osteoporosis, but some studies revealed that height loss was also a factor affecting QOL independently of vertebral fracture. PMID- 18677054 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Anti-diabetic agents and fracture risk]. AB - In the developed countries, three major diseases are diabetes, dyslipidemia, and hypertension. The associations between osteoporosis and the latter two conditions are unclear, however, diabetes has been linked to an increased fracture risk, independent of bone mineral density. Recently, thiazolidinediones (TZD) , a new class of anti-diabetic agent which has promising anti-atherogenic effects, has been linked to an increased fracture risk. In this review, effects of TZD on bone metabolism and fracture risk will be updated. PMID- 18677055 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Effectiveness of exercise for preventing and treating osteoporosis]. AB - There is increasing evidence that exercise is an effective strategy for the prevention and treatment of osteoporosis. The randomized controlled trials and their meta-analyses to date, evaluating the effects of exercise on osteoporosis reveal that the exercise is effective in preserving bone mass, preventing fractures and falls, and improving quality of life in patients with osteoporosis. Emphasis is also given to the importance of the specific protocols of exercises needed to achieve positive effects safely, keeping in view the age and general physical condition of the person. PMID- 18677056 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. The effects of anti-TNF therapy on bone and joint manifestations in rheumatoid arthritis]. AB - Patients diagnosed as RA should be started with disease modifying anti-rheumatic drugs (DMARDs) as early as possible. Among the DMARDs, methotrexate (MTX) is considered the anchor drug and should be used first in patients as risk of developing persistent disease. The main goal of DMARDs treatment is to achieve remission and monitoring of disease activity and adverse events should guide decisions on choice and changes in treatment strategies. However, treatment of RA with DMARDs including MTX often fails to control disease activity and prevent structural damage in a some population of patients. Therefore, more effective treatment strategies are needed. Tumor necrosis factor-alpha (TNF-alpha) , a representative pro-inflammatory cytokine, plays a pivotal role in the pathological process of RA by mediating initiation to autoimmunity, lymphocyte accumulation and angiogenesis in the inflamed synovium, and joint destruction. Treatment of RA patients with TNF inhibitors have been efficacious for a) a prolonged improvement in disease activity and an induction to remission, b) inhibition of radiographic progression, when they are used in combination with MTX. Thus, the combined use of the TNF-inhibitor and MTX has brought about a paradigm shift in the treatment goal of RA. However, their small but definite risks of serious infections are also clear and their occurrence mandates that TNF inhibitors should be supervised by physicians with experience in their use. PMID- 18677057 TI - [Daily practice using the guidelines for prevention and treatment of osteoporosis. Evolution of bisphosphonate therapy. Intravenous injection]. AB - Injectable formulation of bisphosphonate (pamidronate, alendronate, incadronate, and zoledronate) is available only for therapy of malignancy in Japan. Ibandronate and zoledronate are permitted to use by injection in the treatment of osteoporosis in US and Europe and have shown a significant effect in fracture prevention. Recently, it was reported that an annual infusion of zoledronate was associated with improved survival in addition to a reduction in the rate of new fractures. Injectable formulation of ibandronate is now in the 3rd phase of clinical trial for fracture prevention in Japan. It is expected that patients with a high fracture risk who could not take bisphosphonate orally receive the benefit of bisphosphonate through injection. PMID- 18677058 TI - [Bone disease with Pain. Non-traumatic osteonecrosis of the femoral head]. AB - The etiology and pathogenesis of non-traumatic osteonecrosis of the femoral head is still unknown although many studied has been performed. Using Magnetic resonance imaging, decision of the extent and location of the necrotic lesion, exist of bone marrow oedema has been developed. The revised criteria for diagnosis, classification, and staging of the disease from specific disease investigation committee of Japanese ministry of health, labor and welfare was established. As joint preservation operation, transtrochanteric rotational osteotomy, curved varus osteotomy were proved to be effective. Hip arthroplasty for maintaining bone stock has been developed for the treatment of young patients. PMID- 18677059 TI - [Bone mass in puberty]. AB - Bone mass increases as the pubertal sex hormones secretion increases, and it reaches a peak of a maximal bone mass at approximately the age of 18 years in females and 20 years in males. We developed standard values for speed of sound (SOS) with ultrasonic measurement of the tibia in children and adolescents, a method to assess cortical bone. As a result, SOS of the tibia increased in an age dependent manner in both sexes. The maximal increase was observed approximately 1 year after the age of peak height velocity, which was reported to be 11 years and 13 years in females and males, respectively. During the time when body height increases rapidly, there is a period of relative bone fragility in which bone strength gain is delayed against an increase in length of tibia. Appropriate physical activity and nutrient intake during puberty can have an excessive influence on the health of bone. PMID- 18677060 TI - Structural inequalities among social groups continue to result in unequal rates of health and mortality, and these inequalities increasingly disfavor middle income as well as low-income people. A note from the editor. PMID- 18677061 TI - International medical graduates contribute to the diversity of the U.S. physician workforce. PMID- 18677062 TI - Responding to domestic violence: a student's perspective. PMID- 18677063 TI - Who speaks for the health consumer? AB - Although consumer-directed health care has become a fashionable concept in recent years, stories abound asking whether the so-called free market in health care can provide adequate access to quality health care at an affordable price. In spite of these concerns, consumer-directed health care continues as the face of legitimacy behind an industry-driven campaign to limit regulatory protections of the consumer in the market and encourage the growth of health insurance products that place spending options closer to the consumer, whether or not these options are available, affordable, or easily understood. Understanding whether this empowerment is real begins with first asking what it now means to be a health consumer. This commentary offers perspective on the dilemma faced by millions of Americans in navigating our health care system under the assumption that market driven choices foster consumer empowerment in health care, and suggests approaches for expanding the true consumer voice. PMID- 18677064 TI - Diabetes mellitus medication assistance program: relationship of effectiveness to adherence. AB - This retrospective study examines the effect of a medication assistance program (MAP) on HbA1c levels in an uninsured, low-income, type 2 diabetes population. It also examines the degree to which improvement in HbA1c level varied with adherence to medication regimens among those patients using the MAP. The MAP was found to have a mean effect of -0.60% on HbA1c levels. However, MAP users differed in how strictly they adhered to medication regimens, as measured by number of refill opportunities taken. The MAP's effect on HbA1c varied monotonically with adherence level, with greater adherence leading to greater HbA1c improvement. Never refilling the prescription (complete nonadherence) led to no change in HbA1c, while complete adherence led to an estimated -0.88% improvement in HbA1c. Further study is needed to investigate factors related to non-adherence within medication assistance programs and the effect of such programs on other patient outcomes. PMID- 18677065 TI - Access to appointments based on insurance status in Washington, D.C. AB - In the U.S., emergency departments see millions of patients requiring timely and adequate outpatient follow-up. Using a hypothetical patient presenting with a scripted presentation of hypertension evaluated in the emergency department and requiring close outpatient follow-up, randomly selected providers in the District of Columbia were queried by phone to evaluate their accessibility based on insurance status. Seventy one percent of calls for privately insured hypothetical patients resulted in a successful appointment. Medicaid fee-for-service was associated with a significantly lower rate of successful appointments (36.6%) than private insurance (p=.002.) Uninsured "patients" had only a 13% success rate when considering out-of-pocket payment limitations of less than $50 (p<.001 compared with private). Access to primary care follow-up is poor in the District of Columbia for all types of insurance; however people with Medicaid fee-for service and the uninsured face especially strong barriers. PMID- 18677066 TI - Perceptions of race/ethnicity-based discrimination: a review of measures and evaluation of their usefulness for the health care setting. AB - BACKGROUND: To assess discrimination in health care, reliable, valid, and comprehensive measures of racism/discrimination are needed. OBJECTIVE: To review literature on measures of perceived race/ethnicity-based discrimination and evaluate their characteristics and usefulness in assessing discrimination from health care providers. METHODS: Literature review of measures of perceived race/ethnicity-based discrimination (1966-2007), using MEDLINE, PsycINFO, and Social Science Citation Index. RESULTS: We identified 34 measures of racism/discrimination; 16 specifically assessed dynamics in the health care setting. Few measures were theoretically based; most assessed only general dimensions of racism and focused specifically on the experiences of African American patients. Acceptable psychometric properties were documented for about half of the instruments. CONCLUSIONS: Additional measures are needed for detailed assessments of perceived discrimination in the health care setting; they should be relevant for a wide variety of racial/ethnic groups, and they must assess how racism/discrimination affects health care decision making and treatments offered. PMID- 18677067 TI - Access to health care and voting behavior in the United States. AB - This study examines the relationship between difficulties accessing health care and voting behavior, in order to assess the possible impact that increasing constraints on access to care will have on future voting behavior. Using data from the American National Election Study we found that the proportion of people with difficulty accessing care increased significantly from 27% in 2000 to 35% in 2004. A larger proportion of those with difficulties in accessing care voted in 2004 than in 2000 and their preferences also changed to more heavily favor the Democratic candidate. If the number of those experiencing difficulties accessing care continues to grow, access to care could become a more salient campaign issue. In 2004, Democratic candidates were favored by this group, which is likely to be a constituency that both parties will try to capture in future elections. PMID- 18677068 TI - Primary care for low-income populations: comparing health care delivery systems. AB - Amidst recent policy discussions about the health care safety net there has been relatively little information about whether the actual site of care affects care quality. We therefore used National Health Interview Survey data to describe low income adults seeking primary care at different types of sites and the quality of access and preventive care at these sites. After adjusting for sociodemographic characteristics and illness burden, hospital-outpatient- department patients were more likely to receive vaccinations for influenza (adjusted odds ratio [AOR] 1.3, 95% confidence interval [CI] 1.0-1.6) and pneumococcus (AOR 1.4, 95% CI 1.1-1.8) than were those at clinics or health centers. Hospital-clinic patients were more likely to report delays in care due to office administrative difficulties (AOR 1.3, 95% CI 1.1-1.7) and more likely to have more than one emergency room visit (AOR 1.9, 95% CI 1.5-2.3). Physicians' office or HMO patients were less likely to report administrative delays in care than those at clinics or health centers, but there were no other differences in quality between these two site types. Policymakers and health care services analysts and providers must monitor quality as they decide how best to deliver care to vulnerable populations. PMID- 18677069 TI - Poverty and sexual concurrency: a case study of STI risk. AB - This paper is about sexual concurrency, or maintaining multiple sexual partnerships that overlap in time. Sexual concurrency is a concept that is used in the field of public health to explain the spread of sexually transmitted diseases. Sexual concurrency has also been proposed as a site of intervention to reduce sexually transmitted infection (STI) rates, particularly among those populations who carry the heaviest STI burden: adolescents and African Americans. In this paper, we use ethnographic data collected from a group of African American adolescents living in Baltimore to examine the socially produced configurations of risks and relationships that are obscured by the term sexual concurrency. The data we present show the limits of this concept, and suggest that structural reforms, including improvements to education, drug treatment, and work opportunities, are necessary to reduce racial disparities in STI rates. PMID- 18677070 TI - The impact of publicly funded family planning clinic services on unintended pregnancies and government cost savings. AB - Publicly funded family planning clinics serve millions of low-income women each year, providing a range of critical preventive services and enabling women to avoid unintended pregnancies. It is important to quantify the impact and cost effectiveness of such services, in addition to these health benefits. Using a methodology similar to prior cost-benefit analyses, we estimated the numbers of unintended pregnancies prevented by all U.S. publicly funded family planning clinics in 2004, nationally (1.4 million pregnancies) and for each state. We also compared the actual costs of providing these services ($1.4 billion) with the anticipated public-sector costs for maternity and infant care among the Medicaid eligible women whose births were averted ($5.7 billion) to calculate net public sector savings ($4.3 billion). Thus, public expenditures for family planning care not only help women to achieve their childbearing goals, but they also save public dollars: Our calculations indicate that for every $1 spent, $4.02 is saved. PMID- 18677071 TI - Breast and cervical cancer screening among low-income women in Nebraska: findings from the Every Woman Matters program, 1993-2004. AB - OBJECTIVE: To examine the likelihood of enrollees in the Nebraska Every Woman Matters program being screened for breast and cervical cancer. METHODS: We explored the relationship between sociodemographic characteristics and receiving cancer screening services. RESULTS: Older and Native American women were more likely than younger and White women to have mammograms ordered [adjusted odds ratio (OR)=1.41, 95% confidence interval (CI) 1.08, 1.85]. African American [OR=0.54, 95% CI 0.46, 0.64] and Native American women [OR=0.47, 95% CI 0.39, 0.55] were less likely than White women to have clinical breast exams performed. Native American [OR=0.19, 95% CI 0.16, 0.23] and African American women [OR=0.56, 95% CI 0.46, 0.68] were less likely than White women to have a Pap test performed. CONCLUSION: Receiving cancer screening services was related to race; thus, understanding barriers for screening for minority women is warranted. PMID- 18677072 TI - Factors associated with zidovudine receipt and prenatal care among HIV-infected pregnant women in New Jersey. AB - Despite reductions in perinatal HIV transmission, cases continue to occur. To determine factors associated with zidovudine (ZDV) receipt among HIV-infected pregnant women we merged three data sets for women in New Jersey in 1995-1997, identifying 395 HIV-infected pregnant women. Half received two arms of ZDV prophylaxis. Attendance at five or more prenatal visits was the strongest independent factor related to ZDV receipt (OR 6.37, 95% CI 3.84, 10.57). Half (49.0%) had limited prenatal care. AIDS diagnosis, race/ethnicity, and drug use were also independently related to ZDV receipt. Post hoc analysis revealed that being unmarried, Black, multiparous, having no insurance, and illegal drug use were associated with limited prenatal care. Although the U.S. has seen reductions in HIV perinatal transmission, our research showed that HIV-infected women who did not get prenatal care were less likely to receive two arms of ZDV prophylaxis. A wide public health net that brings all women into care is necessary to reduce perinatal transmission further. PMID- 18677073 TI - HIV prevention for Black women: structural barriers and opportunities. AB - Black women bear a disproportionate burden of HIV/AIDS in North America. The purpose of this investigation was to explore Black Canadian women's perspectives on HIV risk and prevention. Four 90-minute focus groups (n=26) and six key informant interviews were conducted in Toronto with Black women of African and Caribbean descent and low socioeconomic status. Data analysis revealed a number of potent barriers to existing HIV preventive interventions: stigma, cultural disconnections, lack of engagement of Black religious institutions, and multiple intersecting forms of discrimination. Recommended HIV prevention opportunities included the Black church, mainstreaming, health care providers, and ethno specific agencies. HIV prevention strategies for North American Black women, rather than focusing on HIV and individual risk behaviors, may benefit from a primary focus on social and structural factors (e.g., promoting gender equality, economic opportunity, women-controlled prevention technologies and combating racism in health care) thereby integrating HIV prevention into the larger context of community health and survival. PMID- 18677074 TI - The role of health insurance coverage in cancer screening utilization. AB - INTRODUCTION: Although previous studies have shown a correlation between health insurance coverage and cancer screening, underinsurance and cancer screening among racial/ethnic minorities has not been examined. METHODS: Data from the 2000 and 2003 National Health Interview Surveys are used in this analysis. Cross tabulations, age, and racial/ethnic group stratified regression analyses are used to examine associations between health insurance status and receipt of mammography, Pap testing, prostate specific antigen tests, fecal occult blood test (FOBT) and colorectal endoscopy. RESULTS: In overall models, uninsurance was associated with lower receipt of all tests except FOBT among participants ages 65 85 years. Underinsurance was associated with lower receipt of mammography among women under 65 years only. CONCLUSION: These findings show age variation in the association between cancer screening and health insurance coverage. In addition, health insurance appears to act similarly across racial/ethnic groups. Further examination of underinsurance in cancer screening utilization and other health behaviors is needed. PMID- 18677075 TI - Mexican immigrant women's perceptions of health care access for stigmatizing illnesses: a focus group study in Albuquerque, New Mexico. AB - This study examines attitudes of Mexican female immigrants to Albuquerque, New Mexico, regarding barriers to health care access in the United States and Mexico for stigmatizing and non-stigmatizing illnesses and moderating effects of social support. Native Spanish speakers conducted three focus groups (in Spanish) lasting two hours with seven to eight participants. Focus groups were transcribed, translated, and coded. Frequency data were calculated by number of times concepts or themes were raised. Comparisons of barriers to health care access were made between U.S. and Mexican cultures. The majority (86%) of comments on barriers for non-stigmatizing illnesses implicated U.S. culture; the majority (90%) for stigmatizing illnesses implicated Mexican culture. Social support for stigmatizing illnesses was discussed. Participants discussed important issues of health care access for stigmatizing illnesses that may have implications for this population's health status. Greater attention should be paid to stigma and social support in future empirical studies. PMID- 18677076 TI - The experience of stigma among Black mental health consumers. AB - Little is known about how stigma affects Black people receiving mental health treatment. For a project to develop a consumer-based stigma intervention, qualitative interviews were conducted with public-sector Black mental health consumers (N=34). Primary themes from the interviews regarding stigma concerns, experiences, and coping strategies were examined. Concerns about stigma prompted most consumers initially to avoid or delay treatment; once in treatment, consumers commonly faced stigmatizing reactions from others. Consumers identified numerous strategies to deal with stigma, including seeking support from accepting members of their existing social networks, and viewing their own health as more important than the reaction of others. These consumer perspectives may be valuable to Black individuals who are contemplating seeking mental health treatment. PMID- 18677077 TI - The association between perceived discrimination and underutilization of needed medical and mental health care in a multi-ethnic community sample. AB - This study examines the association between perceived discrimination and underutilization of needed medical and mental health care, in a representative, multi-ethnic community sample. Data were derived from a cross sectional survey of 10,098 White, U.S.-born Black, African-born Black, American Indian, Hispanic, and Southeast Asian adults in Hennepin County, Minnesota. Even after controlling for socio-demographic characteristics, health care access, and physical and mental health, perceived discrimination was associated with underutilization of medical care among Whites, U.S.-born Blacks, and American Indians and was associated with underutilization of mental health care among Whites, U.S.-born Blacks, Southeast Asians, and American Indians. Correlates of different types of discrimination (major, everyday, health care) on underutilization of care varied among ethnic groups. The higher prevalence of discrimination among racial and ethnic minorities may contribute to their underutilization of health care services. Future research is needed to understand the impacts of different types of discrimination on different groups. PMID- 18677078 TI - Barriers to cervical cancer screening among low-income HIV-positive African American women. AB - This study elucidates the perspective of low-income HIV-positive African American women who have not received cervical cancer screening for five or more years, on the barriers they face in accessing and using reproductive health care. We focused on how women who live in a severely economically depressed and racially segregated neighborhood experience barriers to cervical cancer screening. Andersen's Behavioral Model of Health Services Use, which allows for the organization of conditions and situations that bar utilization of health services, served as the theoretical framework. Findings from individual semi structured interviews with 35 participants revealed the importance of psychological and emotional barriers as well as the more commonly reported economic, social, and health care system barriers. We suggest how access to care for this population can be increased by including psychological and emotional components in intervention efforts. PMID- 18677079 TI - Personal review and justice analysis of nursing experiences in Haiti. AB - Health care providers frequently struggle to develop wise applications of theoretical knowledge. As a nurse studying traditional approaches in ethics while simultaneously volunteering in Haiti, I wrestled with the notion of justice in the midst of radical material inequities. Paul Farmer, physician and anthropologist, provides an analytic perspective for health care providers who work in poor and underserved countries by expanding the liberation theology of the 1970s to include social applications. Analyzing my past experiences in Haiti using Farmer's methodology provided insight into my successes and failures and prompted me to search for personal and professional reasons to provide care for the poor in the future. This type of reflection is essential for health care providers who work with the underserved, regardless of their religion or the country in which they work. PMID- 18677080 TI - The Promise Clinic: a service learning approach to increasing access to health care. AB - The goal of the Promise Clinic (a project of an academic medical center and a local social services group) is to increase access to primary care for an underserved population while addressing deficiencies in medical education. Students manage common primary care problems, creating access for this mostly uninsured population. PMID- 18677081 TI - Delivering care out of the box: the evolution of an HIV harm reduction medical program. AB - Disparities in HIV health care continue to exist among New York City's marginalized populations. We describe the evolution and development of a unique collaborative program that blends harm reduction and medical care. This program addresses disparities and needs of a particularly marginalized population: unstably housed substance users with HIV infection. PMID- 18677082 TI - Dietary intake of homeless women residing at a transitional living center. AB - Women who are homeless experience health problems due to many factors, including poor nutrition. This paper describes a nutritional assessment of women who are homeless living at a transitional living center in an urban setting. A rapid food screener was used to assess fat, fruit and vegetable, and fiber intake, and focus group analysis was used to assess nutritional attitudes and dietary behaviors. We found that the dietary intake of shelter residents does not meet the USDA recommendations in several key areas. We also found that shelter residents considered shelter cafeteria food to be inadequate in terms of taste, nutritional quality, and choices, and they believed the shelter diet contributed to chronic diseases and their symptoms. We conclude that addressing these barriers to good nutrition may help people who are homeless prevent and manage chronic illness. PMID- 18677083 TI - A systematic review of COPC: evidence for effectiveness. AB - PURPOSE: This systematic review was conducted to assess the evidence for effectiveness of the community oriented primary care (COPC) model and discuss alternative approaches to community medicine practice and education. METHODS: A literature search for all articles referring to COPC was conducted. Articles were categorized by type, and the extent of the use of elements of the COPC model, and the level of community involvement. RESULTS: The majority of articles on COPC (60% of those reviewed) describe the general theory or educational use of the methodology. Many published studies are project descriptions not utilizing the complete model as initially described. Few of project descriptions include all elements of COPC or document community participation. Evidence for or against the effectiveness of the COPC model in improving community health outcomes (using the Strength of Recommendation Taxonomy (SORT) classification system as evidence) was found to be limited. CONCLUSIONS: Most publications related to COPC do not use the complete COPC model as originally described and evidence for its effectiveness is lacking. Further research with evaluation of community health outcomes and community participation is needed. Diverse models of community health intervention can be considered for training and collaborative practice with underserved populations. PMID- 18677084 TI - Medical and support service utilization in a medical program targeting marginalized HIV-infected individuals. AB - BACKGROUND: One strategy to facilitate HIV health care services utilization is to incorporate support services with medical services. We developed a program that delivers HIV medical care and support services to marginalized people, and evaluated the association between support and medical services utilization. METHODS: We extracted data on 218 newly enrolled program participants 3 months prior to through 6 months after program enrollment, and analyzed associations between support and medical services. RESULTS: Case management visits (AOR=1.95, 95% CI 1.04-3.67) and group visits (AOR=2.59, 95% CI 1.30-5.16) were associated with greater odds of quarterly medical visits. Outreach visits were associated with greater odds of having a medical visit in a traditional medical setting (AOR=2.31, 95% CI 1.15-4.67). CONCLUSION: Case management, support groups, and outreach were associated with HIV medical visits. Further research exploring how integration of support services into HIV medical programs can improve health care delivery is crucial for health policy and program development. PMID- 18677085 TI - A Taxonomy of medical comorbidity for veterans who are homeless. AB - Homeless veterans have numerous medical and behavioral health problems. Grouping homeless people based on comorbidity patterns may assist in determining severity of illness and triaging health care more effectively. We sought to determine if a finite number of profiles could be identified related to demographic characteristics, living situation, length of homelessness, and referral areas using interview data from 2,733 veterans who were presently or recently homeless. We considered 12 disorders: eye problems, hypertension, cardiovascular problems, COPD/emphysema, tuberculosis, gastrointestinal problems, hepatic disease, neurologic disorders, orthopedic problems, skin problems, and trauma. Ratings were evaluated using cluster analysis. Comparison statistics were used to compare intercluster differences in demographics, homeless situation, and referral recommendations. A four-cluster solution is proposed: generalized illness, hepatic disease, lung disease, and neurologic disorder. Medical health problems are common and heterogeneous in homeless individuals. Classifications of these problems may be useful in planning treatment and predicting outcome. PMID- 18677087 TI - Advances in genomic research on hepatitis C virus with a useful tool, replicon system. AB - The research for hepatitis C virus (HCV) has long delayed by missing of in vitro culture system. Since the development of replicon system, a replication system of subgenomic HCV RNAs in a hepatoma cell line, has been reported, many virological and clinical findings have been discovered. Recently, in addition of subgenomic replication system, hepatitis C virus full-length RNA replication has been possible, and a few cell culture systems producing viral particles have been produced. These developments enabled us to investigate the life cycle or intracellular circumstance of HCV production. By screening of newly synthesized drugs with this replicon system, several possible medicines have been established and clinical researches are now running. Among them, VX950 and SCH503034 are nearest to clinical use. Other possible agents for reducing viral replication such as cyclophyllin inhibitors, inhibitors of sphingomyelin synthesis, HMG-CoA reductase inhibitors, or RNA-dependent RNA polymerase inhibitors have been also investigated. Furthermore the mechanism for development of hepatocellular carcinoma in the HCV infected liver has been vigorously studied using the HCV replicon system. PMID- 18677088 TI - Clinical neurophysiology in the diagnosis of peroneal nerve palsy. AB - Peroneal neuropathy is one of the common focal mononeuropathies in the lower extremities occurring in both adults and children. Foot drop due to weakness of ankle dorsiflexion is the most common presentation of a peroneal neuropathy. It may also result from other causes involving the upper or lower motor neurons. Disorders that must be distinguished from peroneal neuropathy include sciatic mononeuropathy, lumbosacral plexopathy, motor neuron disease, polyneuropathy, and an L5 radiculopathy. To establish a diagnosis, electrodiagnostic studies have been used to localize the level of the abnormality and to establish prognosis. The most common site of injury is the fibular head, but focal neuropathies have also been reported at the level of the calf, ankle, and foot. In this article, we overviewed the peroneal nerve palsy, and its diagnosis by neurophysiologic evaluation, conduction study and needle EMG. The neurophysiologic information gives us the underlying pathophysiology and its prognosis. Therefore the neurophysiologic evaluation must be performed not only for the differential diagnosis, but also for planning the treatment strategy. PMID- 18677089 TI - Relationship between low evaluation by recipients under 40 years old of health interviews by occupational health nurses and deterioration in health checkup results over one year. AB - Improvement of an unhealthy lifestyle decreases the risk of incidence and mortality from lifestyle-related disease. One method for the improvement of an unhealthy lifestyle is interviewing for health and consultation regarding a healthy lifestyle to recipients of health checkups by public health nurses. Original articles in occupational health for recipients' evaluation in health checkups as longitudinal studies are rare. The purpose of the present study was to investigate in a longitudinal study the relationship between recipients' evaluation of interviews by public health nurses in a health checkup and variations in health checkup results for one year. Data on 750 male white collar workers who underwent a company health checkup in 1999 and 2000 were analyzed. The variation in health checkup results for one year from 1999 to 2000 was studied by computing odds ratios in a conditional logistic analysis of the recipients who set a low value on the interview for health by occupational health nurses in the health checkup in 1999 and those who did not. It was found that among the subjects who set a low value on the usefulness of the interview by the occupational health nurses, the numbers of the subjects whose body mass index (BMI) and triglyceride value (TG) were healthy (normal) in 1999 and unhealthy (abnormal) in 2000 were significantly greater than those of the subjects whose values were unhealthy in 1999 and healthy in 2000, while such a finding was not detected in the subjects who set a high value on the interview. The differences for BMI and TG between (i) differences between deterioration and improvement of health checkup results in the subjects who evaluated the interview lowly and (ii) those in the subjects who did not were statistically significant as interactions. It can be supposed that some of the subjects who set a low value on the usefulness of the interview by the occupational health nurses may be indifferent to their health status, indicating that low recipient evaluation of the interview for health in health checkup may be regarded as a risk factor of metabolic syndrome from the viewpoint of prevention. PMID- 18677090 TI - Primary ligament repair for elbow dislocation. AB - Few have been reported on the role of early primary ligament repair for acute unstable elbow dislocation. The purpose of this study is to evaluate the clinical outcome of early primary ligament repair for unstable elbow dislocation followed by protected early joint mobilization exercise. Thirteen patients who underwent operative treatment due to unstable elbow dislocation without associated fracture were retrospectively reviewed. There were 11 male and 2 female with average age of 37 years (range; 18-72 years). Surgical indications of the unstable elbow were subluxation or non-congruent elbow joint on the radiographs following closed reduction. Elbow dislocation which required extension block splint over 45 degrees to maintain reduction was considered as unstable elbow dislocation. In ten patients, full stability was restored only after both medial and lateral structures were re-attached. In two patients, only lateral structure (LCLC and common extensor tendon) was repaired and in one patient, only medial structure (MCL and flexor pronator tendon) was repaired for stability. Overall mean functional Mayo Elbow Performance Score was 93.5 (range; 70-100). No elbow was dislocated or subluxated after operation. All ten patients returned to previous work level in average of 3.5 months. Heterotopic calcification was noted in six patients and three had mild ulnar nerve symptoms after operation. In conclusion, primary ligament repair in acute unstable elbow dislocation were successful, because surgery achieved sufficient stability to allow early exercise. PMID- 18677091 TI - Phosphorylation of delta2 glutamate receptors at serine 945 is not required for cerebellar long-term depression. AB - Long-term depression (LTD) of synaptic transmission at parallel fiber (PF) Purkinje cell synapses is thought to regulate motor learning and memory formation in the cerebellum. Neuronal activity-evoked protein kinase C (PKC) activation is required for the induction of LTD. In addition, the delta2 glutamate receptor (GluRdelta2), which is predominantly expressed at PF-Purkinje cell synapses, is indispensable for the induction of LTD; however, the mechanisms by which GluRdelta2 regulates LTD and its relationship with PKC activation remain elusive. Interestingly, GluRdelta2 is phosphorylated by PKC on serine 945 (Ser945) near its C-terminus and a postsynaptic protein S-SCAM, which could potentially regulate glutamate receptor trafficking and synaptic plasticity, binds to the extreme C-terminus of GluRdelta2 in a phosphorylation-dependent manner on Ser945. Here, using a Sindbis-based virus expression approach, we show that a mutant GluRdelta2, in which alanine replaced Ser945 and did not undergo PKC phosphorylation, was normally localized at the postsynaptic sites of PF-Purkinje cell synapses. In addition, like wild-type GluRdelta2, the phosphorylation disrupted GluRdelta2 successfully rescued abrogated LTD in GluRdelta2-null Purkinje cells. These results indicate that Ser945, a major PKC phosphorylation site of of GluRdelta2, may not play a crucial role in induction of LTD in the cerebellum. PMID- 18677092 TI - Aortic root re-replacement with cryopreserved aortic homograft in a patient with active composite valve-graft endocarditis. AB - We describe a repeat aortic root replacement using a cryopreserved aortic homograft in a patient with active composite valve-graft endocarditis and an aortic root abscess. At the second surgery, infected prosthetic material was removed and surrounding tissue was radically debrided. The cryopreserved aortic homograft was positioned using the full root technique with a concomitant coronary bypass graft. The postoperative course was uneventful and the patient continues to thrive 2 years after the repeat operation. PMID- 18677093 TI - Region- and neuronal phenotype-specific expression of NELL2 in the adult rat brain. AB - NELL2, a neural tissue-enriched protein, is produced in the embryo, and postembryonically in the mammalian brain, with a broad distribution. Although its synthesis is required for neuronal differentiation in chicks, not much is known about its function in the adult mammalian brain. We investigated the distribution of NELL2 in various regions of the adult rat brain to study its potential functions in brain physiology. Consistent with previous reports, NELL2 immunoreactivity (ir) was found in the cytoplasm of neurons, but not in glial fibrillary acidic protein (GFAP)-positive glial cells. The highest levels of NELL2 were detected in the hippocampus and the cerebellum. Interestingly, in the cerebellar cortex NELL2 was observed only in the GABAergic Purkinje cells not in the excitatory granular cells. In contrast, it was found mainly in the hippocampal dentate gyrus and pyramidal cell layer that contains mainly glutamatergic neurons. In the dentate gyrus, NELL2 was not detected in the GFAP positive neural precursor cells, but was generally present in mature neurons of the subgranular zone, suggesting a role in this region restricted to mature neurons. PMID- 18677094 TI - A splice variant of the C(2)H(2)-type zinc finger protein, ZNF268s, regulates NF kappaB activation by TNF-alpha. AB - IkappaB kinase (IKK), the pivotal kinase in signal-dependent activation of nuclear factor-kappaB (NF-kappaB), is composed of multiple protein components, including IKK alpha/beta/gamma core subunits. To investigate the regulation of the IKK complex, we immunoaffinity purified the IKK complex, and by MALDI-TOF mass spectrometry identified a splice variant of zinc finger protein 268 (ZNF268) as a novel IKK-interacting protein. Both the full-length and the spliced form of the ZNF268 protein were detected in a variety of mammalian tissues and cell lines. The genes were cloned and expressed by in vitro transcription/translation. Several deletion derivatives, such as KRAB domain (KRAB) on its own, the KRAB/spacer/4-zinc fingers (zF4), and the spacer/ 4-zinc fingers (zS4), were ectopically expressed in mammalian cells and exhibited had different subcellular locations. The KRAB-containing mutants were restricted to the nucleus, while zS4 was localized in the cytosol. TNF-alpha-induced NF-kappaB activation was examined using these mutants and only zS4 was found to stimulate activation. Collectively, the results indicate that a spliced form of ZNF268 lacking the KRAB domain is located in the cytosol, where it seems to play a role in TNF-alpha-induced NF kappaB activation by interacting with the IKK complex. PMID- 18677095 TI - Effects of base changes at the transcription start site on stringent control of rnpB in Escherichia coli. AB - The GC-rich discriminator sequence between the -10 region and the transcription start site of the rnpB promoter is responsible for stringent control of M1 RNA synthesis. The rnpB promoter also contains a G nucleotide at the previously identified transcription start site. In this study, we examined by mutagenesis of G to A whether this +1G nucleotide is involved in the stringent response. We found that the change did not alter the stringent response. Since the +1 mutation might alter transcription initiation, we compared the transcription start sites of the wt and mutant promoters by primer extension analysis. Surprisingly, we found that wild type rnpB transcription starts at both the +1G position (70%) and the -1C position (30%), and that the +1A mutation led to transcription initiation exclusively at the -1C position. We also generated two transversion mutations at the -1 position, both of which led to transcription starting exclusively at that position. The -1G mutant promoter gave a stringent signal similar to the wild type, whereas the -1A mutant generated a significantly less stringent signal. Base on these results, we propose that a short sequence, up to 7 bp downstream of the -10 region, is involved in the stringent response of the rnpB promoter. PMID- 18677096 TI - Macrophage migration inhibitory factor (MIF) interacts with Bim and inhibits Bim mediated apoptosis. AB - The pro-apoptotic Bcl-2 family member Bim acts as a sensor for apoptotic stimuli and initiates apoptosis through the mitochondrial pathway. To identify novel regulators of Bim, we employed the yeast two-hybrid system and isolated the human gene encoding macrophage migration inhibitory factor (MIF), a ubiquitously expressed proinflammatory mediator that has also been implicated in cell proliferation, the cell cycle and carcinogenesis. The interaction between MIF and Bim was confirmed by both in vitro and in vivo protein interaction assays. Intriguingly, protein complexes between MIF and the three major Bim isoforms (BimEL/BimL/BimS) could be detected in HEK293 and K562 cells, especially in cells undergoing apoptosis. Moreover, exogenous expression of MIF partially inhibited Bim-induced apoptosis in HEK293 cells. SiRNA-mediated knockdown of MIF increased apoptosis in K562 cells exposed to the chemical oxidant diamide. Endogenous MIF may regulate the pro-apoptotic activity of Bim and inhibit the release of cytochrome c from mitochondria. PMID- 18677097 TI - Calcitonin gene-related peptide suppresses pacemaker currents by nitric oxide/cGMP-dependent activation of ATP-sensitive K(+) channels in cultured interstitial cells of Cajal from the mouse small intestine. AB - The effects of calcitonin gene-related peptide (CGRP) on pacemaker currents in cultured interstitial cells of Cajal (ICC) from the mouse small intestine were investigated using the whole-cell patch clamp technique at 30 degrees . Under voltage clamping at a holding potential of -70 mV, CGRP decreased the amplitude and frequency of pacemaker currents and activated outward resting currents. These effects were blocked by intracellular GDPbetaS, a G-protein inhibitor and glibenclamide, a specific ATP-sensitive K(+) channels blocker. During current clamping, CGRP hyperpolarized the membrane and this effect was antagonized by glibenclamide. Pretreatment with SQ-22536 (an adenylate cyclase inhibitor) or naproxen (a cyclooxygenase inhibitor) did not block the CGRP-induced effects, whereas pretreatment with ODQ (a guanylate cyclase inhibitor) or L-NAME (an inhibitor of nitric oxide synthase) did. In conclusion, CGRP inhibits pacemaker currents in ICC by generating nitric oxide via G-protein activation and so activating ATP-sensitive K(+) channels. Nitric oxide- and guanylate cyclase- dependent pathways are involved in these effects. PMID- 18677098 TI - ATF2: a transcription factor that elicits oncogenic or tumor suppressor activities. AB - The Activating Transcription Factor (ATF2) gene encodes a transcription factor important for normal cellular development and survival. In addition to its role as a transcription factor, ATF2 functions in the DNA damage response and in control of HAT complex activity. So far genetic changes in ATF2 have not been identified in human tumors. Yet altered expression and subcellular localization of ATF2 are associated with tumor stage and prognosis. Our studies in melanoma tumor models identified oncogenic ATF2 activity correlating with development of this tumor type. Inhibiting ATF2 suffices to impede melanoma development. Yet work in other tumor models, including breast and skin tumors reveals that ATF2, in cooperation with mutated oncogenes or tumor suppressor genes, can also elicit tumor suppressor function. These findings suggest tissue- and tumor-specific function of ATF2. This review summarizes the current understanding of ATF2 regulation and function. PMID- 18677100 TI - FLASH and NPAT positive but not Coilin positive Cajal Bodies correlate with cell ploidy. AB - Cajal Bodies are one of many specialised organelles contained in the eukaryotic cell nucleus, and are involved in a number of functions, including regulation of replication-dependent histone gene transcription. In normal diploid cells their number varies between 0 and 4 depending on the cell cycle phase, although in cancer cell lines their number is extremely variable and it has been suggested that it correlates with cell ploidy. Here we show that in mammalian cells, as in Drosophila, two distinct though functionally related bodies exist: a histone gene locus body and a Cajal Body. The first one can be detected using FLASH or NPAT as markers while the second is labelled using antibodies against Coilin. Only the number of FLASH/NPAT histone gene locus bodies correlates with ploidy and only these organelles appear to be regulated during the cell cycle. Finally, we show that the two organelles completely co-localize during the S phase of the cell cycle. PMID- 18677101 TI - A naturally occurring cancer with molecular connectivity to human diseases. AB - As Jessani et al.,(1) point out development of cell and animal models that accurately depict human tumorigenesis remains a major goal of cancer research. Clam cancer offers significant advantages over traditional models for genotoxic and non-genotoxic preclinical analysis of treatments for human cancers with a similar molecular basis. The naturally occurring clam model closely resembles an outbreeding, human clinical population and provides both in vitro and in vivo alternatives to those generated from inbred mouse strains or by intentional exposure to known tumor viruses. Fly and worm in vivo models for adult human somatic cell cancers do not exist because their adult somatic cells do not divide. Clam cancer is the best characterized, naturally occurring malignancy with a known molecular basis remarkably similar to those observed in several unrelated human cancers where both genotoxic and non-genotoxic strategies can restore the function of wild-type p53. To further emphasize this point of view, we here demonstrate a p53-induced, mitochondrial-directed mechanism for promoting apoptosis in the clam cancer model that is similar to one recently identified in mammals. Discerning the molecular basis for naturally occurring diseases in non traditional models and correlating these with related molecular mechanisms responsible for human diseases is a virtually unexplored aspect of toxico proteomics and genomics and related drug discovery. PMID- 18677099 TI - microRNA control of cell-cell signaling during development and disease. AB - MicroRNAs (miRNAs) are critical post-transcriptional regulators that may collectively control a majority of animal genes. With thousands of miRNAs identified, a pressing challenge is now to understand their specific biological activities. Many predicted miRNA:target interactions only subtly alter gene activity. It has consequently not been trivial to deduce how miRNAs are relevant to phenotype, and by extension, relevant to disease. We note that the major signal transduction cascades that control animal development are highly dose sensitive and frequently altered in human disorders. On this basis, we hypothesize that developmental cell signaling pathways represent prime candidates for mediating some of the major phenotypic consequences of miRNA deregulation, especially under gain-of-function conditions. This perspective reviews the evidence for miRNA targeting of the major signaling pathways, and discusses its implications for how aberrant miRNA activity might underlie human disease and cancer. PMID- 18677102 TI - Emerging genomic technologies and the concept of personalized medicine. AB - With the completion of the Human Genome Project in May 2006, genetic testing for every American is rapidly becoming a reality. As the advanced technology fuels the path towards personalized medicine, genetic nondiscrimination legislation follows closely behind. It seems that the 2007 Genetic Information Nondiscrimination Act (GINA) will finally pass through both chambers of Congress and will be signed by the President, but questions remain. On May 1, 2008, the House passed GINA by a vote of 414 to 1. Why is this the year that genetic nondiscrimination legislation could finally become the reality? Is this the beginning of a new relationship between science and policy, where policy is finally catching up? We examine the answers to these questions through a look at the history of genetic nondiscrimination legislation and where it stands today, including arguments for and against the bill. We conclude by discussing how we can achieve a future of safe personalized medicine for the populous, which would require continuous productive interactions between policymakers and scientists. PMID- 18677103 TI - Cell cycle regulation by the pro-apoptotic gene Scotin. AB - Scotin is a pro-apoptotic mammalian gene, which is induced upon DNA damage or cellular stress in a p53-dependent manner. In this report, we have used Drosophila as a model system to obtain a preliminary insight into the molecular mechanism of Scotin function, which was further validated using the mammalian system. Targeted expression of Scotin in developing Drosophila induced apoptosis and developmental defects in wings and eyes. Co-expression of Scotin with the anti-apoptotic protein p35, while inhibited the apoptosis in both dividing and non-dividing cells, rescued adult wing or eye phenotypes only when Scotin was expressed in non-dividing cells. This suggests that mechanisms of Scotin-induced apoptosis in dividing and non-dividing cells may vary. Suppressor-enhancer screen using cell cycle regulators suggested that Scotin may mediate cell cycle arrest at both G(1)/S and G(2)/M phases. Overexpression of Scotin in mammalian cells resulted in mitotic arrest and subsequently apoptosis. Furthermore, a larger proportion of cells overexpressing Scotin showed sequestration of Cyclin B1 in the cytoplasm. These results suggest that one of the ways by which Scotin induces apoptosis is by causing cell cycle arrest. PMID- 18677104 TI - The multiple facets of the intra-S checkpoint. AB - In response to hydroxyurea treatment or DNA damage the total rate of DNA replication per cell is reduced. This reduction may be due to physical hindrance of the replication forks or to active, checkpoint-dependent processes. Here we review current knowledge about how and to what extent the intra-S checkpoint affects DNA replication. We discuss evidence that some checkpoint proteins are active even in a normal S phase and we suggest a model that resolves the apparent contradiction between different views on checkpoint-dependent slowing of the rate of DNA replication: does the intra-S checkpoint repress or delay the initiation of all origins or late replication origins only, and to what extent does it inhibit fork progression. Finally, the new model is discussed in the context of cancer development. PMID- 18677105 TI - Synchronization of the cell cycle using lovastatin. AB - Synchronization by Lovastatin arrests many cell types reversibly in the G(1) phase of the cell cycle. Here we show that Lovastatin (10 microM) mediates cell cycle arrest in human breast cancer cells, MCF-7 and MDA-MB-231, where 85% of cells accumulate in the G(1) phase of the cell cycle. Addition of mevalonate (at 100X the Lovastatin concentration) releases the cells from the G(1) arrest and allows for synchronous entry into late G(1), S and G(2)/M phases of the cell cycle. The expressions of different cyclins as a marker for different phases of the cell cycle are detected by western blot analysis and indicative of synchronous transition into each of cell cycle phases following the initial G(1) arrest. Due to its level of synchrony and high yield of synchronous populations of cells, Lovastatin method of cell synchronization can be used for examining gene expression patterns in a variety of different cell lines. PMID- 18677106 TI - The eIF2alpha kinases inhibit vesicular stomatitis virus replication independently of eIF2alpha phosphorylation. AB - The eIF2alpha kinases have been involved in the inhibition of vesicular stomatatis virus replication but the contribution of each kinase to this process has not been fully investigated. Using mouse embryonic fibroblasts (MEFs) from knock-out mice we show that PKR and HRI have no effects on VSV replication as opposed to PERK and GCN2, which exhibit strong inhibitory effects. When MEFs containing the serine 51 to alanine mutation of eIF2alpha were used, we found that VSV replication is independent of eIF2alpha phosphorylation. Nevertheless, the kinase domain of the eIF2alpha kinases is both necessary and sufficient to inhibit VSV replication in cultured cells. Induction of PI3K-Akt/PKB pathway by eIF2alpha kinase activation plays no role in the inhibition of VSV replication. Our data provide strong evidence that VSV replication is not affected by eIF2alpha phosphorylation or downstream effector pathways such as the PI3K Akt/PKB pathway. Thus, the anti-viral properties of eIF2alpha kinases are not always related to their inhibitory effects on host protein synthesis as previously thought and are possibly mediated by phosphorylation of proteins other than eIF2alpha. PMID- 18677108 TI - Direct role of nucleotide metabolism in C-MYC-dependent proliferation of melanoma cells. AB - To identify C-MYC targets rate-limiting for proliferation of malignant melanoma, we stably inhibited C-MYC in several human metastatic melanoma lines via lentivirus-based shRNAs approximately to the levels detected in normal melanocytes. C-MYC depletion did not significantly affect levels of E2F1 protein reported to regulate expression of many S-phase specific genes, but resulted in the repression of several genes encoding enzymes rate-limiting for dNTP metabolism. These included thymidylate synthase (TS), inosine monophosphate dehydrogenase 2 (IMPDH2) and phosphoribosyl pyrophosphate synthetase 2 (PRPS2). C MYC depletion also resulted in reduction in the amounts of deoxyribonucleoside triphosphates (dNTPs) and inhibition of proliferation. shRNA-mediated suppression of TS, IMPDH2 or PRPS2 resulted in the decrease of dNTP pools and retardation of the cell cycle progression of melanoma cells in a manner similar to that of C-MYC depletion in those cells. Reciprocally, concurrent overexpression of cDNAs for TS, IMPDH2 and PRPS2 delayed proliferative arrest caused by inhibition of C-MYC in melanoma cells. Overexpression of C-MYC in normal melanocytes enhanced expression of the above enzymes and increased individual dNTP pools. Analysis of in vivo C-MYC interactions with TS, IMPDH2 and PRPS2 genes confirmed that they are direct C-MYC targets. Moreover, all three proteins express at higher levels in cells from several metastatic melanoma lines compared to normal melanocytes. Our data establish a novel functional link between C-MYC and dNTP metabolism and identify its role in proliferation of tumor cells. PMID- 18677109 TI - Direct visualization of the endomitotic cell cycle in living megakaryocytes: differential patterns in low and high ploidy cells. AB - Endomitosis in megakaryocytes (MKs) involves repeated DNA replication in the absence of cytokinesis and is a crucial part of MK development. However, chromosomal dynamics have never been observed in living MKs. We developed a new transgenic mouse model in which the expression of human histone H2B fused in frame to green fluorescent protein is targeted to MKs. Ex vivo time-lapse microscopy analysis indicated that chromosomal condensation occurs at early mitosis in all MKs. In high ploidy MKs (>or=8N), late anaphase was marked by a ring-type alignment of chromosomes with multiple territories formed between them. By contrast, in low ploidy MKs mitotic chromosomes segregated to form two groups separated by a clear space before re-joining to one cluster. This is the first study to document chromosomal segregation patterns during endomitosis ex vivo and to indicate their potential differential regulation in low and high ploidy cells. PMID- 18677107 TI - FERM control of FAK function: implications for cancer therapy. AB - Integrins are transmembrane receptors that bind to extracellular matrix proteins and convey anchorage-dependent signals regulating normal cell proliferation. Integrin signals within the tumor micro-environment also impact cancer cell survival and invasion during tumor progression. These integrin-associated signaling events are transduced in part through the activation of non-receptor protein-tyrosine kinases. Focal adhesion kinase (FAK) is activated by beta subunit integrins in both normal and transformed cells. As genetic inactivation of beta1 integrin or FAK yield early embryonic lethal phenotypes associated with decreased cell proliferation, and dominant-negative inhibition of FAK can cause increased cell apoptosis, there is a concern that FAK inhibition may have cytotoxic effects on cell growth or survival. However, FAK-specific small molecule inhibitors do not directly impact cell growth in culture, but yet show potent anti-tumor growth effects in vivo. Additionally, recent studies have shed new insight into the FAK kinase-independent regulation of cell proliferation and survival mediated by the FAK N-terminal FERM (band 4.1, ezrin, radixin, moesin homology) domain. Herein, we review the role of the FAK FERM domain in both the intrinsic regulation of FAK kinase activity and how FERM-mediated nuclear localization of FAK promotes enhanced cell survival through the inhibition of tumor suppressor p53 activation during development and under conditions of cellular stress. As we find that FAK FERM-mediated regulation of p53 occurs in human carcinoma cells, elevated FAK expression in tumors may promote both kinase dependent and -independent survival mechanisms. We discuss how the pharmacological inhibition of FAK kinase activity may impact tumor progression through combined effects of blocking both tumor- and stromal-associated signaling regulating neo-vascularization. PMID- 18677110 TI - Multiple p53-independent gene silencing mechanisms define the cellular response to p53 activation. AB - The cellular response to Nutlin-3, a small-molecule inhibitor of the p53 repressor MDM2, varies widely among human cancer-derived cell types. Whereas HCT116 colorectal carcinoma cells display sustained cell cycle arrest, BV173 leukemia cells undergo rapid apoptosis and other cell lines show an intermediate response. We found that the expression of the p53 target genes p21, 14-3-3sigma and the microRNA miR-34a correlates tightly with the cell fate choice adopted. All three genes were strongly induced in arresting cells, but silenced in cells undergoing Nutlin-3-induced apoptosis. In contrast, key apoptotic p53 target genes were equally expressed in arresting and apoptotic cells. Interestingly, we establish that miR-34a cooperates with p21 and 14-3-3sigma to override the apoptotic signals generated by p53 activation. Strikingly, p53 binding to chromatin and p53-mediated recruitment of certain coactivators to all three target loci does not vary among cell types. Instead, the cell type-specific silencing of these genes is due to enhanced p21 mRNA degradation, 14-3-3sigma promoter DNA methylation and reduced processing of the miR-34a primary transcript. Thus, p53-independent events regulating expression of protein-coding genes and microRNAs within the network can define the cellular outcome of p53 activation. PMID- 18677111 TI - Directing T cell differentiation and function with small molecule inhibitors. AB - Regulatory T (Treg) cells that express the signature transcription factor Foxp3 safeguard against autoimmunity and immune pathology. Recent studies show that a signaling network with the components phosphatidyl inositol 3 kinase (PI3K), Akt, and the mammalian target of rapamycin (mTOR) regulates the de novo expression of Foxp3 in CD4 T cells. In addition to CD4 T cell differentiation, PI3K/Akt/mTOR signaling also controls T cell migration. Here we review the new data, consider their evolutionary context and discuss their potential implications for immunotherapy. PMID- 18677112 TI - Low-penetrant RB allele in small-cell cancer shows geldanamycin instability and discordant expression with mutant ras. AB - Certain kindreds with low-penetrant (lp) retinoblastoma carry mutant alleles which retain partial tumor suppressor activity and we previously showed that these alleles exhibit defective, temperature-sensitive binding in yeast. To investigate the molecular basis for incomplete penetrance, we studied three recurrent lp alleles and observed approximately 50% of wildtype activity measured by (i) phosphorylation at key regulatory sites, S780, S795, S807/S811, (ii) transcriptional co-activation, and (iii) 'flat-cell' differentiation in mammalian cells in vivo. In addition, we studied a small-cell carcinoma that is homozygous for the R661W allele providing the first analysis of the effect of a naturally occurring lp allele in a human tumor. While we detected abundant expression of the R661W protein, we noted marked instability of both endogenous and recombinant R661W following treatment in vivo with the Hsp90 inhibitor, geldanamycin and stabilization of R661W following heat shock. In addition, we observed a discordant phenotype in the tumor cells with induction of p16 and loss of cyclin D1 consistent with a null RB status combined with homozygous expression of mutant ras which had not been reported previously for RB (-) small-cell cancer. These findings show that a recurrent missense lp allele retains greater functional activity in vivo than predicted from earlier in vitro assays, proposing a role for stabilizing chaperone-like activity in vivo. In addition, these data suggest that reversible protein instability and the requirement for a cooperating mutation may provide a stochastic explanation for the molecular basis of incomplete penetrance in kindreds carrying these alleles. PMID- 18677113 TI - Structure of the human Mdmx protein bound to the p53 tumor suppressor transactivation domain. AB - The Mdmx oncoprotein has only recently emerged as a critical-independent to Mdm2 regulator of p53 activation. We have determined the crystal structure of the N terminal domain of human Mdmx bound to a 15-residue transactivation domain peptide of human p53. The structure shows why antagonists of the Mdm2 binding to p53 are ineffective in the Mdmx-p53 interaction. PMID- 18677114 TI - Coupled alteration of transcription and splicing by a single oncogene: boosting the effect on cyclin D1 activity. AB - In cancer cells, gene expression is altered at the levels of transcription and mRNA maturation, with many splice variants being associated with cancer. Splicing is tightly connected to transcription and can be affected by transcription elongation dynamics. Moreover, various transcriptional coregulators that are altered in cancer, such as the proto-oncogene EWS, are thought to play a role in splicing. A recent study shows that an alteration of EWS in Ewing sarcoma alters the dynamics of RNA polymerase II over the CCND1 proto-oncogene encoding cyclin D1, leading to an increase in its transcription and to an alteration of splicing that results in high levels of the oncogenic cyclin D1b splice isoform. The cyclin D1b isoform is highly expressed in Ewing sarcoma cells and tumors and stimulates Ewing sarcoma cell growth. Thus, alterations of transcriptional regulators in disease may lead to splicing alterations. We review these data and discuss how this concept may apply to various factors that are altered in cancer. PMID- 18677115 TI - Aurora kinase A controls meiosis I progression in mouse oocytes. AB - Aurora kinase A (AURKA), which is a centrosome-localized serine/threonine kinase crucial for cell cycle control, is critically involved in centrosome maturation and spindle assembly in somatic cells. Active T288 phosphorylated AURKA localizes to the centrosome in the late G(2) and also spreads to the minus ends of mitotic spindle microtubules. AURKA activates centrosomal CDC25B and recruits cyclin B1 to centrosomes. We report here functions for AURKA in meiotic maturation of mouse oocytes, which is a model system to study the G(2) to M transition. Whereas AURKA is present throughout the entire GV-stage oocyte with a clear accumulation on microtubule organizing centers (MTOC), active AURKA becomes entirely localized to MTOCs shortly before germinal vesicle breakdown. In contrast to somatic cells in which active AURKA is present at the centrosomes and minus ends of microtubules, active AURKA is mainly located on MTOCs at metaphase I (MI) in oocytes. Inhibitor studies using Roscovitine (CDK1 inhibitor), LY-294002 (PI3K inhibitor) and SH-6 (PKB inhibitor) reveal that activation of AURKA localized on MTOCs is independent on PI3K-PKB and CDK1 signaling pathways and MOTC amplification is observed in roscovitine- and SH-6-treated oocytes that fail to undergo nuclear envelope breakdown. Moreover, microinjection of Aurka mRNA into GV-stage oocytes cultured in 3-isobutyl-1-methyl xanthine (IBMX)-containing medium to prevent maturation also results in MOTC amplification in the absence of CDK1 activation. Overexpression of AURKA also leads to formation of an abnormal MI spindle, whereas RNAi-mediated reduction of AURKA interferes with resumption of meiosis and spindle assembly. Results of these experiments indicate that AURKA is a critical MTOC-associated component involved in resumption of meiosis, MTOC multiplication, proper spindle formation and the metaphase I-metaphase II transition. PMID- 18677116 TI - Signaling function of alpha-catenin in microtubule regulation. AB - Centrosomes control microtubule dynamics in many cell types, and their removal from the cytoplasm leads to a shift from dynamic instability to treadmilling behavior and to a dramatic decrease of microtubule mass (Rodionov et al., 1999; PNAS 96:115). In cadherin-expressing cells, these effects can be reversed:non centrosomal cytoplasts that form cadherin-mediated adherens junctions display dense arrays of microtubules (Chausovsky et al., 2000; Nature Cell Biol 2:797). In adherens junctions, cadherin's cytoplasmic domain binds p120 catenin and beta catenin, which in turn binds alpha-catenin. To elucidate the roles of the cadherin-associated proteins in regulating microtubule dynamics, we prepared GFP tagged, plasma membrane targeted or untargeted p120 catenin, alpha-catenin and beta-catenin and tested their ability to rescue the loss of microtubule mass caused by centrosomal removal in the poorly adhesive cell line CHO-K1. Only membrane targeting of alpha-catenin led to a significant increase in microtubule length and density in centrosome-free cytoplasts. Expression of non-membrane targeted alpha-catenin produced only a slight effect, while both membrane targeted and non-targeted p120 and beta-catenin were ineffective in this assay. Together, these findings suggest that alpha-catenin is able to regulate microtubule dynamics in a centrosome-independent manner. PMID- 18677117 TI - The Saccharomyces cerevisiae checkpoint genes RAD9, CHK1 and PDS1 are required for elevated homologous recombination in a mec1 (ATR) hypomorphic mutant. AB - Specific ataxia telangiectasia and Rad3-related (ATR) mutations confer higher frequencies of homologous recombination. The genetic requirements for hyper recombination in ATR mutants are unknown. MEC1, the essential yeast ATR/ATM homolog, controls S and G(2) checkpoints and the DNA damage-inducibility of genes after radiation exposure. Since the mec1-Delta (null) mutant is defective in both S and G(2) checkpoints, we measured spontaneous and DNA damage-associated sister chromatid exchange (SCE), homolog (heteroallelic) recombination, and homology directed translocations in the mec1-21 hypomorphic mutant, which is defective in the S phase checkpoint but retains some G(2) checkpoint function. We observed a sixfold, tenfold and 30-fold higher rate of spontaneous SCE, heteroallelic recombination, and translocations, respectively, in mec1-21 mutants compared to wild type. The mec1-21 hyper-recombination was partially reduced in rad9, pds1 and chk1 mutants, and abolished in rad52 mutants, suggesting the hyper recombination results from RAD52-dependent recombination pathway(s) that require G(2) checkpoint functions. The HU and UV sensitivities of mec1-21 rad9 and mec1 21 rad52 were synergistically increased, compared to the single mutants, indicating that mec1-21, rad52 and rad9 mutants are defective in independent pathways for HU and UV resistance. G(2)-arrested mec1-21 rad9 cells exhibit more UV resistance than non-synchronized cells, indicating that one function of RAD9 in conferring UV resistance in mec1-21 is by triggering G(2) arrest. We suggest that checkpoint genes that function in the RAD9-mediated pathway are required for either homologous recombination or DNA damage resistance in the S phase checkpoint mutant mec1-21. PMID- 18677119 TI - PTP-Pez: a novel regulator of TGFbeta signaling. AB - The TGFbetas are a family of pleiotropic cytokines that mediate diverse effects including the regulation of cell cycle progression, apoptosis, tissue remodelling and epithelial-mesenchymal transition (EMT). These diverse effects allow the TGFbetas to play multiple and even opposing roles in different contexts during embryonal development, tissue homeostasis and cancer progression. We recently reported that the protein tyrosine phosphatase Pez is a novel inducer of TGFbeta signaling, regulating EMT and organogenesis in developing zebrafish embryos, and leading to TGFbeta-mediated EMT when overexpressed in vitro in epithelial MDCK cells. A number of mutations in Pez have been shown to be associated with breast and colorectal cancers, although the effect of these mutations on Pez function and their contribution to cancer progression remains unclear. Our finding that Pez regulates TGFbeta signaling is therefore of interest not only in the context of identifying a novel upstream regulator of TGFbeta signaling, but also in implicating the dysregulation of TGFbeta signaling as a possible link between Pez mutation and cancer progression. Here we discuss the implications of our research, in the context of dysregulation of TGFbeta signaling in cancer and other human pathologies. PMID- 18677126 TI - The interaction of radiation therapy and antiangiogenic therapy. AB - Since its beginning in the early 1970s, the field of angiogenesis research has grown rapidly and it has now become apparent that the endothelial cell is a critical regulator of the malignant phenotype. Multiple antiangiogenic agents have now been used in the clinic yet a better understanding of the process of angiogenesis is still needed before these agents can be successfully incorporated into clinical practice. Although antiangiogenic agents offer great therapeutic potential, preclinical and clinical studies suggest that these agents will have a delayed onset of activity and may only induce disease stabilization for patients with advanced malignancy. The use of radiation therapy for cancer is also associated with therapeutic challenges that are distinct from those that might be expected with antiangiogenic agents. Thus, the use of angiogenesis inhibitors in combination with radiation therapy should help to overcome the limitations of each leading to enhanced efficacy and diminished toxicity. The goal of this review is to provide an update of ongoing progress and current challenges related to the use of angiogenesis inhibitors with radiation therapy. PMID- 18677118 TI - Oncogene-induced senescence: an essential role for Runx. PMID- 18677127 TI - More than 200,000 women will be diagnosed with breast cancer in 2008. PMID- 18677128 TI - Sentinel lymph node biopsy and axillary dissection: added morbidity of the arm, shoulder and chest wall after mastectomy and reconstruction. AB - Axillary lymph node surgery is essential for the treatment of breast cancer but can produce both short-term and long-term morbidities, including lymphedema, shoulder restriction, numbness, weakness, and pain syndromes which impact on the quality of life of the estimated 2 million breast cancer survivors living in the United States today. They occur with both sentinel node biopsy and axillary dissection, though less frequently with the former. The incidence and etiology of these complications are investigated as are the risk factors, and methods of prevention and treatment. Recommendations for avoiding these morbidities for postoperative rehabilitation are discussed. PMID- 18677129 TI - Timing of breast reconstruction: immediate versus delayed. AB - Breast reconstruction is an integral part of treatment of breast cancer. Immediate reconstruction is breast reconstruction that is done at the same surgery as the mastectomy, whereas delayed reconstruction is done months or years after the mastectomy. Immediate and delayed reconstruction can be accomplished with autologous tissue flaps or prosthetic breast implants. The esthetic result, psychosocial effect, and cost of breast reconstruction are better with immediate reconstruction, but the risk of surgical complications is less with delayed reconstruction. Although immediate reconstruction is oncologically safe and esthetically advantageous, nationwide less than 20% of patients having a mastectomy have immediate breast reconstruction. Radiation treatment before or after mastectomy has a negative impact on the outcome of breast reconstruction and is one important factor to be considered in determining the optimal timing for breast reconstruction. PMID- 18677130 TI - Breast reconstruction with prosthetic implants. AB - For patients who undergo mastectomy for the treatment of breast cancer, the restoration of a normal breast form through breast reconstruction is important to body image and quality of life. Implant-based reconstruction has the capability of producing excellent results in the well selected patient. In addition, compared with autogenous tissue reconstruction, implant reconstruction offers a shorter operative procedure with a quicker convalescence and no donor site morbidity. Reconstruction options for implant-based reconstruction include: single-stage reconstruction with a standard or adjustable implant, tissue expansion followed by placement of a permanent implant, or combined autologous tissue/implant reconstruction. Procedure selection is based on a range of patient variables, including: location and type of breast cancer, availability of local, regional and distant donor tissue, size and shape of the desired breast(s), surgical risk, and most importantly, patient preference. Although satisfactory results can be obtained with single-stage reconstruction, in most of patients, a more reliable approach involves two-stage expander/implant reconstruction. Individualizing selection of a reconstructive technique for each patient will be the predominant factor in achieving a reconstructive success. PMID- 18677131 TI - Optimizing pedicled transverse rectus abdominis muscle flap breast reconstruction. AB - With improved detection and genetic screening for breast cancer, increasing numbers of patients are choosing mastectomy and immediate breast reconstruction. Advances in breast reconstruction and mastectomy techniques over the last 25 years have increased expectations for breast reconstruction. The purpose of this review is to describe the role of the pedicled transverse rectus abdominis muscle (TRAM) flap in modern breast reconstruction.The pedicled TRAM flap is the most common method of autologous breast reconstruction. TRAM flap reconstruction involves the transfer of lower abdominal skin and subcutaneous fat based on the superior epigastric vessels. Common risk factors for complications from pedicled TRAM flap reconstruction include smoking, obesity, and postoperative radiotherapy. Patients with these risk factors are often candidates for a vascular delay procedure whose purpose is to enhance the blood flow within the TRAM flap.Despite advances in free flap breast reconstruction, pedicled TRAM flap breast reconstruction remains an excellent option for unilateral breast reconstructions. Unlike microsurgical breast reconstruction, the pedicled TRAM flap does not require sophisticated postoperative monitoring and can be performed efficiently in any hospital setting. Furthermore, with the addition of a vascular delay procedure, pedicled TRAM reconstructions can be safely performed even in traditionally "high risk" patients. PMID- 18677132 TI - Microsurgical breast reconstruction. AB - Breast cancer, the most common cancer diagnosed in American women, often necessitates mastectomy. Many studies have demonstrated improved quality of life and well-being after breast reconstruction. Numerous techniques are available for breast reconstruction including tissue expander implants and autologous tissues. Microsurgical tissue transfer involves the use of excess skin and fat (flaps) from a remote location to reconstruct the breast. Most often, tissues are transferred from the abdomen and buttocks. Less commonly, thigh flaps are used. These operations can provide durable, esthetic reconstructions. In addition, advances in microsurgical techniques have improved operative success rates to the range of 99%. The selection of an appropriate flap for microsurgical breast reconstruction is multifactorial and is based on patient and oncologic factors. These factors include patient comorbidities, body habitus/availability of donor tissues, cancer stage, and the need for postoperative adjuvant radiation therapy, as well as the risk of cancer in the contralateral breast. Appropriate choice of flap and surgical technique can minimize the risk of operative complications. Additionally, several large series have established that microsurgical breast reconstruction has no impact on survival, or locoregional/distant recurrence rates. PMID- 18677133 TI - Postmastectomy reconstruction of the breast using the latissimus dorsi musculocutaneous flap. AB - Reconstruction of the breast after mastectomy must be both safe and effective in providing long-term results for patients undergoing treatment for breast cancer. By using several strategically designed technical modifications in the elevation and transfer of the latissimus dorsi musculocutaneous flap, outstanding results can be obtained when this method combines with the use of prosthetic devices in the form of tissue expanders and implants. The morbidity associated with the procedure is low, and the operation is well tolerated by even in the presence of adverse risk factors, such a smoking history, obesity, or medical illness. This procedure is presented as an excellent option for those patients who present for breast reconstruction after mastectomy. PMID- 18677134 TI - Nipple areola reconstruction. AB - Nipple areola reconstruction is often considered the "finishing touch" in the process of breast reconstruction after surgery for breast cancer. Attention to detail with respect to patient selection, timing of reconstruction and surgical technique must therefore be paid to avoid an unsatisfactory result that may taint an otherwise successful reconstructive sequence. There are many surgical techniques available to recreate the nipple areola complex, but each of the various techniques is designed to accomplish similar goals. In every case, regardless of the technique chosen, the surgeon seeks to reconstruct a nipple areola complex that is esthetically pleasing in its color, symmetry, position on the breast mound, and projection. PMID- 18677135 TI - Managing the opposite breast: contralateral symmetry procedures. AB - The importance of symmetry after unilateral breast reconstruction is becoming increasingly realized and important. As our reconstructive options have increased and our technical abilities have improved, the expectations of women with breast cancer have been enhanced by virtue of the fact that we are often able to reconstruct a breast with natural volume and contour. When embarking on a unilateral breast reconstruction, the plastic surgeon has several goals and one of which is to obtain breast symmetry. In many circumstances, breast symmetry is achieved without operating on the contralateral breast; however, in some cases, a contralateral operation will be necessary. This is usually based on the volume, contour, and position of the natural and reconstructed breast. Often, the need and/or desire for a contralateral procedure will be evident during the initial consultation. Otherwise, the need for a contralateral procedure will not be evident until after the primary reconstruction has been completed. In either case, it is important for plastic surgeons to be aware of the incidence of these procedures and the options available to them. PMID- 18677136 TI - Breast reconstruction combined with radiation therapy: long-term risks and factors related to decision making. AB - Mastectomy with immediate breast reconstruction is a surgical procedure that addresses both the need to perform a cancer operation, and the desire of the patient to emerge from anesthesia with a replacement breast. An increasing number of these women with invasive breast cancer will benefit from chest wall and regional nodal radiation, in terms of both a decreased risk in local-regional recurrence and an increased chance of overall survival at 10 years and beyond, based on the most recent 2005 Oxford Overview.Indications for recommending radiation are based primarily on the pathologic assessment of the primary tumor and the axillary lymph nodes. More than a decade ago, only women with 10 or more involved nodes were thought to benefit from this treatment; that recommendation shifted to women with 4 or more involved nodes with the publication of the American Society for Clinical Oncology (ASCO) guidelines in 2001 and more recently to considering postmastectomy radiation (PMRT) in women with 1 to 3 positive nodes.In some scenarios, the need for PMRT is recognized before the patient goes to surgery, and reconstruction can be planned with this in mind, as discussed elsewhere in this issue. In other scenarios, the need for radiation is not realized until the final pathology report is back in the surgeon's hands. If that patient has elected an immediate reconstruction, is the radiation feasible and is it effective? Our published experience at Memorial Sloan Kettering (MSK) Cancer Center suggests the answer to both questions is "yes." However, our results differ dramatically from that of other major centers. Both our experience and that of others is discussed in this article. PMID- 18677137 TI - Outcomes of adjuvant chemoradiotherapy after a radical gastrectomy and a D2 node dissection for gastric adenocarcinoma. AB - PURPOSE: Intergroup 0116 (INT-0116) established adjuvant chemoradiation as the standard of care for resected high-risk adenocarcinoma of the stomach in the United States. However, adjuvant chemoradiation remains controversial in many parts of Asia and Europe, where patients tend to undergo a more thorough D2 dissection. In INT-0116, 90% of patients had a limited or inadequate node dissection (D0 or D1). Also, 17% of patients in the chemoradiation arm had to discontinue treatment because of toxicities. The objectives of this retrospective study are to report the clinical outcomes of a cohort of patients who were mostly treated with a D2 node dissection and received adjuvant chemoradiation as per INT 0116, and the toxicities of chemoradiation in the context of more aggressive surgery. METHODS: After the results of INT-0116 became apparent, we adopted an institutional policy whereby patients who would otherwise fit the inclusion criteria of INT-0116 received adjuvant chemoradiation. Between March 1999 and November 2004, 70 consecutive patients with pathologic stage T3, T4, or node positive disease were treated according to the chemoradiation arm of INT-0116. Patients received intravenous 5-fluorouracil 425 mg/m and leucovorin 20 mg/m in cycles 1, 3, and 4. Concurrent chemoradiation was given in cycle 2 and consisted of bolus 5-fluorouracil and leucovorin and radiotherapy (45 Gy over 25 fractions in 5 weeks). All patients were operated on by dedicated Japan-trained Surgical Oncologists. RESULTS: Sixty-seven patients (96%) had a D2 nodal dissection. Sixty five patients (93%) had negative pathologic margins (R0 resection) and 5 (7%) had microscopically involved margins (R1 resection). The median follow-up was 27 months (range, 10.1-60.3). The 3-year overall survival, disease-free survival, and local control were 60.6%, 54.1%, and 84.3%, respectively. Of the 30 patients who relapsed, 5 (17%) had isolated locoregional recurrences only. The National Cancer Institute--Common Terminology Criteria version 3.0 acute grade 3 or 4 gastrointestinal and hematological toxicity rates were 15.7% and 4.3%, respectively. Toxicities led to chemotherapy dose-reductions in 18 patients and dose-delay in 19 patients. Including chemotherapy dose-reductions and delays, 66 patients (94%) completed the entire chemoradiation regimen. There were no toxicity-related deaths. CONCLUSION: In our cohort of 70 patients who had a more thorough D2 node dissection, adjuvant chemoradiation was well tolerated with acceptable toxicities and reasonable tumor control. PMID- 18677138 TI - Secular changes in NSAID use and invasive colorectal cancer incidence: an ecological study. PMID- 18677140 TI - New antibiotics and vaccines in obstetric practice. Foreword. PMID- 18677141 TI - Vaccines for pertussis and influenza: recommendations for use in pregnancy. AB - The active immunization of pregnant women during pregnancy to protect them from disease and protect their neonate with passive antibodies is a biologic fact. Fortunately, many infectious diseases occur infrequently due to excellent pediatric vaccine programs. However, most adults and many physicians are unaware of the risks of not administering vaccines especially to pregnant women. Influenza vaccine (trivalent inactivated influenza vaccine) is recommended by (Advisory Committee on Immunization Practices (ACIP) for pregnant women in any trimester of pregnancy and Tetanus, reduced diphtheria, and pertussis (TdaP) vaccine is recommended by the ACIP to be given before pregnancy, during pregnancy, or in the immediate postpartum period. Only 2% of the adult US population is protected against pertussis and it is estimated that only 25% of pregnant women receive influenza vaccine during the influenza season. This chapter discusses trivalent inactivated influenza vaccine and TdaP use during pregnancy, the diseases they prevent, and the benefit to the neonate. PMID- 18677142 TI - Once-daily dosing of gentamicin in obstetrics and gynecology. AB - Gentamicin, an aminoglycoside with broad antimicrobial activity, is commonly used in both obstetrics and gynecology. Traditional dosing regimens for gentamicin have called for 3 times daily dosing, but recent insights into the pharmacodynamics of the drug have led to multiple studies of once-daily dosing regimens. Many studies have demonstrated efficacy, safety, and economy of the 24 hour dosing interval, resulting in recommendations that this become the standard for aminoglycoside administration. However, because of the unique considerations for drug administration in pregnant and postpartum women, the once-daily dosing regimens have not been widely adopted. Additional studies in pregnant and postpartum women have demonstrated therapeutic noninferiority, no increase in adverse events, and significant cost savings with once-daily dosing versus 3 times daily dosing of gentamicin. We review the literature and present rationale based on multiple controlled studies supporting single-daily dosing of gentamicin, 5 mg/kg/d actual body weight, for many common obstetrics-gynecology infections. PMID- 18677143 TI - Testing women for human immunodeficiency virus infection: who, when, and how? AB - Obstetrician-gynecologists provide comprehensive primary and preventive care for women and are ideally suited to provide human immunodeficiency virus (HIV) screening for their patients. This paper provides a summary and rationale for the current recommendations for HIV testing among women in the United States, emphasizing recommendations from the Centers for Disease Control and Prevention and the American College of Obstetricians and Gynecologists [corrected] Who should receive HIV testing, when and how often testing should be conducted, and how testing should be offered are discussed. These recommendations are described separately for general populations (including nonpregnant women) and for pregnant women and their infants. PMID- 18677144 TI - New approaches to external genital warts and vulvar intraepithelial neoplasia. AB - Human papillomavirus is the most common sexually transmitted disease in the United States. For the majority of affected individuals, the virus remains subclinical. However, human papillomavirus infection may result in a broad spectrum of vulvar disease including genital warts, dysplasia, and invasive carcinoma. We review the evaluation and currently available therapies to assist in patient management. PMID- 18677145 TI - Human papillomavirus vaccines: an update for gynecologists. AB - Human papillomavirus (HPV) infections cause several common gynecologic problems. Oncogenic HPV infections are responsible for abnormal cervical cytology, cervical dysplasia, and cervical cancer. In 2006, a vaccine was introduced to prevent these common problems in women's health. The currently available HPV vaccine is quadrivalent, with HPV types 6, 11, 16, and 18. Recently published clinical trials have consistently shown efficacy approaching 100%. Current clinical recommendations are for universal vaccination of adolescent and young adult women. This review will discuss the natural history and epidemiology of HPV, the diseases associated with these infections and current clinical usage of this vaccine. Future developments in this area will also be discussed. PMID- 18677146 TI - Evidence-based approach to menopause. Foreword. PMID- 18677147 TI - Evidence-based medicine and the management of menopause. AB - Although all clinicians should practice evidence-based medicine to assure the highest quality healthcare for our patients, there have been several misconceptions about what this really is, or what it should be. In some instances, the overreliance on a single randomized clinical trial (RCT), while ignoring a wealth of other literature resulted in erroneous conclusions. In many disease entities, results of RCTs and observational trials have rendered similar conclusions. Several weaknesses of RCTs and observational trials will be reviewed. In the area of menopausal medicine, a more in depth study of RCT data, including an appreciation of the type of population studied, the ages and risk factor status of the women participating, and the hormonal regimen tested, results in findings which are nearly identical to many observational trials. In the final analysis, no single study should be relied on solely to guide clinical practice. Understanding that there is a hierarchy of clinical data, the entire literature should be sought to provide the basis on which medicine is practiced. PMID- 18677148 TI - Symptoms of menopause: hot flushes. AB - Vasomotor symptoms (VMSs) are highly prevalent during the peri- and early postmenopause. They constitute a major driver for patient self-referral for medical care. Although most women will experience an abatement of their VMS by 5 years after their final menses, women with early or surgical menopause may have worse or more persistent symptoms, and up to 16% of naturally menopausal women continue to experience VMS well after their menopause is past. Although estrogen is the most effective known therapy, it is neither appropriate nor desirable for every symptomatic woman, and nonhormonal treatments such as gabapentin, selective serotonin reuptake inhibitors/selective norepinephrine reuptake inhibitors drugs, and the antihypertensives clonidine and alphamethyldopa may be helpful for some women. There is mounting evidence to support the ineffectiveness of many proposed complementary and alternative modalities. This review will highlight the natural history of VMS and the current medical evidence supporting various treatments. PMID- 18677149 TI - Vulvovaginal complaints. AB - Overview of the etiology and diagnostic evaluation of vulvovaginal complaints in the postmenopausal population including both estrogen's role in atrophic vaginitis and other causes of vulvovaginal complaints and how they relate to sexual function. Summary of various management options such as hormonal, nonhormonal, and botanical interventions. PMID- 18677150 TI - Osteoporosis prevention and management: an evidence-based review. AB - Evidence-based guidelines that have been prepared by many professional organizations aimed at assisting the clinician in the initial evaluation of postmenopausal women who should be considered for bone mineral density evaluation by dual-energy x-ray absorptiometry; history, physical examination, and laboratory testing in those women being considered for pharmacologic intervention; and monitoring and management of women for whom therapy is prescribed are discussed. PMID- 18677152 TI - Hormone therapy and stroke. AB - Available biochemical and clinical data are sufficiently compelling to recommend low dose or transdermal hormone therapy for the management of menopausal symptoms when possible. Women initiating low dose or transdermal hormone therapy are not at increased risk of cardiovascular disease, stroke, venous thrombotic disease, and breast cancer. Evidence suggests that these outcomes are favorably affected by low-dose hormone therapy and persist with continuation of therapy. PMID- 18677151 TI - Postmenopausal hormone therapy and cardiovascular disease in perspective. AB - The totality of data indicate that the window of opportunity for reducing mortality and coronary heart disease is initiation of hormone therapy (HT) within 6 years of menopause and/or by 60 years of age and continued for 6 years or more. Additionally, the risks of HT are rare (<1/1000) especially in younger postmenopausal women and comparable with other primary prevention therapies. In fact, as randomized controlled trial results accumulate, the more they look like the consistent observational data. These studies showed that young postmenopausal women with menopausal symptoms who use HT for long periods of time have lower rates of mortality and coronary heart disease than comparable postmenopausal women who do not use HT. PMID- 18677153 TI - Breast cancer and hormonal therapy. AB - Valid evidence from randomized-controlled trials indicates that breast cancer risk is increased with combined estrogen/progestogen use and that such treatment implies a risk greater than that of estrogen alone. Overall, risk estimates from observational studies are somewhat higher than in randomized-controlled trials but remain modest as compared with other risk factors even after long-term treatment. For combined estrogen/progestogen therapy, risk increases gradually to reach statistical significance after 4 to 5 years. Apart from its many beneficial health effects, the safety data for use of estrogen alone are quite reassuring. The only justifications for progestogen addition are for bleeding control and endometrial protection. At present, there are several new therapeutic compounds and concepts in development, which hold promise to provide both endometrial protection and breast safety. PMID- 18677154 TI - Association of ovarian and uterine cancers with postmenopausal hormonal treatments. AB - During hormone treatments for the relief of the symptoms of postmenopausal women a number of side effects may occur. Some may be due to the wrong choice of the steroids used for treatment or to the route of administration. However, the more important ones deserving much attention are the rare occurrences of malignancies of the uterus and ovaries. The risk for ovarian cancer, if it exists, is minimal and clinically irrelevant. Estrogen only treatments are used only in hysterectomized women. Continuous combined estrogen-progestin treatments have a very low risk of association with endometrial cancers compared with sequential regimens. Tibolone may be associated with a very small risk for endometrial cancers and thus must be properly monitored by transvaginal ultrasound. Breast cancer patients being treated with tamoxifen require careful attention to the endometrium to exclude a carcinoma. For the protection of the endometrium, a progestin-releasing intrauterine devise is an attractive choice. Raloxifene used for a long time to prevent osteoporosis is safe for the endometrium. None of the above-mentioned side effects is enough to prevent a physician from using hormone treatment in postmenopausal women if there are no past or current contraindications. PMID- 18677155 TI - Cognitive changes after menopause: influence of estrogen. AB - The natural menopause is not associated with substantial cognitive change. Limited clinical trial evidence suggests that estrogen-containing hormone therapy has little effect on cognition during midlife, but prompt initiation after surgical menopause may improve aspects of memory. Among older postmenopausal women, strong clinical trial evidence demonstrates that hormone initiation does not improve cognition. More limited clinical trial evidence indicates no improvement in Alzheimer symptoms, and the Women's Health Initiative Memory Study found an increase in dementia risk among older women. Observational findings of reduced Alzheimer risk may reflect early hormone use in younger women, or findings may be biased. Cognitive effects of selective estrogen receptor modulators are not yet well studied. PMID- 18677156 TI - Role of exercise and nutrition in menopause. AB - Menopause and the aging process itself cause many physiologic changes, which explain the increased prevalence of chronic diseases observed in postmenopausal women. Exercise and nutrition play important roles in the prevention and treatment of cardiovascular disease, cancer, obesity, diabetes, osteoporosis, and depression. PMID- 18677160 TI - Comorbidity in patients undergoing coronary artery bypass graft surgery: impact on outcome and implications for cardiac rehabilitation. AB - The increasing comorbid disease burden among patients undergoing coronary artery bypass graft surgery (CABG) and the improved operative survival are expanding the number of post-CABG patients living with prognostically significant comorbidities. In a large contemporary database, 29.9% of the patients receiving isolated CABG had diabetes mellitus, 16% peripheral vascular disease, 18.6% chronic obstructive pulmonary disease, and 27.5% renal dysfunction. Patients with comorbidity are more likely to be old and often female, may have special care requirements early after discharge, and are at increased risk for adverse outcomes. Contemporary available evidence indicates that older individuals, women, and patients with comorbidities are significantly less likely to receive cardiac rehabilitation. In addition, compliance with proven atherosclerosis risk reduction strategies for CABG patients is suboptimal. In this article we will review the impact of comorbidity on short-term and long-term outcome after CABG and their implications for cardiac rehabilitation. PMID- 18677161 TI - Effects of drug, biobehavioral and exercise therapies on heart rate variability in coronary artery disease: a systematic review. AB - BACKGROUND: Heart rate variability (HRV) is reported as a surrogate index for clinical outcome in trials of secondary prevention strategies for coronary artery disease (CAD), but a standardized guide for interpreting HRV change is not established. DESIGN: We evaluated HRV change in trials with CAD patients who received conventional medications (beta-blockers, calcium channel blockers, angiotensin converting enzyme inhibitors), biobehavioral treatment (psychotropics, biofeedback, relaxation) or exercise training. METHODS: Medline, Pubmed, Psycinfo, the Cochrane database, and Embase were searched until July 2007, without language restriction. We identified 33 randomized controlled trials. Two reviewers independently abstracted all trials using a standardized form. A hierarchy of frequency and time domain HRV indices defined outcome. RESULTS: A random-effects model yielded an overall pooled standardized mean difference (SMD) between treatment and control groups of moderate magnitude across treatment classes, based on a composite of time and frequency domain indices (SMD=0.40, P<0.0001), or only time or frequency indices (SMD=0.37 and 0.43, respectively, both P<0.0001). This change was equivalent to an increase in standard deviation of all normal-to-normal RR intervals of 9.0 ms (95% Confidence Interval, CI, 7.3, 10.7 ms) or a relative increase of 15.9% (95% CI, 13.2, 18.6%). To detect HRV change of this magnitude, a hypothetical trial would require a sample size of 660 patients for conventional medications or 1232 patients for all treatment classes. CONCLUSION: Pharmacologic, biobehavioral and exercise strategies for secondary prevention of CAD significantly increase HRV. This review provides a framework to assist efforts to evaluate the contribution of HRV change to CAD prognosis. PMID- 18677162 TI - Elevated total cholesterol: its prevalence and population attributable fraction for mortality from coronary heart disease and ischaemic stroke in the Asia Pacific region. AB - BACKGROUND: About half of the world's cases of cardiovascular disease occur in the Asia-Pacific region. The contribution of serum total cholesterol (TC) to this burden is poorly quantified. DESIGN: The most recent nationally representative data on TC distributions for countries in the region were sought. Individual participant data from 380,483 adults in the Asia Pacific Cohort Studies Collaboration were used to estimate associations between TC and cardiovascular disease. METHODS: High TC was defined as > or =6.2 mmol/l, and nonoptimal TC as > or =3.8 mmol/l. Hazard ratios for fatal coronary heart disease (CHD) and ischaemic stroke (IS) were found from Cox models. Sex-specific population attributable fractions for high TC and nonoptimal TC were estimated for each country. The former used conventional methods, based on single measures of TC and a fixed dichotomy of risk strata; the latter took account of the continuous positive association between TC and both CHD and IS and regression dilution. RESULTS: Data were available from 16 countries. Where reported, the prevalence of high TC ranged from 4 to 27%. The fraction of fatal CHD and IS attributable to high TC ranged from 0 to 14% and 0 to 15%, respectively. Although leaving the relative ranking of countries much the same, the fractions estimated for nonoptimal TC were typically at least twice as big, ranging from 0 to 47% and 0 to 35%, respectively. CONCLUSION: Conventional methods for estimating disease burden severely underestimate the effect of TC. Cholesterol-lowering strategies could have a tremendous effect in reducing cardiovascular deaths in this populous region. PMID- 18677163 TI - Predictive accuracy and usefulness of calibration of the ESC SCORE in Switzerland. AB - BACKGROUND: Prevention of cardiovascular disease (CVD) at the individual level should rely on the assessment of absolute risk using population-specific risk tables. OBJECTIVE: To compare the predictive accuracy of the original and the calibrated SCORE functions regarding 10-year cardiovascular risk in Switzerland. DESIGN: Cross-sectional, population-based study (5773 participants aged 35-74 years). METHODS: The SCORE equation for low-risk countries was calibrated based on the Swiss CVD mortality rates and on the CVD risk factor levels from the study sample. The predicted number of CVD deaths after a 10-year period was computed from the original and the calibrated equations and from the observed cardiovascular mortality for 2003. RESULTS: According to the original and calibrated functions, 16.3 and 15.8% of men and 8.2 and 8.9% of women, respectively, had a 10-year CVD risk > or =5%. Concordance correlation coefficient between the two functions was 0.951 for men and 0.948 for women, both P<0.001. Both risk functions adequately predicted the 10-year cumulative number of CVD deaths: in men, 71 (original) and 74 (calibrated) deaths for 73 deaths when using the CVD mortality rates; in women, 44 (original), 45 (calibrated) and 45 (CVD mortality rates), respectively. Compared to the original function, the calibrated function classified more women and fewer men at high-risk. Moreover, the calibrated function gave better risk estimates among participants aged over 65 years. CONCLUSION: The original SCORE function adequately predicts CVD death in Switzerland, particularly for individuals aged less than 65 years. The calibrated function provides more reliable estimates for older individuals. PMID- 18677164 TI - Evaluation of a recalibrated Systematic Coronary Risk Evaluation cardiovascular risk chart: results from Systematic Coronary Risk Evaluation Germany. AB - BACKGROUND: A recalibrated cardiovascular disease (CVD) risk chart for the German population, Systematic Coronary Risk Evaluation (SCORE) Germany, was developed in 2005. We evaluated the risk prediction by SCORE Germany in two large population based surveys. STUDY POPULATIONS AND METHODS: We applied the SCORE Germany risk function to men and women, aged 40-65 years, who participated in surveys of the Study of Health in Pomerania (SHIP), north-east Germany or the Cooperative Health Research in the Augsburg Region (KORA) study (southern Germany). The prevalence of single risk factor combinations and of the SCORE Germany risk prediction categories was assessed. The 10-year risk of fatal CVD predicted by SCORE Germany (P) was evaluated against the risk observed in the general population using official mortality data (O). RESULTS: Less than one in 25 women in the two studies had a high-predicted 10-year risk of fatal CVD (> or =5%), whereas one in three men in SHIP and about one in four men in KORA belonged to this risk group. The age-specific predicted risk of fatal CVD was consistently higher for men and women from SHIP than from KORA. On comparison, the P/O ratio of 10-year CVD risks was close to unity and tended to decline with higher age in the both studies. CONCLUSION: Overestimations of risks with the original SCORE model are reduced mostly with SCORE Germany. Differences in risk factors and CVD risk between populations within Germany are reflected properly by risk predictions with SCORE Germany. The recalibrated risk chart seems to be suited for use in German clinical practice. PMID- 18677165 TI - Increasing physical education in high school students: effects on concentration of circulating endothelial progenitor cells. AB - AIMS: Levels of endothelial progenitor cells (EPCs), that can be increased by regular exercise, correlate with vascular function. In the context of primary prevention, the impact of regular physical activity on the amount and function of EPC has not yet been investigated in school children. METHODS: Four sixth grade classes of high school students (n=92) were randomly assigned to either the intervention group (IG) with daily physical exercise (45 min) at school or to the control group (CG) with conventional physical education (PE) (2 h/week). In addition, one sixth grade class at a high school focused on competitive sports (PE) served as a reference group. After 1 school year, exercise capacity and the amount and function of EPCs were evaluated. RESULTS: After 1 year, a significantly higher Vo2max was evident in the intervention group. Nevertheless, exercise capacity did not reach the level of children from PE. In addition, exercise intervention was successful in increasing the amount of EPCs but failed to increase the migratory capacity of the cells. CONCLUSION: The result of this study shows, that intensified, supervised school sports leads to an increase in exercise capacity and EPCs in children. Nevertheless, its effect on primary prevention in cardiovascular disease has to be proven in further longitudinal studies. PMID- 18677166 TI - Comparative prognostic value of glomerular filtration rate estimating formulas in ischaemic heart disease. AB - BACKGROUND: Chronic kidney disease is a major risk factor for cardiovascular death and may be detected by measuring serum creatinine levels (SCr). Alternatively, renal function may be evaluated by estimating glomerular filtration rate (GFR) with formulas based on SCr and variables such as age, sex and body weight. The purpose of the study was to compare the predictive value of GFR, calculated with the Cockcroft-Gault (CG) and revised Modification of Diet in Renal Disease formulas, in a population with ischaemic heart disease (IHD). DESIGN: Retrospective observational study. METHODS: Renal function was analysed in 1705 patients (mean age 55+/-9.2 years, 80% male) who had been admitted to our Institute for IHD. The relationship between reduced GFR and outcome was examined through Kaplan-Meier curves and Cox regression analysis. RESULTS: Ten-year mortality was 20% and had as predictors both an abnormal (> or =1.10 mg/dl) SCr and reduced (<60 ml/min/1.73 m2) Modification of Diet in Renal Disease and CG GFR (P<0.0001 for each). Using Cox regression analysis, significant coronary atherosclerosis, defined as a greater than 50% stenosis in at least one major vessel, turned out to be the factor most closely linked to increased mortality [HR corresponds to hazard ratio 4.40, 95% (confidence interval) CI 2.78-6.97, P<0.001], followed by reduced CG GFR (HR 2.08, 95% CI 1.55-2.79, P<0.001) and left bundle branch block (HR 2.00, 95% CI 1.10-3.61, P<0.001). CONCLUSION: GFR, especially if calculated with the CG formula, is an important prognostic indicator for IHD among hospitalized patients, and predicts survival more accurately than SCr and conventional coronary risk factors. PMID- 18677167 TI - Truncal fat determined by dual-energy X-ray absorptiometry is an independent predictor of coronary artery disease extension. AB - BACKGROUND: Controversy prevails regarding the existence of a correlation between the severity of coronary artery disease (CAD) and the extent and distribution of obesity. PURPOSE: To assess the correlation between total fat, truncal fat (TF), and lean mass, obtained with dual-energy X-ray absorptiometry (DEXA) and standard anthropomorphic indices (body mass index, waist circumference, waist-to-hip ratio) and to verify whether DEXA indices can predict the extent and severity of CAD. MATERIALS AND METHODS: Fifty-eight patients (19 females) consecutively referred for coronary angiography underwent physical examination and DEXA assessment of body composition. RESULTS: Of the 58 patients enrolled, 22 were overweight and 13 were obese. Significant CAD was found in 39 (67%) patients. DEXA-derived total mass and fat mass enabled us to distinguish overweight from obese patients (P<0.005), whereas just TF mass correlated with the number of diseased vessels after adjusting for body mass index, sex, age, and smoking habit (odds ratio, 8.68; 95% confidence interval: 1.02-74.10). CONCLUSION: TF determined by DEXA is independently related with CAD extension. PMID- 18677168 TI - Anxiety and depression after coronary artery bypass graft surgery: most get better, some get worse. AB - BACKGROUND: To target interventions, patients at risk for poor outcomes after a cardiac event need to be identified. We investigated trajectories of anxiety and depression after coronary artery bypass graft surgery (CABGS) and identified patients at risk of persistent or worsening anxiety and depression. METHODS: A consecutive sample of 184 patients on the waiting list for CABGS at The Royal Melbourne Hospital completed self-report questionnaires before surgery, and at 2 and 6 months postsurgery. Anxiety and depression were measured using the Hospital Anxiety and Depression Scale. Growth mixture modelling identified trajectories of anxiety and depression. RESULTS: Two possible trajectories emerged for anxiety, whereas three trajectories emerged for depression. Most patients (92%) followed a trajectory of minor presurgical anxiety that remitted in 6 months after CABGS, with the remainder (8%) following a trajectory of major anxiety that remitted in the same period. Minor remitted depression was also common (72% patients). Two less common depression trajectories indicated worsening or unresolved depression. One trajectory began with major presurgical depression that partially remitted by 6 months (14% patients) and the other began with minor presurgical depression that worsened by 6 months (14% patients). Unpartnered patients, smokers, those with presurgical anxiety, high cholesterol, angina, more severe disease or having repeat CABGS were at increased risk for a poor depression trajectory. CONCLUSION: Although initial anxiety and depression resolved or lessened for most patients, some patients experienced persistent or worsening depression after CABGS. Interventions can be targeted toward 'at risk' patients. PMID- 18677169 TI - Perioperative and postoperative predictors of outcome in patients with low ejection fraction early after coronary artery bypass grafting: the additional value of left ventricular remodeling. AB - BACKGROUND: Scant data exist on factors that may identify outcome in patients with severe left ventricular (LV) dysfunction early after coronary artery bypass graft surgery (CABG). DESIGN: This study was designed to determine the prognostic value of clinical, operative, and postoperative factors in patients with LV dysfunction early after CABG. METHODS: In 333 consecutive patients with ejection fraction < or =35% on admission to residential cardiac rehabilitation after isolated CABG, potential preoperative, perioperative, and postoperative predictors of outcome, including 6-month LV remodeling, were recorded and patients followed up for a median of 3 years. The study end points were cardiovascular (CV) mortality and the combination of CV mortality and nonfatal CV events requiring hospitalization. RESULTS: The 3-year CV mortality-free survival and survival free of nonfatal CV event rates were 87 and 73%, respectively. Independent predictors of CV mortality were history of congestive heart failure [hazard ratio, HR: 2.8; 95% Confidence Interval (CI): 1.51-5.21], low ejection fraction on admission to cardiac rehabilitation (HR: 0.9; 95% CI: 0.87-0.96), and early complications after CABG (HR: 2.5; 95% CI: 1.23-5.15). When the combined end points were considered, postoperative left atrial size (HR: 1.07; 95% CI: 1.01-1.11), New York Heart Association class III or IV (HR: 1.69; 95% CI: 1.04 2.74), and 6-month remodeling (HR: 2.12; 95% CI: 1.33-3.36) were independent predictors. CONCLUSION: Simple preoperative and postoperative variables may help identify patients with LV dysfunction early after CABG who are still at risk of major CV events. In this setting, 6-month LV remodeling is a strong predictor of a poor prognosis. PMID- 18677170 TI - The role of sex in health-related quality of life after cardiac surgery: a prospective study. AB - BACKGROUND: To assess health-related quality of life (HRQOL) in patients after cardiac surgery with emphasis on sex differences. DESIGN AND METHODS: Between September 2004 and September 2005, 534 patients (413 males and 121 females) were consecutively included. HRQOL was measured by the short-form 36 (SF-36) before surgery with follow-up 6 and 12 months after surgery. RESULTS: Five hundred and twenty-one patients were alive after 12 months, 462 (89%) and 465 (89.4%) responded after 6 and 12 months, respectively. Female patients had less favorable scores than male patients on most subscales of the SF-36 both before and after surgery. Both male and female patients improved substantially after surgery, but female patients reported significantly less improvement on two of eight subscales of the SF-36; role emotional and bodily pain. CONCLUSION: The study demonstrates that there are sex differences concerning HRQOL both before and after cardiac surgery. A clear overall improvement in HRQOL over the first year after cardiac surgery, more specifically during the first 6 months for both sexes was found. PMID- 18677171 TI - Importance of exercise training session duration in the rehabilitation of coronary artery disease patients. AB - BACKGROUND: In cardiac rehabilitation, 40-60-min exercise training sessions are advised. However, because of the increasing coronary artery disease (CAD) prevalence and higher workload for cardiac rehabilitation centres, it remains unclear whether 40-min exercise training sessions are equally effective as 60-min exercise training sessions. DESIGN: Prospective randomized clinical trial. METHODS: One hundred and thirty-four CAD patients were included in a 7-week rehabilitation programme. All patients exercised 3 days per week, at a heart rate corresponding to 65% of baseline peak oxygen uptake (VO2peak). Patients were randomized in two groups: 40 versus 60-min exercise training sessions. Changes of body anthropometrics, resting haemodynamics, exercise capacity and ventilatory threshold, blood plasma lipid profile and C-reactive protein level were assessed. RESULTS: As a result of rehabilitation, exercise capacity, ventilatory threshold, and blood plasma lipid profile improved significantly in the total population (P<0.05), without differences between subgroups (P>0.05). Body weight and waist circumference decreased significantly in total population (P<0.01), but with a greater magnitude in the 40 versus 60-min exercise session group (P<0.05). CONCLUSIONS: In the early rehabilitation of CAD patients, 40-min exercise training sessions seem to be at least as effective for improving body anthropometrics, blood plasma lipid profile and exercise capacity, as compared with 60-min exercise training sessions. PMID- 18677172 TI - Plasma B-type natriuretic peptide predicts atrial fibrillation during rehabilitation after cardiac surgery. AB - BACKGROUND: B-type natriuretic peptide (BNP) levels are known to predict atrial fibrillation (AF) occurrence short-term after cardiac surgery, but no information is available on their predictive potential at a later time point. We evaluated whether BNP levels predict postcardiac surgery AF events occurring during rehabilitation program. AF impact on hospitalization length and rehabilitation program have also been evaluated. METHODS AND RESULTS: One hundred and forty-nine patients who underwent cardiac surgery were monitored for 'late' AF, defined as AF occurring during the rehabilitation period (20+/-5 days) in contrast to 'early' AF defined as AF documented in the surgical department soon after surgery. BNP was determined at rehabilitative hospital admission (10+/-5 days after surgery). Late AF was observed in 17% of patients. AF patients had higher BNP levels than event-free patients (459+/-209 vs. 401+/-449 pg/ml, P=0.01). Lower kaliemia values (P=0.048), early AF (P<0.001), and combined surgery (coronary artery by pass graft and valve replacement; P=0.016) were also associated with late AF. At multivariate analysis, BNP levels more than 322 pg/ml (P=0.02), and early AF (P=0.003) showed an independent association with late AF occurrence, which did not interfere with the physical training program but prolonged hospitalization (22+/-5 vs. 20+/-5 days, P=0.062) and telemetry monitoring (6+/-5 vs. 1+/-3 days, P<0.001). CONCLUSION: BNP levels measured at the beginning of the rehabilitation program are independent predictors of late AF after cardiac surgery. These results suggest a more aggressive therapeutical approach during the rehabilitation period in patients with elevated BNP levels who have already experienced AF in the surgical department. PMID- 18677173 TI - Changes in lipid profile of patients referred to a cardiac rehabilitation program. AB - BACKGROUND: Cardiac rehabilitation program (CRP) has been demonstrated to have beneficial effects on physical and mental functioning as well as on mortality of patients with cardiovascular diseases, but its exact effect on lipid profile of these patients is still vague. In this study we aimed to evaluate the effect of comprehensive CRP on lipid profile of the Iranian population. DESIGN: Self controlled descriptive study. METHODS: We evaluated 547 patients with documented coronary heart disease before and after a 24-session CRP between 1998 and 2003. Some of them received antilipid drugs. RESULTS: Except for diastolic blood pressure, all other biophysical, biochemical [total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglyceride (TG)], functional, and psychosocial parameters had significant response to CRP. We can attribute the normalization of lipid in these patients to CRP in combination with medications 34.9% for TC, 17.7% for LDL, and 27.2% for TG (P<0.001 for all). Antilipid drugs had some more effect in normalizing the TC (9.7%), LDL (1.8%), and TG (7.3%). CONCLUSION: Comprehensive CRP improves TC, TG, LDL, and HDL in Iranian patients with cardiovascular disease even without antilipid drugs, and antilipid therapy can boost this effect. PMID- 18677174 TI - Exercise training but not rosiglitazone improves endothelial function in prediabetic patients with coronary disease. AB - BACKGROUND: Impaired glucose tolerance (IGT) is associated with endothelial dysfunction and upregulation of inflammatory markers, which is potentially reversible by adequate treatment. It was our aim to compare the impact of exercise training with that of rosiglitazone on endothelial function and inflammatory markers in patients with IGT and coronary artery disease (CAD). METHODS: Patients with IGT and CAD were randomly assigned to either exercise training (n=13), rosiglitazone (8 mg; n=11), or a control group (n=10). During the first week, exercise training consisted of 6 x 15 min/d followed by three weeks of 30 min/d submaximal ergometer exercise. In addition, group exercise training of 1 h was performed twice per week. RESULTS: After 4 weeks, triglycerides and uric acid were significantly lower in the exercise group whereas fasting glucose, HbA1c, low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol, C-reactive protein, fibrinogen, and body mass index did not differ between groups. In the exercise group, exercise capacity (123+/-33 vs. 144+/-31 W; P=0.006) and endothelium-dependent, flow-mediated vasodilatation (P<0.01) increased significantly, whereas in the rosiglitazone group and in the control group (P=n.s.) no changes were seen. CONCLUSION: In patients with IGT and CAD, 4 weeks of exercise training exert significant and superior improvement of endothelium-dependent vasodilatation as compared with rosiglitazone therapy or usual care. This finding should be seen as an even further encouragement to recommend and, where available, prescribe exercise training to our patients. PMID- 18677175 TI - Usefulness of a single-item general self-rated health question to predict mortality 12 months after an acute coronary syndrome. AB - BACKGROUND: A single-item general self-rated health (GSRH) question consistently predicts mortality in community cohort studies, but has not been examined in patients with acute coronary syndrome (ACS). We investigated whether a single item GSRH question predicted mortality 12 months post-discharge in 800 ACS patients. METHODS: Logistic regression was used to assess the relationship of the single-item GSRH question with mortality, controlling for cardiac risk factors, including depressive symptoms. RESULTS: The single-item GSHR question was associated with mortality on a bivariable basis (odds ratio=0.50, 95% confidence interval=0.28-0.92, P=0.027), but was not significant after controlling for other risk factors (odds ratio=0.80, 95% confidence interval=0.40-1.60, P=0.522). PMID- 18677176 TI - 'Lone' increase in C-reactive protein after cardiac surgery: prevalence, clinical characteristics, in-hospital course, and prognostic value. AB - BACKGROUND: Serum C-reactive protein (CRP) is involved in the acute phase reaction after surgery, even though its clinical significance remains a matter of debate. We evaluated CRP levels in cardiac surgery patients without clinical or laboratory signs of infection. METHODS: We screened 737 consecutive patients referred to our center 8+/-5 days after cardiac surgery. Patients with fever (>37.2 degrees C), elevated white blood cell count (>11,000/ml), neutrophilia (>70%), or any inflammatory, infective or malignant disease were excluded. CRP levels were measured on admission and at discharge and the values were related to the following variables: age, sex, diabetes mellitus, renal failure, type of surgery, postoperative atrial fibrillation, pericardial or pleural effusion, and length of hospital stay. Follow-up (mean: 23+/-8.5 months) was available for 175 patients (94%). RESULTS: In the 187 patients enrolled in the study, the CRP values were significantly elevated (median: 4.23 mg/dl, interquartiles range: 2.68-6.64) independent of any variable analyzed. At discharge, CRP levels were significantly reduced compared with values on admission (median: 1.55 mg/dl, interquartiles range: 0.84-2.37, P<0.001). At follow-up, 19 events (10.8%) occurred (two noncardiac deaths, 17 hospital readmissions for cardiac reasons); nonetheless, no correlation was found with CRP values either on admission or at discharge. CONCLUSION: Early after cardiac surgery, in patients without clinical or laboratory signs of acute infection, CRP levels are significantly elevated, do not correlate with clinical variables, and decrease at discharge. These findings suggest a systemic inflammatory response to surgery-related stress, which carries a favorable prognosis at follow-up. PMID- 18677177 TI - Intensified preventive care to reduce cardiovascular risk in healthcare centres located in deprived neighbourhoods: a randomized controlled trial. AB - BACKGROUND: We examined the effectiveness of a structured collaboration in general practice between a practice nurse, a peer health educator, the general practitioner (GP) and a GP assistant in providing intensified preventive care for patients at high risk of developing cardiovascular diseases. DESIGN: A randomized controlled trial in three healthcare centres (18 GPs) in deprived neighbourhoods of two major Dutch cities. METHODS: Two hundred seventy-five high-risk patients (30-70 years) from various ethnic groups were randomized to intervention (n=137) or usual care group (n=138). We determined group differences in outcomes [10-year absolute risk (Framingham risk equation), blood pressure, lipids and body mass index] at 12-month follow-up. RESULTS: The 10-year absolute risk was reduced by 1.76% (standard error: 0.81) in intervention and by 2.27% (standard error: 0.69) in usual care group; the difference in mean change was 0.88% [95% confidence interval: -1.16 to 2.93]. In both groups significant reductions were observed in the following individual risk factors: total cholesterol, total cholesterol/high density lipoprotein cholesterol, and low-density lipoprotein cholesterol, with no relevance between group differences. CONCLUSION: The cardiovascular risk profile of intervention and control patients improved after 1-year follow-up. However, no extra effect of the structured preventive care on the risk for cardiovascular diseases was achieved. PMID- 18677178 TI - Diarrhoea in childhood and cause-specific mortality in older age: analyses of 5642 deaths in 33,261 individuals from the Hertfordshire studies. AB - The objective of these analyses were to examine if episodes of diarrhoea in childhood are related to adult coronary heart disease (CHD) mortality. Data were pooled from two prospective UK cohort studies (participants born 1923--1939). Reported diarrhoea in infancy (3.1%) and between 1 and 5 years (1.1%) was rare. No relationship between diarrhoea in infancy and CHD mortality or mortality from all-causes and cardiovascular disease was observed. Although diarrhoea between 1 and 5 years was also not related to CHD risk, slightly lower rates of total and cardiovascular disease mortality were apparent in this group relative to the diarrhoea-free. In the largest study to date to examine the relation, there was no evidence that diarrhoea in early life had an influence on later CHD mortality. PMID- 18677179 TI - Green tea: beneficial effects on cholesterol and lipid metabolism besides endothelial function. PMID- 18677181 TI - Detection and treatment of refractory status epilepticus in the intensive care unit. AB - Status epilepticus (SE) is not rare in critically ill intensive care unit (ICU) patients, but its diagnosis is often delayed or missed, in part because it is mistaken for other causes of altered mental status. Even once diagnosed, SE in the ICU can be refractory to treatment. We sought to determine the causes, clinical features, and difficulties in diagnosis of SE in the ICU, and the effects of antiepileptic drugs (AEDs) on its course. We reviewed the course of ICU patients with both clinical and EEG evidence of SE, attempting to determine which patients are at risk for unsuspected SE, what was the typical delay in diagnosis, and whether AED treatment made a difference in their clinical courses. By clinical and EEG evidence, 91 ICU patients with SE were identified, all with abnormal mental status: 74 were comatose. Vascular disease (in 24) and anoxia (22) were the most common causes; most had multiple medical problems. Although 76 patients had clinically evident seizures earlier (and 56, clinical SE) only 20 were thought to be in SE at the time of the diagnostic EEG. There was a median delay of 48 hours from clinical deterioration until diagnosis in patients with earlier clinical seizures and 72 hours without seizures. Among the 68 nonanoxic patients treated with AEDs, 38 (56%) seemed to improve in alertness, including 25 who were comatose. Although patients who were stuporous or confused (vs. comatose) improved more often on AEDs, they were less often realized to be in SE before the EEG. Patients with earlier seizures were also more likely to improve, but no more likely to be diagnosed before the EEG. Patients who responded to AEDs were more likely to survive. ICU patients with altered mental status and EEG evidence of SE often have severe medical and surgical illnesses, refractory SE, and a high mortality. The delay to diagnosis is substantial, but a significant subset of patients improves on AEDs once SE is discovered. This diagnosis should be sought more often in ICU patients with abnormal mental status, especially after clinical seizures or SE without full recovery. PMID- 18677182 TI - Resting-state oscillatory brain dynamics in Alzheimer disease. AB - Altered oscillatory brain activity in Alzheimer disease (AD) may reflect underlying neuropathological changes, and its characterization might lead to new diagnostic possibilities. The present study using quantitative magnetoencephalography was set up to examine power spectrum changes in AD patients, and their diagnostic strength. Whole-head 151-channel magnetoencephalography was recorded during an eyes-closed resting state. Magnetoencephalography channels were grouped in 10 cortical regions, and both global and regional relative power was analyzed for the commonly used frequency bands. Eighteen AD patients [mean age 72.1 years +/- 5.6 (SD); 7 women; mean Mini Mental State Examination score 19.2, range: 13-25] and 18 healthy controls [mean age 69.1 +/- 6.8 (SD), 11 women; mean Mini Mental State Examination score 29, range: 27-30] were recruited, controls being mainly spouses of patients. Relative power analysis showed significant differences in most frequency bands, particularly in the temporo-parietal regions, with some relation to Mini Mental State Examination scores. Greatest diagnostic accuracy was found in the beta band, especially in the right occipital area (sensitivity 94%, specificity 78%). Quantitative relative power analysis of magnetoencephalography recordings is able to show widespread abnormalities in oscillatory brain dynamics in AD patients. By analyzing distinct cortical regions, this study provides a more detailed topographical view of abnormal brain activity in AD. PMID- 18677183 TI - The impact of stimulus properties on low- and high-frequency median nerve somatosensory evoked potentials. AB - Questioning whether stimulation properties in median nerve somatosensory evoked potentials show interactions multichannel recordings were performed in a three factorial repeated measures design with the parameters (i) eyes opened versus eyes closed, (ii) stimulation intensity above motor threshold versus intensity sub motor threshold, (iii) stimulation rate 0.5 Hz versus 9 Hz resulting in somatosensory evoked potentials recorded during eight different conditions. Varying the stimulation intensity revealed an impact on the amplitude and the latency of the N20 source activity and on the amplitude, the duration and the number of peaks of the high frequency oscillatory (HFOs) sources. Modifying the stimulation rate lead to an effect on the amplitude and latency of the N20 and on the amplitude of the high-frequency oscillatory sources. The condition opened/closed eyes had an impact on the duration and number of high-frequency oscillatory. No relevant interactions between the stimulation properties were found. In consequence, varying one stimulus parameter already leads to a saturation of the low as well as high-frequency somatosensory evoked potentials components. Thus, the careful choice of stimulation parameters is a condition precedent for reasonable data interpretation. PMID- 18677184 TI - Corticospinal facilitation following prolonged proprioceptive stimulation by means of passive wrist movement. AB - The purpose of this study was to evaluate the delayed effects of repetitive sensory stimulation with passive wrist movement on corticospinal excitability of the forearm and hand musculature. Motor evoked potential responses to single and double pulse transcranial magnetic stimulation were recorded from the flexor carpi radialis, extensor carpi radialis, and the first dorsal interosseous muscles of the right limb. Data were collected before and after a 1 hour session of passive wrist movement (intervention group, n = 11) or after a same period of rest (control group, n = 9). Motor evoked potential size and area were analyzed to evaluate corticospinal excitability and short interval intracortical inhibition and facilitation. Training with passive movement resulted in a prolonged increase in corticospinal excitability in the flexor carpi radialis and extensor carpi radialis (until at least 1 hour postintervention), but did not evoke significant changes in the levels of short interval intracortical inhibition and facilitation. No such effects were noted in the control group or first dorsal interosseous muscle. Prolonged proprioceptive stimulation with passive wrist movement induces a delayed increase in corticospinal excitability of the forearm muscles. Accordingly, this intervention may promote motor cortical reorganization in the targeted muscles. Results show induced effects from passive movement training that may prove useful for neurorehabilitation therapies. PMID- 18677185 TI - Ankle paresis in incomplete spinal cord injury: relation to corticospinal conductivity and ambulatory capacity. AB - There is limited data on the relation of corticospinal tract conductivity to clinical measures in incomplete spinal cord injury. This study examined the relationship of muscle strength to corticospinal tract input assessed by motor evoked potentials (MEPs) during static and dynamic conditions and to gait. Dorsiflexor strength was established by manual muscle test, maximal voluntary contraction, and maximal movement velocity, the latter being acquired during auditory-paced ankle movements. MEPs were elicited during isometric contractions with constant or continuously increasing ankle joint torque. Gait was assessed by quantifying its speed and independence. Linear regression analyses showed that maximal movement velocity was related to the MEP latencies and amplitudes in the dynamic condition (R(2)(adj.) = 0.62) and to the MEP latencies in the static condition (R(2)(adj.) = 0.45). Maximal voluntary contraction was only related to the MEP latencies in the static (R(2)(adj.) = 0.45) and the dynamic condition (R(2)(adj.) = 0.21), whereas manual muscle test did not show any relationship to the MEPs. In incomplete spinal cord injury patients, the dynamic measure maximal movement velocity might be a useful clinical assessment of corticospinal tract function. Clinical studies on recovery and repair of corticospinal tract function in spinal lesions could substantially benefit from implementing dynamic measures in the clinical assessment protocol. PMID- 18677186 TI - Interrater and intrarater reliability in multiple sleep latency test. AB - Multiple sleep latency test (MSLT) remains the golden standard for the clinical diagnosis and management of excessive daytime sleepiness. However, there was limited data on the reliability measurement of MSLT. Forty-four (M/F ratio: 26/18, mean age of 43.4 +/- 13.9 years) subjects of an ongoing family study of narcolepsy underwent a standard polysomnogram and MSLT. Three trained staff of various level of experiences independently and blindly scored each nap (n = 219). To test for intrarater reliability, 100 naps (n = 20 subjects) were re-scored half a year later. The interrater reliability for the mean sleep latency of 5 naps, sleep latency, and rapid eye movement (REM) sleep latency of individual nap, presence and numbers of sleep onset REM periods varied from the range of 0.668 to 0.964. The mean interrater reliability for the clinical diagnosis of narcolepsy was 0.883 (range, 0.824-0.938), whereas it was 0.750 (range, 0.674 0.858) for the diagnosis of narcolepsy spectrum disorder (shortened mean sleep latency and/or the presence of sleep onset REM periods) and 0.796 (range, 0.697 0.846) for normal cases. The intrarater reliability for the sleep latency and sleep onset REM periods varied from the range of 0.625 to 0.991. Our study demonstrated excellent inter- and intrarater reliability in scoring the sleep latency and sleep onset REM periods of MSLT. They also had excellent agreement on the diagnosis of narcolepsy and an "excellent to good" agreement on the diagnosis of narcolepsy spectrum disorder and normal cases. Our study lent further support that MSLT was an objective measurement with high inter- and intrarater reliability across various sleep disorders and healthy controls among different sleep centers. PMID- 18677187 TI - Electromyographic findings in central nervous system disorders: case series and literature review. AB - We reviewed electrodiagnostic findings in patients whose electromyograph pointed toward a disorder of the central nervous system. Of the 34 patients identified, 17 did not have a clinical diagnosis of a central nervous system disorder before electromyogram. Signs suggesting a central nervous system disorder included poor activation, poor relaxation, extensor plantar responses, and flexor spasms on needle electromyogram. Ultimate central nervous system diagnoses included myelopathy, demyelinating disease, parkinsonism, tumor, and stroke. PMID- 18677188 TI - Spontaneous electromyographic activity during microvascular decompression in trigeminal neuralgia. AB - Intraoperative monitoring of spontaneous facial nerve electromyographic activity during surgery for microvascular decompression in trigeminal neuralgia was evaluated. Fifteen patients with trigeminal neuralgia underwent surgery for microvascular decompression. During the entire operation, free-running facial nerve electromyographic signals were recorded. The data were analyzed with respect to waveform patterns known from vestibular schwannoma-surgery. Special regard was given to the occurrence of A-trains that are associated with postoperative paresis in patients operated on vestibular schwannoma. The spectrum of the observed activities matched patterns known from surgery of vestibular schwannoma; even A-trains, a pattern known to be an indicator of postoperative deterioration of facial nerve function (Romstock et al., J Neurosurg 2000;93:586 593), were seen in 3 of the 15 patients with trigeminal neuralgia. The quantity of A-trains observed was much less than it is known from patients operated on tumors of the cerebellopontine angle. None of the trigeminal neuralgia-patients experienced postoperative deterioration of facial nerve function. The present study shows that A-trains do not only occur during tumor surgery, but also during procedures with indirect manipulation of the facial nerve. They do not necessarily lead to postoperative paresis as long as certain thresholds concerning amount and length of these A-trains are not exceeded. PMID- 18677189 TI - Neurophysiological findings of the late-onset, dominant, proximal spinal muscular atrophies with dysautonomia because of the VAPB PRO56SER mutation. AB - The vesicle-associated membrane protein/synaptobrevin-associated membrane protein B (VAPB) Pro56Ser mutation has been identified in Brazilian families showing various motor neuron syndromes. However, the neurophysiological characteristics of these patients have not been detailed, and some questions still need to be solved, such as the possible presence of myotonia and the origin of the abdominal protrusion seen in most patients. The eventual finding of suggestive electrophysiological characteristics would be helpful not only for clinical diagnosis but also to selection of the appropriate DNA test. To clarify these questions we carried out sensory and motor conduction studies, including sympathetic skin response, and needle examination in six genetically proven affected members. The electromyographic findings were those of a slowly progressive motor neuron disorder. Topographically, the abdominal muscles were severely affected, but the facial and laryngeal muscles were preserved or very mildly involved. Sensory conduction studies and sympathetic skin responses were normal. No myotonic discharge was recorded. These findings are indistinguishable from those of other motor neuron disorders, although the predominant involvement of the proximal limbs and of the abdominal muscles may be of some help in the appropriate clinical setting. PMID- 18677190 TI - Could electrical needle stimulation reduce diagnostic errors in electromyography? AB - During the electromyographic examination there is a risk of wrongly inserting the needle in a muscular area contiguous to the muscle of investigation. The objective of the work is to verify the mistakes chance during the needle placement into the muscle target. We have investigated on 45 patients the identification of the insertion site by electrical stimulation of the muscle, where the needle electrode is inserted, through the same recording needle and calculated how many times the needle can be inserted incorrectly in a muscle close to muscle target. The results showed that during standard needle electromyographic there are a significant error prevalence (33.3%) in the examination of some deeper muscles such as pronator teres (26.3%) and tibialis posterior (40.5%). In conclusion, a correct identification of the muscles tested may a useful practice for all patients especially in patients with motor recruitment problems avoiding other serial examinations and reducing patients discomfort. In other hand, this kind of method can be helpful during a training for a neurophysiologist to improve their technical skill. PMID- 18677192 TI - Early detection of periodic sharp wave complexes on EEG by independent component analysis in patients with Creutzfeldt-Jakob disease. PMID- 18677194 TI - Overview of the STAndards for BipoLar Excellence (STABLE) project. AB - The goal of the STAndards for BipoLar Excellence (STABLE) project was to advance quality of care for bipolar disorder by promoting improved recognition and increased use of evidence-based practices. This article presents a brief overview of the project's objectives, the make-up of the STABLE National Coordinating Council, and the development and field testing of a series of evidence-based clinical performance measures. Findings from field testing of these measures are summarized and highlight the continuing gap between current care and evidence based recommendations in many key clinical areas in psychiatric and primary care. PMID- 18677195 TI - Quality improvement in psychiatry: why measures matter. AB - Increasing attention has been directed in healthcare today to the importance of performance measurement, (i.e., the implementation of measurable methods to demonstrate that practitioners are engaged in high-quality, evidence-based medicine). Many medical specialties, as well as many state medical licensing boards, now require that candidates submit performance measurement data, to be eligible for maintenance of board certification or medical licensure. National organizations such as the National Quality Forum and the Physicians Consortium for Performance Improvement of the American Medical Association are active collaborators with federal, state, and medical specialty initiatives to improve healthcare. These developing efforts are summarized here, with a specific focus on the status of these efforts in the field of psychiatry. PMID- 18677196 TI - Development of evidence-based performance measures for bipolar disorder: overview of methodology. AB - The STAndards for BipoLar Excellence (STABLE) Project was organized in 2005 to improve quality of care for bipolar disorder by developing and testing a set of evidence-based clinical process performance measures related to identifying, assessing, managing, and coordinating care for bipolar disorder. This article first briefly reviews the literature on the science of performance measurement and the use of performance measures as a tool for quality improvement. It then presents a detailed overview of the methodology used to develop the STABLE performance measures. Steps included choosing a clinical area to be measured, selecting key aspects of care for measurement, designing specifications for the measures, developing a data collection strategy, testing the scientific strength (validity, reliability, feasibility) of the measures, and obtaining, analyzing, and reporting conformance findings for the measures. Five of the STABLE measures have been endorsed by the National Quality Forum as part of their Standardizing Ambulatory Care Performance Measures project: screening for bipolar mania/hypomania in patients diagnosed with depression, assessment for risk of suicide, assessment for substance use, screening for hyperglycemia when atypical antipsychotic agents are prescribed, and monitoring change in level of functioning in response to treatment. Additional STABLE measures will be submitted to appropriate national organizations in the future. It is hoped that these measures will be used in quality assessment activities and that the results will inform efforts to improve care for individuals with bipolar disorder. PMID- 18677197 TI - Clinical and economic effects of unrecognized or inadequately treated bipolar disorder. AB - The authors review the literature on the clinical and economic impact of unrecognized and inadequately treated bipolar disorder, highlighting the need to improve identification and treatment of this disabling disorder. Epidemiologic data on prevalence, diagnosis, and treatment of bipolar disorder (including subthreshold conditions) are presented, including data from the recent National Comorbidity Survey Replication. Clinical factors that contribute to misdiagnosis and resulting inappropriate treatment of bipolar disorder are reviewed as well as negative clinical consequences of such misdiagnosis and inappropriate treatment. The economic impact of underrecognized and inadequately treated bipolar disorder is discussed. The data provide empirical support for screening all patients diagnosed with depression for evidence of bipolar disorder before initiating treatment, to ensure that bipolar illness is not misdiagnosed and treated as unipolar mood disorder. Readers are referred to performance measures and treatment resources assembled by the STAndards for BipoLar Excellence (STABLE) Project to help clinicians screen more accurately for bipolar disorder. PMID- 18677198 TI - Psychosocial interventions as adjunctive therapy for bipolar disorder. AB - Evidence suggests that adjunctive psychosocial interventions in bipolar disorder are clinically beneficial and cost effective when used in conjunction with pharmacotherapy. Appropriate adjunctive psychosocial interventions in bipolar disorder have been found to be associated with improved treatment adherence, greater stability, fewer hospitalizations, fewer days hospitalized, less need for crisis interventions, decreased relapse risk, and fewer acute episodes. Specific types of therapy that have shown efficacy include bipolar-specific cognitive behavioral therapy, family-focused therapy, interpersonal and social rhythm therapy, and systematic care management. A positive impact on medication adherence is a major goal of these adjunctive treatments. Studies show variable efficacy depending on patient characteristics, phase of illness, and presence of comorbid conditions so that therapies should be selected and administered on an individualized basis that takes into account each specific patient's current presentation and treatment history. For example, psychosocial interventions have been found to be more effective in patients with depressive than manic symptoms and during maintenance treatment. Given findings showing benefits of psychosocial interventions in the treatment of bipolar disorder, the STAndards for BipoLar Excellence (STABLE) project has developed and tested a performance measure to evaluate the use of this critical treatment component. PMID- 18677199 TI - The recovery concept: clinician and consumer perspectives. AB - This article presents an overview of the concept of recovery in serious and persistent mental illness from the perspective of both clinicians and consumers. Dr. Stotland, a psychiatrist, first highlights how treatment goals for bipolar disorder have changed in recent years, moving beyond symptomatic recovery to also encompass functional recovery (return to the level of functioning the person enjoyed before onset of the illness). She then discusses factors that play an important role in the recovery process, including resilience, the consumer's understanding of and participation in the treatment and recovery process, and collaboration between clinician and patient in setting specific functional goals as treatment progresses. She also focuses on the need for policy and system changes to facilitate recovery, including improved funding for recovery-oriented care, implementation of recovery-oriented, collaborative care models that bring together psychiatrists and primary care providers, and dissemination of improved tools for monitoring symptoms and functioning over time. Two relevant performance measures for monitoring changes in symptoms and level of functioning are discussed, with results of their field testing. Matthew Mattson, Director of Training for the Depression and Bipolar Support Alliance (DBSA), and Sue Bergeson, President of DBSA, then present the consumer's perspective on recovery oriented care. Drawing on recommendations from the Institute of Medicine, the President's New Freedom Commission, the U.S. Surgeon General, and the Annapolis Coalition on the Behavioral Health Workforce, they stress that the ultimate goal of treatment must be recovery; that, to the greatest extent possible, care should based on consumers' needs and values; that consumers should take an active role in the design and delivery of their own care; and that a priority of all care delivery should be to engender hope. Promising research on peer support groups and the use of peer sup-port specialists as consumer-providers is then reviewed. The article concludes with 20 specific recommendations to help mental health professionals move beyond a focus on symptom reduction alone to more recovery oriented care. PMID- 18677200 TI - Collaborative mental health and primary care for bipolar disorder. AB - This article discusses the use of integrated care models, in particular, collaborative care, in the treatment of bipolar disorder. Dr. Williams first discusses how care delivered via a collaboration between primary care and psychiatric providers has the potential to improve both mental health and general medical outcomes for patients with bipolar disorder. He describes promising findings from studies of the use of collaborative care in the treatment of depression, an area where this model has received the most study. Dr. Williams then discusses how such collaborative care models might best be implemented in the treatment of bipolar disorder. In the second half of the article, Dr. Manning focuses on five key issues that are an especially appropriate focus for collaborative care for bipolar disorder and for which the STAndards for BipoLar Excellence (STABLE) Project developed quality improvement performance measures: assessment for risk of suicide, assessment for substance use/abuse, monitoring for extrapyramidal symptoms, monitoring of metabolic parameters (e.g., monitoring for weight gain, hyperglycemia, hyperlipidemia), and provision of bipolar specific psychoeducation. PMID- 18677201 TI - Randomized, single-blind, factorial design study of the interaction of food and time on intestinal activity in 99mTc-tetrofosmin stress myocardial perfusion scintigraphy. AB - PURPOSE: Intestinal activity at the inferior myocardial wall represents an issue for assessment of myocardial perfusion imaging (MPI) with 99mTc-labelled tracers. The aim of this study was to investigate the effect of time and food on upper abdominal activity in 99mTc-tetrofosmin MPI. METHODS: The study population consisted of 152 consecutive patients referred for routine MPI. All patients underwent 2-day stress-rest 99mTc-tetrofosmin single-photon emission computed tomography MPI. Before stress testing, patients were randomized in a factorial design to four different regimens. Group A: early scan (image acquisition initiated within 15 min after injection of the tracer) and no food; group B: early scan and food (two pieces of white bread with butter and a minimum of 450 ml of water); group C: late scan (image acquisition 30-60 min after injection of the tracer) and no food; and group D: late and scan with food. Patients underwent standard bicycle exercise or pharmacological stress test. The degree of upper abdominal activity was evaluated by trained observers blinded to the randomization code. The primary endpoint was the proportion of accepted scans in the intention-to-treat population in stress MPI. RESULTS: The results showed statistical significant impact on both time and food on upper abdominal activity. The primary endpoint showed that the acceptance rate improved from 55% in group A to 100% success rate in group D. An early scan reduced the acceptance rate by 30% versus a late scan [hazard ratio 0.70, 95% confidence interval 0.58-0.84; P<0.0001], whereas the addition of food improved the success rate versus no food by 27% (hazard ratio 1.27, 95% confidence interval 1.07-1.51; P=0.006). No significant interaction between food and time was observed. An analysis of accepted scans according to the actual scan time and food consumption confirmed the findings of the intention-to-treat analysis. In addition, similar findings were seen in 116 of 152 patients with a rest MPI (success rate of 53% in group A vs. 96% in group D). CONCLUSION: A combination of solid food and water administered after injection of the tracer and delayed image acquisition led to significant and clinically relevant decrease of interfering upper abdominal activity in 99mTc-tetrofosmin MPI. PMID- 18677202 TI - Early uptake of 99mTc-C2A in the acute phase of myocardial infarction as a prognostic indicator for follow-up cardiac dysfunction. AB - OBJECTIVE: The C2A domain of Synaptotagmin I is a molecular probe for the specific imaging of cell death. Here we test the hypothesis that the uptake of 99mTc-C2A in the acute phase of an infarction is associated with cardiac dysfunction in follow-ups. METHODS: The left coronary artery was occluded in Sprague-Dawley rats for 0, 10, 20, and 30 min. 99mTc-C2A was injected intravenously at 2 h of reperfusion. Anterior planar images were acquired with one million counts on a gamma camera 3 h after injection. 99mTc-C2A uptake was calculated as the total counts in the left ventricle region minus blood pool signal. The in-vivo signal detected was correlated with wall motion score index at 1 and 3 weeks follow-ups measured by echocardiography. RESULTS: 99mTc-C2A uptake was higher with increased ischemic time (2244+/-852, 4054+/-1223, and 6178+/-1451 for 10, 20, and 30 min ischemia, analysis of variance P<0.001). A significant correlation was found between 99mTc-C2A uptake and wall motion score index at 1 week (R=0.800, P=0.0006) and 3 weeks (R=0.810, P=0.0008). CONCLUSION: In this ischemia/reperfusion model, 99mTc-C2A uptake in the acute phase was associated with functional abnormality at 1 and 3 weeks. This demonstrates the potential diagnostic and prognostic value of 99mTc-C2A as a novel imaging agent. PMID- 18677203 TI - Quantitative regional blood flow measurements in exercising leg skeletal muscle based on 99mTc-pertechnetate clearance. AB - BACKGROUND AND AIM: Skeletal muscle blood flow (SMBF) is a crucial indicator of impaired peripheral circulation. Although 133Xe has long been used for estimation of regional SMBF, its lipophilic and volatile nature hampers precise measurement. Therefore, we established an improved method based on 99mTc-pertechnetate ion (99mTcO-4) clearance. METHODS: Ten healthy male volunteers including five rugby players and five non-athletes (each group aged 25-35 years) received injection of 99mTcO-4 into the bilateral tibialis anterior muscles and gastrocnemius muscles (GCMs). The radioactivity of 99mTcO-4 before, during, and after toe-up or treadmill exercise was traced using a gamma-camera. Regional SMBF in absolute values (in ml/min/100 g muscle) was then calculated based on the half-time obtained from the time-activity curve. RESULTS: In both the groups, SMBF in tibialis anterior muscle changed similarly with values at the same levels. In contrast, SMBF in GCM showed marked difference between the two groups: in rugby players, SMBF in GCM returned to the baseline level (6.5+/-1.7, n=10) immediately after treadmill exercise, whereas that in non-athletes remained high (16.2+/-3.2, n=10). CONCLUSION: Regional SMBF measured by 99mTcO-4 clearance can indicate effectiveness of exercise training, and would be a diagnostic tool and prognostic indicator for use in patients with impaired peripheral circulation. PMID- 18677205 TI - Delayed gastric emptying in patients with caustic ingestion. AB - INTRODUCTION: Patients with caustic ingestion may develop esophageal and/or gastric cicatrization. OBJECTIVE: Scintigraphic assessment of liquid gastric emptying time in patients with caustic ingestion. MATERIALS AND METHODS: Consecutive patients with caustic-induced esophageal cicatrization attending the gastroenterology clinic of our institute were studied (patients with age >or=60 years, earlier gastric surgery, vagotomy, peptic ulcer disease, diabetes, systemic sclerosis, and those on gastrointestinal motility-altering drugs were excluded). Gastric emptying time was assessed by radionuclide scintigraphy (ingestion of 200 ml mango juice containing of 18.5 MBq 99mTc sulfur colloid after an overnight fast by static imaging in anterior and posterior projections in supine posture at 10 min intervals each for 1 h). A time-activity curve was generated and gastric half-emptying time (GET 1/2) was calculated. Results were compared with GET 1/2 values estimated for normal individuals in our laboratory (mean+/-2 SD). The emptying study protocol for normal individuals was the same as in patients. RESULTS: Twenty patients (males 10) median age 32 years, 16 with acid ingestion and four with alkali ingestion, were studied. No patient had symptoms suggestive of gastric outlet obstruction or gastroparesis. Eight patients had evidence of gastric cicatrization in the form of straightening of the lesser curvature and pulling of incisura and duodenal bulb medially and loss of parallelism between the fundus and left dome of diaphragm. Gastric distensibility was however normal in them. As compared with values for normal controls (25+/-9 min), GET 1/2 was significantly prolonged in the study group as a whole (53.2+/-27.77 min, P=0.000). No significant difference was observed between different age groups, sex, or type of caustic agent consumed. GET 1/2 differed in patients (n=10) with stricture involving lower-third of esophagus (72.2+/-27.67 min) when compared with those (n=10) who had a stricture involving upper and/or middle-third of esophagus (34.3+/-8.02 min, P=0.000). In the former, GET 1/2 was maximally prolonged in patients (n=6) with involvement of the lower esophagus and reduced stomach capacity (84.6+/-27.03 min), followed by patients (n=4) with lower esophageal involvement with normal stomach capacity (53.7+/ 17.41 min), but the difference did not reach statistical significance (P=0.078). Patients (n=10) without lower esophageal involvement did not have statistically significant altered GET 1/2 compared with normal controls (P>0.05). CONCLUSION: Our results show that patients with caustic ingestion have prolonged liquid gastric emptying even in the absence of any gastric symptoms. PMID- 18677204 TI - Evaluation of 99mTcN-15C5 as a new myocardial perfusion imaging agent in normal dogs and canines with coronary stenosis. AB - OBJECTIVES: This study was designed to evaluate the biodistribution and blood clearance characteristics of 99mTcN-15C5 and its potential as a myocardial perfusion radiotracer. METHODS: Five normal mongrel dogs were injected with 99mTcN-15C5 intravenously. Blood samples were collected at 0.5, 1, 2, 3, 4, 5, 10, 15, and 30 min postinjection (p.i.). Anterior planar images were acquired at 5, 15, 30, 60, 90, and 120 min p.i. Regions of interest (ROIs) were drawn around heart, liver, and lungs. The raw activity in each ROI was expressed as counts/pixel/min. Heart/liver and heart/lung ratios were calculated by dividing the mean counts in heart ROI by the mean counts in liver and lung ROI, respectively. For comparison, 99mTc-sestamibi was also evaluated in the same five dogs. In dogs with coronary stenoses, single photon emission computed tomography images were acquired at 30, 60, and 120 min after administration of 99mTcN-15C5 with/without adenosine. RESULTS: 99mTcN-15C5 and 99mTc-sestamibi had very similar blood clearance characteristics during the first 30 min p.i. The heart/liver ratio of 99mTcN-15C5 increased from 0.48+/-0.05 at 5 min p.i. to 1.85+/-0.11 at 120 min p.i., whereas the heart/liver ratio of 99mTc-sestamibi was improved only slightly from 0.45+/-0.04 at 5 min p.i. to 0.74+/-0.15 at 120 min p.i. SPECT imaging studies in canines with coronary stenoses indicated that good visualization of the perfusion defect could be achieved at 30 min after administration of 99mTcN-15C5 with the adenosine stress. CONCLUSION: The combination of high heart uptake and rapid liver clearance makes 99mTcN-15C5 a promising new radiotracer for myocardial perfusion imaging. PMID- 18677206 TI - 99mTc-MIBI imaging of AIDS-related Kaposi's sarcoma in the lungs. AB - OBJECTIVE: Pulmonary Kaposi's sarcoma (KS) occurs in more than 10% of patients with acquired immunodeficiency syndrome (AIDS) and has a high mortality rate. Prompt detection, diagnosis, and treatment reduce patient morbidity and mortality. The objective of this study was to determine the efficacy of 99mTc hexakis-2-methoxy isobutyl isonitrile (99mTc-MIBI) imaging in detecting pulmonary AIDS-related KS. METHODS: 99mTc-MIBI imaging was performed on 72 human immunodeficiency virus-seropositive patients with bronchoscopic diagnosis of pulmonary KS (36 patients), pneumonia (22), normal tracheo-bronchial tree (11), lymphoma (2), and bronchogenic carcinoma (1). Lung uptake and lymph node detection in KS were compared on planar and single photon emission computed tomography (SPECT) scans. RESULTS: The lung/myocardium ratios on the 1-h planar images were significantly higher in KS and normal lungs than opportunistic infection. Using the lung/myocardium ratio of 1 as cutoff, the sensitivity, specificity, and accuracy of the 1-h planar images for detecting pulmonary KS were 75, 57.58, and 66.67%, respectively. Abnormal lymph node uptake, pleural/pericardial effusions, and ascites were detected more readily on SPECT. CONCLUSION: Planar 99mTc-MIBI imaging has moderate sensitivity, specificity, and accuracy for detecting pulmonary KS. SPECT is more effective in detecting abnormal lymph nodes, pleural/pericardial effusions, and ascites. 99mTc-MIBI SPECT followed by planar imaging at 40-60 min can be useful in assessing pulmonary KS. PMID- 18677207 TI - Role of fluorodeoxyglucose-PET versus fluorodeoxyglucose-PET/computed tomography in detection of unknown primary tumor: a meta-analysis of the literature. AB - The objectives of this study were to evaluate the diagnostic accuracy of fluorodeoxyglucose (FDG)-PET and FDG-PET/computed tomography (CT) in the detection of primary tumors in patients presenting with carcinoma of unknown primary (CUP) unidentified by conventional workup, and to compare the statistical difference between the FDG-PET and FDG-PET/CT. Twenty-eight studies (involving a total of 910 patients) published between 1990 and 2007 were reviewed. These studies evaluated the role of FDG-PET and FDG-PET/CT in the detection of unknown primary tumors after physical examination and conventional workup failed to detect a primary tumor. Systematic methods were used to identify, select, and evaluate the methodological quality of the studies as well as to summarize the overall findings of sensitivity, specificity, and detection capacity of the primary tumor. The overall sensitivity and specificity of FDG-PET in detecting unknown primary tumors were 0.78 [95% confidence interval (CI): 0.72-0.84)] and 0.79 (95% CI: 0.74-0.83), respectively. Furthermore, FDG-PET detected 28.54% of tumors that were not apparent after CUP failed to be detected by conventional workup. Data were collected on the locations of primary tumors detected by FDG PET in 17 studies and detected by FDG-PET/CT in seven studies. Tumors from the base of the tongue accounted for 20.7% (six of 29) of all false-positive FDG-PET scans, corresponding to a false-positive rate of 28.6% (six of 29), much higher than tumors from the others. FDG-PET exhibited a lower sensitivity with respect to the tumors at the base of the tongue and tonsils, which was 68.2 and 76.7%, respectively. In the eight studies with 430 patients diagnosed with CUP by FDG PET/CT, 31.4% (n=135) of primary tumors were detected. The pooled sensitivity and specificity were 0.81 (95% CI: 0.74-0.87) and 0.83 (95% CI: 0.78-0.87), respectively. FDG-PET and FDG-PET/CT can detect primary tumors that went undetected by physical examination and conventional workup. FDG-PET exhibited lower sensitivity with respect to the tumors at the base of the tongue and the tonsils. PMID- 18677208 TI - Diagnostic accuracy of 99mTc-MIBI-SPECT in the detection of lymph node metastases in patients with carcinoma of the tongue: comparison with computed tomography and MRI. AB - AIM: To compare the diagnostic efficacy of computerized tomography (CT), magnetic resonance imaging (MRI), and Tc-methoxyisobutylisonitrile-single photon emission computed tomography (MIBI-SPECT) for the detection of lymph node (LN) metastases in clinically neck-negative patients (T1-T2N0) with carcinoma of the tongue. MATERIALS AND METHODS: Twenty adult patients (aged 54.75+/-14.57 years) with histopathologically proven carcinoma of the tongue and clinically neck-negative (T1-T2N0) status were included in this study. Before the wide local excision of the primary tumor followed by modified functional neck dissection, all the patients underwent contrast-enhanced CT and MRI and 99mTc-MIBI-SPECT. The SPECT data acquisition (3600 arc, 128 x 128 matrix, 64 projections, 20 s/projection) was performed 15 min after an intravenous injection of 555-740 MBq of 99mTc-MIBI under the dual head gamma-camera (E Cam, Siemens, Erlangen, Germany) using the low-energy, high-resolution, parallel hole collimator. SPECT data reconstruction was achieved by filtered back projection using the Butterworth filter with a critical frequency of 0.5 and power 10.0. Only visual assessment was used for defining a positive lymph nodes (LNs) on reconstructed SPECT images and maximum intensity projection images. Histopathological examination of LNs was performed using hematoxylin and eosin staining. RESULTS: A total of 351 LNs were excised surgically (functional neck dissection) from 20 tongue cancer patients having N0 neck disease. Only nine of 351 LNs were documented as being positive for malignancy on histopathological examination. These nine LNs were distributed in eight of 20 patients. CT, MRI, and MIBI-SPECT could accurately identify one, three, and five LNs, respectively. In one of the eight patients, 99mTc-MIBI-SPECT could not distinguish between two LNs present at the same level, having size of 6.0 and 8.0 mm. In this study, 99mTc-MIBI-SPECT was shown to have better sensitivity/positive predictive values (55.5%/71%) than either CT (11%/33%) or MRI (33%/60%). The histopathologically documented metastatic LNs picked up by CT or MRI were also identified by 99mTc-MIBI-SPECT. CONCLUSION: In this preliminary prospective study, we observed that 40% (8/20) of the clinically neck-negative carcinoma tongue patients harbored metastatic LNs. 99mTc-MIBI-SPECT is a more effective imaging modality in the staging of clinically neck-negative LN metastases in carcinoma tongue as compared with CT or MRI. PMID- 18677209 TI - Usefulness of 99mTc-pertechnetate single-photon emission computed tomography in remnant mass estimation of postsurgical patients of differentiated thyroid cancer during internal dosimetry. AB - OBJECTIVE: No satisfactory methods exist for postsurgical remnant mass estimation required for dosimetric calculations in differentiated thyroid cancer patients undergoing I-131 remnant ablation. We estimated the remnant mass by Tc-99m pertechnetate single-photon emission computed tomography to calculate the radiation dose delivered. MATERIALS AND METHODS: Thyroid phantoms of known volumes (1.1-6 cm3) were fabricated, filled with different concentrations of Tc 99m pertechnetate activity, and single-photon emission computed tomography acquired on a dual head camera and processed with Hanning filter. Area of the organ on coronal slices was calculated at different thresholds and the sum was multiplied by slice thickness to get the volume. A threshold of 50% was found to distinguish the organ from the background activity and give the most accurate results. Remnant mass was calculated by this method in 50 patients of differentiated thyroid cancer after thyroid surgery. Effective half-life (EHL) of I-131 and 24 h radioactive iodine uptake (RAIU) were calculated by repeated neck count measurements for 4 days. After administering I-131 for therapy, 39 patients were followed up at 6 months and the effects of various parameters in achieving complete ablation were analyzed. RESULTS: Mean values of remnant mass, RAIU, and EHL were 6.6+/-4.3 g, 8.8+/-6.1%, and 4.5+/-0.9 days, respectively. Type of surgery, remnant mass, and RAIU were the most important predictors of complete ablation (P<0.05). Radiation-absorbed dose, EHL, and the administered activity had no significant effect on complete ablation. CONCLUSION: Remnant thyroid mass estimation by Tc-99m pertechnetate single-photon emission computed tomography is a simple technique and allows overcoming the dependence on structural imaging techniques or making empirical assumptions during dosimetry. PMID- 18677210 TI - Dosimetric evaluation and therapeutic response to internal radiation therapy of hepatocarcinomas using iodine-131-labelled lipiodol. AB - OBJECTIVE: Vectorized internal radiation therapy using lipiodol-labelled with iodine-131 (131 I-lipiodol) is an effective treatment for inoperable hepatocellular carcinomas. However, few dosimetric data are available based on this approach. We have developed a dosimetric protocol based on scintiscan imaging and that is designed to calculate the tumoural absorbed dose during the treatment of hepatocarcinoma by 131 I-lipiodol. METHODS: This concept was developed on a gamma-camera coupled to a computed tomography scanner. It integrates corrections for attenuation phenomena, scattering and dead time. The tumoural absorbed dose calculation was carried out according to the Medical Internal Radiation Dose Committee formalism. This protocol was applied to a series of 41 patients in the framework of a retrospective study. RESULTS: The mean tumoural absorbed dose with the first treatment is 248 Gy (+/-176), as opposed to 152 Gy (+/-122) during the second. We highlighted a correlation between the tumoural absorbed dose, calculated in tomographic mode, and the morphological response to the first treatment (P=0.0071). Moreover, a tumoural absorbed dose of 280 Gy seems to be an effective absorbed dose threshold in our population. Above this absorbed dose, 84% of the patients are responders after the first treatment, whereas no responses are recorded below this threshold. CONCLUSION: These results are promising because, for the first time, they allow us to predict the effectiveness of a treatment by 131 I-lipiodol. They are required to be validated on a broader exploratory trial, including a dosimetric study of the critical organs, so an individualized dosimetry can be defined for each patient. PMID- 18677211 TI - Tc-99m-methoxy isobutyl isonitrile scintimammography: imaging postexcision biopsy for residual and multifocal breast tumor. AB - OBJECTIVES: The interpretation of mammogram in a postsurgical breast can be extremely complex and difficult because masses, calcifications, and architectural distortion can mimic cancer. Scintimammography has been proposed because it is not affected by these morphological changes and can potentially be used in patients after excision biopsy to assess any residual tumor, other foci of disease (multifocal disease). MATERIALS AND METHODS: The population comprised of 21 patients (mean age: 47.80 years, median age: 50 years, and age range: 26-77 years) evaluated for suspected residual breast cancer after excision biopsy. All patients received a 740-1000 MBq bolus IV injection of Tc-99m-methoxy isobutyl isonitrile (MIBI) preferably in pedal vein. At 5-10 min after injection, planar images were obtained in prone lateral and supine anterior positions using dual head gamma camera. MIBI uptake was scored as follows: 1 - as normal uptake (compared with contralateral side), 2 - focal low-intense uptake (equivocal), and 3 - focal high-intense uptake (positive). All patients had histopathology for tissue diagnosis. RESULTS: Of the 21 patients evaluated, scintimammography planar was found true positive in 13 and true negative in six. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 92.85, 85.71, 92.85, 85.71, and 90.47%, respectively (P<0.001). Scintimammography detected 2/2 multifocal disease. CONCLUSION: In patients after excision biopsy, scintimammography with Tc-99m-MIBI is valuable in assessing residual or multifocal disease. Scintimammography has high positive predictive value and may influence planning further management. PMID- 18677212 TI - Scintigraphic imaging using technetium-99m-labeled ceftizoxime in an experimental model of acute osteomyelitis in rats. AB - PURPOSE: To investigate inflammatory (zymosan) and infectious (Staphylococcus aureus) processes in experimental models in rats using technetium-99m-labeled ceftizoxime (CFT). METHODS: Male Wistar rats were used for the development of the inflammatory (zymosan) and infectious (S. aureus) processes in the medullary cavity of the left tibia. Sterile saline was used for the control group. At 48 h after induction of the processes, the animals were anesthetized and scintigraphic images were acquired at 1, 2, 4, and 6 h after intravenous injection of 0.1 ml of 99mTc-CFT (55 MBq). Quantitative analysis of the scintigraphic images was performed by counting the radioactivity in the regions of interest. Samples of tibia were taken for histopathological examination. RESULTS: The images showed that 99mTc-CFT presented higher tropism to infectious foci than with the inflammatory site. The average value of the target/nontarget ratio of the 99mTc CFT was significantly higher in the infected (2.40+/-0.22) than in the inflamed tibia (1.50+/-0.05) and the control group (1.05+/-0.04) for all of the investigated times. The histological data showed a similar inflammatory response for both the S. aureus and zymosan groups. CONCLUSION: The 99mTc-CFT presented a high tropism and retention for an infected region in this model of osteomyelitis, thereby constituting an interesting strategy to distinguish aseptic from septic sites. PMID- 18677213 TI - On the effect of sample size of the normal database on statistical power of single subject analysis. PMID- 18677214 TI - 18F-fluorodeoxyglucose-positron emission tomography imaging in idiopathic inflammatory myositis. PMID- 18677215 TI - Quantification of pertechnetate in 99mTc-methoxy isobutyl isonitrile. PMID- 18677216 TI - Scintigraphic assessment of recurrent glioma with focus on proliferation. PMID- 18677217 TI - Added value of PET and PET-computed tomography in oesophageal cancer: a review of current practice. PMID- 18677222 TI - Mucins in contact lens wear and dry eye conditions. AB - Ocular mucins are thought to play integral roles in ocular surface lubrication, anchoring of the aqueous, stabilizing the lipid components of the tear film, eliminating foreign bodies and pathogens, and with potential involvement in cell cycle mediation and apoptotic activity of ocular surface epithelia. Ocular mucins are of secreted and membrane-associated types. Secreted mucins may be of large gel-forming type or small soluble mucins (e.g., MUC5AC and MUC7). Membrane associated mucins such as MUCs 1 and 4 are a major component of the glycocalyx. They are thought to render structural support to the microplicae and mediate epithelial cell cycle and apoptotic activity. The alterations in ocular mucins with contact lens wear are unclear. Recent work shows mucin expression may be up regulated during the early years of contact lens wear, and with long-term lens wear, mucin expression may return to normal levels or sub-normal levels, although this is not well understood. Further, the polar nature of mucins may be associated with their affinity for contact lens surfaces making them a component of contact lens deposition. This has potential implications in the wettability and tolerability of contact lenses, and may be impacted by surface coatings, polymer characteristics, or care solutions. Conjunctival mucin gene expression and secretion may be deficient in several ocular surface disorders associated with dry eye. Deficiency and alterations in glycosylation characteristics of MUC5AC and MUC2 have been reported in both Sjogren and non-Sjogren dry eye types. Decreased binding of the membrane-associated mucin MUC16 to the conjunctival epithelium has been reported in Sjogren dry eye while MUC1 alterations have been reported in Sjogren and non-Sjogren dry eye states. In view of the mucin involvement in dry eye conditions, stimulation of mucus secretion pathways may hold promise in the pharmaceutical treatment of dry eye. PMID- 18677223 TI - Antibody protein array analysis of the tear film cytokines. AB - PURPOSE: Many bioactive proteins including cytokines are reported to increase in dry eye disease although the specific profile and concentration of inflammatory mediators varies considerably from study to study. In part, this variability results from inherent difficulties in quantifying low abundance proteins in a limited sample volume using relatively low sensitivity dot ELISA methods. Additional complexity comes with the use of pooled samples collected using a variety of techniques and intrinsic variation in the diurnal pattern of individual tear proteins. The current study describes a recent advance in the area of proteomics that has allowed the identification of dozens of low abundance proteins in human tear samples. METHODS: Commercially available stationary phase antibody protein arrays were adapted to improve suitability for use in small volume biological fluid analysis with particular emphasis on tear film proteomics. Arrays were adapted to allow simultaneous screening for a panel of inflammatory cytokines in low volume tear samples collected from individual eyes. RESULTS: A preliminary study comparing tear array results in a small population of Sjogren's syndrome patients was conducted. The multiplex microplate array assays of cytokines in tear fluid present an unanticipated challenge due to the unique nature of tear fluid. The presence of factors that exhibit an affinity for plastic, capture antibodies and IgG and create a complex series of matrix effects profoundly impacting the reliability of dot ELISA, including with elevated levels of background reactivity and reduction in capacity to bind targeted protein. CONCLUSIONS: Preliminary results using tears collected from patients with Sjogren's syndrome reveal methodological advantages of protein array technology and support the concept that autoimmune-mediated dry eye disease has an inflammatory component. They also emphasize the inherent difficulties one can face when interpreting the results of micro-well arrays that result from blooming effects, matrix effects, image saturation and cross-talk between capture and probe antibodies that can greatly reduce signal-to-noise and limit the ability to obtain meaningful results. PMID- 18677224 TI - Tear film with "Orgahexa EyeMasks" in patients with meibomian gland dysfunction. AB - PURPOSE: To evaluate the efficacy of a new Orgahexa eye warmer mask for patients with simple meibomian gland dysfunction (MGD) in a prospective comparative study. METHODS: Twenty right eyes of 20 patients with simple MGD, and 22 right eyes of 22 healthy controls were studied. Subjects were allocated to Orgahexa or conventional eye mask wear for 10 min (short-term study), and for 2 weeks (long term study). Eyelid temperature measurements, slit lamp examination, tear film break-up time, Schirmer test, vital staining, tear film lipid layer interferometry, and dry eye symptomatology scoring with visual analog scales were performed. RESULTS: The Orgahexa eye warmer improved both tear function and ocular surface status, and decreased symptoms significantly without any complications. CONCLUSIONS: The Orgahexa eye warmer is a simple, safe, and convenient method, which seems to improve the ocular surface status and tear functions in patients with simple MGD. PMID- 18677225 TI - Dry eye symptoms assessed by four questionnaires. AB - PURPOSE: To establish the relationships between commonly used questionnaires including Dry Eye Questionnaire, McMonnies Questionnaire, and Ocular Surface Disease Index, and to test the construct and face validity of the simple Subjective Evaluation of Symptom of Dryness. METHODS: Ninety-seven non-contact lens wearing subjects were enrolled in the study and classified into either a "dry" and "non-dry" group using a single score from an initially applied subjective evaluation of symptom of dryness. The four questionnaires were then completed in a random order. The unidimensionality and accuracy of the responses was assessed using Rasch and receiver (or relative) operating characteristics curve analysis and the characteristics of and association between symptoms were compared using non-parametric statistics. RESULTS: The responses from the Dry Eye Questionnaire, McMonnies Questionnaire, and Ocular Surface Disease Index met the Rasch analysis criterion of unidimensionality. Each test separated the symptomatic and asymptomatic groups well [all receiver (or relative) operating characteristics area-under-the-curve statistics at least 0.88] and there were significant associations between the results from each questionnaire (all Spearman rho at least 0.64). CONCLUSIONS: The results illustrate that different questionnaire-based instruments examining symptoms in controls and symptomatic subjects derive unidimensional data that are similar inasmuch as the overall scores are highly correlated. The data also point to the utility of a quick, three-question screening tool in dry eye research. PMID- 18677226 TI - Clinically important difference in dry eye: change in IDEEL-symptom bother. AB - PURPOSE: The impact of dry eye on everyday living (IDEEL) is a valid, reliable questionnaire with 3 modules; symptom bother (SB), quality of life, and treatment satisfaction. This study tests the utility of the 20-item IDEEL-SB to discrimate self-assessed severity in dry eye subjects and to determine the clinically important difference (CID) in IDEEL-SB score that relates to a self-report of global change in dry eye condition after treatment. METHODS: After randomization to 1 of 3 marketed tear replacements, dry eye subjects completed the IDEEL-SB at baseline, 1 and 4 wk and global change questionnaire on status of general health and dry eye at 1 and 4 wk (5-point Likert scale; "much better" to "much worse"). The IDEEL-SB score was the unweighted mean score x 25. CID was initially estimated by receiver-operator curve analysis (ROC) and global change questionnaire. Various IDEEL-SB change criteria were tested to maximize % agreement, kappa score, and effect size. RESULTS: The 74 subjects were 54.0 +/- 17.2 years old and 52 (70.3%) were female. At baseline, 56.3% rarely/never used replacement tears. Subjects rated their dry eye severity as mild (41%), moderate (50%), or severe (9%). IDEEL-SB discriminated dry eye severity well; average baseline scores were mild: 40.0 (SD = 7.5), moderate: 50.6 (SD = 11.0) and severe 64.3 (SD = 8.0) (p = 0.001). After 4 wk of drops usage, IDEEL-SB dropped among "improved" subjects by -13.3 (SD = 10.9), "same" shifted by -4.7 (SD = 9.4), "worsened" changed by 1.4 (SD = 11.1). ROC results show that a 12-point change in IDEEL-SB is a clinically significant change in symptoms. CONCLUSIONS: 12-Point shift in the IDEEL-SB module is a CID based on ROC and the distribution across self-assessed severity groups compared with their global assessment of change in condition with treatment. PMID- 18677227 TI - Effect of viscosity on tear drainage and ocular residence time. AB - PURPOSE: An increase in residence time of dry eye medications including artificial tears will likely enhance therapeutic benefits. The drainage rates and the residence time of eye drops depend on the viscosity of the instilled fluids. However, a quantitative understanding of the dependence of drainage rates and the residence time on viscosity is lacking. The current study aims to develop a mathematical model for the drainage of Newtonian fluids and also for power-law non-Newtonian fluids of different viscosities. METHODS: This study is an extension of our previous study on the mathematical model of tear drainage. The tear drainage model is modified to describe the drainage of Newtonian fluids with viscosities higher than the tear viscosity and power-law non-Newtonian fluids with rheological parameters obtained from fitting experimental data in literature. The drainage rate through canaliculi was derived from the modified drainage model and was incorporated into a tear mass balance to calculate the transients of total solute quantity in ocular fluids and the bioavailability of instilled drugs. RESULTS: For Newtonian fluids, increasing the viscosity does not affect the drainage rate unless the viscosity exceeds a critical value of about 4.4 cp. The viscosity has a maximum impact on drainage rate around a value of about 100 cp. The trends are similar for shear thinning power law fluids. The transients of total solute quantity, and the residence time agrees at least qualitatively with experimental studies. CONCLUSIONS: A mathematical model has been developed for the drainage of Newtonian fluids and power-law fluids through canaliculi. The model can quantitatively explain different experimental observations on the effect of viscosity on the residence of instilled fluids on the ocular surface. The current study is helpful for understanding the mechanism of fluid drainage from the ocular surface and for improving the design of dry eye treatments. PMID- 18677228 TI - Efficacy of punctum plug treatment in short break-up time dry eye. AB - PURPOSE: To evaluate the efficacy of punctum plug insertion on tear and dynamic visual function in dry eye patients with short break-up time (s-BUT) of the tears. METHODS: A prospective comparative case series design was used. Twenty seven eyes of 27 dry eye patients with a s-BUT seen at Keio University School of Medicine, Department of Ophthalmology, were studied. Functional visual acuity (FVA) measurements, tear function examinations and ocular surface evaluations including Schirmer test, tear break-up time, fluorescein and Rose Bengal vital staining scores were performed before and 1 month after insertion of punctum plugs (PP) in dry eye patients with s-BUT. Degree of satisfaction with the PP treatment was also graded. RESULTS: Nineteen out of 27 eyes (70.4%) showed a satisfactory outcome with the PP treatment because of decreased subjective dry eye symptoms. On the other hand, six out of eight eyes with an unsatisfactory outcome had epiphora. The visual maintenance ratio value was observed to significantly increase from 0.87 +/- 0.09 to 0.91 +/- 0.07 in eyes with no epiphora, whereas the visual maintenance ratio value significantly decreased from 0.98 +/- 0.10 to 0.86 +/- 0.13 in the group of eyes with epiphora after PP treatment. (p < 0.05) Likewise, FVA significantly improved after PP insertion in the group with no epiphora and decreased in eyes with epiphora. Visual acuity in the conventional Landolt visual testing remained significantly unchanged in both group of eyes with or without epiphora. CONCLUSIONS: FVA was useful in assessing and quantifying vision related symptomatology in s-BUT type of dry eye treated with PP occlusion. PMID- 18677229 TI - The changing times in dry eye research. PMID- 18677230 TI - Contributions of evaporation and other mechanisms to tear film thinning and break up. AB - PURPOSE: To evaluate the contribution of three mechanisms-evaporation of the tear film, inward flow of water into the corneal epithelium or contact lens, and "tangential flow" along the surface of epithelium or contact lens-to the thinning of the tear film between blinks and to tear film break-up. In addition to a discussion of relevant studies, some previously unpublished images are presented illustrating aspects of tear film break-up. CONTRIBUTIONS OF THREE MECHANISMS TO TEAR FILM BREAK-UP: Inward flow of water into the epithelium or contact lens is probably unimportant, and a small flow in the opposite direction may actually occur. Tangential flow is probably important in certain special cases of tear film break-up-at the black line near the tear meniscus, over surface elevations, after partial blinks, and from small thick lipid spots in the tear film. In all these special cases it is argued that tangential flow is important initially, but evaporation may be needed for final thinning to break-up. It is argued that most of the observed tear film thinning between blinks is due to evaporation, rather than tangential flow, and that large "pool" break-up regions are the result of evaporation over an extended area. CONCLUSION: Evaporation in our "free-air" conditions may be four to five times faster than the average of the values reported in the literature when air currents are prevented by preocular chambers. However, recent evaporation measurements using "ventilated chambers" give higher values, which may correspond better to free-air conditions. Thus evaporation may be fast enough to explain many cases of tear film break-up, and to give rise to considerable increases in the local osmolarity of the tear film between blinks. PMID- 18677232 TI - Tear lipocalin and lysozyme in Sjogren and non-Sjogren dry eye. AB - PURPOSE: To evaluate the concentration of tear lipocalin, lysozyme, and total protein in Sjogren's Syndrome (SS), non-Sjogren's keratoconjunctivitis sicca (KCS), and non-dry-eyed (NDE) individuals. METHODS: Seventy-six subjects were recruited for this study: 25 SS subjects; 25 KCS subjects, and 26 NDE individuals. Symptoms were measured with a visual analogue scale. Tear flow was measured by the Schirmer I test without anesthesia. Tears were collected using an eye wash technique. Total tear protein was quantified using the DC Protein Assay Kit. Tear lipocalin and lysozyme were quantified via Western blotting performed on a Phast System. RESULTS: By definition, the SS and KCS groups both had significantly lower mean Schirmer scores (5.12 +/- 5.96 mm and 7.84 +/- 7.35 mm) compared with the NDE group (23.83 +/- 7.85 mm; p < 0.0001). There was no difference in mean Schirmer scores between SS and KCS groups (p = 0.19). The tear film of the SS group was characterized by significantly reduced (p < 0.0001) total protein and lipocalin concentrations compared with both KCS and NDE groups. No difference between the KCS and NDE groups was found in total protein (p = 0.92) or lipocalin (p = 0.19) concentration. In contrast, the concentration of tear film lysozyme was found to be statistically similar in all three groups examined. No statistically significant correlation was found in any group between mean Schirmer values compared with total protein, lipocalin or lysozyme concentration. CONCLUSION: Our data demonstrate a biochemical distinction between the Sjogren's group compared with both KCS and control groups, in that both tear lipocalin and total tear protein were significantly reduced. Although correlations were not found between protein measurements and tear flow, a combination of tests including Schirmer I and quantitation of tear film biomarkers may allow for the identification of SS patients without the need for invasive testing. PMID- 18677231 TI - Dry eye and designer ophthalmics. AB - Expressed sequence tag (EST), proteomic, and antibody capture assays are revealing a level of tear film protein complexity far greater than previously appreciated. A systems biology approach will be needed to fully appreciate function as tear protein doses fluctuate in time through different conditions. Although consensus is growing on what fully constitutes the human tear proteome, questions remain about the source and significance of the approximately 256 tear proteins designated as "intracellular." Many of these may derive from normal cellular turnover and could therefore be informative. A further >183 are designated as "extracellular." Surprisingly, only 4 to 5% of these appear to be dysregulated in the three forms of dry eye preliminarily examined to date. Some differ and a couple overlap, suggesting that disease-specific signatures could be identified. Future dry eye treatment might include recombinant tear protein rescue as a personalized ophthalmic approach to ocular surface disease. PMID- 18677233 TI - Long-term incidence of dry eye in an older population. AB - PURPOSE: To estimate the ten-year incidence of dry eye in an older population and examine its association with various risk factors. METHODS: The 43 to 86 year old population of Beaver Dam, WI, was examined in 1988 to 1990 (n = 4926) and 1993 to 1995 (n = 3722). Dry eye data were first collected in 1993 to 1995. Subsequent examinations or interviews occurred in 1998 to 2000 (n = 2827) and 2003 to 2005 (n = 2124). The incidence cohort comprised 2414 subjects not reporting dry eye in 1993 to 1995. Risk factor information, ascertained in 1993 to 1995, included demographics, medical history, cardiovascular disease risk factors, medications, and life-style factors. Ten-year cumulative incidence was estimated by the product-limit method. RESULTS: Over the 10-year period, 482 subjects developed a history of dry eye for an incidence of 21.6% (95% confidence interval, 19.9 to 23.3%). Incidence increased significantly (p < 0.001) with age. Incidence was greater in women (25.0%) than men (17.2%, p < 0.001). After adjusting for age, incidence was greater (p < 0.05) in subjects with arthritis, allergy or thyroid disease not treated with hormone, using antihistamines, antianxiety medications, antidepressants, oral steroids or vitamins, and poorer self-rated health. Incidence was less (p < 0.05) in subjects consuming alcohol. It was not significantly associated with blood pressure, hypertension, serum total or high density lipoprotein cholesterol, body mass, diabetes, gout, osteoporosis, cardiovascular disease, smoking, caffeine use, or taking calcium channel blockers or anticholesterol medications. In a multivariable model with time-varying covariates, increased incidence was associated with age, female gender, poorer self-rated health, antidepressant or oral steroid use, and thyroid disease untreated with hormone. It was lower for those using angiotensin-converting enzyme inhibitors or with a sedentary lifestyle. CONCLUSIONS: Dry eye incidence is substantial. However, there are few associated risk factors. Some drugs (antihistamines, antianxiety drugs, antidepressants, oral steroids) are associated with greater risk, while angiotensin-converting enzyme inhibitors may be associated with lower risk. PMID- 18677234 TI - Inner eyelid surface temperature as a function of warm compress methodology. AB - PURPOSE: To (1) determine an optimal method of warm compress (WC) application to maximize heating meibomian glands in minimal time, (2) determine the maximum inner eyelid temperature achievable during 30 min of routine WC application, (3) examine the cooling curve for 10 min after WCs have been discontinued. METHODS: Subjects were randomly assigned to one of three WC methodologies. Group A (GrA), (n = 10): 15 min WC application without reheating. GrB, (n = 10): 30 min WC application with reheating every 2 min. GrC, (n = 12): 30 min WC application optimizing contact with the lower lid and reheating every 2 min. WCs were heated to 45 +/- 0.5 degrees C. Outer and inner lower eyelid surface temperatures were measured at baseline and regular intervals. RESULTS: GrA: the maximum outer and inner lower eyelid surface temperatures = 41.2 +/- 0.3 degrees C at 1 min and 38.8 +/- 0.2 degrees C after 4 min, respectively. GrB: the maximum outer upper eyelid temperature, 43.3 +/- 0.5 degrees C, was reached after 6 min while it required 30 min to reach the maximum inner lower eyelid temperature, 40.4 +/- 0.3 degrees C. GrC: it required 4 min to reach the maximum outer lower eyelid temperature, 42.2 +/- 0.4 degrees C, while it required 20 min to reach the maximum inner lower eyelid temperature, 40.8 +/- 0.3 degrees C. CONCLUSIONS: To optimize WC efficiency, patients should (1) heat the WC to approximately 45 degrees C, (2) optimize contact between the WC and outer eyelid surfaces, (3) reheat the WC frequently and have a replacement heated WC on hand for exchange, and (4) perform the activity for at least 4 min in order to achieve an inner lower eyelid temperature > or = 40 degrees C. Longer therapy may be necessary for more severe obstructions. These data suggest that precise, customized, labor intensive WC procedure is necessary to optimize treating meibomian gland dysfunction and obstruction using WCs. PMID- 18677235 TI - Menisci and fullness of the blink in dry eye. AB - PURPOSE: This study was to investigate the role of the upper meniscus in tear film formation and blinking. METHODS: One microliter of 2% fluorescein was instilled under the upper lid of 15 dry eye (DE) and 15 control subjects. Subjects were instructed to blink partially and hold the eye open as long as possible, and analysis of tear breakup dynamics was used to quantify the area of breakup. This procedure was repeated following a full blink. Meniscus height was measured from digital videos. RESULTS: Both menisci were significantly decreased in DE compared with controls (p < 0.02, t test). Tear breakup dynamics analysis showed that significantly greater areas of breakup occurred with full compared with incomplete blinks in DE (p < 0.003 Mann Whitney U test), but not in controls. CONCLUSIONS: A stable tear film can be deposited by the upper meniscus alone following a partial blink, without contribution from the lower meniscus. The increased tear stability of partial blinks in DE may be due to less stretching of the already fragile tear film compared with a full blink, which covers more surface area. PMID- 18677236 TI - Precorneal residence time of artificial tears measured in dry eye subjects. AB - PURPOSE: The purpose of this investigation was to measure the precorneal residence time of saline and five marketed artificial tears in dry eye subjects using fluorometry. METHODS: FITC-dextran, 70 kDa molecular weight, was admixed under sterile conditions (0.1% wt/vol) into buffered saline and the marketed artificial tear formulations of varying viscosity. Precorneal residence time (RT) was measured directly in 16 mild to moderate dry eye subjects, classified by sub type, in a six-way cross-over, masked and randomized study. FITC-dextran tracer decay with a scanning fluorometer was used to estimate the gross RT (i.e., the time in minutes for the signal to return to baseline). RESULTS: All subjects were classified as having non-inflammatory meibomian gland dysfunction except one, who had a mixture of aqueous deficiency and meibomian gland dysfunction. In two separate determinations, the saline RTs were 19.1 +/- 7.4 and 17.6 +/- 8.2 min. The RTs for the formulations varied to some degree by viscosity, with two higher viscosity formulations demonstrating the longest RTs of 36 to 41 min, approximately twice that of saline (p < 0.001 for both 0.4% polyethylene glycol/0.3% propylene glycol, and 1.0% carboxymethylcellulose). An oil emulsion, low viscosity carboxymethylcellulose and moderate viscosity hydroxypropylmethylcellulose-containing formulation were not statistically different from saline (RTs of 18, 22 and 24 min, p values = 0.983, 0.818 and 0.099, respectively). CONCLUSIONS: More than two-fold RT differences were found for the higher viscosity, more muco-adhesive formulations compared to saline. However, other formulations provided RTs close to saline, suggesting that RT is influenced by factors other than simple viscosity. Future studies should examine the interplay of spreading characteristics, pseudoplasticity and muco-adhesion relative to RT to determine the individual and cumulative effects on formulation retention. PMID- 18677237 TI - Effect of artificial tears on tear stress test. AB - PURPOSE: To determine the effect of two artificial tears of different viscosities in the relief of environmental dry eye induced with a novel tear stress test (TST). METHODS: A novel TST was developed and validated. The following four test conditions were evaluated in 12 healthy normal subjects in a cross-over and subject masked study; unprotected (no test solution used), relief (test solution instilled just after application of TST), immediate protection (test solution instilled just before application of TST), and chronic protection (1 week prophylactic use of the solution). The test solutions were Cellumed, with high viscosity, and Refresh Contacts, with low viscosity. Low contrast visual acuity, symptoms with analogue scales (symptom scoring), non-invasive tear break-up time, and tear evaporation were measured before (prestress) and 2 min after (poststress) application of TST. Two weeks of washout period was allowed after first test solution. RESULTS: In unprotected test condition, there was no significant difference between pre- and poststress visual acuity (p = 0.102), tear evaporation (p = 0.530), and non-invasive tear break-up time (p = 0.878), however, poststress total symptom score was significantly higher than prestress (p = 0.002). No significant differences were seen between pre- and post-total symptom score at relief (p = 0.241 for Cellumed and 0.114 for Refresh Contacts) and immediate protection (p = 0.890 for Cellumed and 0.136 for Refresh Contacts) for both test solutions, whereas postsymptoms total score was significantly higher than prestress at chronic protection (p = 0.003 for both). No significant differences were seen in the effect of the two solutions in all test conditions. CONCLUSIONS: There is an increase in dry eye symptomatology after ocular stress. The use of artificial tears just before or after ocular stress is helpful in relieving resultant symptoms in normals. PMID- 18677238 TI - Subjective and objective variation of the tear film pre- and post-sleep. AB - PURPOSE: To date, few studies have correlated the overnight effects of the preocular tear film (POTF) with subjective symptoms. This study investigates the POTF volume and stability, bulbar hyperemia (BH), tear ferning (TF) and the participant's subjective symptoms, pre- and post-sleep. METHODS: Thirty subjects were recruited, consisting of two evenly distributed groups who were symptomatic of dry eye (DE) and those that were asymptomatic dry eye, determined using the McMonnies questionnaire. Subjects were evaluated at 10 p.m. (baseline), on waking at 7 a.m., and then hourly until 10 a.m. At each visit, tear meniscus height (TMH), various subjective factors, BH and POTF stability by non-invasive break-up time (NIBUT) were assessed. Tear collection was performed at 10 p.m, 7 and 10 a.m. for TF analysis. RESULTS: With the exception of burning, all other symptoms (comfort, dryness, clarity of vision, and grittiness) revealed an overnight change (p < 0.05) within each group, but not between the two groups (p > 0.05). Both the tear meniscus height and BH were elevated upon waking and differed significantly between test times for each group (p < 0.05), but not between groups (p > 0.05). NIBUT was lower for the DE group (p < 0.001). The non-dry eye (NDE) group did not significantly alter over time (p > 0.05), but the DE group did (p = 0.004), with a longer NIBUT in the morning. TF demonstrated a degraded pattern upon waking for both groups (p < 0.05). Most of the changes returned to baseline within an hour after waking. CONCLUSIONS: The properties of the POTF undergo a change during extended periods of eye closure and the human POTF is different upon waking to that present immediately before sleep. Most of the parameters determined rapidly revert to baseline levels once the POTF is allowed to refresh. PMID- 18677239 TI - Carbomer and sodium hyaluronate eyedrops for moderate dry eye treatment. AB - PURPOSE: This randomized, double-masked study compared the effectiveness of two commercially available ocular lubricants containing either 0.3% Carbomer 934 or 0.18% sodium hyaluronate (SH) in treating moderate dry eye. METHODS: Sixty-five subjects with dry eye were recruited and supplied with eyedrops containing either Carbomer or SH to use for a month. Principle outcome measures were the severity of symptoms of ocular irritation, tear break-up time without (NIBUT) and with (TBUT) fluorescein, and corneal and conjunctival staining with fluorescein and lissamine green, respectively. At the end of the experiment, subjects were also asked, on average, how many times a day they used the treatment and the duration of any post-instillation blur. RESULTS: Both Carbomer and SH reduced the symptom severity and ocular surface staining, but neither had a lasting effect on NIBUT or TBUT. The treatment effects of Carbomer and SH were equivalent for symptoms, NIBUT and TBUT. However, for both corneal and conjunctival staining, SH outperformed Carbomer in improving the integrity of the ocular surface. There was no difference in the average instillation frequency of the two products. Visual disturbance after instillation of either formulation was generally short, but lengthy periods of blur were significantly more common after the use of Carbomer. CONCLUSIONS: Both of the eyedrops trialled are suitable for patients with moderate dry eye, but of the two, the SH-containing treatment has marginal benefits in therapeutic efficacy and has less propensity to cause visual disturbance. PMID- 18677240 TI - Treatment, material, care, and patient-related factors in contact lens-related dry eye. AB - PURPOSE: To examine the effect of general contact lens and material characteristics, care solutions, treatment, and patient-related factors on contact lens-related dry eye. METHODS: The data were derived from the Contact Lens and Dry Eye Study, designed as a cross-sectional and nested case-control study including 360 subjects. In separate statistical models, logistic regression was used to examine general contact lens characteristics, specific hydrogel lens materials, care solutions, and patient-related factors associated with dry eye status (controlled for age, gender, and current treatments). RESULTS: Several factors were significantly associated with dry eye, including treatment factors such as a recent contact lens refitting (odds ratios [OR] = 5.75, 95% confidence intervals [CI] = 2.14 to 15.46) and use of artificial tears/rewetting drops (OR = 1.09, 95% CI = 1.02 to 1.16), in addition, currently worn materials including Food and Drug Administration (FDA) group II (OR = 2.98, 95% CI = 1.14 to 6.19) and IV (OR = 1.87, 95% CI = 1.08 to 3.24). Significant patient-related factors included decreased overall satisfaction (OR = 3.57, 95% CI = 2.08 to 5.88,), dry eye in the absence of contact lens wear (OR = 6.54, 95% CI = 2.57 to 16.62), reduced daily lens wear duration (OR = 1.16, 95% CI = 1.06 to 1.26), and reduced ability to wear lenses as long as desired (OR = 2.44, 95% CI = 1.30 to 4.54). Care solutions were not associated with contact lens-related dry eye. CONCLUSIONS: The strong association of common treatment factors with dry eye status in contact lens wearers suggests that these treatments are not entirely effective. The use of high water content materials was strongly related to dry eye in lens wearers, whereas care solutions were not. Contact lens-related dry eye was also associated with several patient-related factors such as greater ocular discomfort (without lenses), dissatisfaction, and inability to wear lenses for desired durations. PMID- 18677242 TI - Improving contact-lens related dryness symptoms with silicone hydrogel lenses. AB - PURPOSE: To determine dryness symptoms attributable to hydrogel contact lens (HCL) wear by comparing symptoms from age-matched HCL wearers and non-wearers in a cross-sectional study, and to compare that difference to the change in proportion of subjects reporting frequent dryness among HCL wearers after refitting with lotrafilcon A or B silicone hydrogels (SHCLs). METHODS: Prevalence of frequent dryness symptoms was compared between HCL and non-wearers from a cross-sectional, historical dataset of Dry Eye/Contact Lens Dry Eye Questionnaires using an age-matched subset of 259 HCL and 246 non-wearers. Prospective change in prevalence of frequent dryness from non-randomized studies (n = 1036), in which daily wear (DW) HCL wearers were refit with lotrafilcon A or B SHCLs, was then compared to the cross-sectional difference between HCL wearers and non-wearers. RESULTS: In the Dry Eye/Contact Lens Dry Eye Questionnaires dataset, 47% fewer non-wearers reported frequent dryness than HCL wearers (p = 0.0001). In the lotrafilcon A DW refitting trials, frequent dryness was reported by 67 and 62% fewer subjects for during the day (DD) and end of the day (EOD) dryness (p = 0.0001, both) after refitting. In continuous wear lotrafilcon A trials, there were 63 and 41% reductions in the proportion reporting frequent DD and EOD dryness (p = 0.014 and p = 0.02). Refitting to DW lotrafilcon B yielded a 48 and 46% reduction in the number of subjects reporting frequent DD and EOD dryness (p < 0.0001, both). CONCLUSIONS: After refitting DW HCL wearers with either lotrafilcon A or B SHCLs, the proportion of symptomatic wearers was approximately half; an amount similar to the difference in proportion between HCL and non-wearers. This raises the possibility that refitting HCL wearers with SHCLs eliminates the component of dryness that is induced by HCL wear. Further research with more robust experimental design is recommended to test this hypothesis. PMID- 18677241 TI - Lubricant effects on low Dk and silicone hydrogel lens comfort. AB - PURPOSE: To investigate the influence of three lubricants of varying viscosity, on postinsertion and 6 h comfort with contact lens wear. METHODS: Comfort and associated symptoms of dryness were assessed in 15 experienced contact lens wearers. Subjects wore a low Dk lens in one eye and a silicone hydrogel in the other and participated in four separate trials involving no lubricant (baseline), saline, and two commercially available lubricants of differing viscosity. The in eye lubricants were used immediately following lens insertion and every 2 h postinsertion for a 6 h wear period. RESULTS: Postlens insertion comfort was significantly better for both lens types when lubricants or saline were used compared with no lubricant use. After 6 h lens wear, comfort was influenced by lens type and not by in-eye lubricant or saline use. Also after 6 h lens wear, less dryness sensation was reported for silicone hydrogel lenses when using lubricants but not saline. DISCUSSION: Although lubricant use does help reduce dryness symptoms with silicone hydrogel lens wear, there appears to be minimal longer-term benefit to comfort. Furthermore, increased lubricant viscosity did not lead to improved longer-term comfort. PMID- 18677243 TI - Staying connected. PMID- 18677244 TI - Changing practice demands...changing educational opportunities. PMID- 18677246 TI - A roadmap to the disease specific care certification process. AB - The Joint Commission's Disease Specific Care Certification (DSC) recognizes disease management programs for superior quality. Programs are awarded DSC that meets established criteria for managing chronic conditions. Awarding of DSC is based on a program's compliance with disease-specific standards, evidence-based clinical practice guidelines, and performance improvement measures. Programs will be evaluated during a site visit by a Joint Commission reviewer; once certification is awarded, it is valid for 2 years. Programs are required to submit an Intra-Cycle Evaluation Report after 1 year. PMID- 18677248 TI - Venous thromboembolic prophylaxis: the use of aspirin. AB - Venous thromboembolism (VTE) is a term used collectively for deep vein thrombosis (DVT) and pulmonary embolism. Without prophylaxis, the incidence of documented DVT in the orthopaedic surgery patient is reported in the range of 50%-60%. A multimodal approach to DVT prophylaxis is the standard of care for all patients undergoing total hip arthroplasty and total knee arthroplasty. At our local hospital, low-risk patients are being sent home with aspirin as the medication for VTE prophylaxis. This article will provide an overview of the pathophysiology of VTE and the current prevention guidelines including the use of aspirin. PMID- 18677250 TI - Proactive patient rounding to increase customer service and satisfaction on an orthopaedic unit. AB - Customer service and patient satisfaction have become increasingly important in the healthcare industry. Given limited resources and a myriad of choices, on which facets of patient satisfaction should healthcare providers focus? An analysis of 40,000 observations across 4 hospitals found 1 important intervention: timely staff responsiveness. Using the Plan-Do-Check-Act (PDCA) quality methodology, the goal was set to improve staff responsiveness to orthopaedic patient needs and requests, thus improving patient satisfaction. A model to improve staff responsiveness was systematically developed and implemented. The I Care Rounding model places the emphasis on proactively meeting patient needs through hourly rounding, rather than caregivers providing care in a reactionary mode. After full implementation, positive improvement was demonstrated. PMID- 18677252 TI - All-terrain vehicle safety: a long-standing issue! AB - An increasing number of children younger than 16 are being killed or severely injured from riding adult-sized all-terrain vehicles. Parents and communities need the education regarding the danger of these vehicles, which are not toys! NAON has been consistent with attempting to get the word out and prevent these injuries and deaths so we can improve the safety of our children. PMID- 18677253 TI - Evaluation and management of joint pain. AB - The purpose of this article was to provide a review for orthopaedic nurses and nurse practitioners who evaluate, manage, and care for patients with joint pain. Joint pain is a common complaint evaluated by primary care providers. The causation of joint pain is complicated to identify because of an extensive range of differential diagnosis. The history and physical examination are crucial components in evaluating and managing joint pain. The primary care provider uses clinical factors such as patient demographics, presence of inflammation, acute/chronic duration, extra-articular manifestations, pattern of joint involvement, and disease chronology. Many rheumatologic laboratory tests are nonspecific, but aspiration of the joint with synovial fluid analysis may provide diagnostic clues, especially to differentiate infection versus inflammation. Primary care providers utilize both pharmacologic and nonpharmacologic regimens to manage acute and chronic joint pain. PMID- 18677259 TI - Agency for Healthcare Research and Quality (AHRQ) Web site. PMID- 18677271 TI - Postexposure interventions to prevent infection with HBV, HCV, or HIV, and tetanus in people wounded during bombings and other mass casualty events--United States, 2008: recommendations of the Centers for Disease Control and Prevention and Disaster Medicine and Public Health Preparedness. AB - People wounded during bombings or other events resulting in mass casualties or in conjunction with the resulting emergency response may be exposed to blood, body fluids, or tissue from other injured people and thus be at risk for bloodborne infections such as hepatitis B virus, hepatitis C virus, human immunodeficiency virus, or tetanus. This report adapts existing general recommendations on the use of immunization and postexposure prophylaxis for tetanus and for occupational and nonoccupational exposures to bloodborne pathogens to the specific situation of a mass casualty event. Decisions regarding the implementation of prophylaxis are complex, and drawing parallels from existing guidelines is difficult. For any prophylactic intervention to be implemented effectively, guidance must be simple, straightforward, and logistically undemanding. Critical review during development of this guidance was provided by representatives of the National Association of County and City Health Officials, the Council of State and Territorial Epidemiologists, and representatives of the acute injury care, trauma, and emergency response medical communities participating in the Centers for Disease Control and Prevention's Terrorism Injuries: Information, Dissemination and Exchange project. There recommendations contained in this report represent the consensus of US federal public health officials and reflect the experience and input of public health officials at all levels of government and the acute injury response community. PMID- 18677272 TI - Steroids in otolaryngology. AB - Steroids are increasingly being used to treat a wide spectrum of otolaryngological disorders. It is important for ear-nose-throat surgeons to understand the short- and long-term complications associated with steroid use. The aim of this study was to review the role of steroids in common ear-nose throat disorders. Corticosteroid therapy has been shown to be effective in the management of idiopathic facial nerve palsy, allergic rhinitis, acute sinusitis, sinonasal inflammatory polyposis, and croup. The therapeutic efficacy of steroids in the management of Meniere's disease, sudden idiopathic sensorineural HL, chronic otitis media, and vestibular neuronitis remain controversial. PMID- 18677273 TI - Treatment responsiveness of the Singing Voice Handicap Index. AB - OBJECTIVES/HYPOTHESIS: To examine the responsiveness of the previously validated Singing Voice Handicap Index (SVHI) to treatment related changes in patients' singing voice. STUDY DESIGN: Prospective. METHODS: Thirty singing patients at a tertiary voice clinic prospectively completed the Voice Handicap Index and SVHI at their initial presentation, pretreatment, and posttreatment. RESULTS: Patients had lower SVHI scores posttherapy compared with the initial SVHI scores (P or=2 years (P = .013), and every patient with a duration of symptoms >2 years recurred (P = .002). There was a trend for more local recurrences after limited resection (P = .061). CONCLUSIONS: ACC of the EAC often recurs many years after definitive treatment. Although our sample size was too small to make definitive conclusions, we recommend aggressive local therapy with lateral temporal bone resection and adjuvant postoperative radiotherapy. In addition to successful local therapy, early diagnosis may be the only other effective means of preventing distant metastases. PMID- 18677278 TI - Topical therapy in anosmia: relevance of steroid-responsiveness. AB - OBJECTIVES/HYPOTHESIS: The use of steroids either systemically or topically is known as a common therapy in patients with anosmia. Nevertheless, investigations giving proof for the benefit of a topical therapy are very rare, and no prognostic factors are known. In our study, we for the first time evaluated the additional effect of a topical therapy not only with steroids but also with antibiotics after conventional pretreatment with oral steroids and propose the steroid-responsiveness of an anosmia as a prognostic factor. STUDY DESIGN: Retrospective design. METHODS: We analyzed the data of 299 patients with olfactory dysfunction. Eighty-nine underwent initial pretreatment with systemic steroids and presented data over a sufficient follow-up time. In a second step all these patients were given a topical treatment in a head down forward position, namely either budesonid alone or in combination with neomycin. Primary outcome parameter was the threshold, discrimination and identification (TDI) score. RESULTS: Initial therapy with oral steroids changed the TDI from 15.5 to 18.7 in the means (P < .001). In general, leaving away systemic steroids while applying local therapy did not led to a reduction of the TDI (P < .001). Dividing up the patients into those suffering from a steroid-responsive anosmia (SRA) and those without benefit from initial systemic steroids (non-SRA), the topical treatment led to a significant difference between the two groups with benefit toward the non-SRA group (P < .001). Regarding only the nonchronic rhinosinusitis anosmics, these findings became even more apparent. Furthermore, in non-SRA patients we found even better results with steroids in combination with neomycin as a topical therapy. In this group, the combined topical therapy elevated the TDI for +2.1 points whereas topical steroids alone raised it only for +1.0 point. CONCLUSIONS: The steroid-responsiveness of anosmia seems to be a relevant prognostic indicator for a significant benefit of a topical therapy in general. Within all patients, the effect of an initial systemic therapy could be maintained by the adjacent topical treatment whereas in non-SRA patients a topical therapy has a significant greater impact. Furthermore, antibiotics even seem to have an additional effect in this group. Different reasons, first of all an overwhelmed steroid resistance by additional antiinflammatory effects of antibiotics, e.g., the inhibition of apoptosis might play a role and are discussed. PMID- 18677279 TI - Disease-associated novel CD46 splicing variants and pathologic bone remodeling in otosclerosis. AB - OBJECTIVE/HYPOTHESIS: Otosclerotic bone is supposed to show unique CD46 expression pattern because otosclerosis is an organ-specific disease with viral etiology. STUDY DESIGN: Otosclerosis is a complex bone remodeling disorder of the human otic capsule, which is associated with persisting measles virus infection. The general cellular receptor of measles virus is the CD46, which has 14-known splicing isoforms. METHODS: Nucleic acid was extracted from ankylotic stapes footplates (N = 99) removed during stapedectomies. Consecutive histological, CD46 specific immunohistologic analysis, and multiple polymerase chain reaction (PCR) amplifications were performed. Measles virus was detected by seminested reverse transcriptase-PCR. Splicing variants of CD46 were identified by nested reverse transcriptase-PCR and finally determined by mass sequencing of complementary DNA. RESULTS: Measles virus RNA was detectable only in histologically otosclerotic stapes footplates. Virus negative-fixed stapes represent degenerative disorders of variable histopathology. Otosclerosis is featured by an increased number of osteoclasts showing strong CD46 immunoreaction in contrast to nonotosclerotic stapes fixations. Normal and nonotosclerotic stapes footplates show consistent expression of "c," "d," "e," "f," and "l" CD46 splicing isoforms. In contrast, four novel CD46 splicing variants were additionally detected in otosclerosis: os1, os2, os3, and os4. CONCLUSIONS: Newly described CD46 isoforms have shorter or missing transmembrane domain and a rare cytoplasmic tail with pathological or uncommon signal transduction; however, virus binding ability remains equal and invariable. These changes may be responsible for the smooth virus replication. A special expression pattern and altered functions of CD46 could explain the organ specific and virus-associated pathogenesis of otosclerosis. PMID- 18677280 TI - Salvage laryngectomy for failed conservative treatment of laryngeal cancer. AB - OBJECTIVES/HYPOTHESIS: To determine the rate of salvage laryngectomy after failure of radiation or chemoradiation over 10-year course, identify factors associated with increased risk of treatment failure, assess survival after salvage laryngectomy. STUDY DESIGN: Retrospective review. METHODS: Twenty patients who underwent salvage total laryngectomy were studied. Overall interval to recurrence (ITR) was 9.9 months, overall postsalvage-laryngectomy survival (PSL-OAS) 25 months, and interval to death (PSL-ITD) 15.8 months. Mortality was 85%. RESULTS: Four patients failed chemoradiation and 16 failed radiation alone. Based on treatment, there was no difference in ITR (12.8 vs. 8.8 months, P = .27) or PSL-OAS (27.5 vs. 24.4 months, P = .86). PSL-ITD was significantly higher after chemoradiation (27.5 vs. 12.2 months, P = .02). Advanced T-stage patients were significantly younger than early-stage (53 vs. 64 years, P = .02), but had no significant difference in ITR (12.7 vs. 6.8 months, P = .08), PSL-OAS (20.1 vs. 33.6 months, P = .35), or PSL-ITD (12 vs. 21.6 months, P = .1). Analysis by site and differentiation showed no significant differences. Heavy alcohol use/alcoholism was reported in >50%. Rate of fistulization was 20%. Three patients currently survive, with average postsalvage-laryngectomy disease-free survival (PSL-DFS) 77 months. CONCLUSIONS: Salvage laryngectomy is an uncommon procedure (7% over 10-year period). There is no difference in survival after salvage laryngectomy for radiation or chemoradiation failure. Patients failing chemoradiation have longer PSL-ITD than those failing radiation alone. Patients with T3-4 tumors tend to be younger. T-stage of tumor does not affect rate of salvage procedure. Fistulae formed in 20%. Neither location nor differentiation affects survival. Surgical salvage after primary conservative treatment is associated with an approximately 2-year survival. PMID- 18677281 TI - Endoscopic arytenoid lateropexy for isolated posterior glottic stenosis. AB - OBJECTIVES/HYPOTHESIS: A posterior glottic stenosis (PGS) may limit the abduction of the arytenoid cartilages. One option for the treatment of dyspnea in lower grade stenoses is endoscopic laterofixation of the vocal cords after scar excision. In our prospective study, we assess a refined method for effective endoscopic mobilization and lateropexy of the arytenoid cartilages. STUDY DESIGN AND METHODS: Thirty-two consecutive patients with PGS underwent surgery. Endoscopically, the scar between the arytenoid cartilages was transected with a CO2 laser. The scars that had spread into the cricoarytenoid joint were transected with a right-angled endolaryngeal scythe designed for this purpose. The lateropexy of the adequately mobilized arytenoid cartilages was performed with a reinforced Lichtenberger's needle carrier instrument, with consideration of the real abduction of the cricoarytenoid joint. RESULTS: Twenty-eight patients achieved an excellent breathing ability, only effort dyspnea remained in three cases. One patient could not be decannulated due to aspiration. The early postoperative improvement in the airway function test results showed no relationship with the grade of stenoses. However, in cases of higher grade stenoses with bilateral joint damage, the later postoperative airway function results had decreased slightly. In 25 cases, phonation significantly improved after the removal of the fixing sutures. CONCLUSIONS: After proper mobilization, endoscopic arytenoid lateropexy can be considered as a minimally invasive function-preserving procedure even for severe PGS. This treatment option provides stable improvements in breathing ability and good voice quality without the need for tracheostomy. PMID- 18677282 TI - Galectin-3 upregulation during tumor progression in head and neck cancer. AB - OBJECTIVES/HYPOTHESIS: To examine the level of expression of galectin-3 in relation to neoplastic progression of hypopharyngeal squamous cell carcinomas (HSCCs) and laryngeal squamous cell carcinomas (LSCCs). STUDY DESIGN: Retrospective study. METHODS: Using a polyclonal antibody against galectin-3 without cross-reactivity to other galectins, we analyzed the presence of galectin 3 using quantitative immunohistochemistry in i) a series of 79 HSCCs compared with 16 normal epithelia, 20 low-grade dysplasia (Low_D) and 25 high-grade dysplasia (High_D) and in ii) a series of 58 LSCCs compared with 34 normal epithelia, 12 Low_D, and 18 High_D. In parallel, galectin-3 expression was studied using Western blotting on a series of 19 fresh biopsies from patients presenting a head and neck tumor. RESULTS: Western blotting excluded a notable degree of proteolytic truncation of galectin-3 in situ. Immunohistochemical galectin-3 positivity expressed as percentage of cells was significantly higher in LSCCs and HSCCs than in Low_D (P = .01) or High_D (P = .0002), respectively. Increased expression of galectin-3 in HSCCs was accompanied by a shift from the cytoplasmic compartment to the nucleus (P = .007). In intertumor-type comparison, laryngeal carcinomas presented nuclear presence of galectin-3 only rarely (1 of 58 cases in laryngeal cancer vs. 27 of 79 cases in hypopharyngeal cancer, P = .00006) and a comparatively low labeling index (P < 10(-6)). CONCLUSIONS: Our data reveal an association between level of presence of galectin-3 and neoplastic progression of HSCCs and LSCCs. PMID- 18677283 TI - Audiological application criteria for implantable hearing aid devices: a clinical experience at the Nijmegen ORL clinic. AB - OBJECTIVES/HYPOTHESIS: To define audiological application criteria for different implantable hearing aid devices. STUDY DESIGN: Retrospective study. METHODS: Comparisons were made between aided speech recognition scores obtained at conversational level (65 dB) in patients with the Vibrant Soundbridge (VSB) (n = 22), the Otologics middle ear transducer (MET) (n = 10), conventional hearing aids (behind-the-ears) (n = 47), and cochlear implants (CIs) (n = 123). RESULTS: In relation to hearing loss, only for mild hearing loss, speech recognition scores with VSB were comparable to that with conventional hearing aids. In the Otologics MET users, speech recognition scores were comparable with those of the conventional hearing aid users until a mean hearing loss of about 75 dB HL. At a sensorineural hearing loss of about 65 dB HL or more, the Otologics MET users have better speech recognition scores than the VSB users. For comparison with CI users, we followed a more conservative approach. In 90% of the users of a CI, speech recognition scores were better than those in: 1) patients with a conventional hearing aid and a mean hearing loss of about 95 dB HL or worse; 2) patients with an Otologics MET and a mean hearing loss of 85 dB HL or worse. CONCLUSIONS: Patients fitted with a VSB or an Otologics MET middle ear implant do not demonstrate better speech recognition scores than patients fitted with today's conventional hearing aids. Results might even been worse. However, the VSB and Otologics MET are a good option in patients with moderate (VSB) to severe (Otologics MET) sensorineural hearing loss and external otitis. PMID- 18677284 TI - Is there a relationship between Eagle Syndrome and cervicofacial painful soft tissue rheumatisms? AB - OBJECTIVES/HYPOTHESIS: To investigate the incidence of locomotor system pathologies such as myofacial pain syndrome (MPS), fibromyalgia syndrome (FMS), and temporomandibular dysfunction in patients with Eagle Syndrome. STUDY DESIGN: Prospective study. PATIENTS AND METHODS: Fourteen patients with Eagle Syndrome, who were treated surgically, were enrolled in the study. Etiologic factors for cervicofacial pain were assessed, pain status was compared preoperatively and postoperatively using visual analogue scale. Palpation of tonsillar fossa, neck and shoulder examination, neurologic examination, evaluation of occlusion status, trigger points, and painful zones were done. Plain anteroposterior and lateral radiographs of the cervical spine were obtained and the lengths of the transverse processes of the seventh cervical vertebrae were measured bilaterally. RESULTS: MPS, FMS, and temporomandibular dysfunction were diagnosed in 9 (64.3%), 3 (21.4%), and 2 (14.3%) patients, respectively. Visual analogue scale scores decreased significantly after the surgical excision of elongated styloid processes (from 6.7 +/- 2.3 to 2.1 +/- 1.8), and all the complaints except for headache had diminished (P < .05). Lengths of transverse processes of seventh cervical vertebra were found to be correlated with the length of styloid process (right; r = 0.644, P = .024, left; r = 0.616, P = .033). CONCLUSIONS: Cervicofacial pain is a common complaint in patients with Eagle Syndrome. It frequently coexists with rheumatic disorders resulting in chronic pain such as MPS and FMS. Even though Eagle Syndrome is a rare condition, it should be kept in mind in patients suffering from chronic cervicofacial pain that is refractory to treatment. Clinicians should be alert to diagnose and treat coexisting locomotor system disorders. PMID- 18677286 TI - Adult subglottic stenosis: management with laser incisions and mitomycin-C. AB - OBJECTIVES/HYPOTHESIS: To assess the efficacy of endoscopic laser radial incisions with mitomycin-C application (ELRM) in managing adult subglottic stenosis (SGS). STUDY DESIGN: Retrospective case series review. METHODS: Fifteen consecutive cases of adult SGS treated with ELRM at a single tertiary referral center over three years were reviewed. Subjects with SGS secondary to Wegener's granulomatosis (WG) and idiopathic SGS were included. Patients with cartilaginous SGS were excluded. The primary outcome measure was postoperative reduction in symptoms. Secondary outcome measures included total number of procedures required to relieve symptoms, interval between procedures, and improvement in pulmonary function tests when available. In addition to surgery, 14 of 15 patients were treated medically for reflux. RESULTS: Ten women and five men with average age 48 years were identified. Ten patients had idiopathic SGS and five had WG. The predominant presenting symptom was dyspnea on exertion in all patients. All subjects reported at least a temporary postoperative reduction in symptoms. Six patients (40%) required only one ELRM and nine patients (60%) required repeat ELRM at an average interval of 9 months. The average interval for the six patients with idiopathic etiology requiring a second procedure was 9 months. One subject with WG required four procedures. His interval improved from 2.5 to 7 months between procedures. Evidence of extrathoracic airway obstruction resolved in three of four patients with pre and postoperative pulmonary function tests. CONCLUSIONS: ELRM is an effective method of managing SGS associated with idiopathic causes. In patients with WG, ELRM reduced airway associated symptoms and avoided need for tracheotomy. PMID- 18677285 TI - Characteristics of age-related changes in cultured human vocal fold fibroblasts. AB - OBJECTIVES/HYPOTHESIS: Normal human vocal fold fibroblast (hVFF) primary cell lines are unavailable commercially and are very difficult to acquire, subsequently little is known about their characteristics. The purpose of this study was to compare the morphological and proliferation characteristics and gene expression of hVFFs from different aged donors. STUDY DESIGN: In vitro. METHODS: We developed three normal hVFF primary cell lines from donors aged 21 (21T), 59 (59T) and 79 (79T) years. We characterized their morphological features, proliferative abilities, telomere lengths, and their functional gene expression by quantitative real-time PCR. RESULTS: The 21T line maintained a typical spindle shape until passage 14 whereas 59T and 79T changed morphology to wider, shorter cells at passage 7. Proliferation rates were constant for the 21T through passage 14; 59T's proliferative half-life was passage 9, whereas 79T maintained lower proliferation rates from passage 4. Gene expression levels for fibronectin, collagen I, collagen VI, procollagen I and elastin demonstrated similar patterns for all lines, however, relative amounts decreased with the age of donor. Telomere lengths did not show differences related with donor age. CONCLUSIONS: hVFF primary cultures have limited proliferative capacity. The morphology, proliferation, differentiation, and gene expression levels of VFF can be affected by age, but senescence patterns were similar across the ages. PMID- 18677287 TI - Refining the use of warfarin through genetic testing. PMID- 18677288 TI - Does the preventive effect of different drugs depend on location of the atherosclerotic process? AB - Atherosclerosis can affect nearly any part of the arterial system. Therefore, it is considered as a generalized disease. As most probably similar or identical etiopathogenetic mechanisms are involved in different atherosclerotic diseases, a different effect of treatment of risk factors on atherosclerotic lesions in different parts of the vascular system is expected. Until now, great emphasis has been placed on the aggressive pharmacological management of coronary artery disease, less attention has been devoted to the management of cerebrovascular and much less to peripheral arterial disease, despite their significant morbidity and mortality. The data from recent trials have indicated that treatment of patients with antiplatelet drugs, statins, antihypertensive and antidiabetic drugs prevents the progression of coronary atherosclerosis, reduces cardiovascular events and improves prognosis of coronary patients. Subgroup analyses from large studies have also shown that treatment of risk factors for atherosclerosis with drugs reduces cardiovascular events and improves prognosis of cerebrovascular and peripheral arterial occlusive disease. Although some studies indicate that the effects of distinct preventive procedures are to some extent dependent on the locations of atherosclerotic disease, it seems that the success of preventive measures is mostly related to the progression of the disease or the risk of treated population and not on the treated vascular bed. PMID- 18677289 TI - Autologous intra-arterial infusion of bone marrow mononuclear cells in patients with critical leg ischemia. AB - AIM: The injection of bone marrow mononuclear cells (BMMC) into the gastrocnemius muscle has given promising results in patients with critical limb ischemia (CLI). In this article, we have assessed whether a less invasive procedure, i.e. intravascular BMMC infusion, could be effective in this population of patients. METHODS: A total of 28 limbs in 24 patients with CLI were treated. An amount of 276-700 mL of marrow blood was harvested from posterior iliac crests and BMMC were obtained by standard procedure used for bone marrow transplantation. After performance of digital subtraction angiography, BMMC were injected laterally through a 4 Fr sheet. Primary outcome was efficacy of the procedure measured as healing of defects, frequency of high amputations and change of ischemia grade; among secondary outcomes were safety of the procedure, angiographic changes and changes in quality of life. RESULTS: One year after treatment, all patients were alive and only 2 patients have undergone high amputation. Eleven of 14 defects have healed (78%) and Fontaine grade of ischemia has changed from median grade 3.5 to median grade 2 (P<0.0001). Collateral vessel development has improved by mean 1.13 and 1.3 points on a four-point semiquantitative scale in calf and foot, respectively (P<0.0001). There were no grade III-IV adverse events. According to the SF-36 quality of life questionnaire, 1 year after the procedure patients have reported significant improvement in all measured items. CONCLUSION: Intra arterial infusion of BMMC can lead to significant and long-lasting subjective and objective improvements in patients with CLI. The results merit validation by randomized controlled studies in patients with less critical limb ischemia. PMID- 18677290 TI - Infrainguinal endovascular procedures based upon the results of duplex scanning. AB - AIM: The aim of this study was to report the role of duplex scanning in selection of patients with lower limb ischemia for infrainguinal endovascular revascularization. METHODS: From January 2002 to December 2005, 95 patients (66 male, 29 female) underwent infrainguinal endovascular revascularization based on duplex scanning. The indications for surgery were severe claudication (11%), rest pain (15%), gangrene (40%), and non-healing ulcer (34%). RESULTS: Duplex procedure time averaged 60+/-30 min. A total of 120 arterial hemodynamic relevant lesions were treated with endovascular therapy, 47 were localized in the aorto iliac segment, 55 in the femoro-popliteal segment and 18 were infrapopliteal. Out of a total of 120 lesions, 107 (89%) were successfully dilated; 105 lesions (88%) predicted by preoperative duplex scanning were confirmed by contrast arteriography (CA) at the time of surgery. Additional lesions were revealed by intraoperative arteriography in 15 cases (12%). The accuracy and sensitivity of duplex scanning in the selection of aorto-iliac lesions for endovascular procedures was 86%, 91% for femoro-popliteal lesions, and 78% for infrapopliteal lesions. CONCLUSION: The results of this experience show that duplex scanning may be a safe alternative to CA for patients with chronic limb ischemia. Adequate training and experience is necessary to utilize this technique for the selection of patients for infrainguinal endovascular procedures. PMID- 18677291 TI - Tissue factor, tissue pathway factor inhibitor and risk factors of atherosclerosis in patients with chronic limbs ischemia: preliminary study. AB - AIM: Thrombus formation plays a critical role in pathogenesis of cardiovascular complications in atherosclerotic peripheral arterial occlusive disease (PAOD). Tissue factor (TF) initiates the clotting cascade and is considered an important regulator of hemostasis and thrombosis. TF activity is regulated by TF pathway inhibitor (TFPI). The aim of our study was to evaluate plasma levels of the TF, TFPI and their relation to coagulation system and various other risk factors of atherosclerosis in patients with chronic limbs ischemia. METHODS: Plasma TF, total TFPI, truncated TFPI, full-length TFPI were assessed by ELISA using commercially available kits (IMUBIND Tissue Factor; Total TFPI; Truncated TFPI ELISA Kit; American Diagnostica Inc. Stamford) in 62 claudicant patients with PAOD and 20 healthy controls. RESULTS: We observed statistically higher levels of TF (94+/-52 pg/mL), total TFPI (43+/-8 ng/mL), and truncated TFPI (22+/-7 ng/mL) in patients with PAOD compared to healthy individuals (TF: 66+/-15 pg/mL; total TFPI: 36+/-4 ng/mL; truncated TFPI: 14+/-5 ng/mL). Full-length TFPI (20+/-4 ng/mL) is lower in patients with PAOD than in controls (23+/-5 ng/mL). The study indicated a positive correlation between TF and truncated TFPI (r=0.34), total TFPI and full TFPI (r=0.5), total TFPI and truncated TFPI (r=0.83) in patients with PAOD, and negative correlation between full TFPI and truncated TFPI (r= 0.65) in the control. CONCLUSION: Elevated levels of TF, disorders of balance between full-length TFPI and truncated TFPI as well as significantly increased truncated TFPI level in patients with PAOD can be independent risk factors of atherosclerotic complications. PMID- 18677292 TI - The relationship of endothelin-1 and inducible nitric oxide synthase at several stages of the anastomotic healing process. AB - AIM: The aim of this study was to investigate the interaction between the endothelin-1 (ET-1) and inducible NO synthase (iNOS) in anastomotic healing. METHODS: The expression of ET-1 and iNOS were investigated by immunohistochemistry in a rat end-to-end arterial anastomotic model. The aorta of 50 male Wistar rats was exposed, then transversely divided and re-anastomosed. The animals were sacrificed immediately after the operation (group A, control group), after 24 h (group B), on 7th postoperative day (group C), on 30th day (group D) and at 6 months (group E). Intima and media thickness and their ratio of the anastomotic segments in each group were calculated from computer digitized images of the individual sections. ET-1 and iNOS expression were measured on a semiquantitative scale ranging from 0 to 3. RESULTS: ET-1 was expressed from endothelial and smooth muscle cells (SMCs), while iNOs was expressed from SMCs and inflammatory cells. An intense expression of ET-1 was demonstrated mainly at 1 week and to a lesser degree at 1 month. Yet, at 6 months this expression was significantly weakened (P<0.001). In contrast, an intense iNOS expression was identified at 24 h, substantially regressing at statistical significant lower levels after 1 week (P<0.001). Bivariate correlation test showed a positive correlation between ET-1 and iNOS expression. CONCLUSION: ET-1 appears to play an important role in intimal thickening during anastomotic healing, especially in the late period of the process. Although there is a positive correlation between ET-1 and iNOS production, the activity of the latter is relatively limited after the first postanastomosis week. PMID- 18677293 TI - Levels of adhesion molecules bear a relationship to triglyceride levels in type 2 diabetic subjects with proven silent ischemia. AB - AIM: The aim of the study was to examine the levels of adhesion molecules, high sensitivity C-reactive protein (hs-CRP) and lipid spectrum of type 2 diabetic subjects with proven silent myocardial ischemia. METHODS: We included in the study 19 patients with ischemia (Group 1) and 16 patients without ischemia (Group 2). We documented silent ischemia by an exercise-myocardial single photon emission computed tomography. We examined the levels of total cholesterol, high density lipoprotein, low-density lipoprotein, triglycerides, vascular cell adhesion molecule 1, intercellular adhesion molecule 1, E-selectin, HbA1c, microalbuminuria (MAU), hs-CRP and carotid intima-media thickness. RESULTS: The differences among the values of lipids, adhesion molecules, HbA1c, hs-CRP, MAU between the groups were not statistically significant. E-selectin levels positively correlated with triglyceride levels in the group 1 (Spearman correlation, P<0.05). This correlation was not proven in the Group 2. CONCLUSION: Statistically differences between the study groups were not significant. Levels of E-selectin positively correlated with high triglyceride levels in type 2 diabetic subjects with silent ischemia. This correlation documents a disturbance of the reverse cholesterol transport system. PMID- 18677295 TI - Percutaneous bronchial artery embolization in the management of massive hemoptysis in chronic lung diseases. Immediate and long-term outcomes. AB - AIM: Bronchial artery embolization (BAE) is a well-established, non-surgical procedure in the emergency treatment of massive hemoptysis. This study aims to evaluate the immediate and long-term prognosis of BAE for the management of massive hemoptysis in our center. METHODS: Twenty consecutive patients (mean age: 59+/-14 years) with massive hemoptysis, underwent BAE with microspheres (Embospheres BioSphere Medical SA, Paris, France), polyvinyl alcohol particles (PVA, Ivalon, Cathmed Science; Paris, France) or/and steel coils (Cook, Denmark) after thoracic aortography and diagnostic selective and superselective catheterization of bronchial arteries and systemic collateral vessels in the bleeding lung area. Hemoptysis was due to bronchiectasis (55%), non-operable aspergillomas (15%), active tuberculosis (15%), malignancy (10%) and cystic fibrosis (5%). Mean duration of follow-up was 29+/-18 months. The recurrent-free time was calculated with Kaplan-Meier analysis. RESULTS: Immediate control of bleeding was achieved in all patients. Recurrent cases of hemoptysis were observed in 6/20 patients (30%) within 3 years and 4 of them (66.6%) occurred early in the first 3 months. Recurrent-free time was 9 months (standard error: 4) (95% confidence interval: 0-17). Repeated interventions were required in all early recurrences, due to either recanalization of the occluded arteries or non bronchial systemic artery supply. Combined use of PVA and coils was proved effective in these cases. No serious complications were observed. CONCLUSION: BAE is an effective and safe intervention in cases of massive hemoptysis. However, recurrences are common and long-term follow-up is considered important with a view to perform repeated interventions with combination of embolic materials. PMID- 18677294 TI - Fibrinolytic intra-arterial therapy in treatment of arterial occlusion in femoropopliteal segment. AB - AIM: Local intra-arterial thrombolysis is one of the methods of treatment of acute ischemia of extremities. Its aim is rapid restoration of arterial patency and the reperfusion of the ischemic extremity. It allows the identification of the cause of arterial occlusion. Identification and description of arterial wall lesions informs decisions optimizing further therapy, either surgical or endovascular. METHODS: From 2003 to 2005, 17 patients with femoro-popliteal occlusion were treated with local intra-arterial thrombolysis. All patients underwent color Doppler examination followed by angiography, which confirmed the diagnosis. A catheter was introduced through a femoral artery puncture and its tip was wedged against the thrombus or within its mass. All patients received streptokinase. The progress of thrombolysis was monitored by repeated angiography and treatment was continued for up to 6 h after the radiographically confirmed restoration of patency of the artery. RESULTS: In 15 (88%) treated patients we restored patency of the previously occluded artery and obtained complete remission of symptoms of lower limb ischemia. CONCLUSION: Local intra-arterial thrombolysis is a safe and effective method of treating lower limb ischemia in selected patients. It allows one to decrease the total dose of thrombolytic agent and it is a good alternative for surgical procedure. PMID- 18677296 TI - Relationship between the diameter of the vein graft and postoperative ankle brachial pressure index following femoro-popliteal bypass. AB - AIM: We studied whether the diameter of the saphenous vein graft affects the result of femoro-popliteal bypass surgery. METHODS: Thirty-eight patients with bypasses from the femoral artery to the above knee popliteal artery were studied. Bypasses without a patent anterior or posterior tibial artery were excluded. The great saphenous vein was used as a bypass graft in 20 extremities and Dacron grafts (6 mm or 8 mm diameter) were used in 18 arteries. The smallest diameter of the saphenous vein was measured preoperatively with ultrasonography. Vein grafts were divided into two groups: small vein graft (< or =3 mm) and large vein graft (> or =3.5 mm). The ankle brachial pressure index (ABI) was measured at 1 week and 3 months after operation. RESULTS: The diameter of the vein graft (2.5 to 4 mm, 3.4+/-0.5 mm) was positively correlated with postoperative ABI (R2 0.607, P<0.0001). The postoperative ABI at 1 week was significantly lower in the small vein graft group (0.72+/-0.09) than in the large vein graft group (0.95+/-0.11) and in the Dacron graft group (1.05+/-0.16). The ABI at 3 months was still significantly lower in small vein graft group (0.78+/-0.07). CONCLUSION: The diameter of the vein graft was positively correlated with postoperative ABI after femoro-popliteal above knee bypass. Postoperative ABI was lower using a vein graft with a diameter of < or =3 mm than that using a bigger vein graft or a Dacron graft. PMID- 18677297 TI - Guidelines for antithrombotic therapy: periprocedural management of antithrombotic therapy and use of bridging anticoagulation. PMID- 18677298 TI - Writing a case report: polishing a gem? AB - Case reports describe patient cases which are of particular interest due to their novelty and their potential message for clinical practice. While there are several types of case reports, originality and clinical implications constitute the main virtues by which case reports are judged. Defining the educational message and choosing the right audience are vital steps in the process of writing. Generally, a case report is structured, its main components being the abstract, the introduction, the case description and the discussion. Guidelines and tips for writing case reports are not enough for making a successful author, but they help, especially less experienced doctors, to exercise and improve their writing. If properly prepared, case reports can still communicate new observations in an interesting and pleasant way, thereby enriching our knowledge, even in the era of evidence-based medicine. PMID- 18677299 TI - Endovascular repair of post-surgical pseudoaneurysm of suprarenal abdominal aorta in a patient with Behcet's disease. AB - Vascular Behcet's disease has a poor prognosis. This poor prognosis is due to postoperative complications including pseudoaneurysm with 50% occurrence after previous surgical repair of aneurysm especially at the site of surgery. It is suggested that the fragility of the vascular wall may play a major role in this recurrence. Recent studies have demonstrated the effectiveness of endovascular stent-grafting for recurrent aortic aneurysm in patients with Behcet's disease. We present a recurrent aneurysm of the abdominal aorta at a previously scarred operative site in a known case of Behcet's disease. We attempted an endovascular treatment. PMID- 18677300 TI - S-shaped ilio-mesenteric bypass in a young high risk patient. AB - Symptomatic chronic mesenteric ischemia is a rare condition. Several surgical and endovascular techniques have been described, but treatment is individualized according to the conditions of each patient. We report a successful superior mesenteric artery revascularization by using an S-shaped retrograde polytetrafluoroethylene ilio-mesenteric bypass graft in a young overweight patient with a history of two abdominal vascular operations and several comorbidities. PMID- 18677302 TI - Potentially important microRNA cluster on chromosome 17p13.1 in primary peritoneal carcinoma. AB - MicroRNAs are a group of small non-coding RNAs approximately 22 nucleotides in length. Recent work has shown differential expression of mature microRNAs in human cancers. We characterized the alteration in expression of a select group of microRNAs in primary peritoneal carcinoma relative to matched cases of ovarian serous carcinoma. MicroRNA expression was analysed using semi-quantitative stem loop RT-PCR on a set of 34 formalin-fixed paraffin-embedded samples. Protein expression of p53 and bcl-2 was quantified in the corresponding tissue microarray. We provide definitive evidence that there is downregulation of a select group of microRNAs in tumours meeting Gynaecological Oncology Group criteria for primary peritoneal carcinoma relative to ovarian serous carcinoma. Specifically, we show decreased p53 expression and downregulation of miR-195 and miR-497 from the microRNA cluster site at chromosome 17p13.1 in primary peritoneal carcinoma relative to ovarian serous carcinoma. miR-195 and miR-497 may have potential roles as tumour-suppressor genes in primary peritoneal tumourigenesis. PMID- 18677305 TI - Biomimetic synthesis of the IDO inhibitors exiguamine A and B. AB - Biomimetic synthesis is an attempt to assemble natural products along biosynthetic lines without recourse to the full enzymatic machinery of nature. We exemplify this with a total synthesis of exiguamine A and the newly isolated natural product exiguamine B. The most noteworthy feature of this work is an oxidative endgame drawing from the complex chemistry of catecholamines, which allows for ready access to a new class of nanomolar indoleamine-2,3-dioxygenase inhibitors. PMID- 18677311 TI - Identification of loci associated with schizophrenia by genome-wide association and follow-up. AB - We carried out a genome-wide association study of schizophrenia (479 cases, 2,937 controls) and tested loci with P < 10(-5) in up to 16,726 additional subjects. Of 12 loci followed up, 3 had strong independent support (P < 5 x 10(-4)), and the overall pattern of replication was unlikely to occur by chance (P = 9 x 10(-8)). Meta-analysis provided strongest evidence for association around ZNF804A (P = 1.61 x 10(-7)) and this strengthened when the affected phenotype included bipolar disorder (P = 9.96 x 10(-9)). PMID- 18677313 TI - ADAMTSL2 mutations in geleophysic dysplasia demonstrate a role for ADAMTS-like proteins in TGF-beta bioavailability regulation. AB - Geleophysic dysplasia is an autosomal recessive disorder characterized by short stature, brachydactyly, thick skin and cardiac valvular anomalies often responsible for an early death. Studying six geleophysic dysplasia families, we first mapped the underlying gene to chromosome 9q34.2 and identified five distinct nonsense and missense mutations in ADAMTSL2 (a disintegrin and metalloproteinase with thrombospondin repeats-like 2), which encodes a secreted glycoprotein of unknown function. Functional studies in HEK293 cells showed that ADAMTSL2 mutations lead to reduced secretion of the mutated proteins, possibly owing to the misfolding of ADAMTSL2. A yeast two-hybrid screen showed that ADAMTSL2 interacts with latent TGF-beta-binding protein 1. In addition, we observed a significant increase in total and active TGF-beta in the culture medium as well as nuclear localization of phosphorylated SMAD2 in fibroblasts from individuals with geleophysic dysplasia. These data suggest that ADAMTSL2 mutations may lead to a dysregulation of TGF-beta signaling and may be the underlying mechanism of geleophysic dysplasia. PMID- 18677316 TI - Human microRNAs regulate stress-induced immune responses mediated by the receptor NKG2D. AB - MICA and MICB are stress-induced ligands recognized by the activating receptor NKG2D. A microRNA encoded by human cytomegalovirus downregulates MICB expression by targeting a specific site in the MICB 3' untranslated region. As this site is conserved among different MICB alleles and a similar site exists in the MICA 3' untranslated region, we speculated that these sites are targeted by cellular microRNAs. Here we identified microRNAs that bound to these MICA and MICB 3' untranslated region sequences and obtained data suggesting that these microRNAs maintain expression of MICA and MICB protein under a certain threshold and facilitate acute upregulation of MICA and MICB during cellular stress. These microRNAs were overexpressed in various tumors and we demonstrate here that they aided tumor avoidance of immune recognition. PMID- 18677318 TI - Direct fabrication of metavanadate phosphor films on organic substrates for white light-emitting devices. AB - White-light-emitting materials have attracted considerable attention because of their applications, such as large-surface emitting devices. Inorganic phosphor films are expected to be applied to these devices because of good chemical stability; however, a substantial reduction of fabrication temperature is required for future industrial uses such as lighting materials fabricated onto flexible organic substrates. Here we show the optical properties of white-light emitting metavanadate phosphors, AVO3 (A: K, Rb and Cs), and we report a new direct fabrication process for RbVO3 films onto flexible polyethylene terephthalate (PET) substrates by means of a vacuum ultraviolet irradiation using an excimer lamp. In addition, the (Ca,Sr,Pr)TiO3/a-Al2O3/RbVO3/PET heterostructure prepared by an excimer-laser-assisted metal-organic deposition process has demonstrated the possibility of colour modification for RbVO3 films on PET. Our findings suggest new possibilities for further development of large surface emitting lighting devices. PMID- 18677317 TI - The kinase p38 alpha serves cell type-specific inflammatory functions in skin injury and coordinates pro- and anti-inflammatory gene expression. AB - The mitogen-activated protein kinase p38 mediates cellular responses to injurious stress and immune signaling. Among the many p38 isoforms, p38 alpha is the most widely expressed in adult tissues and can be targeted by various pharmacological inhibitors. Here we investigated how p38 alpha activation is linked to cell type specific outputs in mouse models of cutaneous inflammation. We found that both myeloid and epithelial p38 elicit inflammatory responses, yet p38 alpha signaling in each cell type served distinct inflammatory functions and varied depending on the mode of skin irritation. In addition, myeloid p38 alpha limited acute inflammation via activation of anti-inflammatory gene expression dependent on mitogen- and stress-activated kinases. Our results suggest a dual function for p38 alpha in the regulation of inflammation and show mixed potential for its inhibition as a therapeutic strategy. PMID- 18677319 TI - Caenorhabditis elegans mutant allele identification by whole-genome sequencing. AB - Identification of the molecular lesion in Caenorhabditis elegans mutants isolated through forward genetic screens usually involves time-consuming genetic mapping. We used Illumina deep sequencing technology to sequence a complete, mutant C. elegans genome and thus pinpointed a single-nucleotide mutation in the genome that affects a neuronal cell fate decision. This constitutes a proof-of-principle for using whole-genome sequencing to analyze C. elegans mutants. PMID- 18677321 TI - eSGA: E. coli synthetic genetic array analysis. AB - Physical and functional interactions define the molecular organization of the cell. Genetic interactions, or epistasis, tend to occur between gene products involved in parallel pathways or interlinked biological processes. High throughput experimental systems to examine genetic interactions on a genome-wide scale have been devised for Saccharomyces cerevisiae, Schizosaccharomyces pombe, Caenorhabditis elegans and Drosophila melanogaster, but have not been reported previously for prokaryotes. Here we describe the development of a quantitative screening procedure for monitoring bacterial genetic interactions based on conjugation of Escherichia coli deletion or hypomorphic strains to create double mutants on a genome-wide scale. The patterns of synthetic sickness and synthetic lethality (aggravating genetic interactions) we observed for certain double mutant combinations provided information about functional relationships and redundancy between pathways and enabled us to group bacterial gene products into functional modules. PMID- 18677325 TI - Screening the hard to reach: improving morbidity and mortality from cervical cancer in New Zealand. PMID- 18677326 TI - Demographic predictors of cervical cancer screening in Chinese women in New Zealand. AB - OBJECTIVE: This pilot study examined the cervical cancer screening practices of Chinese women living in Auckland and the association with social demographic factors. METHODS: A community-based survey was conducted and 234 questionnaires were administrated to ascertain the uptake of cervical screening. Participants were asked whether they had ever been screened in New Zealand and whether it had occurred in the previous 3 years. RESULTS: One hundred and fifty-two (65.0%; 95% CI: 58.5-71.1) respondents reported having been screened in New Zealand and 56.0% (95% CI: 49.4-62.4) reported they were screened in the last 3 years. Factors independently associated with cervical cancer screening practice included age and duration of residence in New Zealand. The most frequently cited reason for never having had a smear test was that "thought it is unnecessary" (39%), followed by "don't know where to go" (36.6%). CONCLUSION: The uptake of cervical cancer screening is lower among women migrants from Mainland China living in New Zealand than that of the national New Zealand average. In addition, it is lower than that of Chinese women living in North America. The study highlights the information needs of new immigrants and older or younger women. PMID- 18677327 TI - Prospective 10-year study of postmenopausal women with asymptomatic primary hyperparathyroidism. AB - AIM: There are few prospective studies of people with asymptomatic primary hyperparathyroidism (PHPT) who have not had parathyroidectomy. We followed a group of postmenopausal women with asymptomatic PHPT for up to 10 years to determine whether they could be managed conservatively without parathyroidectomy. METHODS: A 10-year, prospective, longitudinal study of 23 postmenopausal women with asymptomatic PHPT initially enrolled into a 4-year randomised controlled trial of hormone replacement therapy. Serum total and ionised calcium, biochemistry, urine calcium, and bone mineral density were measured every 6-12 months. RESULTS: Serum ionised calcium, creatinine, and urine calcium:creatinine remained stable throughout follow-up. In contrast, there was a steady increase in the total and adjusted serum calcium and a small rise in serum PTH. Only one woman had an adjusted serum calcium >3.0 mmol/L during follow-up. There were few other clinical events possibly related to PHPT (1 possible episode of nephrolithiasis, 4 fractures, 1 severe osteoporosis). Three women underwent parathyroidectomy, although 19/23 women met the updated criteria for parathyroidectomy from the 2002 NIH-sponsored workshop during follow-up. CONCLUSIONS: Many postmenopausal women with asymptomatic PHPT do not develop symptoms or complications of PHPT, and their biochemical parameters remains stable. Therefore, such asymptomatic women with PHPT can often be managed conservatively without parathyroidectomy. PMID- 18677328 TI - The maternal outcome in placenta accreta: the significance of antenatal diagnosis and non-separation of placenta at delivery. AB - AIM: To evaluate the effects of antenatal diagnosis and subsequent placental non separation at delivery on the maternal outcome in confirmed cases of placenta accreta. METHOD: The perinatal database and medical records for women who delivered in the period 2000-6 in a teaching hospital in New Zealand with a diagnosis of placenta accreta or postpartum haemorrhage or hysterectomy were reviewed. In confirmed placenta accreta cases, the amount of blood loss and blood transfused at delivery and subsequent emergency hysterectomy were analysed in respect to the presence/absence of antenatal diagnosis and the management at delivery. RESULTS: 16 women had placenta accreta confirmed (15 histologically and 1 visually). Antenatal diagnosis was made in 7 women, elective Caesarean delivery planned in all, hysterectomy to follow in 5 (4 elective, 1 emergency preterm), and elective placental separation in 2 women. When an antenatal diagnosis was not made (n=9), attempted placental separation led to emergency hysterectomy for all (p=0.001). Antenatal diagnosis and placental non-separation resulted in less mean blood loss (1.4 L vs 3.6 L, p=0.003; 1.0 L vs 3.4 L, p<0.001) and mean units of blood transfused (1.2 vs 5.1, p=0.005) in the latter. CONCLUSION: In placenta accreta, antenatal diagnosis and avoidance of placental separation may result in better maternal outcome. PMID- 18677329 TI - Changes in cause of neonatal death over a decade. AB - AIMS: To classify neonatal deaths at Wellington Hospital (Wellington, New Zealand) over a 10-year period and assess changes in cause of death over time. METHODS: Retrospective audit from 1995-2004 of live-born infants > or = 20 weeks gestation dying before 28 days of age. Deaths were classified according to the PSANZ-NDC Classification guideline. The years 1995-1999 and 2000-2004 were compared to analyse for changes in cause of death. RESULTS: There were 219 neonatal deaths: 67(31%) of these were term infants and 154 preterm; 109 infants from 1995-1999 and 110 from 2000-2004. The autopsy rate was 62% and highest in term infants (76%). Deaths due to congenital anomaly and extreme prematurity decreased over time and deaths due to infection increased. CONCLUSIONS: Use of the PSANZ-NDC death classification system enables an accurate cause of death to be established for most neonatal deaths and allows monitoring of mortality rates over time. PMID- 18677330 TI - Unusual primary manifestations of multiple sclerosis. AB - OBJECTIVES: The aim of this study is to describe patients with unusual symptoms that were primary manifestations of multiple sclerosis (MS). PATIENTS AND METHODS: We report 21 multiple sclerosis patients who presented unusual initial pictures (acute brachial pain n=4, headache n=6, ptosis n=1, oculomotor nerve palsy n=1, peripheral facial palsy n=1, throat pain n=1, hypoglossal nerve palsy n=1, visual field defect n=2, epilepsia n=2, and coma n=2) as the first manifestations in the absence of other obvious symptoms or signs. RESULTS: Investigations demonstrated changes highly suggestive of multiple sclerosis on magnetic resonance imaging, cerebrospinal fluid analysis and electrophysiological tests. All cases completely or partially recovered after high-dose corticosteroid therapy. These patients have been followed up for 5 years. CONCLUSION: In this study, we discuss possible correlations between clinical disturbances and neuroradiological abnormalities and show some rare or previously undescribed manifestations in multiple sclerosis. PMID- 18677331 TI - Cystic lesions of the liver: 6 years of surgical management in New Zealand. AB - INTRODUCTION: Cysts are a common radiological finding in the liver. Many affected patients do no require treatment. However a minority require further investigation and treatment for symptoms or risk of underlying malignancy. METHODS: A computerised database of patients presenting to Auckland Hospital for the management of liver lesions was established in 2000. Details of demographics, presentation, investigations, management, and follow-up are entered prospectively. RESULTS: Forty-seven patients (36 female, median age 61, range 37 86 years) requiring surgical treatment of cystic liver lesions were identified from a total of over 800 patients enrolled in the database. Twenty-five patients presented with simple cysts, of whom 12 had radiological evidence of polycystic liver and kidney disease. All 25 patients were symptomatic and all were managed successfully with laparoscopic fenestration. Nine patients presented with complex cysts which were treated with liver resection. Of these patients, four had benign cysts, three had underlying biliary cystadenomas, and two had biliary cystadenocarcinomas. One patient with a biliary cystadenocarcinoma is dead of disease. The median follow-up for all 47 patients was 26 months. CONCLUSIONS: Cystic liver lesions can represent a spectrum of underlying conditions. All cysts require investigation and complex cysts or symptomatic simple cysts require further treatment. PMID- 18677333 TI - Accessory breast tissue presenting as a large pendulous mass in the axilla: a diagnostic dilemma. AB - Accessory breasts are an uncommon entity. They may present as asymptomatic masses or cause symptoms such as pain or restriction of arm movements. They may prove to be a diagnostic challenge if found in locations along or outside the mammary line. We report a case of an ectopic accessory breast presenting as a large pendulous mass in the axilla which proved to be a diagnostic dilemma. Excision biopsy was diagnostic. PMID- 18677332 TI - Risk and severity of injury in a population of BASE jumpers. AB - AIM: To determine the frequency and severity of accidents in a population of BASE jumpers (people who jump with parachute from a fixed object such as from a cliff). METHODS: To determine the frequency and characteristics of BASE jumping accidents, a cross-sectional survey of experienced BASE jumpers was completed. RESULTS: 35 BASE jumpers enrolled in the study. Findings revealed that there were approximately 9914 jumps made and the estimated rate of injury was 0.4%. 39 accidents involving 21 (60%) BASE jumpers were found. 28 accidents (72%) predominantly involved the lower limbs, 12 (31%) involved the back/spine, 7 (18%) the upper limbs, and 1 (3%) was a head injury. CONCLUSION: BASE jumping is associated with a high risk of serious injury and appears to be significantly more dangerous than skydiving. PMID- 18677334 TI - Aorto-enteric fistula: recurrence of a fistula to the sigmoid stump following Hartmann's procedure. PMID- 18677335 TI - Hyaline vascular-type Castleman's disease of the retroperitoneum. PMID- 18677336 TI - Medical image. Genital lupus. PMID- 18677337 TI - Medical image. A sweaty lady with large digits. Acromegaly with pituitary macroadenoma. PMID- 18677338 TI - Pharmaceutical industry spending. PMID- 18677339 TI - The frequency of medicines advertising on New Zealand television. PMID- 18677340 TI - Psychotropic medicines in residential care facilities. PMID- 18677341 TI - Screening of Cushing's syndrome in patients with Type 2 diabetes. PMID- 18677342 TI - Deaths caused by diabetes mellitus. PMID- 18677343 TI - Ciprofloxacin-induced seizures in a healthy patient. PMID- 18677344 TI - Having more knowledge of cervical cancer does not increase uptake of screening in Chinese women: findings from a New Zealand study. PMID- 18677345 TI - On the generalized Wertheim law in integrated photoelasticity. AB - Two-dimensional photoelasticity is based on the classical Wertheim law. The integral Wertheim law can be used in integrated photoelasticity only in the case when the directions of the secondary principal stresses are constant on the light beam. We generalize the integral Wertheim law for the case when a slight rotation of the secondary principal directions takes place. PMID- 18677346 TI - Modified diffraction theory of Kirchhoff. AB - The diffraction theory of Kirchhoff is reinterpreted and a new form of a surface diffraction integral is developed by using the axioms of the modified theory of physical optics, which leads to the exact scattered fields by conducting bodies. The new integral is arranged according to the interpretation of Young, and the diffracted waves are expressed in terms of a line integral. The method is applied to the diffraction problem by a semi-infinite edge contour. PMID- 18677347 TI - Appropriate interface termination to improve imaging resolution of multilayered structures in the infrared and optical canalization regime. AB - Appropriate surface termination is used to improve dramatically the subwavelength imaging resolution of a multilayered positive-negative permittivity structure operating in the infrared or optical canalization regime. The imaging resolution of the improved multilayered structure resists practical material loss well, and it is not sensitive to the thickness of the interface layers, the total thickness, nor the period of the multilayered structure. Such a structure can be used to transfer a subwavelength image to a far distance through a thick structure. PMID- 18677348 TI - Application of Mie theory to assess structure of spheroidal scattering in backscattering geometries. AB - Inverse light scattering analysis seeks to associate measured scattering properties with the most probable theoretical scattering distribution. Although Mie theory is a spherical scattering model, it has been used successfully for discerning the geometry of spheroidal scatterers. The goal of this study was an in-depth evaluation of the consequences of analyzing the structure of spheroidal geometries, which are relevant to cell and tissue studies in biology, by employing Mie-theory-based inverse light scattering analysis. As a basis for this study, the scattering from spheroidal geometries was modeled using T-matrix theory and used as test data. In a previous study, we used this technique to investigate the case of spheroidal scatterers aligned with the optical axis. In the present study, we look at a broader scope which includes the effects of aspect ratio, orientation, refractive index, and incident light polarization. Over this wide range of parameters, our results indicate that this method provides a good estimate of spheroidal structure. PMID- 18677349 TI - Unitary transformation between Cartesian- and polar-pixellated screens. AB - A unitary transformation between Cartesian and polar pixellations of finite two dimensional images is obtained from the su(2) model for discrete and finite signals. This transformation analyzes the original image into its finite Cartesian "Laguerre-Kravchuk" modes (involving Wigner little-d functions) and synthesizes it back using a polar mode basis with the same set of mode coefficients. The polar basis is derived from the quantum angular momentum theory, and its modes are given by Clebsch-Gordan coefficients. PMID- 18677350 TI - Analysis of the influence of the passive facet of blazed transmission gratings in the intermediate diffraction regime. AB - Blazed diffraction gratings are of enormous practical importance for imaging and analyzing hybrid optical systems. The intermediate diffraction regime is characterized by the transition from the scalar to the rigorous electromagnetic theory. An effect known as shadowing occurs and reduces the diffraction efficiency. Based on rigorous calculations for optimized sawtooth-shaped and binary-multilevel blaze profiles, we deduce a semianalytical model describing the shadowing phenomenon for the general case of oblique incidence. We discuss illumination both from air and from the substrate. Though a multilevel blaze possesses a discrete substructure, our shadowing model remains valid, if substructural effects are neglected. We find that electromagnetic effects due to the passive blaze facet lead to the efficiency reduction, and the blazing efficiency shows a linear dependence on the ratio of blaze wavelength to grating period. Our shadowing model is applied to predict the performance of a Littrow like blazing condition in transmission geometry as, e.g., for a diffractive solid immersion lens. PMID- 18677351 TI - Holographic features of spatial coherence wavelets. AB - The behavior of the marginal power spectrum as a two-channel-multiplexed hologram is analyzed. Its "negative energies" make it quite different from the conventional holograms, i.e., it is not recordable in general and the objects to be reconstructed (the cross-spectral densities at both the aperture and the observation planes) are virtual. The holographic reconstruction results from the superposition of the spatial coherence wavelets that carry the marginal power spectrum. These features make the marginal power spectrum a powerful tool for analysis and synthesis of optical fields, for instance, in optical information processing (signal encryption) and beam shaping for microlithography. PMID- 18677352 TI - Equivalence between optimum Young's fringe visibility and position-independent stochastic behavior of electromagnetic fields. AB - Stochastic electromagnetic fields characterized by optimized fringe visibility in a Young interferometric arrangement are shown to be those whose random character is position independent. The optimization procedure involves local unitary transformations, which can be implemented by using reversible anisotropic polarization devices placed at the two pinholes. It is also shown that the local degree of polarization in the optimized interferometer is constant across the superposition region and coincides with the degree of polarization at the two pinholes. PMID- 18677353 TI - Reconstruction of periodic structures from optical scattering measurements using adjoint equations. AB - A new numerical approach to efficiently reconstruct the profile of a grating from measurements of reflection coefficients is demonstrated. The problem is posed in the mathematical framework of an inverse scattering problem and solved using gradient-based algorithms. The gradient is computed efficiently using adjoint equations, which amounts to an extra scattering computation per iteration. For symmetric profiles it is shown that only knowledge of the scattered field is sufficient to compute the gradient. As a result, complex profiles can be reconstructed rapidly, and the method can be potentially used in metrology applications in semiconductors. The technique is demonstrated for the case of TE polarization. PMID- 18677354 TI - Simulation of subwavelength metallic gratings using a new implementation of the recursive convolution finite-difference time-domain algorithm. AB - We introduce a new implementation of the finite-difference time-domain (FDTD) algorithm with recursive convolution (RC) for first-order Drude metals. We implemented RC for both Maxwell's equations for light polarized in the plane of incidence (TM mode) and the wave equation for light polarized normal to the plane of incidence (TE mode). We computed the Drude parameters at each wavelength using the measured value of the dielectric constant as a function of the spatial and temporal discretization to ensure both the accuracy of the material model and algorithm stability. For the TE mode, where Maxwell's equations reduce to the wave equation (even in a region of nonuniform permittivity) we introduced a wave equation formulation of RC-FDTD. This greatly reduces the computational cost. We used our methods to compute the diffraction characteristics of metallic gratings in the visible wavelength band and compared our results with frequency-domain calculations. PMID- 18677355 TI - Numerical simulations of global approach for photon scanning tunneling microscopy: coupling of finite-element and boundary integral methods. AB - An accurate global model is proposed for a two-dimensional probe-sample system of photon scanning tunneling microscopy in near-field optics. A coupling of a finite element method in the inhomogeneous sample and a boundary integral method on the artificial boundary of the truncated domain is developed. Numerical experiments are included to demonstrate the effectiveness of the proposed method and to show the features of wave propagation in photon scanning tunneling microscopy. PMID- 18677356 TI - Why a harmonic solution for lossless, perfectly homogeneous, left-handed material cannot exist. AB - In a preceding paper [J. Opt. Soc. Am. A21, 122 (2004)], we proposed proof of the nonexistence of harmonic solutions for a perfectly homogeneous left-handed material with both relative permittivity and relative permeability equal to -1 using the theorem of analytic continuation of an analytic function. The use of this theorem of analyticity has been questioned in a recent paper [Phys. Rev. E73, 046608 (2006)], arguing the possible inadequacy of the conditions of application of the theorem. We avoid the use of the analyticity theorem and propose a direct and simple proof of the nonexistence of such solutions. Furthermore, this proof is extended to any left-handed material with negative permeability and permittivity. PMID- 18677357 TI - Generalized Stokes parameters of a stochastic electromagnetic beam propagating through a paraxial ABCD optical system. AB - On the basis of the generalized diffraction integral formula for an ABCD optical system in the spatial domain, a propagation law for the generalized Stokes parameters of a stochastic electromagnetic beam passing through an ABCD optical system is obtained. We describe the Stokes parameters of the source as linear combinations of the elements of the cross-spectral density matrix, and study the changes in the spectral degree of polarization and in the state of the polarization ellipse of a stochastic electromagnetic Gaussian Schell-model beam propagating through a gradient-index fiber with the help of generalized Stokes parameters and the cross-spectral density matrix. The medium has significant effect on the change of the spectral degree of polarization. However, when the correlation coefficients of the source satisfy the relation delta(xx)=delta(yy)=delta(xy)=delta(yx), the medium does not influence the spectral degree of polarization. PMID- 18677358 TI - Laser beam projection with adaptive array of fiber collimators. I. Basic considerations for analysis. AB - We present a mathematical model and provide an analysis of optical beam director systems composed of adaptive arrays of fiber collimators (subapertures), referred to here as conformal optical systems. Performances of the following two system architectures are compared: A conformal-beam director with mutually incoherent output laser beams transmitted through fiber collimators (beamlets), and a corresponding coherent system whose beamlets can be coherently combined (phase locked) at a remote target plane. The effect of the major characteristics of the conformal systems on the efficiency of laser beam projection is evaluated both analytically and through numerical simulations. The characteristics considered here are the number of fiber collimators and the subaperture and conformal aperture fill factors, as well as the accuracy of beamlet pointing. PMID- 18677359 TI - Laser beam projection with adaptive array of fiber collimators. II. Analysis of atmospheric compensation efficiency. AB - We analyze the potential efficiency of laser beam projection onto a remote object in atmosphere with incoherent and coherent phase-locked conformal-beam director systems composed of an adaptive array of fiber collimators. Adaptive optics compensation of turbulence-induced phase aberrations in these systems is performed at each fiber collimator. Our analysis is based on a derived expression for the atmospheric-averaged value of the mean square residual phase error as well as direct numerical simulations. Operation of both conformal-beam projection systems is compared for various adaptive system configurations characterized by the number of fiber collimators, the adaptive compensation resolution, and atmospheric turbulence conditions. PMID- 18677360 TI - Curvature-based perceptual singularities and texture saliency with early vision mechanisms. AB - Recent work has shown that salient perceptual singularities occur in visual textures even in the absence of feature gradients. In smoothly varying orientation-defined textures, these striking non-smooth percepts can be predicted from two texture curvatures, one tangential and one normal [Proc. Natl. Acad. Sci. USA103, 15704 (2006)]. We address the issue of detecting these perceptual singularities in a biologically plausible manner and present three different models to compute the tangential and normal curvatures using early cortical mechanisms. The first model relies on the response summation of similarly scaled even-symmetric simple cells at different positions by utilizing intercolumnar interactions in the primary visual cortex (V1). The second model is based on intracolumnar interactions in a two-layer mechanism of simple cells having the same orientation tuning but significantly different scales. Our third model uses a three-layer circuit in which both even-symmetric and odd-symmetric receptive fields (RFs) are used to compute all possible directional derivatives of the dominant orientation, from which the tangential and normal curvatures at each spatial position are selected using nonlinear shunting inhibition. We show experimental results of all three models, we outline an extension to oriented textures with multiple dominant orientations at each point, and we discuss how our results may be relevant to the processing of general textures. PMID- 18677361 TI - Evolution of the polarization state in material media with uniform linear birefringence and an applied nonhomogeneous external magnetic field: application to bulk-type Faraday current sensors. AB - In this work we find the exact solution for the evolution of the polarization state of a light wave that propagates in a material medium with uniform linear birefringence and a nonuniform external magnetic field. The obtained results could be used to improve the precision of existing Faraday current sensors. PMID- 18677362 TI - Experimental observation of truncated fractional Fourier transform for a partially coherent Gaussian Schell-model beam. AB - The truncated fractional Fourier transform (FRT) is applied to a partially coherent Gaussian Schell-model (GSM) beam. The analytical propagation formula for a partially coherent GSM beam propagating through a truncated FRT optical system is derived by using a tensor method. Furthermore, we report the experimental observation of the truncated FRT for a partially coherent GSM beam. The experimental results are consistent with the theoretical results. Our results show that initial source coherence, fractional order, and aperture width (i.e., truncation parameter) have strong influences on the intensity and coherence properties of the partially coherent beam in the FRT plane. When the aperture width is large, both the intensity and the spectral degree of coherence in the FRT plane are of Gaussian distribution. As the aperture width decreases, the diffraction pattern gradually appears in the FRT plane, and the spectral degree of coherence becomes of non-Gaussian distribution. As the coherence of the initial GSM beam decreases, the diffraction pattern for the case of small aperture widths gradually disappears. PMID- 18677363 TI - Single-scatter vector-wave scattering from surfaces with infinite slopes using the Kirchhoff approximation. AB - This paper presents a new formulation of the 3D Kirchhoff approximation that allows calculation of the scattering of vector waves from 2D rough surfaces containing structures with infinite slopes. This type of surface has applications, for example, in remote sensing and in testing or imaging of printed circuits. Some preliminary calculations for rectangular-shaped grooves in a plane are presented for the 2D surface method and are compared with the equivalent 1D surface calculations for the Kirchhoff and integral equation methods. Good agreement is found between the methods. PMID- 18677364 TI - Signal-subspace method approach to the intensity-only electromagnetic inverse scattering problem. AB - This paper investigates the signal-subspace method approach to solve the electromagnetic inverse scattering problem using intensity-only (phase-free) data. Due to the polarization of electromagnetic fields, the relationship between the rank of the multistatic matrix and the number of small scatterers is different from that associated with the scalar wave. Multiple scattering between scatterers is considered, and the inverse scattering problem of determining the polarization tensors is nonlinear, which, however, is solved by the proposed analytical approach where no associated forward problem is iteratively evaluated. PMID- 18677366 TI - Diffraction by a circular aperture: an application of the vectorial theory of Huygens's principle in the near field. AB - A vectorial theory that brings new insight into the nature of diffraction is used to obtain mathematical expressions that evaluate diffraction patterns in the near field. The equations allow us to discriminate between the contributions of the vectorial and the scalar approaches. In the near field we studied the pattern of light diffracted through a circular aperture, and it was proved that the vectorial approach is significant in a region very near the circular aperture. In spite of the obvious differences between the circular aperture and other obstacles, the present theory may also be used with other geometries. PMID- 18677365 TI - Efficiency of capturing a phase image using cone-beam x-ray Talbot interferometry. AB - We assesses the efficiency of x-ray Talbot interferometry (XTI), a technique based on the Talbot effect for measuring a wavefront gradient, in terms of how quickly it can capture a high-quality phase image with a large signal-to-noise ratio for a given incident photon number. Photon statistics cause errors in the phase of the moire fringes and impose a detection limit on the wavefront gradient. The relation between the incident photon number and the detection limit is determined, and a figure of merit of XTI for a monochromatic cone beam is then defined. The dependence of the figure of merit on optical system parameters, such as grating pitch and position, is then discussed. The effects of varying the pattern height and linewidth of the second grating are shown for rectangular and trapezoidal teeth. Finally, we show how to design a practical cone-beam Talbot interferometer for certain boundary conditions. PMID- 18677367 TI - Cephalopod coloration model. II. Multiple layer skin effects. AB - A mathematical model of multiple layer skin coloration in cephalopods, a class of aquatic animals, is presented. The model incorporates diffuse and specular reflection from both pigment and structural photonic components found in the skin of these animals. Specific physical processes of this coloration are identified and modeled utilizing available biological materials data. Several examples of combination spectra are calculated to illustrate multiple layer and incident light effects as well as the potentially rich repertoire of color schemes available to these animals. A detailed understanding of the physical principles underlying cephalopod coloration is expected to yield insights into their possible functions. PMID- 18677368 TI - Predicting range performance of sampled imagers by treating aliased signal as target-dependent noise. AB - This paper presents a new theory to predict the impact of sampling on target acquisition. The aliased signal that results from sampling is treated as noise. The aliased signal is different from detector noise in two ways. First, aliasing disappears as the target contrast decreases. Second, the image corruption due to aliasing gets worse with increased range. This is because sampling is constant in angle space, and targets become poorly sampled as range increases. The theory is presented, along with the results of three experiments. The match between model and experiment is excellent. PMID- 18677369 TI - Spectral prediction model for piles of nonscattering sheets. AB - The present paper investigates the reflection and transmission properties of piles of nonscattering sheets. Using a spectral prediction model, we perform a detailed analysis of the spectral and color variations induced by variations of the number of superposed sheets, the absorbance of the sheet material, the refractive index of the medium between the sheets, and the reflectance of the background. The spectral prediction model accounts for the multiple reflections and transmissions of light between the interfaces bounding the layers. We describe in detail the procedure for deducing model parameters from measured data. Tests performed with nonscattering plastic sheets demonstrate the excellent accuracy of the predictions. A large set of predicted spectra illustrate the different evolutions of reflected and transmitted spectra as well as the corresponding colors for various types of piles. PMID- 18677370 TI - Use of a customized vision model to analyze the effects of higher-order ocular aberrations and neural filtering on contrast threshold performance. AB - Customized optical filtering and light-dependent neural filtering were implemented in an ideal-observer model for an L-alternative forced-choice visual task. The model was applied to a contrast threshold visual task with adaptive optics correction of ocular higher-order (HO) aberrations under different light regimes, for which experimental data have previously been obtained (J. Mod. Opt.55, 791, 2008). A separability measure was used to assess the model-observer performance and to investigate the joint effect of optical and neural filtering. The numerical results were consistent with the experimental data in the assessment of the effect of HO aberrations as a function of light level. PMID- 18677372 TI - Superresolution along extended depth of focus with binary-phase filters for the Gaussian beam. AB - In the paraxial Debye regime, simple and power-efficient pupil filters are designed to break the diffraction limit along a large depth of focus (DOF) for the Gaussian beam. Dependences of the superresolution factor, DOF gain, Strehl ratio, sidelobe strength, and axial intensity nonuniformity on the Gaussian profile in the pupil plane are characterized using the numerical method. Optimal filter designs are proposed for either high-resolution or ultra-large-DOF applications followed by experimental verifications. PMID- 18677371 TI - Can laser speckle flowmetry be made a quantitative tool? AB - The ultimate objective of laser speckle flowmetry (and a host of specific implementations such as laser speckle contrast analysis, LASCA or LSCA; laser speckle spatial contrast analysis, LSSCA; laser speckle temporal contrast analysis, LSTCA; etc.) is to infer flow velocity from the observed speckle contrast. Despite numerous demonstrations over the past 25 years of such a qualitative relationship, no convincing quantitative relationship has been proven. One reason is a persistent mathematical error that has been propagated by a host of workers; another is a misconception about the proper autocorrelation function for ordered flow. Still another hindrance has been uncertainty in the specific relationship between decorrelation time and local flow velocity. Herein we attempt to dispel some of these errors and misconceptions with the intent of turning laser speckle flowmetry into a quantitative tool. Specifically we review the underlying theory, explore the impact of various analytic models for relating measured intensity fluctuations to scatterer motion, and address some of the practical issues associated with the measurement and subsequent data processing. PMID- 18677373 TI - Robust determination of the anisotropic polarizability of nanoparticles using coherent confocal microscopy. AB - A coherent confocal microscope is proposed as a means to fully characterize the elastic scattering properties of a nanoparticle as a function of wavelength. Using a high numerical aperture lens, two-dimensional scanning, and a simple vector-beam shaper, the rank-2 polarizability tensor is estimated from a single confocal image. A method for computationally efficient data processing is described, and numerical simulations show that this algorithm is robust to noise and uncertainty in the focal plane position. The proposed method is a generalization of techniques that provide an estimate of a limited set of scattering parameters, such as a single orientation angle for rodlike particles. The measurement of the polarizability obviates the need for a priori assumptions about the nanoparticle. PMID- 18677374 TI - Multiconjugate adaptive optics results from the laboratory for adaptive optics MCAO/MOAO testbed. AB - We report on the development of wavefront reconstruction and control algorithms for multiconjugate adaptive optics (MCAO) and the results of testing them in the laboratory under conditions that simulate an 8 meter class telescope. The University of California Observatories (UCO) Lick Observatory Laboratory for Adaptive Optics multiconjugate testbed allows us to test wide-field-of-view adaptive optics systems as they might be instantiated in the near future on giant telescopes. In particular, we have been investigating the performance of MCAO using five laser beacons for wavefront sensing and a minimum-variance algorithm for control of two conjugate deformable mirrors. We have demonstrated improved Strehl ratio and enlarged field-of-view performance when compared to conventional AO techniques. We have demonstrated improved MCAO performance with the implementation of a routine that minimizes the generalized isoplanatism when turbulent layers do not correspond to deformable mirror conjugate altitudes. Finally, we have demonstrated suitability of the system for closed loop operation when configured to feed back conditional mean estimates of wavefront residuals rather than the directly measured residuals. This technique has recently been referred to as the "pseudo-open-loop" control law in the literature. PMID- 18677375 TI - Effect of dispersion on the spectrum of partially coherent beams. AB - Taking the Gaussian Schell-model (GSM) beam as a typical example of partially coherent beams, the analytical expressions of the spectrum of GSM beams propagating in dispersive media are derived, and the spectral properties are studied in detail. It is shown that, in comparison with propagation in free space and in turbulence, whether or not GSM beams satisfy the scaling law, the normalized spectrum of GSM beams in dispersive media changes on propagation in general, because the dispersive medium affects different spectral components differently. As compared with the free-space propagation, for the scaling-law GSM beams the dispersion results in spectrum change, and for the nonscaling-law GSM beams the dispersion gives rise to a further increase in spectral changes. The structure constant of the dispersive property of the media, the transverse coordinate of the observation point, the spatial correlation length of the source, and the propagation distance affect the spectral behavior of GSM beams; this effect is illustrated numerically. PMID- 18677376 TI - Multiple-viewpoint projection holograms synthesized by spatially incoherent correlation with broadband functions. AB - We present a theoretical framework for recording and reconstructing incoherent correlation holograms of real-existing three-dimensional scenes observed from multiple viewpoints. This framework is demonstrated by generating and reconstructing a modified Fresnel hologram as well as a new correlation hologram called a protected correlation hologram. The reconstructed scene obtained from the protected correlation hologram has a significantly improved transverse resolution for the far objects in the scene compared to the modified Fresnel hologram. Additionally, the three-dimensional information encoded into the protected correlation hologram is scrambled by a secretive point spread function and thus the hologram can be used for encrypting the observed scene. The proposed holography methods are demonstrated by both simulations and experiments. PMID- 18677377 TI - 4x4 ray matrix for a curved chirped grating at oblique incidence. AB - The general 4x4 ray matrix of a grating is presented. This matrix suits the cases of transmission and reflection by a chirped grating with curved lines lying on a curved interface. The matrix presented applies to the general oblique incidence of the optical axis and therefore to nonorthogonal cases and to conical diffraction. PMID- 18677378 TI - [Nucleolar localization of human acetyltransferase-like protein and its expression in tumors]. AB - OBJECTIVE: To confirm the nucleolar localization of telomerase-regulation associated protein-human N-acetyltransferase-like protein (hALP) and its associated functions. METHODS: Immunofluoresent staining and immunoelectron microscopy were used to detect the distribution of hALP in HeLa and Saos2 cells, and the co-localization of hALP and rDNA was analyzed by fluorescence in situ hybridization and immunofluoresence. RNAi was performed to further verify the nucleolar localization of hALP. A series of eukaryotic expression plasmids carrying various portions of hALP sequence were constructed and transiently transfected to HeLa and Saos2 cells. The expression of hALP in tumor tissues was stained by immunohistochemistry. RESULTS: hALP distributed predominantly in the nucleoli of HeLa and Saos2 cells, and colocalized with rDNA. Granular component was the precise distribution of hALP in the nucleolus under electron microscope. Nucleolar signals for hALP reduced significantly in cells transfected with hALP siRNA. The carboxy terminus of hALP including residues 549-834 was necessary for its nucleolar localization. hALP could be detected in the nucleoli of many kinds of tumor cells, including leiomyosarcoma, primitive neuroectodermal tumor, neuroblastoma, melanoma, prostatic cancer, and clear cell renal carcinoma. CONCLUSION: hALP is a nucleolar protein, and the nucleolar localization is mediated by its carboxy terminal domain, and hALP could be detected in the nucleoli of many tumor tissues, which is worthy of further investigation. PMID- 18677379 TI - [Clinicopathologic, immunohistochemical and molecular analysis in 15 cases of angioimmunoblastic T-cell lymphomas]. AB - OBJECTIVE: To evaluate angioimmunoblastic T-cell lymphoma(AITL) completely, we gave injdepth investigation of histopathological features, specific immunochemical markers, antigen receptor gene rearrangements and in situ hybridization for Epstein-Barr virus (EBV). METHODS: 15 cases of typical AITL displayed effacement of the normal lymph node architecture partially or completely, abundance of arborizing high endothelial vessels, infiltration of polymorphic cells and hyperplastic atypical T lymphocytes with or without clear cytoplasm. Clinical characteristics, histological manifestations, and immunohistochemical staining for CD3, CD20, CD4iCD21, CXCL13, CD10, and BCL6 were analyzed. Polymerase chain reaction for immunoglobulin heavy chain (IgH) and T cell receptor gamma (TCRgamma) rearrangements and in situ hybridization for Epstein-Barr virus encoded RNA (EBER-1) were performed. RESULTS: Histologically, we found eight cases with regressed lymphoid follicles, six with absence of follicles and one with hyperplastic follicles with interfollicular lesions. We also found eight cases displaying aggregation of clear cells, four infiltration of large lymphoid cells, five abundant epithelioid histiocytes. CD20 staining showed hyperplasia of large B cells in four cases. CD21 expression exihibited extrafollicular expansion of follicular dendritic cell meshworks in 11 cases (73.3%), partially with a tendency of perivascular distribution. Positive rate for CXCL13 and CD10 are 73.3% and 6.7% respectively. Monoclonal rearrangements of TCRgamma were detected in 6/15 (40%) of cases, IgH rearrangements in 7/15 (46.7%), of which five were monoclonal, while two oligoclonal. 8 out of 15 cases (53h3%) contained EBV-positive cells. Among the four cases with large B cell proliferation, three were EBV-positive. CONCLUSION: AITL display great complexity and diversity clinicopathologically. Only when we recognize such diversity, can we reasonably apply and properly evaluate immunochemical markers and molecular techniques, and thus give a correct diagnosis. PMID- 18677380 TI - [Application of the in situ hybridization with EWS dual-color break-apart fluorescence probe and anti-CD99 and anti-FLI-1 antibodies in the diagnosis of Ewing's sarcoma/primitive neuroectodermal tumor]. AB - OBJECTIVE: To investigate the value of using EWS dual-color, break-apart rearrangement probe fluorescence in situ hybridization (FISH) technique, CD99 and FLI-1 antibodies immunohistochemistry in the diagnosis of Ewing's sarcoma/primitive neuroectodermal tumor (EWS/PNET). METHODS: Thirty-five cases of EWS/PNET and 24 cases of non-EWS/PNET small round cell tumor were analyzed by FISH and immunohistochemically detected with FLI-1 and CD99 antibodies. Comparison between FISH and immunohistochemical results was carried out and their diagnostic value was evaluated. RESULTS: The sensitivity, specificity, positive predictive value and negative predictive value of FISH in EWS/PNET were 93.8%(30/32), 81.8%(18/22), 88.2%(30/34) and 90%(18/20); those of CD99 were 100%(35/35), 58.3% (14/24), 77.8%(35/45) and 100%(14/14); those of FLI-1 were 71.4%(25/35), 62.5%(15/24), 73.5%(25/34), 60%(15/25), respectively. The sensitivity, specificity, positive predictive value and negative predictive value of combined use of CD99 and FLI-1 were 71.4%(25/35), 75%(18/24), 80.6%(25/31) and 64.3%(18/28), those of combined use of FLI-1 and FISH were 68.7%(22/32), 86.4%(19/22), 88%(22/25) and 65.5%(19/29), those of combined use of CD99 and FISH were 93.7%(30/32), 95.5%(21/22), 96.8%(30/31) and 91.3%(21/23), respectively. CONCLUSION: EWS dual-color, break-apart rearrangement probe FISH is a highly sensitive and specific technique in the diagnosis of EWS/PNET. The combination of CD99 and FISH is the method of choice for the diagnosis of EWS/PNET. The combination of CD99 and FLI-1 can improve the specificity in EWS/PNET without the data of FISH. PMID- 18677381 TI - [Correlation of telomere length and the expression of its regulating proteins in mesenchymal sarcomas]. AB - OBJECTIVE: To explore the significance in the change of telomere length in mesenchymal sarcomas, through analyzing telomere length and expression of its associated proteins, including TRF1, POT1, hTERT, P53 and c-myc. METHODS: The telomere length in 20 cases of osteosarcomas, 25 of chondrosarcomas, 19 of rhabdomyosarcomas, 26 of liposarcomas was measured by telomere fluorescence in situ hybridization (Telo-FISH), and the expression of TRF1, POT1, hTERT, p53 or c myc was analyzed by immunohistochemistry, respectively. RESULTS: The telomere length in osteosarcomas was significantly shorter than that of either chondrosarcomas or liposarcomas (P<0.05). Similarly, the telomere length of rhabdomyosarcoma was shorter than that of chondrosarcoma (P<0.05). Meanwhile, telomere shortening was positively correlated with down expression of telomere binding proteins TRF1 and POT1 (P<0.05), but trends were detected more frequently in positive expression of hTERT (P<0.05) and in nuclear accumulation of P53 or expression of c-myc. With advancing in histological grading, telomere length was shortened markedly in chondrosarcomas, especially in liposarcomas (P<0.05). CONCLUSION: The shortening of telomere could prevail in mesenchymal sarcoma and reflect the malignant potential. Telomere attrition usually correlated with down expression of POT1, TRF1 and with increased levels of hTERT, P53 and c-myc. PMID- 18677382 TI - [Correlation of TFRC polymorphism with the susceptibility and clinicopathologic phenotypes of IgA nephropathy]. AB - OBJECTIVE: To explore the association of its polymorphism of TFRC with the susceptibility, clinical and pathologic phenotypes of IgA nephropathy. METHODS: A total of 380 patients with IgA nephropathy and 250 normal controls were enrolled in the study. The regions with 424G/A and -5184C/T polymorphism sites of TFRC were amplified by PCR from genomic DNA and then the PCR-RFLP were performed by restriction enzymes, BanI and BsmA I, respectively. The genetic association of genotypes with the clinical and pathologic phenotypes was analyzed. RESULTS: The distribution of frequency in TFRC was consistent with Hardy-Weinberg equilibrium; however, we found no significant difference in genotypes distribution between patients and controls. There were no differences between genotypes in age, blood pressure, 24 h urine protein excretion, serum creatinine, creatinine clearance and serum IgA. 424G/A and -5184C/T polymorphisms were associated with immunofluorescent intensity of IgA deposit in mesangial area, though there was no difference in pathological lesions evaluated by HAAS grade. CONCLUSION: The polymorphisms of TFRC in 424G/A and -5184C/T sites were not associated with susceptibility to IgA nephropathy, but associated with density of immunofluorescence of IgA in mesangium in our large population based Chinese patients. The association of IgA nephropathy and other polymorphism sites, as well as interaction between TFRC polymorphism and other geneso polymorphisms, neededs to be further investigated. PMID- 18677383 TI - [Reassessment of the pathological diagnosis in 33 cases of malignant fibrous histiocytoma]. AB - OBJECTIVE: Since malignant fibrous histiocytoma (MFH) may be taken as an undifferentiated pleomorphic sarcoma (UPS), this study was conducted to reassess 33 previously diagnosed MFH cases in the past 10 years based on the latest WHO concept. And then to search for the clinicopathological features, probably tumorigenesis, and the line of differentiation of the remaining MFH/UPS cases. METHODS: Thirty-three cases in tissue microarray were studied by immunohistochemistry with panels of neurogenic, myogenic, and lipogenic antibodies. Three expertise pathologists reevaluated the slides separately. RESULTS: Among the 33 cases, 17 cases (51.5%) of MFH had their diagnoses changed, including 5 leiomyosarcomas, 3 malignant peripheral nerve sheath tumors, 1 fibrosarcoma, 1 inflammatory myofibrosarcoma, 1 giant cell tumor and 1 angiomatoid fibrous histiocytoma. The remaining 16 cases (48.5%) were finally diagnosed as MFH/UPS, among which patients were mainly old adults (median age: 63 years; range: 38 to 76 years). The median tumor size was 6.0 cm (range: 3.0 to 14.0 cm), 8 cases (50%) located in lower limb and 5 cases (31.3%) located in thigh. These tumors had marked cytological and nuclear pleomorphism. Immunohistochemistry showed that Vimentin was strongly positive in all 16 MFH/UPS (100%), Muscle-specific actin was variously positive in 8 cases (50%) and 1 case focally expressed Desmin. Eleven cases (68.8%) variously expressed CD68 (KP1) and 7 cases (43.8%) expressed CD68 (PG-M1), which were much higher than leiomyosarcoma, malignant peripheral nerve sheath tumor and liposarcoma with significant difference. Moreover, Ki67 expression rates were from 10% to 100%, including 14 cases more than 50% and 11 cases more than 70%. However, only 2 cases (12.5%) showed P53 positive. CONCLUSION: MFH/UPS often show marked histological pleomorphism, and the diagnosis must be made by exclusion of other definitive sarcomas, especially myogenic and neurogenic sarcoma. Only Vimentin was always expressed in MFH/UPS, while some of the tumors were positive for myogenic antigen and CD68. It was suggested that MFH/UPS might arise from primary mesenchymal cells, and some cases exhibited fibroblastic and/or myofibroblastic features. In addition, histiocytic phenotypic marker did have more expression in MFH/UPS than in other sarcomas. MFH/UPS still had certain clinicopathological characteristics. PMID- 18677385 TI - [Clinicopathologic study of hepatosplenic T-cell lymphoma]. AB - OBJECTIVE: To explore the clinicopathologic features and diagnostic criteria of hepatosplenic T-cell lymphoma (HSTCL). METHODS: Three cases of HSTCL were searched for morphology, immunophenotypings, Epstein-Barr virus (EBV) in situ hybridization and T-cell receptorgamma (TCRgamma) gene rearrangement. RESULTS: In the bone marrow, the infiltrative pattern of tumour cells was interstitial and sinusoidal infiltration in one caseiwhile the other two cases showed diffuse infiltration. In the liver of one case and the spleen of another case, tumour cells respectively showed sinusoidal infiltration. The immunophenotyping: three cases showed strongly positive for CD3 and TIA-1, but negative for Granzyme B, CD56 and TCRbeta. EBV in situ hybridization was not detected in all the cases. TCRgamma monoclonal rearrangements were detected in two cases. CONCLUSION: HSTCL is a rare entity which is classified into peripheral T cell lymphomas. And it is regarded as a subset of unactived cytotoxic T-cell lymphomas. The negative results of EBV in situ hybridization and the presentation of TCRgamma gene monoclonal rearrangements may be helpful in diagnosis and differential diagnosis of HSTCL. These three cases showed similar characters to those of international cases reported. PMID- 18677384 TI - [Anaplastic lymphoma kinase gene abnormality and the expression of its fusion protein in primary systemic anaplastic large cell lymphoma]. AB - OBJECTIVE: To study the expression of anaplastic lymphoma kinase (ALK) and chromosome breakage of the anaplastic lymphoma kinase (ALK) gene retrospectively and to investigate their possible value as indicators of prognosis in primary systemic anaplastic large cell lymphomas (S-ALCL). METHODS: Twenty-eight cases of S-ALCL were collected from the Lymphoma Lab, the Department of Pathology, Peking University Health Science Center and Beijing Children's Hospital. The morphologic characteristics were studied under light microscope, and essential immunohistochemical staininings (IHC) were performed and reviewed to confirm the diagnosis of S-ALCL. ALK-1 monoclonal antibody was used to assess ALK fusion protein expression, and EnVision method was used in IHC. Locus specific interphase fluorescence in situ hybridization (LSI-FISH) was also performed on the neoplastic cells using paraffin-embedded tissues to detect ALK gene abnormality. RESULTS: ALK-1 protein was expressed in 19 of the 28 cases. In 14 ALCL cases, ALK gene breakage was detected by LSI-FISH, using a dual-color break apart ALK gene DNA (LSI-ALK) probe. Of the other 14 cases which did not show ALK gene breakage, 5 showed 2 copies of ALK gene as normal, and 9 showed multi-copies of ALK gene. Of all the 28 cases, 22 had complete follow-up materials. Sixteen survived and 6 died, their survival time ranged from 0.5 to 36.0 months, and the survival time on average was 12.8 months, cumulative proportion survival rate was 73.9% in the 1st year. Those cases showing multi-copies of ALK gene might have the worst outcome, with only 47.6% of cumulative proportion survival rate in the 1st year. CONCLUSION: IHC detection for ALK fusion protein is important to the diagnosis of S-ALCL. ALK gene breakage detected by interphase LSI-FISH might not be always consistent with abnormal expression of ALK fusion protein. Complex abnormalities of ALK gene exist in S-ALCL cases, and different types of ALK gene might lead to different clinical outcome. Those cases with multi-copies of ALK gene probably have the poorest prognosis. PMID- 18677386 TI - [Kidney injury in transplantation-associated thrombotic microangiopathy]. AB - OBJECTIVE: To study clinicopathological features and mechanism of renal injury in transplantation-associated thrombotic microangiopathy (TA-TMA). METHODS: The renal biopsies obtained from patients after hematopoietic cell transplantation or kidney transplantation were observed by immunofluorescence, light microscopy and electron microscopy (EM). RESULTS: The patients presented hypertension, massive proteinuria, intravascular haemolysis and renal insufficiency some time after transplantation. Various immunoglobulins and complements were negative in immunofluorescent staining. Light microscopy showed endothelial cell proliferation and swelling of glomerular capillary and small arteries, irregular thickened glomerular basement membrane and microthrombosis. EM revealed endothelial cell proliferation and swelling with thickened lamina rara interna of glomerular basement membrane. CONCLUSION: TA-TMA is a rare complication after organ or tissue transplantation. Kidney is often involved in TA-TMA with massive proteinuria, anemia and renal failure. Differential diagnosis should be made between TA-TMA and rejection or other renal diseases. Renal biopsy is a major method for accurate diagnosis. TA-TMA correlates to endothelial cell injury caused by viral infection and/or immunosuppressive drugs for anti-rejection. PMID- 18677387 TI - [Differences of the molecular phenotypes and the histogenesis between dermatofibroma and dermatofibrosarcoma protuberans]. AB - OBJECTIVE: To explore the histogenesis and differentiation of dermatofibroma (DF) and dermatofibrosarcoma protuberans (DFSP). METHODS: Clinical information and microscopic characteristics of 26 cases of DF and 26 cases of DFSP were investigated. The immunohistochemical study was performed on microarray sections by a panel of antibodies including Factor XIIIa, HLA-DR, CD34, CD14, S-100, MSA, and Ki67. Probe was labeled by in vitro transcription. The mRNA expression levels of TGF-beta and bFGF were investigated by in situ hybridization. RESULTS: All cases showed positive for Factor XIIIa, HLA-DR and CD34 to different extent. The medians of positive rates in DF were Factor XIIIa 90%, HLA-DR 70%, and CD34 5%, and in DFSP were Factor XIIIa 10%, HLA-DR 5%, and CD34 80%. CD14 was positive in 3 cases of DF and 1 case of DFSP. S-100 was positive in 6 cases of DFSP and 2 cases of DF. MSA was positive in 5 cases of DFSP and 3 cases of DF. In all cases, positive rate of Ki67 was less than 5%. The mRNA expression levels of TGF-beta was elevated in DF in comparison with DFSP. CONCLUSION: Both DF and DFSP can differentiate to dendritic cells (DC) in different degree. Considering the character of microscopic features and immunohistochemical phenotype, cells of DF are much similar to mature DC, while those of DFSP much similar to immature dermal reserve cell (DRC). The differences of cell differentiation between DF and DFSP result in different prognosis. DF is a benign tumor, while DFSP a low grade malignant tumor. The different expression of Factor XIIIa and CD34 may be helpful to differential diagnosis of DF and DFSP. PMID- 18677388 TI - [Clinicopathologic characteristics and outcomes of IgA nephropathy in elder patients]. AB - OBJECTIVE: To investigate the clinical, pathological characteristics and outcomes of IgA nephropathy in the elderly. METHODS: Seventy patients over age 60 with IgA nephropathy were studied and compared with 82 patients under 60 years in the clinical and pathological features as well as the outcomes. RESULTS: Compared with non-elderly group, elder group patients had higher blood pressure [systolic pressure (142.0+/-20.4) mmHg vs (124.2+/-16.9) mmHg (1mmHg=0.133 kPa), diastolic pressure (83.1+/-11.8) mmHg vs (78.9+/-12.3) mmHg], serum creatinine [(172.7+/ 125.8) micromol/L vs (94.4+/-42.5) micromol/L], serum cholesterol[(5.7+/-1.6) mmol/L vs (5.1+/-1.6) mmol/L], 24 hj urinary protein excretion rate [(3.4+/-2.9) g/d vs (1.8+/-2.0) g/d], and the incidence of CKD stages 3-5(64.0% vs 14.6%)(P<0.05). No significant differences were seen in the disease courses, rate of gross hematuria, serum triglyceride and serum IgA level between two groups(P>0.05).Renal pathological investigation showed the chronic lesions were dominated in elder group. There was significant difference in portion of glomerular sclerosis [(19.7+/-20.1)% vs (13.4+/-17.8)%], renal tubule atrophy (>1 score,34.2% vs 25.6%), interstitial fibrosis (>1,score 34.2% vs 18.2%), and arteriolosclerosis (>2 score,20.0% vs 8.5%) between two groups (P<0.05). However, there were no significant difference in proportion of mesangial proliferation, crescent and interstitial inflammatory cell infiltration (P>0.05). After a mean post-biopsy follow-up of (34.6+/-33.3) months, the 3-year and the 5-year renal survival rates for elder group were 74.6% and 62.2%, respectively, which were lower than those of non-elder group (100% and 92.9%, P=0.002). CONCLUSION: Elder patients with IgA nephropathy were more likely to suffer from hypertension, hyperlipidemia, renal insufficiency and chronic pathologic lesions, which might be the risk factors for the patient's unfavorable prognosis. PMID- 18677389 TI - [One patient with Kimura's disease and angiolymphoid hyperplasia with eosinophilia also suffers from kidney injury]. AB - OBJECTIVE: Reporting a rare case with Kimura's disease and angiolymphoid hyperplasia with eosinophilia (ALHE), who also has nephrotic syndrome resulting from minimal change glomerulopathy. METHODS: Histological manifestations, immunohistochemical staining, immunofluorescence staining, and electron microscope were performed. RESULTS: One 37-year-old male patient with recurrent erythra, cutaneous node, painless cervical lymph node enlargement and nephrotic syndrome in 5 years. It's confirmed through biopsies that the subcutaneous node is ALHE, the cervical lymph node is kimura disease, and renal lesion is minimal change glomerulopathy. CONCLUSION: Kimura disease and ALHE may be different features of the same disease, and may complicate glomerulopathy. PMID- 18677390 TI - [A clinicopathologic study on complications of orthotopic liver transplantation in 54 patients with chronic hepatitis B virus infection]. AB - OBJECTIVE: To study the complication incidence of 54 patients with chronic HBV infection following their orthotopic liver transplantation (OLT), and factors associated with HBV recurrence and hepatocellular carcinoma (HCC) recurrence or metastasis post-OLT. METHODS: The light-microscopic appearance of hepatic allograft biopsies in 54 patients with chronic HBV infection following OLT was examined. The related clinical data were analyzed. The incidence and occurrence time of post-OLT complications were studied. Furthermore, the relationship between recurrent type B viral hepatitis and acute rejection and the relationship among HCC recurrence/metastasis, acute rejection, the tumor diameter, and the portal vein invasion were particularly studied. RESULTS: The frequent complications of patients with chronic HBV infection following OLT were acute rejection [38(70.4%); occurrence time: 5-365 d], chronic rejection [ 1(1.9%); occurrence time: 10.7 months],bile duct complications [24(44.4%); occurrence time: 7-940 d], HBV recurrence [7(13.0%); occurrence time: 1-540 d], HCV infection [3(5.6%); occurrence time: 60 d, 60 d, 33 months], CMV infection [8(14.8%); occurrence time: 67-90 d], and HCC recurrence or metastasis [17(31.5%); occurrence time: 2-41 months]. At the end of 1 year post-OLT, 95% of patients with post-hepatitis B cirrhosis were alive; At the end of 3 years post OLT, 85% of patients with post-hepatitis B cirrhosis were alive. However, at the end of 1 year post-OLT, 67.6% of patients with post-hepatitis B HCC were alive; At the end of 3 years post-OLT, 50% of patients with post-hepatitis B HCC were alive. The number of acute rejection episodes in patients with recurrent HBV infection and that without recurrent HBV infection was (0.86+/-1.46) time/patient and (1.07+/-0.90) time/patient respectively(P>0.05); the number of moderate acute rejection episodes(RAI>or=score 4) in patients with recurrent HBV infection and that without recurrent HBV infection was (0.29+/-0.49) time/patient and (0.50+/ 0.63) time/patient(P>0.05); Incidence of patients with >or=3 episodes of acute rejection in patient with recurrent HBV infection and that without recurrent HBV infection was 14.3% and 10.6%(P>0.05). Furthermore, the number of acute rejection episodes in patients with HCC recurrence or metastasis and that without HCC recurrence or metastasis was (1.12+/-0.93) time/patient and (1.06+/-1.39) time/patient respectively(P>0.05); the number of moderate acute rejection episodes(RAI>or=score 4) in patients with HCC recurrence or metastasis and that without HCC recurrence or metastasis was (0.65+/-0.79) time/patient and (0.65+/ 1.06) time/patient respectively(P>0.05); Incidence of patients with >or=3 episodes of acute rejection in patient with HCC recurrence or metastasis and that without HCC recurrence or metastasis was 5.9% and 17.6% respectively(P>0.05). The tumor diameter in patients with HCC recurrence or metastasis was (6.72+/-3.40)cm, however, that in patients without HCC recurrence or metastasis was (3.55+/ 2.17)cm(P=0.004 7).The incidence of Portal vein invasion in patients with HCC recurrence or metastasis and that without HCC recurrence or metastasis was 68.75% and 33.3% respectively(P=0.006). CONCLUSION: There was no significant difference among HBV recurrence and acute rejection post-liver transplantation in patients with chronic HBV infection. There was no significant difference between HCC recurrence and acute rejection. The tumor diameter of patients with HCC recurrence or metastasis was significantly greater than that of no HCC recurrence or metastasis. The portal vein invasion of patients with HCC recurrence or metastasis was significantly frequent than that of no HCC recurrence or metastasis. PMID- 18677391 TI - [Pathological characteristics mixed epithelial and stromal tumor of kidney]. AB - OBJECTIVE: To study the clinicopathological features of mixed epithelial and stromal tumor of the kidney(MESTK). METHODS: Clinical and pathological characteristics of one case of MESTK was studied. RESULTS: A case of MESTK which uncommonly occurred in a 16-year old adolescent male presented with dysuria and a large mass in the right renal region without a history of estrogen/progestogen treatment and/or obesity or urogenital surgery. Radiology revealed a large cystic/solid mass within the right kidney. Grossly, it was well demarcated and had a solid and cystic appearance on sectioning. Microscopically, the tumor was composed of a mixture of stromal and epithelial components. The epithelial component was composed of flat to columnar cells forming glands or tubules. The stromal components essentially consisted of bland, loosely packed spindle cells in an edematous and myxoid background. In some areas, there were smooth muscle cells forming bands and fascicles but no ovarian-type stroma was present. Immunohistochemical staining revealed that the epithelial components were positive for AE1/AE3 and focally positive for estrogen receptor (ER),progesterone receptor(PR), CD10 and Vimentin, whereas the stromal components were positive for ER, PR, Desmin and smooth muscle actin(SMA). Both epithelial and stromal components were negative for HMB-45, S-100, alpha-inhibin and WT-T. Five months after resection, the patient was well without evidence of recurrence. CONCLUSION; MESTK occurred in a pubertal male, as in the current case, supports the hypothesis that proliferation of remnants of the primitive mesenchyme in the kidney in situation of sex-steroid abnormity may play an important role in the pathogenesis of male MESTK. PMID- 18677392 TI - [Immunological characteristics of chronic graft versus host diseases murine model with lupus nephritis]. AB - OBJECTIVE: To establish the murine systemic lupus erythematosus (SLE) model of chronic graft versus host diseases(cGVHD). To analyze the pathological changes and serological and immunological features in the animals. METHODS: Female (C57BL/10 x DBA/2)F1 hybrids aged 6-8 weeks were randomly divided into model group and healthy controls (HC). Lymphocytes from female DBA/2 were injected intravenously to the model group on days 0, 3 and 8, while PBS were injected to the HC under the same condition as a control group. Bradford was applied to monitor the development of albuminuria quantitively. Sera were tested by enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF) for the presence of autoantibodies. To compare the differences of CD4+ CD25+ Treg cells between the two groups by flow cytometry (FCM) and the differences in the expression of Foxp3 by real time polymerase chain reaction(RT-PCR). The kidneys of model mice were removed in the 12th week and were made frozen sections for direct immunofluorescence(DIF)and paraffin imbedding for PASM staining. RESULTS: The titers of proteinuria in model group in the 6th week, 8th week, 10th week, and 12th week were significantly higher than those of the HC groups(P=0.004, 0.005,respectively). The titers of anti-dsDNA and anti-nucleosome antibodies were significantly increased in the model group compared with the HC (P<0.05). And the positive rates curves of ANA, anti-dsDNA Abs and anti-nucleosome Abs in model group were significantly different from those of control group (P<0.05). And The proportions of CD4+CD25+ regulatory T cells from peripheral blood of model group were significantly lower than those of control group (P=0.002), while the expression of Foxp3, one of the most important biomarkers of Treg cells, was not significant. There were mesangial matrix expansion and mesangial cell proliferation in the nephritic pathology in model group and the depositions of IgG along the glomerular capillary walls and in the mesangium were observed in model group. There were no pathological changes and depositions in HC group. CONCLUSION: It has been proved that there are not only protienuria and autoantibodies, but also decrease of the regulatory T lymphocytes in murine model of cGVHD. All of these results suggest that the cGVHD murine SLE models were successfully established. PMID- 18677394 TI - [A system for quantitatively measurement of alveolar bone density and the evaluation of its precision and validity]. AB - OBJECTIVE: To establish a system for quantitatively measuring alveolar bone density and to test its precision and validity. METHODS: With the aid of computer technique, the system measured the average gray level intensity of regions of interest (ROIs) on standardized exposed direct digital periapical radiograph. To correct the variation resulting from exposure condition, an aluminum stepwedge was exposed simultaneously. By referring the image of aluminum stepwedge on each radiograph the equivalent aluminum thickness(EAT)and corrected gray level intensity(CGL)of the ROIs could be calculated as indicators of alveolar bone density. Nineteen specimens containing different amounts of hydroxy-phosphorite were radiographed. The precision of the system was tested by repeatedly measuring 2 of the specimens. The set of specimens was exposed under two different exposure time in order to test the system's validity to correct the gray level difference caused by different exposure time. RESULTS: After repeated measurement of specimens, we found the Relative Standard Deviation(RSD) of EAT and CGL was between 0.83%-2.15%; At the level of 95%, if the difference of EAT between the two ROIs was larger than 0.05 mm or the difference of CGL was larger than 3, the content of hydroxy-phosphorite in them was different; The two sets of data during different exposure time were processed with Wilcoxon Signed Ranks Test, and the result showed effective correction of the variation caused by different exposure time. CONCLUSION: The precision and validity of the quantitative alveolar bone density measuring system are acceptable. The system can be used to compare alveolar bone density longitudinally and cross-sectionally. PMID- 18677393 TI - [Activation of endoplasmic reticulum stress proteins in light-induced retinal degeneration]. AB - OBJECTIVE: To investigate the expression of endoplasmic reticulum stress proteins in photoreceptor apoptosis in light-induced retinal degeneration. METHODS: Exposure to excessive levels of light induced photoreceptor apoptosis and had been previously used as a model for the study of retinal degeneration. Photoreceptor apoptosis was detected by terminal dUTP transferase nick end labeling (TUNEL). The protein expression levels of ER stress sensors including glucosejregulated protein-78 (GRP78/BiP), caspase-12, phospho-eukaryotic initiation factor 2alpha (eIF2alpha) and phospho- double-stranded RNA-activated protein kinase-like endoplasmic reticulum kinase (PERK) were examined by immunojfluorescence and Western blot analysis. RESULTS: Following light exposure, the protein expression levels of GRP78/BiP, caspase-12, phospho-eIF2alpha and phospho-PERK were up-regulated in a time dependent manner. The up-regulation of these proteins coincided with or preceded the photoreceptor apoptosis. At the peak of their expression, they were mainly located in the photoreceptor inner segments and/or outer nuclear layers (ONL). CONCLUSION: Activation of endoplasmic reticulum stress proteins appears to play an important role in light-induced retinal degeneration. Therefore endoplasmic reticulum stress modulators could become a strong candidate for a therapeutic agent in treatment of these diseases. PMID- 18677395 TI - [Item selection for postnatal quality of life scale]. AB - OBJECTIVE: To select items to compose the final scale from 101-item pool for assessment of postnatal quality of life which was developed by the authors before. METHODS: Ten experts in maternal and child health scored the importance of every item. A total of 200 women at 0-12 months after childbirth who took their babies for vaccination or clinic services were investigated by the item pool, and 10% of them were retested in one month. Based on these data, items in the item pool were screened by following principles respectively: score of item importance, factor loading, variation coefficient, Cronbach Alpha, correlation coefficient of item with domains (2 types) and test-retest correlation coefficient. Then we summarized the screening results of these methods, and retained the items which could be selected in at least five methods. RESULTS: The numbers of items deleted in these seven screening methods respectively were: 39 items in experts scoring method; 43 items in factor analysis method; 49 items in variation analysis method; 50 items in Cronbach Alpha method; 26 items in correlation coefficient method 1; 9 items in correlation coefficient method 2; and 11 items in test-retest method. After summarizing these screening results, 60 items were selected. In order to simplify the scale,only kept 2 items in each aspect which had more frequency retained. Finally, 40 items remained in the postnatal quality of life scale. CONCLUSION: This is the first Chinese scale for evaluating postnatal quality of life of women. It needs to establish the validity and reliability in the future. PMID- 18677396 TI - [Synthesis of diethyl N-[p-(methylamino)benzoyl]-L-glutamate]. AB - OBJECTIVE: To find a best synthesis method of diethyl N-[p-(methylamino)benzoyl] L-glutamate(1). METHODS: Using diethyl N-(p- aminobenzoyl)-L-glutamate (2) as a starting material via a one-pot process of benzylation and methylation, the diethyl N-[p-(methylamino)benzoyl ]-L-glutamate (3) is prepared in high yield. Then compound 1 was conveniently obtained from debenzylation of compound 3 by Pd/C. RESULTS: The target compound was obtained through 2-step reaction in 88% overall yield in only 6 h, and its structure was identified by 1HNMR and MS. CONCLUSION: It is a convenient and efficient method of preparation of diethyl N [p-(methylamino)benzoyl]-L-glutamate. PMID- 18677397 TI - [Progress in the study of the chemical synapse in glioma-associated epileptogenesis]. AB - Seizures may be the first or sometimes the only manifestation of patients with glioma in clinics. The aim of operation is to eliminate epilepsy far beyond mere resection of tumor mass. The underlying mechanisms of glioma-associated epileptogenesis are poorly understood. Recently the theory of amino-acid like neurotransmitters in chemical synapse is gradually accepted. However, the molecular mechanisms remain to be further investigated on how glutamate release is regulated and how synaptic homeostasis in peripheral neurons is kept or disturbed. So detailed studies are needed to clarify specific molecular target and provide proper evidence for optimal antiepileptic drugs in glioma-associated epileptogenesis. PMID- 18677398 TI - Native and recombinant bovine placental lactogens. AB - The bovine placenta produces a wide variety of proteins that are structurally and functionally similar to the pituitary proteins from the GH/PRL gene family. Bovine placental lactogen (bPL) is a 200-amino acid long glycoprotein hormone that exhibits both lactogenic and somatogenic properties. The apparent molecular masses of purified native (n) bPL molecules (31-33 kDa) exceed 23 041 Da, which is the theoretical molecular mass of the protein core. At least six isoelectric variants (pI: 4.85-6.3) of bPL were described in cotyledonary extracts and three different bPL isoforms (pI: 4.85-5.25) were found in fetal sera. The bPL molecules that are detected in higher concentrations in peripheral circulation exhibit a more acidic pI than those present in placental homogenates. This may reflect an important glycosylation process occurring just prior to the bPL secretion. The bPL mRNA is transcribed in trophectoderm binucleate cells starting from Day 30 of pregnancy until the end of gestation. In mothers, bPL is involved in the regulation of ovarian function, mammogenesis, lactogenesis, and pregnancy stage-dependent adaptation of nutrient supplies to the fetus. Due to the higher fetal, compared to maternal concentrations of circulating hormone, it has been suggested that bPL primarily targets fetal tissues. PMID- 18677399 TI - Decreased expression of pigment epithelium derived factor (PEDF), an inhibitor of angiogenesis, in placentas of unexplained stillbirths. AB - Normal placental vascular development depends upon the complex interactions between angiogenic inducers and inhibitors within the placenta. Alterations within the placental microenvironment can promote an imbalance in angiogenic mediators which may be associated with adverse perinatal outcomes. The purpose of this study was to investigate the placentas of infants with unexplained stillbirth as compared to live-born infants and to determine whether alterations in angiogenic inducer vascular endothelial growth factor (VEGF) or inhibitor pigment epithelium-derived factor (PEDF) are associated with altered angiogenesis, vascular remodeling and stillbirth. Placentas of 22 unexplained stillbirths and 44 age-matched live-born controls were scored for microvascular density (MVD), vasculopathy and microvascular permeability. A subset was scored for expression of angiogenic inducer VEGF and inhibitor pigment epithelium derived factor. Stillborn placentas demonstrated higher MVD than controls (mean+SD: 116.6+/-46.3 v. 60.8+/-13.5, respectively, p<0.001). Vasculopathy was present in 10/22 (45%) stillbirths compared to 0/44 (0%) controls (p<0.001); increased vascular permeability was present in 15/22 (68%) cases and 5/44 (11%) controls (p<0.001). PEDF expression was significantly lower in stillborn placentas (1.7+/-0.3) than live-born controls (3.6+/-0.8, p<0.01) while VEGF expression was similar (3.3+/-0.7 v. 3.7+/-0.4, respectively, p>0.05). In conclusion, we found that unexplained stillbirth is associated with loss of angiogenic inhibitor PEDF, vasculopathy and heightened angiogenesis in the placenta. PMID- 18677400 TI - The influence of steroid hormones on in vitro NOx production by porcine fetal membranes. AB - The aim of the study was to examine: 1/ allantochorial concentrations of nitrate/nitrite (NOx) and 2/ plasma concentration of NOx in pigs on days 25, 35, 40 and 60 of pregnancy as well as 3/ the influence of estradiol-17beta (E(2)) and/or progesterone (P(4)) on NOx production by porcine fetal membranes on the studied days of pregnancy. Total NOx concentration was determined using a microplate assay method based on the Griess reaction. Fetal membrane NOx content gradually increased from day 25 to day 60 of gestation. Blood plasma NOx concentration decreased from day 25 to 40, and then plasma NOx concentration significantly increased on day 60. In addition, the stimulatory effect of E(2), P(4) and E(2)+P(4) on NO in vitro production by porcine fetal membranes was demonstrated. The stimulatory effect of steroid hormones on NOx release depended on steroid dose and day of pregnancy. It is possible that the observed differences in the strength of the stimulatory action of E(2), P(4) and E(2)+P(4) on fetal membrane NOx production are associated with an activation of different isoforms of nitric oxide synthase. PMID- 18677401 TI - Immune response to lytic peptides conjugated to a betaCG fragment in treated BALB/C mice. AB - Hecate-betaCG and Phor14-betaCG(ala) are relatively short, amphipathic alpha helical cationic peptides with the ability to destroy selectively breast, prostate and ovarian cancer cells. Treatment with proteins and peptides frequently initiated antibody formation. Short peptides may minimize the risk of the immune system mobilization after treatment but it is necessary to investigate whether Hecate-betaCG and Phor14-betaCG(ala) induce the immune system to produce antibody and whether they affect the reproductive organs in normal wild-type mice. The results of our experiments showed that specific antibodies, tested by the enzyme-immunoassay, were not detected in the group treated with Hecate-betaCG and Phor14-betaCG(ala). The blood concentrations of both peptides begun to decrease from 60 minutes after injection and after 240 minutes its levels were undetectable. Histopatho-logical examination exhibited degenerative changes in the prostate glands and testes in males and in the ovaries and uteri of females treated with both peptides. In conclusion, our results indicate that both relatively small and rapidly metabolized peptides are not immunogenic and can be used for further investigation as a potential cancer treatment. PMID- 18677402 TI - Changes in the GnRH mRNA and GnRH receptor (GnRH-R) mRNA levels in the hypothalamic-anterior pituitary unit of anestrous ewes after infusion of GnRH into the third cerebral ventricle. AB - In the present paper the role of GnRH in the ultrashort loop of the negative feedback action on GnRH secretion was evaluated on the molecular level by the Real-time PCR technique. Specifically, the effect of GnRH infused into the third cerebral ventricle on the expression of GnRH and GnRH receptor (GnRH-R) genes was analyzed in the hypothalamic-pituitary unit of anestrous ewes. GnRH did not significantly affect GnRH mRNA levels in the preoptic/anterior hypothalamic area but drastically increased its level in the ventromedial hypothalamus. In addition, GnRH infusion augmented GnRH-R mRNA level in the entire hypothalamus. In the GnRH-treated animals, anterior pituitary GnRH-R mRNA level and plasma LH concentration were also elevated. The changes in GnRH mRNA and GnRH-R mRNA levels in the hypothalamus in response to treatment with GnRH suggest that GnRH acts differently on the stability of these transcripts. On the basis of presented results it seems that GnRH may affect GnRH and GnRH-R biosynthesis and, consequently, GnRH/LH release. PMID- 18677403 TI - The effect of insulin-like growth factor-I, relaxin and luteinizing hormone on vascular endothelial growth factor secretion by cultured endometrial stromal cells on different days of early pregnancy in pigs. AB - The effect of insulin-like growth factor-I (IGF-I), relaxin (RLX) and luteinizing hormone (LH) on vascular endothelial growth factor (VEGF) in vitro secretion by endometrial stromal cells in pigs was investigated on days 10-12 and 20-22 of gestation. LH-stimulated stromal cell secretion of VEGF did not differ among tested days of early pregnancy. However, IGF-I- and RLX-mediated release of VEGF depended on the day of pregnancy. It seems that IGF-I and RLX may be considered as potent activators of VEGF-mediated angiogenesis in porcine endometrium, and their action may be more pronounced during maternal recognition of pregnancy. PMID- 18677404 TI - Glycosylated polyacrylate nanoparticles by emulsion polymerization. AB - A selection of glycosylated polyacrylate nanoparticles has been prepared by radical-initiated emulsion polymerization in aqueous media. Using ethyl acrylate as a co-monomer, carbohydrate acrylates were incorporated into the poly(ethyl acrylate) framework to give stable emulsions of glyconanoparticles with an average particle size of around 40 nm. Using this technique a variety of glyconanoparticles were prepared from 3-O-acryloyl-1,2:5,6-di-O-isopropylidene alpha-D-glucofuranose, 1-O-acryloyl-2,3:5,6-di-O-isopropylidene-alpha-D mannofuranose, 6-O-acryloyl-1,2:3,4-di-O-isopropylidene-alpha-D-galactopyranose, 2-N-acryloyl-1,3,4,6-tetra-O-acetyl-beta-D-glucosamine, 5-O-acryloyl-2,3 isopropylidene-1-methoxy-beta-D-ribofuranose and 4-N-acetyl-5'-O-acryloyl-2',3'-O isopropylidene cytidine. Scanning electron microscopy, dynamic light scattering and proton NMR analysis of the emulsions indicated essentially 100% incorporation of the carbohydrate acrylate monomer into the polymer with the exception of O benzyl- and O-benzoyl-protected carbohydrate acrylates, which gave incomplete incorporation. Formation of larger glyconanoparticles of ~80nm with (unprotected) 3-O-acryloyl-D-glucose and 5-O-acryloyl-1-methoxy-beta-D-ribofuranose revealed the influence of free hydroxyl groups in the monomer on the particle size during polymerization, a feature which is also apparently dependent on the amount of carbohydrate in the matrix. This methodology allows for a new, simple route to the synthesis of polymeric glyconanoparticles with potential applications in targeted drug delivery and materials development. PMID- 18677405 TI - HHMI to support young physician-scientists. PMID- 18677406 TI - Paraspeckle protein p54nrb links Sox9-mediated transcription with RNA processing during chondrogenesis in mice. AB - The Sox9 transcription factor plays an essential role in promoting chondrogenesis and regulating expression of chondrocyte extracellular-matrix genes. To identify genes that interact with Sox9 in promoting chondrocyte differentiation, we screened a cDNA library generated from the murine chondrogenic ATDC5 cell line to identify activators of the collagen, type II, alpha 1 (Col2a1) promoter. Here we have shown that paraspeckle regulatory protein 54-kDa nuclear RNA-binding protein (p54nrb) is an essential link between Sox9-regulated transcription and maturation of Sox9-target gene mRNA. We found that p54nrb physically interacted with Sox9 and enhanced Sox9-dependent transcriptional activation of the Col2a1 promoter. In ATDC5 cells, p54nrb colocalized with Sox9 protein in nuclear paraspeckle bodies, and knockdown of p54(nrb) suppressed Sox9-dependent Col2a1 expression and promoter activity. We generated a p54nrb mutant construct lacking RNA recognition motifs, and overexpression of mutant p54nrb in ATDC5 cells markedly altered the appearance of paraspeckle bodies and inhibited the maturation of Col2a1 mRNA. The mutant p54nrb inhibited chondrocyte differentiation of mesenchymal cells and mouse metatarsal explants. Furthermore, transgenic mice expressing the mutant p54nrb in the chondrocyte lineage exhibited dwarfism associated with impairment of chondrogenesis. These data suggest that p54nrb plays an important role in the regulation of Sox9 function and the formation of paraspeckle bodies during chondrogenesis. PMID- 18677407 TI - Inflammatory macrophage migration requires MMP-9 activation by plasminogen in mice. AB - Inflammation plays a critical role in the development of cardiovascular diseases. Infiltration of leukocytes to sites of injury requires their exit from the blood and migration across basement membrane; this process has been postulated to require remodeling of the ECM. Plasminogen (Plg) is a protease that binds to the ECM and, upon conversion to plasmin, degrades multiple ECM proteins. In addition, plasmin directly activates MMPs. Here, we used Plg(-/-) mice to investigate the role of Plg in inflammatory leukocyte migration. After induction of peritonitis by thioglycollate injection, we found that Plg(-/-) mice displayed diminished macrophage trans-ECM migration and decreased MMP-9 activation. Furthermore, injection of the active form of MMP-9 in Plg(-/-) mice rescued macrophage migration in this model. We used periaortic application of CaCl2 to induce abdominal aortic aneurysm (AAA) and found that Plg(-/-) mice displayed reduced macrophage infiltration and were protected from aneurysm formation. Administration of active MMP-9 to Plg(-/-) mice promoted macrophage infiltration and the development of AAA. These data suggest that Plg regulates macrophage migration in inflammation via activation of MMP-9, which, in turn, regulates the ability of the cells to migrate across ECM. Thus, targeting the Plg/MMP-9 pathway may be an attractive approach to regulate inflammatory responses and AAA development. PMID- 18677408 TI - Resistance of human glioblastoma multiforme cells to growth factor inhibitors is overcome by blockade of inhibitor of apoptosis proteins. AB - Multiple receptor tyrosine kinases (RTKs), including PDGFR, have been validated as therapeutic targets in glioblastoma multiforme (GBM), yet inhibitors of RTKs have had limited clinical success. As various antiapoptotic mechanisms render GBM cells resistant to chemo- and radiotherapy, we hypothesized that these antiapoptotic mechanisms also confer resistance to RTK inhibition. We found that in vitro inhibition of PDGFR in human GBM cells initiated the intrinsic pathway of apoptosis, as evidenced by mitochondrial outer membrane permeabilization, but downstream caspase activation was blocked by inhibitor of apoptosis proteins (IAPs). Consistent with this, inhibition of PDGFR combined with small molecule inactivation of IAPs induced apoptosis in human GBM cells in vitro and had synergistic antitumor effects in orthotopic mouse models of GBM and in primary human GBM neurospheres. These results demonstrate that concomitant inhibition of IAPs can overcome resistance to RTK inhibitors in human malignant GBM cells, and suggest that blockade of IAPs has the potential to improve treatment outcomes in patients with GBM. PMID- 18677409 TI - Hepatic CB1 receptor is required for development of diet-induced steatosis, dyslipidemia, and insulin and leptin resistance in mice. AB - Diet-induced obesity is associated with fatty liver, insulin resistance, leptin resistance, and changes in plasma lipid profile. Endocannabinoids have been implicated in the development of these associated phenotypes, because mice deficient for the cannabinoid receptor CB1 (CB1-/-) do not display these changes in association with diet-induced obesity. The target tissues that mediate these effects, however, remain unknown. We therefore investigated the relative role of hepatic versus extrahepatic CB1 receptors in the metabolic consequences of a high fat diet, using liver-specific CB1 knockout (LCB1-/-) mice. LCB1(-/-) mice fed a high-fat diet developed a similar degree of obesity as that of wild-type mice, but, similar to CB1(-/-) mice, had less steatosis, hyperglycemia, dyslipidemia, and insulin and leptin resistance than did wild-type mice fed a high-fat diet. CB1 agonist-induced increase in de novo hepatic lipogenesis and decrease in the activity of carnitine palmitoyltransferase-1 and total energy expenditure were absent in both CB1(-/-) and LCB1(-/-) mice. We conclude that endocannabinoid activation of hepatic CB1 receptors contributes to the diet-induced steatosis and associated hormonal and metabolic changes, but not to the increase in adiposity, observed with high-fat diet feeding. Theses studies suggest that peripheral CB1 receptors could be selectively targeted for the treatment of fatty liver, impaired glucose homeostasis, and dyslipidemia in order to minimize the neuropsychiatric side effects of nonselective CB1 blockade during treatment of obesity-associated conditions. PMID- 18677410 TI - Leukemia-associated NOTCH1 alleles are weak tumor initiators but accelerate K-ras initiated leukemia. AB - Gain-of-function NOTCH1 mutations are found in 50%-70% of human T cell acute lymphoblastic leukemia/lymphoma (T-ALL) cases. Gain-of-function NOTCH1 alleles that initiate strong downstream signals induce leukemia in mice, but it is unknown whether the gain-of-function NOTCH1 mutations most commonly found in individuals with T-ALL generate downstream signals of sufficient strength to induce leukemia. We addressed this question by expressing human gain-of-function NOTCH1 alleles of varying strength in mouse hematopoietic precursors. Uncommon gain-of-function NOTCH1 alleles that initiated strong downstream signals drove ectopic T cell development and induced leukemia efficiently. In contrast, although gain-of-function alleles that initiated only weak downstream signals also induced ectopic T cell development, these more common alleles failed to efficiently initiate leukemia development. However, weak gain-of-function NOTCH1 alleles accelerated the onset of leukemia initiated by constitutively active K ras and gave rise to tumors that were sensitive to Notch signaling pathway inhibition. These data show that induction of leukemia requires doses of Notch1 greater than those needed for T cell development and that most NOTCH1 mutations found in T-ALL cells do not generate signals of sufficient strength to initiate leukemia development. Furthermore, low, nonleukemogenic levels of Notch1 can complement other leukemogenic events, such as activation of K-ras. Even when Notch1 participates secondarily, the resulting tumors show "addiction" to Notch, providing a further rationale for evaluating Notch signaling pathway inhibitors in leukemia. PMID- 18677413 TI - Hybrid zones as a tool for identifying adaptive genetic variation in outbreeding forest trees: lessons from wild annual sunflowers (Helianthus spp.). AB - The identification and study of adaptively important genes in forest trees represents a formidable challenge because of their long generation spans. In annual or perennial herbs, formal genetic studies can be employed to identify the quantitative trait loci (QTLs) and/or candidate genes that underlie important traits, and the segregating populations can be transplanted into natural populations to measure the strength and direction of selection. However, the application of these methods to forest trees is difficult, because the creation of appropriate genetic material is extremely time-consuming in long-lived, woody plants, and lifetime fitness estimates are difficult or impossible to obtain. Although QTL mapping should in principle be feasible in wild intraspecific populations (as an alternative to artificial crosses), this approach is less likely to be successful in trees because LD (linkage disequilibrium) will decay quickly in large outbreeding plant populations. Within the present paper, we discuss a modified approach based on natural hybrid zones. We describe the use of wild annual sunflowers (Helianthus spp.) as a model for exploring the hybrid zone approach. Transplanted experimental hybrids allowed us to assess the adaptive value of individual chromosomal blocks in nature, and data on natural Helianthus hybrids suggest that similar approaches are possible in natural hybrid zones. Our results allowed us to test the role of hybridization in the origin of ecological divergence in wild sunflowers. In addition, they have practical implications for identifying adaptively important genes or QTLs in trees. This is exemplified by three temperate forest taxa, Populus (poplars, aspens, cottonwoods), Fraxinus (ash), and Quercus (oak). All three are diploid and important genomic tools are under development. Moreover, all three offer extensive hybrid zones whose likely age can be inferred from fossil data. Age data enables estimates of the size and frequency of chromosomal blocks in hybrids, thereby providing guidance in designing marker-based experiments. We predict that natural hybrid zones will be valuable tools for identifying the QTLs and/or candidate genes responsible for adaptive traits in forest trees. PMID- 18677414 TI - Reconstructing patterns of reticulate evolution in plants. AB - Until recently, rigorously reconstructing the many hybrid speciation events in plants has not been practical because of the limited number of molecular markers available for plant phylogenetic reconstruction and the lack of good, biologically based methods for inferring reticulation (network) events. This situation should change rapidly with the development of multiple nuclear markers for phylogenetic reconstruction and new methods for reconstructing reticulate evolution. These developments will necessitate a much greater incorporation of population genetics into phylogenetic reconstruction than has been common. Population genetic events such as gene duplication coupled with lineage sorting and meiotic and sexual recombination have always had the potential to affect phylogenetic inference. For tree reconstruction, these problems are usually minimized by using uniparental markers and nuclear markers that undergo rapid concerted evolution. Because reconstruction of reticulate speciation events will require nuclear markers that lack these characteristics, effects of population genetics on phylogenetic inference will need to be addressed directly. Current models and methods that allow hybrid speciation to be detected and reconstructed are discussed, with a focus on how lineage sorting and meiotic and sexual recombination affect network reconstruction. Approaches that would allow inference of phylogenetic networks in their presence are suggested. PMID- 18677415 TI - Making Sense of Qualitative and Quantitative Findings in Mixed Research Synthesis Studies. AB - The synthesis of qualitative and quantitative research findings is increasingly promoted, but many of the conceptual and methodological issues it raises have yet to be fully understood and resolved. In this article, we describe how we handled issues encountered in efforts to synthesize the findings in forty-two reports of studies of antiretroviral adherence in HIV-positive women in the course of an ongoing study to develop methods to synthesize qualitative and quantitative research findings in common domains of health-related research. Working with these reports underscored the importance of looking past method claims and ideals and directly at the findings themselves, differentiating between aggregative syntheses in which findings are assimilated and interpretive syntheses in which they are configured, and understanding the judgments involved in designating relationships between findings as confirmatory, divergent, or complementary. PMID- 18677417 TI - Speciation analysis of arsenic in biological matrices by automated hydride generation-cryotrapping-atomic absorption spectrometry with multiple microflame quartz tube atomizer (multiatomizer). AB - Analyses of arsenic (As) species in tissues and body fluids of individuals chronically exposed to inorganic arsenic (iAs) provide essential information about the exposure level and pattern of iAs metabolism. We have previously described an oxidation state-specific analysis of As species in biological matrices by hydride-generation atomic absorption spectrometry (HG-AAS), using cryotrapping (CT) for preconcentration and separation of arsines. To improve performance and detection limits of the method, HG and CT steps are automated and a conventional flame-in-tube atomizer replaced with a recently developed multiple microflame quartz tube atomizer (multiatomizer). In this system, arsines from As(III)-species are generated in a mixture of Tris-HCl (pH 6) and sodium borohydride. For generation of arsines from both As(III)- and As(V)-species, samples are pretreated with L-cysteine. Under these conditions, dimethylthioarsinic acid, a newly described metabolite of iAs, does not interfere significantly with detection and quantification of methylated trivalent arsenicals. Analytical performance of the automated HG-CT-AAS was characterized by analyses of cultured cells and mouse tissues that contained mono- and dimethylated metabolites of iAs. The capacity to detect methylated As(III)- and As(V)-species was verified, using an in vitro methylation system containing recombinant rat arsenic (+3 oxidation state) methyltransferase and cultured rat hepatocytes treated with iAs. Compared with the previous HG-CT-AAS design, detection limits for iAs and its metabolites have improved significantly with the current system, ranging from 8 to 20 pg. Recoveries of As were between 78 and 117%. The precision of the method was better than 5% for all biological matrices examined. Thus, the automated HG-CT-AAS system provides an effective and sensitive tool for analysis of all major human metabolites of iAs in complex biological matrices. PMID- 18677418 TI - Tuberculosis Infection: Insight from Immunogenomics. AB - Tuberculosis continues to be one of the most important global infectious causes of morbidity and mortality. Development of a more effective vaccine is a high worldwide priority and depends on a thorough understanding of the host response to infection. In this review, we highlight recent advances in our understanding of the innate immune response to MTb infection. We also describe recent discoveries in immunogenetics that are generating insight into the potential development of immunomodulatory therapies. PMID- 18677419 TI - On diffusion processes with variable drift rates as models for decision making during learning. AB - We investigate Ornstein-Uhlenbeck and diffusion processes with variable drift rates as models of evidence accumulation in a visual discrimination task. We derive power-law and exponential drift-rate models and characterize how parameters of these models affect the psychometric function describing performance accuracy as a function of stimulus strength and viewing time. We fit the models to psychophysical data from monkeys learning the task to identify parameters that best capture performance as it improves with training. The most informative parameter was the overall drift rate describing the signal-to-noise ratio of the sensory evidence used to form the decision, which increased steadily with training. In contrast, secondary parameters describing the time course of the drift during motion viewing did not exhibit steady trends. The results indicate that relatively simple versions of the diffusion model can fit behavior over the course of training, thereby giving a quantitative account of learning effects on the underlying decision process. PMID- 18677420 TI - HUMAN TISSUE LEGISLATION AND MEDICAL PRACTICE: A BENEFIT OR A BURDEN? AB - The scandals surrounding organ removal and retention throughout the United Kingdom provoked several Inquiries and ultimately led to law reform. Although the medical professions were well represented at the Inquiries, little was heard of the voices of those at the 'coal face'. In this scoping study, funded by the Wellcome Trust, we interviewed a number of doctors and others engaged in the uses of human tissue and organs to explore their hopes, concerns and fears about the role of the law in their practices. We found that those involved in transplantation were more aware of, and more actively involve with, the law, whereas others, such as pathologists, had less direct engagement with the law. Most of those we interviewed expressed the hope that law reform would provide much-needed clarity. Although some expressed concern that the law might be over intrusive, most felt that the placing of authority firmly in the hands of the person him or her self to decide what should happen to their bodies was to be welcomed. PMID- 18677421 TI - Empirical assessments of clinical ethics services: implications for clinical ethics committees. AB - The need to evaluate the performance of clinical ethics services is widely acknowledged although work in this area is more developed in the United States. In the USA many studies that assess clinical ethics services have utilized empirical methods and assessment criteria. The value of these approaches is thought to rest on their ability to measure the value of services in a demonstrable fashion. However, empirical measures tend to lack ethical content, making their contribution to developments in ethical governance unclear. The steady increase of clinical ethics committees in the UK must be accompanied by efforts to evaluate their performance. As part of this evaluative work it is important to examine how the practice of clinical ethics committees can be informed by empirical measures. PMID- 18677422 TI - FTIR Microspectroscopy Coupled with Two-Class Discrimination Segregates Markers Responsible for Inter- and Intra-Category Variance in Exfoliative Cervical Cytology. AB - Infrared (IR) absorbance of cellular biomolecules generates a vibrational spectrum, which can be exploited as a "biochemical fingerprint" of a particular cell type. Biomolecules absorb in the mid-IR (2-20 mum) and Fourier-transform infrared (FTIR) microspectroscopy applied to discriminate different cell types (exfoliative cervical cytology collected into buffered fixative solution) was evaluated. This consisted of cervical cytology free of atypia (i.e. normal; n = 60), specimens categorised as containing low-grade changes (i.e. CIN1 or LSIL; n = 60) and a further cohort designated as high-grade (CIN2/3 or HSIL; n = 60). IR spectral analysis was coupled with principal component analysis (PCA), with or without subsequent linear discriminant analysis (LDA), to determine if normal versus low-grade versus high-grade exfoliative cytology could be segregated. With increasing severity of atypia, decreases in absorbance intensity were observable throughout the 1,500 cm(-1) to 1,100 cm(-1) spectral region; this included proteins (1,460 cm(-1)), glycoproteins (1,380 cm(-1)), amide III (1,260 cm(-1)), asymmetric (nu(as)) PO(2) (-) (1,225 cm(-1)) and carbohydrates (1,155 cm(-1)). In contrast, symmetric (nu(s)) PO(2) (-) (1,080 cm(-1)) appeared to have an elevated intensity in high-grade cytology. Inter-category variance was associated with protein and DNA conformational changes whereas glycogen status strongly influenced intra-category. Multivariate data reduction of IR spectra using PCA with LDA maximises inter-category variance whilst reducing the influence of intra class variation towards an objective approach to class cervical cytology based on a biochemical profile. PMID- 18677423 TI - The Use of Structural Equation Modeling in Stuttering Research: Concepts and Directions. AB - This article provides a brief introduction to the history and applications of the class of data analytic techniques collectively known as Structural Equation Modeling (SEM). Using an example based on psychological factors thought to affect the likelihood of stuttering, we discuss the issues of specification, identification, and model fit and modification in SEM. We also address points relating to model specification strategies, item parceling, advanced modeling, and suggestions for reporting SEM analyses. It is noted that SEM techniques can contribute to the elucidation of the developmental pathways that lead to stuttering. PMID- 18677424 TI - Memory for Items and Associations: Distinct Representations and Processes in Associative Recognition. AB - In two experiments, participants studied word pairs and later discriminated old (intact) word pairs from foils, including recombined word pairs and pairs including one or two previously unstudied words. Rather than making old/new memory judgments, they chose one of five responses: (1) Old-Old (original), (2) Old-Old (rearranged), (3) Old-New, (4) New-Old, (5) New-New. To tease apart the effects of item familiarity from those of associative strength, we varied both how many times a specific word-pair was repeated (1 or 5) and how many different word pairs were associated with a given word (1 or 5). Participants could discriminate associative information from item information such that they recognized which word of a foil was new, or whether both were new, as well as discriminating recombined studied words from original pairings. The error and latency data support the view that item and associative information are stored as distinct memory representations and make separate contributions at retrieval. PMID- 18677425 TI - A novel panel of mouse models to evaluate the role of human pregnane X receptor and constitutive androstane receptor in drug response. AB - The pregnane X receptor (PXR) and the constitutive androstane receptor (CAR) are closely related orphan nuclear hormone receptors that play a critical role as xenobiotic sensors in mammals. Both receptors regulate the expression of genes involved in the biotransformation of chemicals in a ligand-dependent manner. As the ligand specificity of PXR and CAR have diverged between species, the prediction of in vivo PXR and CAR interactions with a drug are difficult to extrapolate from animals to humans. We report the development of what we believe are novel PXR- and CAR-humanized mice, generated using a knockin strategy, and Pxr- and Car-KO mice as well as a panel of mice including all possible combinations of these genetic alterations. The expression of human CAR and PXR was in the predicted tissues at physiological levels, and splice variants of both human receptors were expressed. The panel of mice will allow the dissection of the crosstalk between PXR and CAR in the response to different drugs. To demonstrate the utility of this panel of mice, we used the mice to show that the in vivo induction of Cyp3a11 and Cyp2b10 by phenobarbital was only mediated by CAR, although this compound is described as a PXR and CAR activator in vitro. This panel of mouse models is a useful tool to evaluate the roles of CAR and PXR in drug bioavailability, toxicity, and efficacy in humans. PMID- 18677427 TI - Tissue distribution and depuration of the extracted hepatotoxic cyanotoxin microcystins in crucian carp (Carassius carassius) intraperitoneally injected at a sublethal dose. AB - An acute toxicity experiment was conducted by intraperitoneal injection with a sublethal dose of extracted microcystins (MCs), 50 microg MC-LR (where L = leucine and R = arginine) equivalent/kg body weight (BW), to examine tissue distribution and depuration of MCs in crucian carp (Carassius carassius). Liver to body weight ratio increased at 3, 12, 24, and 48 h postinjection compared with that at 0 h (p < 0.05). MC concentrations in various tissues and aquaria water were analyzed at 1, 3, 12, 24, 48, and 168 h postinjection using liquid chromatography coupled with mass spectrometry (LC-MS). The highest concentration of MCs (MC-RR + MC-LR) was found in blood, 2-270 ng/g dry weight (DW), followed by heart (3-100 ng/g DW) and kidney (13-88 ng/g DW). MC levels were relatively low in liver, gonad, intestine, spleen, and brain. MC contents in gills, gallbladder, and muscle were below the limit of detection. Significant negative correlation was present between MC-RR concentration in blood and that in kidney, confirming that blood was important in the transportation of MC-RR to kidney for excretion. Rapid accumulation and slow degradation of MCs were observed in gonad, liver, intestine, spleen, and brain. Only 0.07% of injected MCs were detected in liver. The recovery of MCs in liver of crucian carp seemed to be dose dependent. PMID- 18677426 TI - Pathogenic autoantibodies in systemic lupus erythematosus are derived from both self-reactive and non-self-reactive B cells. AB - Previous studies have shown that both murine and human anti-double-stranded DNA (anti-dsDNA) antibodies can develop from non-DNA-reactive B cells and suggest a crucial role for somatic mutation in dsDNA binding. However, since only a limited number of human anti-dsDNA antibodies have been analyzed previously, we could not exclude other mechanisms for the generation of anti-dsDNA antibodies in patients with systemic lupus erythematosus (SLE). Therefore, we isolated IgM anti-dsDNA antibodies from peripheral blood B cells of a patient with SLE. Three somatically mutated IgM anti-DNA antibodies with pathogenic potential (glomerular binding) were reverted to their germline configuration. Although all three IgM anti-dsDNA antibodies came from the same lupus patient, they displayed different profiles. Reversion to the germline sequence of autoantibodies A9 and B5 resulted in decreased dsDNA binding. In contrast, the germline form of G3-recognized dsDNA as well as the mutated counterpart. These results suggest that mutated IgM anti dsDNA antibodies may develop from both DNA- and non-DNA-reactive B cells. The implications are that B cell activation occurs in response to self and non-self antigens, while selection after activation may be mediated by self antigen in SLE. Moreover, ineffective tolerance checkpoints may exist before and after antigen activation in SLE. PMID- 18677428 TI - Synthesis and biological aspects of mycolic acids: an important target against Mycobacterium tuberculosis. AB - Mycolic acids are an important class of compounds, basically found in the cell walls of a group of bacteria known as mycolata taxon, exemplified by the most famous bacteria of this group, the Mycobacterium tuberculosis (M. tb.), the agent responsible for the disease known as tuberculosis (TB). Mycolic acids are important for the survival of M. tb. For example, they are able to help fight against hydrophobic drugs and dehydration, and also allow this bacterium to be more effective in the host's immune system by growing inside macrophages. Due to the importance of the mycolic acids for maintenance of the integrity of the mycobacterial cell wall, these compounds become attractive cellular targets for the development of novel drugs against TB. In this context, the aim of this article is to highlight the importance of mycolic acids in drug discovery. PMID- 18677429 TI - Antileishmanial activity of aldonamides and N-acyl-diamine derivatives. AB - A number of lipophilic N-acyl-diamines and aldonamides have been synthesized and tested for their in vitro antiproliferative activity against Leishmania amazonensis and L. chagasi. Ribonamides, having one amino group, displayed good to moderate inhibition of parasite growth. The best result was obtained for compounds 10 and 15 with IC50 against L. chagasi below 5 microM. PMID- 18677430 TI - Can the spinal cord learn and remember? AB - Learning and memory traditionally have been associated with cellular processes occurring in a specialized region of the brain called the hippocampus. However, recent data have provided strong evidence to suggest that comparable processes are also expressed in the spinal cord. Experiments performed mainly in spinal cord-transected animals have reported that, indeed, spinal-mediated functions, such as the stretch or flexion reflex, pain signaling, micturition, or locomotion, may undergo plasticity changes associated with partial functional recovery that occur spontaneously or conditionally. Many of the underlying cellular mechanisms strikingly resemble those found in the hippocampus. This mini review reports, mainly, animal data that support the idea that other areas of the central nervous system, such as the spinal cord, can also learn and remember. PMID- 18677431 TI - [Prescription reference: combination of drugs--increased risk of incompatibilities]. PMID- 18677433 TI - Effects of dietary benzo[a]pyrene on growth and hematological parameters in juvenile rockfish, Sebastes schlegeli (Hilgendorf). AB - Experiments were conducted to evaluate the effect of dietary benzo[a]pyrene (B[a]P) on the growth and hematological parameters of rockfish, Sebastes schlegeli, after they were fed with subchronic dietary B[a]P for 30 days. The weight growth rate of the rockfish was significantly different from that of the control group at dietary B[a]P concentration of 1.5 and 2.0 mg/kg. Significant reduction in red blood cells, hematocrit, and hemoglobin were observed during 30 days of exposure to the highest concentration. Over 30 days, serum AST increased at a B[a]P concentration of 1.5 mg/kg and LDH increased at a B[a]P concentration of 1.0 mg/kg concentration. However, no changes were observed in ALT, total protein, magnesium or calcium. PMID- 18677434 TI - The aetiology of prolapse. AB - Female pelvic organ prolapse is a common condition that is said to be multifactorial in aetiology. While a number of risk factors such as vaginal childbirth, obesity and ageing are commonly accepted, it is not clear as to how these risk factors affect the development of prolapse-that is, which pathophysiologic mechanisms are responsible for disease manifestation. Measures used in epidemiological studies, such as presentation for surgical treatment, are generally confounded by other conditions, and evidence is lacking for some of the most frequently quoted aetiological factors. In this paper, I will try to summarise the available evidence in order to separate hearsay and hypothesis from available research findings and to suggest a way forward for diagnosis and treatment. PMID- 18677435 TI - Vaginal smooth muscle cell apoptosis is increased in women with pelvic organ prolapse. AB - To compare the smooth muscle content and apoptosis of the vagina in women with and without anterior vaginal wall prolapse. Vaginal tissues were sampled in women with (n = 6) or without (n = 6) anterior vaginal wall prolapse undergoing hysterectomy. Smooth muscle of the vagina was studied by immunohistochemistry. Digital image analysis was used to determine the fractional area of smooth muscle in the histologic cross-sections. Apoptosis was assessed by TUNEL assay. The fractional area of non-vascular smooth muscle in the vagina of women with anterior vaginal wall prolapse was significantly decreased compared to women without prolapse (0.36 +/- 0.12 vs. 0.16 +/- 0.12 P = 0.021) and the apoptotic index was significantly higher compared to women without prolapse (0.04 +/- 0.01 vs. 0.02 +/- 0.03, P = 0.041). The fraction of smooth muscle in the vagina is significantly decreased and the rate of apoptosis is higher in women with anterior vaginal wall prolapse compared to women without prolapse. PMID- 18677436 TI - Clinical, angiographic and cardiovascular magnetic resonance findings in consecutive patients with Takotsubo cardiomyopathy. AB - BACKGROUND: The aim was to assess clinical, angiographic and cardiovascular magnetic resonance (CMR) findings in patients with Takotsubo cardiomyopathy. METHODS: Between 2003 and 2007, 20 consecutive patients admitted to our hospital with suspected acute myocardial infarction and presenting with apical ballooning in the left ventricular (LV) angiogram in the absence of a significant coronary artery disease, were included in the study. Echocardiography and CMR was performed in all patients. RESULTS: The mean age of patients with Takotsubo cardiomyopathy was 62 +/- 8 years (range 43-78 years). Eighteen (90%) were female. Clinical presentations included chest pain (95%) and cardiogenic shock (5%). The mean angiographic LV ejection fraction on admission was 45% +/- 9% (range 26%-60%) and resolved rapidly in all cases. Mean time delay between presentation CMR was 2 +/- 1 days (range 1-6 days). Mean ejection fraction was 51% +/- 15% (range 25%-81%). While 19 (95%) patients showed no evidence of late enhancement or signs of myocarditis in the CMR, 1 (5%) patient who was resuscitated showed hyperenhancement confined to the apex. CONCLUSION: In patients showing the clinical picture of an acute myocardial syndrome and angiographic picture of a TakoTsubo cardiomyopathy, CMR might be helpful in confirming the diagnosis through the exclusion of other causes for the acute LV dysfunction. PMID- 18677437 TI - Lithium treatment of trichotillomania with comorbid bipolar II disorder. PMID- 18677438 TI - Experiences of immigrant new mothers with symptoms of depression. AB - Refugee, asylum seeking, non-refugee, and immigrant new mothers with depressive symptoms, were interviewed in a qualitative study to better understand their (a) experiences and attributions of depressive symptoms, (b) their experiences with health care providers and support services, (c) factors that facilitated or hindered help seeking, (d) factors that aided recovery or (e) were associated with women continuing to experience symptoms of depression. Ten immigrant new mothers who scored 10 or over on the Edinburgh Postpartum Depression Scale shortly after giving birth, participated 12 to 18 months later in postpartum semi structured interviews which were taped, transcribed and analyzed using a constant comparative approach. Many women attributed their depressive symptoms to social isolation, physical changes, feeling overwhelmed and financial worries. They had poor knowledge of community services. Barriers to care included stigma, embarrassment, language, fear of being labeled an unfit mother, or the attitude of some staff. Facilitators to recovery included social support from friends, partners and family, community support groups, "getting out of the house", or personal psychological adjustment. Personal and systematic barriers exist in new immigrant mothers obtaining care for symptoms of depression. Suggestions for improvements are offered. PMID- 18677440 TI - Effect of fluctuations in salinity on anaerobic biomass and production of soluble microbial products (SMP(s)). AB - This study investigated the acclimation potential of batch fed anaerobic biomass with salinities of 0-50 gNaCl l(-1). Anaerobic biomass was acclimatized to salinities up to 20 gNaCl l(-1)over a period of 35 days, with 3 consecutive feedings. After this period the biomass was subjected to non-saline conditions to simulate fluctuating feed compositions. High activity was obtained after the first exposure to non saline conditions for biomass previously exposed to 30 gNaCl l(-1). Short exposure (2-48 h) to high salinity (40 gNaCl l(-1)) did not reduce biomass activity when it was re-subjected to normal conditions. The sensitivity of each anaerobic bacterial group showed that propionate utilisers were the most affected by sudden changes in salinity. Using size exclusion chromatography (SEC) it was found that biomass exposed to concentrations of salinity above 30 gNaCl(-1) produced higher molecular weight soluble microbial products (SMPs) which were present in the culture for longer periods than the control indicating that the effluent was more difficult to degrade. With the sudden removal of salinity anaerobic biomass can easily readapt to normal conditions without any high MW compounds being produced. These findings highlight the fact that anaerobic biomass is able to overcome sharp decreases in salinity in contrast with aerobic biomass as reported in the literature. PMID- 18677441 TI - Effect of continuous Cd feeding on the performance of a nitrification reactor. AB - The inhibitory effect of Cd on nitrification was investigated in a continuous flow system with enriched nitrifying bacteria. The maximum specific ammonium utilization rate and the half-saturation constant were found as 671 mg NH(4)-N/g VSS day and 0.48 mg/l, respectively. In the case of continuous Cd input at 1 and 2.5 mg/l, nitrification was inhibited by 30% and 47%, respectively. Inhibition ranged from 20% to 40% and no further increase in inhibition was exhibited in new runs except at 10 mg/l influent Cd. At 10 mg/l influent Cd, specific ammonium utilization and nitrate production rates were inhibited by 90%. On the contrary, a serious nitrite accumulation was not observed during this period. When Cd feeding was stopped, recovery from inhibition was observed after 37 day which was seen by the improvement in ammonium utilization and nitrate production rates. A shift in microbial population from the initial Nitrosomonas sp. to the Cd tolerant Nitrosospira sp. was observed in the recovery period from severe Cd inhibition. After the domination of Nitrosospira species, redosing at 10 mg/l and then at 15 mg/l did not affect the performance as before. PMID- 18677442 TI - Spotted fever group Rickettsia in ticks from southeastern Spain natural parks. AB - During an 8-years study, we collected from vegetation or domestic and wild mammals 1246 ticks (624 males, 511 females and 111 nymphs) belonging to 13 species in Jaen province (Andalusia) and we analyzed these ticks by PCR and sequencing for the presence of rickettsiae. Specific rickettsiae DNA was detected in 243 (19.5%) of the ticks tested. Sequence analysis of amplicons of gltA, ompA and ompB genes revealed that Ixodes ricinus were infected with R. monacensis, including strain IRS3, and R. helvetica (prevalences of 27.0% and 2.7%, respectively), while in I. ventalloi we found only this last species (12.5%). Moreover, Dermacentor marginatus presents R. slovaca (24.7%) and R. raoultii (59.9%). In Rhipicephalus sanguineus group ticks (Rh. sanguineus, Rh. turanicus and Rh. pusillus) only R. massiliae (15.2%) was found. Haemaphysalis punctata and Ha. sulcata were infected with a Rickettsia sp. near R. hoogstraalii (prevalence of 3.1% and 16.1%, respectively). In addition, Ha. punctata appeared infected with R. monacensis-like Rickettsia (1.0%) and R. raoultii (9.3%). None of I. hexagonus, Hyalomma lusitanicum, Hyalomma sp., Ha. hispanica or Rh. bursa studied ticks contained rickettsiae. PMID- 18677443 TI - Clinical and molecular analysis of 49 patients with X-linked agammaglobulinemia from a single center in Argentina. AB - INTRODUCTION: Argentina has a large number of patients with definite diagnosis of X-linked agammaglobulinemia reported in the Latin-American registry. Forty-nine of them were seen in our referral pediatric hospital, between 1987 and 2005. RESULTS AND DISCUSSION: A retrospective study of clinical, laboratory, and molecular data showed that respiratory tract infections were the most frequent initial clinical presentation and the most common among all manifestations prior to diagnosis (69%). Up to diagnosis, we found a high frequency of severe infections (sepsis, 14% and meningitis, 16%) and a high proportion of patients with chronic lung disease. During follow-up, the development of chronic lung disease was significantly related with age at diagnosis and inappropriate treatment. CONCLUSION: Although molecular diagnosis has been available in our center for the past 10 years, there is no doubt that awareness for early recognition of immunodeficiency should be improved through broader and more comprehensive education programs emphasizing characteristics of patients with immunodeficiencies. PMID- 18677444 TI - Common variable immunodeficiency in horses is characterized by B cell depletion in primary and secondary lymphoid tissues. AB - INTRODUCTION: Common variable immunodeficiency (CVID) in horse patients is characterized by late-onset B cell lymphopenia or depletion, hypo- or agammaglobulinemia, impaired humoral response to tetanus toxoid vaccination, and recurrent fevers and bacterial infections. DISCUSSION: This study describes the clinical and immunologic findings of 14 affected horses (average age 10.7 +/- 4.4 years) of both genders (six females, eight males) and different breeds (eight Thoroughbreds, four Quarter Horses, one Warmblood, one Pony). Serial immunological testing in peripheral blood revealed persistent, severe B cell lymphopenia (mean 1.3 +/- 2.3% positive cells) in all patients. Serum IgG (range <200 to 800 mg/dL) and IgM (0.05). Most patients (88.9%) felt disturbed by their voice problem in a total of 118 different daily situations. CONCLUSIONS: The way patients experience voice disorders does not depend on the origin of the disorder and not on the gender. The data allow the conclusion that the case history of dysphonic patients can be assessed by a structured voice interview. PMID- 18677456 TI - [Pancreatic carcinoma]. AB - Ductal adenocarcinomas represent over 90% of all pancreatic cancers. The high mortality has not changed over the last decades. Most patients are elderly and typically present with dull upper abdominal pain radiating to the back and often report reduced appetite, weight loss, and jaundice. A solid pancreatic tumor should be resected because this is the only chance for a cure. Cystic tumors are usually less malignant and need further diagnostic work-up. There is a benefit from adjuvant chemotherapy after resection. Locally advanced or metastatic tumors are treated symptomatically including pain medication and biliary drainage in the case of jaundice. There is a benefit from systemic chemotherapy mainly in patients with good performance status. PMID- 18677461 TI - Genetic analysis of photoperiod sensitivity in a tropical by temperate maize recombinant inbred population using molecular markers. AB - Photoperiod sensitivity is an important consideration in maize cultivation. Flowering time is affected by photoperiod and sensitivity to it limits the potential for successful exchange of germplasm across different latitudes. For resolving the genetic basis of photoperiod sensitivity in maize, a set of 207 recombinant inbred lines derived from a temperate and tropical inbred line cross was evaluated for 2 years in a long-day and short-day environment. Genetic linkage maps were constructed using 237 SSR markers with a total length 1,974.3 cM, and an average space between two makers of 8.33 cM. Twenty-nine QTL were detected for the five measured photoperiod sensitivity traits using composite interval mapping and multiple interval mapping. QTL for flowering time, plant height and leaf number, under long-day conditions, were found clustered on chromosome 10, while QTL for short-day conditions resided on chromosome 3. The QTL in the bin 10.04 region of chromosome 10 were detected associated with photoperiod sensitivity and related traits during long days. These results indicated that this region might contain an important photoperiod sensitivity element. PMID- 18677462 TI - [Biomechanical issues in patellofemoral joint rehabilitation]. AB - Specific knowledge of the biomechanics of the patellofemoral joint is crucial for successful nonoperative or postsurgical rehabilitation. The biomechanical aspects of different situations should be considered when designing an exercise program. Joint reaction forces, contact area, and contact stress are dependent on flexion angle and exercise situations. In weight-bearing activities, the amount of knee flexion directly influences the magnitude of quadriceps muscle force, which affects the magnitude of patellofemoral joint reaction forces. Open and closed chain exercises should be performed within a safe range of motion to allow quadriceps activation while minimizing patellofemoral joint reaction forces. The isolated knee extension (90 degrees -40 degrees ), the squat (0 degrees -30 degrees -60 degrees ), and the leg press (0 degrees -30 degrees -60 degrees ) are the three main exercise situations in the acute rehabilitation phase. Controlled body positions and low levels of pain and symptoms should also be emphasized to achieve a functional progression, focusing on neuromuscular control. PMID- 18677463 TI - Aryl hydrocarbon receptor-mediated regulation of the human estrogen and bile acid UDP-glucuronosyltransferase 1A3 gene. AB - UDP-glucuronosyltransferases contribute to the detoxification of drugs by forming water soluble beta-D-glucopyranosiduronic acids. The human UGT1A3 protein catalyzes the glucuronidation of estrogens, bile acids and xenobiotics including non-steroidal anti-inflammatory drugs and lipid lowering drugs. Regulation of UGT1A3 by xenobiotic response elements is likely, but the responsible elements are yet uncharacterized. In addition, genetic promoter variants may affect UGT1A3 regulation and potential induction by xenobiotics. The UGT1A3 promoter was analyzed by mutagenesis, reporter gene, and mobility shift analyses. Three hundred and eighty-nine blood donors were genotyped for promoter single nucleotide polymorphisms (SNPs) showing an allelic frequency of 42% of variants at -66 (T to C) and -204 (A to G). A xenobiotic response element regulating aryl hydrocarbon receptor (AhR)-mediated UGT1A3 transcription was identified and characterized. UGT1A3 transcription was reduced in the presence of promoter SNPs. These data demonstrate xenobiotic induced regulation of the UGT1A3 gene by the AhR, which shows genetic variability. PMID- 18677464 TI - Development of saw palmetto (Serenoa repens) fruit and extract standard reference materials. AB - As part of a collaboration with the National Institutes of Health's Office of Dietary Supplements and the Food and Drug Administration's Center for Drug Evaluation and Research, the National Institute of Standards and Technology has developed two standard reference materials (SRMs) representing different forms of saw palmetto (Serenoa repens), SRM 3250 Serenoa repens fruit and SRM 3251 Serenoa repens extract. Both of these SRMs have been characterized for their fatty acid and phytosterol content. The fatty acid concentration values are based on results from gas chromatography with flame ionization detection (GC-FID) and mass spectrometry (GC/MS) analysis while the sterol concentration values are based on results from GC-FID and liquid chromatography with mass spectrometry analysis. In addition, SRM 3250 has been characterized for lead content, and SRM 3251 has been characterized for the content of beta-carotene and tocopherols. SRM 3250 (fruit) has certified concentration values for three phytosterols, 14 fatty acids as triglycerides, and lead along with reference concentration values for four fatty acids as triglycerides and 16 free fatty acids. SRM 3251 (extract) has certified concentration values for three phytosterols, 17 fatty acids as triglycerides, beta-carotene, and gamma-tocopherol along with reference concentration values for three fatty acids as triglycerides, 17 fatty acids as free fatty acids, beta carotene isomers, and delta-tocopherol and information values for two phytosterols. These SRMs will complement other reference materials currently available with concentrations for similar analytes and are part of a series of SRMs being developed for dietary supplements. PMID- 18677465 TI - Electroanalytical study of proflavine intercalation in 5-methyl or inosine containing amplicons. AB - Amplicons corresponding to the GC-rich p53 exon 5 and its analogues, synthesized by substituting 60% of cytosine by 5-methyl-cytosine, or 60% of guanosine by inosine and GC-poor p53 exon 6 were synthesized and investigated electrochemically, in the presence and absence of proflavine, by differential pulse voltammetry (DPV). Incorporation of base analogues and the thermal stability of the resulting amplicons were tested in the presence of a fluorescent probe (Sybr-Green). Peak current at 1.0 V was lower for methylated than for unmethylated PCR amplicons and was similarly affected by proflavine intercalation. In contrast, considerable peak current differences were observed in the presence of proflavine for unmodified exon 5 v.s. exon 6 or inosine containing amplicons. Thermal analysis verified the expected shifts in melting temperature (T (m)) due to the base analogue incorporation and GC-content variations. In conclusion, methylated and unmethylated PCR amplicons could be distinguished in model DNA systems using differential pulse voltammetry (DPV) and use of proflavine could serve as an electrochemical probe for identifying different DNA conformations. PMID- 18677467 TI - A paradoxical signal intensity increase in fatty livers using opposed-phase gradient echo imaging with fat-suppression pulses. AB - With the increase in obese and overweight children, nonalcoholic fatty liver disease has become more prevalent in the pediatric population. Appreciating subtleties of magnetic resonance (MR) signal intensity behavior from fatty livers under different imaging conditions thus becomes important to pediatric radiologists. We report an initially confusing signal behavior-increased signal from fatty livers when fat-suppression pulses are applied in an opposed-phase gradient echo imaging sequence-and seek to explain the physical mechanisms for this paradoxical signal intensity behavior. Abdominal MR imaging at 3 T with a 3 D volumetric interpolated breath-hold (VIBE) sequence in the opposed-phase condition (TR/TE 3.3/1.3 ms) was performed in five obese boys (14+/-2 years of age, body mass index >95th percentile for age and sex) with spectroscopically confirmed fatty livers. Two VIBE acquisitions were performed, one with and one without the use of chemical shift selective (CHESS) pulse fat suppression. The ratios of fat-suppressed over non-fat-suppressed signal intensities were assessed in regions-of-interest (ROIs) in five tissues: subcutaneous fat, liver, vertebral marrow, muscle and spleen. The boys had spectroscopically estimated hepatic fat levels between 17% and 48%. CHESS pulse fat suppression decreased subcutaneous fat signals dramatically, by more than 85% within regions of optimal fat suppression. Fatty liver signals, in contrast, were elevated by an average of 87% with CHESS pulse fat suppression. Vertebral marrow signal was also significantly elevated with CHESS pulse fat suppression, while spleen and muscle signals demonstrated only small signal increases on the order of 10%. We demonstrated that CHESS pulse fat suppression actually increases the signal intensity from fatty livers in opposed-phase gradient echo imaging conditions. The increase can be attributed to suppression of one partner of the opposed-phase pair that normally contributes to the destructive interference between water and fat. The result is a paradoxical increase in signal from fatty liver that will depend on both fat content and the relative longitudinal relaxation times of fat methylene protons and water. PMID- 18677468 TI - High throughput functional screening of an ion channel library for drug safety and efficacy. AB - Development of a large, representative library of ion channel-expressing cell lines is described. Validation on a full range of automated patch clamp and fluorescence illumination platforms is ongoing. Library "books" can be mixed and matched into channel panels according to tissue, therapeutic area and ion channel family. Unexpected results using a cardiac channel panel show that this panel may serve as a biomarker for cardiac risk assessment. PMID- 18677469 TI - Mast cell hyperplasia in the skin of Dsg4-deficient hypotrichosis mice, which are long-living mutants of lupus-prone mice. AB - Desmosomal cadherins are essential cell adhesion molecules expressed in the epidermis. We identified a mutation of a cadherin superfamily member, namely, desmoglein 4 (Dsg4), in early onset of death (EOD)( hage ) mice with hypotrichosis. The mutation was induced by the insertion of an early transposon II-beta into intron 8 of Dsg4. Mast cell hyperplasia was observed in the skin of EOD( hage ) mice. The abnormally expanded population of lpr T cells, i.e., CD4( )CD8(-)B220(+)Thy1.2(+) alphabetaT cells, in the splenocytes of EOD mice was reduced in EOD( hage ) mice. Therefore, it was suspected that the long-living mutant EOD( hage ) mice were selected from lupus-prone EOD mice because of their immunological immaturity. These findings clearly indicate that Dsg4 is an important molecule for the formation of hair follicles and hypothesize that unorganized hyperplastic hair follicles in anagen due to the Dsg4 mutation provide niches for mast cell precursors in the skin. PMID- 18677470 TI - Conjugation of protein antigen to microparticulate beta-glucan from Saccharomyces cerevisiae: a new adjuvant for intradermal and oral immunizations. AB - Immunostimulatory glucose polymers known as beta-glucans have been studied for many years. Our laboratory has prepared and characterized a novel microparticulate beta-glucan (MG) from the budding yeast Saccharomyces cerevisiae. Because MG particles are rapidly phagocytized by murine peritoneal macrophages and induce the expression of B7 costimulatory molecules, we hypothesized that MG could serve as a vaccine adjuvant to enhance specific immune responses. Here, we describe a procedure for conjugating the test vaccine antigen bovine serum albumin (BSA) to MG via water-soluble carbodiimide linkage. Conjugates with up to 0.4 mg of BSA/mg MG were prepared. MG/BSA conjugates were still actively phagocytized by mouse peritoneal macrophages. When used to immunize mice by the intradermal route, these conjugates enhanced the primary IgG antibody response to BSA in a manner comparable to the prototypic complete Freund's adjuvant. Although primary oral immunization with MG/BSA caused no increase in serum anti-BSA antibody titers, booster immunization elicited a significant anti-BSA antibody response. These results suggest that protein antigens can be conjugated to MG via a carbodiimide linkage and that these conjugates provide an adjuvant effect for stimulating the antibody response to the protein antigens. PMID- 18677472 TI - Growth of Salinispora tropica strains CNB440, CNB476, and NPS21184 in nonsaline, low-sodium media. AB - We recently described the development of a potassium-chloride-based salt formulation containing low sodium concentration (5.0 mM) to support the growth of Salinispora tropica strain NPS21184 and its production of salinosporamide A (NPI 0052). In order to determine whether the above low-sodium salt formulation can also support the growth of other S. tropica strains, we examined the growth of the type strain CNB440 and the parent strain CNB476, from which strain NPS21184 was derived as a single colony isolate. We demonstrated that good growth rate and yield of S. tropica strains CNB440 and CNB476, similar to S. tropica strain NPS21184 reported earlier, were detected in both agar and liquid media containing the potassium-chloride-based salt formulation with sodium concentration of 5.0 mM. Furthermore, we also detected good growth rate and yield of all three S. tropica strains on potassium-sulfate-based salt formulation agar medium containing both low-sodium (5.7 mM) and low-chloride (14 mM) content. This finding confirms the observation that the species of S. tropica does not have a seawater growth requirement but requirement for a specific combination of salts to provide a balance of salts and maintain a high enough ionic strength for growth. PMID- 18677471 TI - Reduction of volatile acidity of wines by selected yeast strains. AB - Herein, we isolate and characterize wine yeasts with the ability to reduce volatile acidity of wines using a refermentation process, which consists in mixing the acidic wine with freshly crushed grapes or musts or, alternatively, in the incubation with the residual marc. From a set of 135 yeast isolates, four strains revealed the ability to use glucose and acetic acid simultaneously. Three of them were identified as Saccharomyces cerevisiae and one as Lachancea thermotolerans. Among nine commercial S. cerevisiae strains, strains S26, S29, and S30 display similar glucose and acetic acid initial simultaneous consumption pattern and were assessed in refermentation assays. In a medium containing an acidic wine with high glucose-low ethanol concentrations, under low oxygen availability, strain S29 is the most efficient one, whereas L. thermotolerans 44C is able to decrease significantly acetic acid similar to the control strain Zygosaccharomyces bailii ISA 1307 but only under aerobic conditions. Conversely, for low glucose-high ethanol concentrations, under aerobic conditions, S26 is the most efficient acid-degrading strain, while under limited-aerobic conditions, all the S. cerevisiae strains studied display acetic acid degradation efficiencies identical to Z. bailii. Moreover, S26 strain also reveals capacity to decrease volatile acidity of wines. Together, the S. cerevisiae strains characterized herein appear promising for the oenological removal of volatile acidity of acidic wines. PMID- 18677473 TI - PCR-mediated one-step deletion of targeted chromosomal regions in haploid Saccharomyces cerevisiae. AB - Chromosome rearrangements, especially chromosomal deletions, have been exploited as important resources for functional analysis of genomes. To facilitate this analysis, we applied a previously developed method for chromosome splitting for the direct deletion of a designed internal or terminal chromosomal region carrying many nonessential genes in haploid Saccharomyces cerevisiae. The method, polymerase chain reaction (PCR)-mediated chromosomal deletion (PCD), consists of a two-step PCR and one transformation per deletion event. In this paper, we show that the PCD method efficiently deletes internal regions in a single transformation. Of the six chromosomal regions targeted for deletion by this method, five regions (16 to 38 kb in length) containing 10 to 19 nonessential genes were successfully eliminated at high efficiency. The one targeted region on chromosome XIII that was not deleted was subsequently found to contain sequences essential for yeast growth. While 14 individual genes in this region have been reported to be nonessential, synthetic lethal interactions may occur among these nonessential genes. Phenotypic analysis showed that four deletion strains still exhibited normal growth while possible synthetic growth defects were observed in another strain harboring a 19-gene deletion on chromosome XV. These results demonstrate that the PCD method is a useful tool for deleting genes and for analyzing their functions in defined chromosomal regions. PMID- 18677474 TI - Microbiological analysis of the population of extremely haloalkaliphilic sulfur oxidizing bacteria dominating in lab-scale sulfide-removing bioreactors. AB - Thiopaq biotechnology for partial sulfide oxidation to elemental sulfur is an efficient way to remove H(2)S from biogases. However, its application for high pressure natural gas desulfurization needs upgrading. Particularly, an increase in alkalinity of the scrubbing liquid is required. Therefore, the feasibility of sulfide oxidation into elemental sulfur under oxygen limitation was tested at extremely haloalkaline conditions in lab-scale bioreactors using mix sediments from hypersaline soda lakes as inoculum. The microbiological analysis, both culture dependent and independent, of the successfully operating bioreactors revealed a domination of obligately chemolithoautotrophic and extremely haloalkaliphilic sulfur-oxidizing bacteria belonging to the genus Thioalkalivibrio. Two subgroups were recognized among the isolates. The subgroup enriched from the reactors operating at pH 10 clustered with Thioalkalivibrio jannaschii-Thioalkalivibrio versutus core group of the genus Thioalkalivibrio. Another subgroup, obtained mostly with sulfide as substrate and at lower pH, belonged to the cluster of facultatively alkaliphilic Thioalkalivibrio halophilus. Overall, the results clearly indicate a large potential of the genus Thiolalkalivibrio to efficiently oxidize sulfide at extremely haloalkaline conditions, which makes it suitable for application in the natural gas desulfurization. PMID- 18677475 TI - Targeting 11q23 positive acute leukemia cells with high molecular weight-melanoma associated antigen-specific monoclonal antibodies. AB - BACKGROUND: Acute leukemia with 11q23 aberrations is associated with a poor outcome with therapy. The lack of efficacy of conventional therapy has stimulated interest in developing novel strategies. Recent studies have shown that 11q23 positive acute leukemia cells express the high molecular weight-melanoma associated antigen (HMW-MAA). This tumor antigen represents a useful target to control growth of human melanoma tumors in patients and in severe combined immunodeficient (SCID) mice, utilizing antibody-based immunotherapy. This effect appears to be mediated by inhibition of the HMW-MAA function such as triggering of the focal adhesion kinase/proline-rich tyrosine kinase 2 (Pyk2) pathways. Therefore, in this study we tested whether HMW-MAA-specific monoclonal antibodies (mAb) could inhibit growth of 11q23-positive leukemia cells in SCID mice. METHODS: HMW-MAA-specific mAb were tested for their ability to inhibit the in vitro proliferation of an 11q23-positive acute myeloid leukemia (AML) cell line and blasts from four patients with 11q23 aberrations and their in vivo growth in subcutaneous and disseminated xenograft models. RESULTS: The HMW-MAA-specific mAb did not affect in vitro proliferation although they down-regulated phosphorylated (P) Pyk2 expression. Furthermore, the mAb enhanced the in vitro anti proliferative effect of cytarabine. In vivo the mAb inhibited the growth of leukemic cells in a dose-dependent fashion. However, the difference did not reach statistical significance. No effect was detected on P-Pyk2 expression. Furthermore, HMW-MAA-specific mAb in combination with cytarabine did not improve tumor inhibition. Lastly, the combination of two mAb which recognize distinct HMW MAA determinants had no detectable effect on survival in a disseminated xenograft model. CONCLUSIONS: HMW-MAA-specific mAb down-regulated P-Pyk2 expression and enhanced the anti-proliferative effect of cytarabine in vitro, but had no detectable effect on survival or growth of leukemia cells in vivo. Whether the HMW-MAA-specific mAb can be used as carriers of toxins or chemotherapeutic agents against 11q23-acute leukemia remains to be determined. PMID- 18677476 TI - cDNA clone, prokaryotic expression and purification of human interleukin-13 receptor [alpha]2 chain. AB - Despite advances in surgical technology and radiation therapy, the prognosis in the patients with malignant glioma remains poor. Recent studies show that interleukin-13 receptor [alpha]2 chain (IL-13Ra2), a brain tumor-associated receptor for IL-13, may play a role in immunotherapy for glioblastoma. We thus amplified human IL-13Ra2 gene from the human glioblastoma cell line using RT-PCR and cloned the target gene into the pET-28a, a prokaryotic expressing plasmid. After transformation, the recombinant plasmid expressed a soluble protein induced by IPTG. The purified recombinant protein was shown to be a single band on the SDS-PAGE with a predicated molecular weight of human IL-13Ra2 gene, suggesting that the recombinant protein of human IL-13Ra2 was successfully expressed. Recombinant IL-13Ra2 protein can be used as an anti-tumor vaccine, which may provide a promising new strategy for the treatment of brain malignant gliomas. PMID- 18677477 TI - Targeting of epidermal Langerhans cells with antigenic proteins: attempts to harness their properties for immunotherapy. AB - Langerhans cells, a subset of skin dendritic cells in the epidermis, survey peripheral tissue for invading pathogens. In recent functional studies it was proven that Langerhans cells can present exogenous antigen not merely on major histocompatibility complexes (MHC)-class II molecules to CD4+ T cells, but also on MHC-class I molecules to CD8+ T cells. Immune responses against topically applied antigen could be measured in skin-draining lymph nodes. Skin barrier disruption or co-application of adjuvants was required for maximal induction of T cell responses. Cytotoxic T cells induced by topically applied antigen inhibited tumor growth in vivo, thus underlining the potential of Langerhans cells for immunotherapy. Here we review recent work and report novel observations relating to the potential use of Langerhans cells for immunotherapy. We investigated the potential of epicutaneous immunization strategies in which resident skin dendritic cells are loaded with tumor antigen in situ. This contrasts with current clinical approaches, where dendritic cells generated from progenitors in blood are loaded with tumor antigen ex vivo before injection into cancer patients. In the current study, we applied either fluorescently labeled protein antigen or targeting antibodies against DEC-205/CD205 and langerin/CD207 topically onto barrier-disrupted skin and examined antigen capture and transport by Langerhans cells. Protein antigen could be detected in Langerhans cells in situ, and they were the main skin dendritic cell subset transporting antigen during emigration from skin explants. Potent in vivo proliferative responses of CD4+ and CD8+ T cells were measured after epicutaneous immunization with low amounts of protein antigen. Targeting antibodies were mainly transported by langerin+ migratory dendritic cells of which the majority represented migratory Langerhans cells and a smaller subset the new langerin+ dermal dendritic cell population located in the upper dermis. The preferential capture of topically applied antigen by Langerhans cells and their ability to induce potent CD4+ and CD8+ T cell responses emphasizes their potential for epicutaneous immunization strategies. PMID- 18677478 TI - Recognition of NY-ESO-1+ tumor cells by engineered lymphocytes is enhanced by improved vector design and epigenetic modulation of tumor antigen expression. AB - The therapeutic use of T cell receptor (TCR)-transduced peripheral blood lymphocytes (PBL) targeting tumor-associated antigens is emerging as a promising investigational treatment for patients with cancer. Initial response rates to therapy were low, suggesting the need to improve the function of TCR-transduced PBL. We constructed standard bicistronic retroviral vectors using an internal promoter or internal ribosomal entry site element as well as vectors incorporating coding sequences for 2A linker peptides between coding sequences for alpha and beta chains targeting the cancer-testis (CT) antigen, NY-ESO-1. Incorporation of coding sequences for 2A linker peptides in the bicistronic TCR expression cassette resulted in up to a fourfold increase in TCR expression and a significant improvement in effector function as measured by interferon-gamma release following co-culture with peptide-pulsed targets and NY-ESO-1+ tumors. We also sought to enhance reactivity of TCR-transduced PBL against tumor targets by modulation of tumor antigen expression on target cells. Induction of NY-ESO-1 expression on tumor targets using the demethylating agent 5-aza-2'-deoxycytidine (alone or in combination with the histone deacetylase inhibitor depsipeptide) resulted in enhanced interferon-gamma secretion by the TCR-transduced PBL on culture with treated targets. Taken together, these results indicate that design of TCR-based vectors incorporating 2A linker peptides improves TCR expression and effector function of transduced PBL. Furthermore, induction of CT antigen expression through treatment of tumor targets with chromatin-remodeling agents may augment TCR-based immunotherapy targeting these antigens. These results have relevance for TCR-based gene therapies targeting common epithelial malignancies. PMID- 18677479 TI - Generation of a human IgM monoclonal antibody directed against HLA class II molecules: a potential agent in the treatment of haematological malignancies. AB - Major histocompatibility complex (MHC) class II molecules have been considered as a good target molecule for use in immunotherapy, because of the high expression in some lymphoma and leukaemia cells and, also, because of their restricted expression on human cells (monocytes, dendritic, B lymphocytes, thymic epithelial cells, and some cytokine-activated cells, such as T lymphocytes). We have obtained a human IgM monoclonal antibody directed against human leukocyte antigen (HLA) class II molecules, using transgenic mice carrying human Ig genes. The antibody BH1 (IgM/kappa isotype) recognises HLA-class II on the surface of tumour cells from patients suffering from haematological malignancies, such as chronic and acute lymphocytic leukaemias, non-Hodgkin lymphomas and myeloid leukaemias. Interestingly, functional studies revealed that BH1 mAb recognises and kills very efficiently tumour cells from several leukaemia patients in the presence of human serum as a source of complement. These results suggest that this human IgM monoclonal antibody against HLA-class II could be considered as a potential agent in the treatment of several malignancies. PMID- 18677480 TI - Leg length discrepancy in total hip arthroplasty: comparison of two methods of measurement. AB - Measurement of leg length discrepancy is an important part in planning a successful total hip arthroplasty (THA). Many clinical and radiological methods with variable degrees of accuracy have been advocated to carry out this measurement. We studied the accuracy of a commonly used clinical method by comparing it to a well-known and reliable radiological method. A total of 139 patients aged 44-89 (mean: 67.5 years) scheduled to undergo THA were examined for clinical and radiological leg length discrepancy measurements before and after the operation by the same observers. There was a poor correlation between the clinical and radiological methods preoperatively [r = 0.21, intra-class correlation coefficient (ICC) = 0.33]. The correlation was better postoperatively (r = 0.45, ICC = 0.62). The clinical method used is not recommended for leg length discrepancy measurement preoperatively. Caution should even be taken when using this method postoperatively. The authors recommend using the radiological method when measuring leg length discrepancy as a part of planning for THA. PMID- 18677481 TI - Surgical management of unstable elbow dislocation without intra-articular fracture. AB - The purpose of this study was to describe soft tissue injury patterns and report the clinical results of primary ligament repair with use of protected early mobilisation in unstable elbow dislocations with pure capsulo-ligamentous injuries. Twenty-four patients who presented with traumatic unstable elbow dislocation without associated intra-articular fracture were reviewed. Anatomical repair was performed using metal anchor screws and the bone tunnel method. Ligament avulsion was noted in 55% for the medial collateral ligament, 80% for the lateral collateral ligament, 60% for the flexor tendon and 80% for the extensor tendon. The overall mean Mayo Elbow Performance Score was 93.2. Brachial artery injuries occurred in two elbows. Heterotopic calcification was noted in 14 patients and there was one severe traumatic arthrosis. This study showed a high incidence of reattachable avulsion injuries to ligaments, tendon/muscle and capsule in unstable elbow dislocations. Primary ligament repair coupled with early rehabilitation provided satisfactory outcomes at two to four years postoperatively. PMID- 18677482 TI - Non-nasal natural killer cell lymphoma: not non-nasal after all. PMID- 18677483 TI - Protective effect of aged garlic extract (AGE) on the apoptosis of intestinal epithelial cells caused by methotrexate. AB - PURPOSE: Methotrexate (MTX) causes intestinal damage, resulting in diarrhea. The side effects often disturb the cancer chemotherapy. We previously reported that AGE protected the small intestine of rats from the MTX-induced damage. In the present paper, the mechanism of the protection of AGE against the MTX-induced damage of small intestine was investigated, using IEC-6 cells originating from rat jejunum crypt. METHODS: The viability and apoptosis of IEC-6 cells were examined in the presence of MTX and/or AGE. RESULTS: The viability of IEC-6 cells exposed to MTX was decreased by the increase of MTX concentration. The MTX induced loss of viable IEC-6 cells was almost completely prevented by the presence of more than 0.1% AGE. In IEC-6 cells exposed to MTX, the cromatin condensation, DNA fragmentation, caspase-3 activation and cytochrome c release were observed. These were preserved to the control levels by the presence of AGE. MTX markedly decreased intracellular GSH in IEC-6 cells, but the presence of AGE in IEC-6 cells with MTX preserved intracellular GSH to the control level. IEC-6 cells in G2/M stage markedly decreased 72 h after the MTX treatment, which was preserved to the control level by the presence of AGE. These results indicated that AGE protected IEC-6 cells from the MTX-induced damage. CONCLUSIONS: The MTX induced apoptosis of IEC-6 cells was shown to be depressed by AGE. AGE may be useful for the cancer chemotherapy with MTX, since AGE reduces the MTX-induced intestinal damage. PMID- 18677485 TI - Genes involved in carotene synthesis and mating in Blakeslea trispora. AB - Mating of Blakeslea trispora and other molds of the order Mucorales requires the interaction of mycelia of opposite sex, (+) and (-), leading to the development of specialized structures and to an enhanced accumulation of beta-carotene. Industry obtains beta-carotene by co-cultivating appropriate strains of Blakeslea ("mated cultures"). Gene transcription in single and mated cultures was assayed by cDNA-AFLP, a technique to observe the differential expression of subsets of mRNA fragments. Overexpression in mated cultures is about ten times more frequent than underexpression. We obtained and sequenced fragments of 97 candidate genes that appeared to be overexpressed during mating and confirmed four of them by reverse transcription and real-time PCR. Comparisons with gene sequences from other organisms suggest functions in carotene biosynthesis (4 genes), energy metabolism (8), cell wall synthesis (1), transfer of acetyl groups (1), and regulatory processes (10). Sodium acetate inhibited sexual overexpression in about two-thirds of the candidate genes and acted as a signal with broad effects on the metabolism and the morphology of mated cultures. Our work offers new materials for the study of carotene biosynthesis and its regulation and for the improvement of carotene production with Mucorales. PMID- 18677484 TI - The Werner's syndrome 4330T>C (Cys1367Arg) gene variant does not affect the in vitro cytotoxicity of topoisomerase inhibitors and platinum compounds. AB - PURPOSE: Werner's syndrome (WS) is a recessive disorder of premature onset of processes associated with aging. Defective DNA repair has been reported after exposure of cells isolated from WS patients to DNA-damaging agents. The germline 4330T>C (Cys1367Arg) variant in the WS gene (WRN) has been associated with protection from age-related diseases, suggesting it has a functional role. We studied whether the 4330T>C variant confers altered drug sensitivity in vitro. METHODS: 4330T>C was genotyped in 372 human lymphoblastoid cell lines (LCLs) from unrelated healthy Caucasian individuals using a TaqMan-based method. The study was powered to detect the effect of the 4330T>C genotypes after exposure to camptothecin (based upon preliminary data). The effect of the 4330T>C variant on the cytotoxicity of etoposide, carboplatin, cisplatin and daunorubicin was also tested. WRN expression in 57 LCLs was measured by microarray. RESULTS: No significant difference between the IC50 of the cells was observed among genotypes (P = 0.46) after exposure to camptothecin. No association was also observed for etoposide, carboplatin, cisplatin, and daunorubicin (ANOVA, P > 0.05). WRN expression also did not vary across genotypes (ANOVA, P = 0.37). CONCLUSION: These results suggest that this nonsynonymous variant has relatively normal function at the cellular level. PMID- 18677486 TI - Characterization of microcalcification: can digital monitor zooming replace magnification mammography in full-field digital mammography? AB - The aim of this study was to compare the diagnostic accuracy and image quality of microcalcifications in zoomed digital contact mammography with digital magnification mammography. Three radiologists with different levels of experience in mammography reviewed 120 microcalcification clusters in 111 patients with a full-field digital mammography system relying on digital magnification mammogram (MAG) images and zoomed images from contact mammography (ZOOM) using commercially available zooming systems on monitors. Each radiologist estimated the probability of malignancy and rated the image quality and confidence rate. Performance was evaluated by sensitivity, specificity, positive predictive value, negative predictive value, and receiver operating characteristic (ROC) analysis. All three radiologists rated MAG images higher than ZOOM images for sensitivity with statistical significance (average value, 92% vs. 87%, P<0.05) and performance by ROC analysis improved with MAG imaging. The confidence rate for diagnosis decision and the assessment of lesion characteristics were also better in MAG images than in ZOOM images with statistical significance (P<0.0001). Digital magnification mammography can enhance diagnostic performance when characterizing microcalcifications. Images zoomed from digital contact mammography cannot serve as an alternative to direct magnification digital mammography. PMID- 18677487 TI - Dual energy CT: preliminary observations and potential clinical applications in the abdomen. AB - Dual energy CT (DECT) is a new technique that allows differentiation of materials and tissues based on CT density values derived from two synchronous CT acquisitions at different tube potentials. With the introduction of a new dual source CT system, this technique can now be used routinely in abdominal imaging. Potential clinical applications include evaluation of renal masses, liver lesions, urinary calculi, small bowel, pancreas, and adrenal glands. In CT angiography of abdominal aortic aneurysms, dual energy CT techniques can be used to remove bones from the datasets, and virtual unenhanced images allow differentiation of contrast agent from calcifying thrombus in patients with endovascular stents. This review describes potential applications, practical guidelines, and limitations of dual energy CT in the abdomen. PMID- 18677488 TI - Osteoblastic response as a healing reaction to chemotherapy mimicking progressive disease in patients with small cell lung cancer. AB - The osteoblastic response (OR) phenomenon as a healing reaction during effective chemotherapy-defined by the appearance of new osteoblastic bone lesions while disease response in other tumor sites was well documented-has previously been described for breast and prostate cancer. The purpose of this study was to investigate this phenomenon that could erroneously be interpreted as progressive disease in patients with small cell lung cancer (SCLC) and to establish guidelines for interpretation of follow-up computed tomography (CT) examinations in this situation. Twenty-four patients with newly diagnosed SCLC and bone metastases were retrospectively included in this study. The characteristics of bone lesions in CT examinations were correlated with bone scintigraphy and magnetic resonance imaging, if available. In target lesions the CT density quantified in Hounsfield units (HU) was evaluated at baseline and during follow up. New osteoblastic lesions occurred during follow-up in 17 of 24 patients. OR was proven in 4 patients and considered most likely in 11 patients; mean density increase in target lesions was 153 HU. The study indicates that osteoblastic response as a healing reaction seems to occur in the majority of patients with SCLC and bone metastases and should not be misinterpreted as progressive disease. PMID- 18677489 TI - "Tenomalacia": a new sonographic sign of tendinopathy? AB - The purpose of this study was to assess tendon compressibility with sonography in extensor tendinopathy and in asymptomatic extensor tendons of the elbow. Sonography of both elbows was performed in eight patients with a clinical diagnosis of unilateral lateral epicondylitis. Tendons were assessed for compressibility by measuring their thickness before and after compression with the transducer. The same manoeuvre was performed while tendon vascularity was assessed with colour Doppler. All eight cases showed increased compressibility of the common extensor tendon on the painful side compared to the asymptomatic side, as well as increased vascularity with compressible vessels on colour Doppler. Other signs of tendinopathy were hypoechogenicity (n = 8), loss of fibrillar pattern (n = 8), intratendinous calcifications (n = 1), partial tears (n = 3), and enthesophytes (n = 5). Increased tendon compressibility indicative of tendon softening or "tenomalacia" is a new sonographic sign of common extensor tendinopathy. PMID- 18677490 TI - Healing of colonic anastomoses after immediate postoperative intraperitoneal administration of oxaliplatin. AB - AIM: The aim of this experimental study was to investigate the effect of intraperitoneal administration of oxaliplatin on the healing of colonic anastomoses when injected immediately after colon resection. MATERIALS AND METHODS: Thirty male Wistar rats were used. During the operation, the rats were randomized to two groups of 15 rats each. Immediately after colonic anastomoses were performed, the rats were injected intraperitoneally with either 3 ml of 0.9% NaCl solution or oxaliplatin (2.4 mg/kg body weight) depending on their group. All rats were killed on the eighth postoperative day. The anastomoses were examined macroscopically. The anastomotic bursting pressures were recorded, the anastomoses graded histologically, and the hydroxyproline tissue contents determined. RESULTS: Anastomotic leakage was noted in four rats (26.7%) of the oxaliplatin group, whereas no anastomotic dehiscence was detected among rats of the control group (p = 0.016). The adhesion formation at the anastomotic sites and the inflammatory cell infiltration were significantly higher in the oxaliplatin group than in the control group (p = 0.001). The bursting pressures (p = 0.001), the hydroxyproline tissue content (p = 0.001), the neoangiogenesis (p = 0.033), the fibroblast activity (p = 0.001), and the collagen deposition (p = 0.001) were significantly lower in the oxaliplatin group in comparison to the control group. CONCLUSION: The immediate postoperative intraperitoneal administration of oxaliplatin seems to impair healing of colonic anastomoses in rats. PMID- 18677491 TI - Efficacy of an aiming device for the placement of distal interlocking screws in trochanteric fixation nailing. AB - INTRODUCTION: Locked intramedullary nailing continues to be the surgical treatment of choice for most long bone fractures. Performing distal interlocks can be a technical challenge. Free hand (FH) technique remains to be most popular. Radiation exposure, particularly to the surgeon still remains a concern with this technique. METHOD: A prospective randomized analysis of 20 patients undergoing operative fixation with long trochanteric fixation nailing for intertrochanteric or subtrochanteric fractures was performed. The groups were randomized into (1) aiming arm group (AA) and (2) FH group by computer generated randomization technique. Two distal interlocking screws were placed in every case. Various parameters were analyzed including total operating time, distal interlocking time, total fluoroscopy time, distal fluoroscopy time and nail dimensions. The variables in two groups were compared to each other using Fischer's exact test. RESULT: The mean distal interlock time was 7.1 +/- 2.4 and 12.1 +/- 3.2 min for AA and FH techniques respectively. There was a 41.3% decrease in the distal interlock time with the device, which was statistically significant (P < 0.001). The distal interlock fluoroscopy time was 9.2 +/- 4.9 and 28.9 +/- 16.4 s with AA technique and the FH technique respectively. This 68.2% reduction in time taken for distal fluoroscopy was statistically significant (P < 0.001). However, the reduction in the total fluoroscopy time was statistically not significant. CONCLUSION: The AA is very efficient and user friendly and also reduces the radiation exposure. PMID- 18677492 TI - Intramedullary screw fixation in proximal fifth-metatarsal fractures in sports: clinical and biomechanical analysis. AB - INTRODUCTION AND PURPOSE: Intramedullary screw fixation (ISF) of proximal fifth metatarsal fractures is known as first treatment option in young, sports active patients. No study analyzed functional and biomechanical outcome before. Hypothetically ISF leads to (1) a high bony union rate within 12 weeks, (2) normal hindfoot eversion strength, and (3) normal gait and plantar pressure distribution. METHODS: Fourteen out of 22 patients were available for follow-up with an average follow-up of 42 months; clinical and radiological follow-up, and biomechanical evaluation by isometric muscular strength measurement (inversion, eversion strength) and dynamic pedobarography, comparing to the non-affected contralateral foot. Level of significance: 0.05. RESULTS: Subjective result: Excellent or good result in 14 patients, none fair or poor. AOFAS midfoot score: 100 points in 13 patients and 87 points in 1 patient. The same sports activity level (0-4) was reached in 13 out of 14 patients. Radiologic examination: consolidation after 6 weeks in 9 patients and after 12 weeks in another 4 patients, one partial union. Average maximal eversion strength 59 N (ratio to the contralateral foot: 0.92, not significant). Dynamic pedobarography showed ratios of 0.99-1.01 to the contralateral side for ground reaction force, ground peak time, peak pressure and contact area (not significant). INTERPRETATION: A very high patient-satisfaction, a fast bony healing and complete return to sports were documented. Muscular strength measurement and dynamic pedobarography showed complete functional rehabilitation. Therefore, ISF in proximal fifth-metatarsal fractures can be recommended as a secure procedure. PMID- 18677493 TI - Nonunion of distal radius fracture and distal radioulnar joint injury: a modified Sauve-Kapandji procedure with a cubitus proradius transposition as autograft. AB - The Sauve-Kapandji (SK) procedure is indicated in distal radius nonunion or malunion and distal radioulnar joint (DRUJ) instability. It can also be used to treat the rheumatoid wrist with severe degenerative changes in the DRUJ. The main objective is to allow a pain-free range of movement. We present a patient with rheumatoid arthritis and distal radius nonunion who, after three operations, was treated with the SK procedure. The clinical and radiological results were excellent. A 53-year-old woman diagnosed with rheumatoid arthritis fell on her forearm at home 2 years ago. Examination at an outpatient clinic revealed a distal radius fracture classified as type V according to the Frykman classification. She had been operated three times with open reduction internal fixation using a plate, screws, and bone allograft. She came to our institution with a distal radius nonunion, positive post-traumatic ulnar variance, and ulnar nerve paresis. The range of movements was 20 degrees -10 degrees flexion extension and 40 degrees -30 degrees pronation-supination, and she needed daily fentanyl. We performed a modified SK procedure with an autologous iliac crest bone graft and ulnar bone graft from the osteotomy area (cubitus proradius), bone morphogenetic protein, and a low profile distal radius plate. After 1 year of follow-up, the distal radius fracture has healed and the wrist is pain-free with a complete range of movement in flexion-extension and pronation-supination. The main indication for the SK procedure is post-traumatic positive ulnar variance and associated ulnocarpal impaction. The cubitus proradius bone graft transposition is an interesting technical note that makes this case a challenge for skilled orthopedic hand surgeons. PMID- 18677495 TI - The efficacy of Biobon and Ostim within metaphyseal defects using the Gottinger Minipig. AB - INTRODUCTION: To compare bio, osteocompatibility, rate of resorption and remodeling dynamics of two clinically used bone substitutes. MATERIALS AND METHODS: In a randomized fashion Biobon and Ostim were implanted bilaterally into the proximal metaphyseal tibiae of 18 Gottinger Minipigs in a direct right versus left "intra-individual" comparison. Fluorescent labelling was used. Microradiographic, histological and morphometric evaluation was carried out at 6, 12 and 52 weeks. RESULTS: Both bone substitutes showed good biocompatibility, bioactivity and osteoconductivity. The degradation dynamics of both materials differed. Degradation of Ostim stopped after 6 weeks postoperatively, whereas Biobon was degraded slowly but evenly over the time intervals. Only at 6 weeks a significant (P < 0.05) difference in resorption rate was detected. Both Biobon and Ostim showed incomplete resorption after a year. CONCLUSION: After 1 year no "restitutio ad integrum" could be observed in either group. Similar to ceramics, a thorough osseous incorporation seemed to inhibit further degradation of both bone substitute materials. PMID- 18677496 TI - Surgical management of a double-layered patella: a case report. AB - The double-layered patella (DLP) is a rare intra-articular disorder that in most cases is associated with multiple epiphyseal dysplasia (MED). The DLP usually consists of an anterior and a posterior layer with a cartilaginous interface between the two parts. In this article, we present a case of bilateral DLP of a 17-year-old female adolescent with MED, suffering clinical symptoms on her left knee. Upon clinical examination, a painful "snapping" of the patella at 30-40 degrees flexion of the left knee was evident and two osseous layers could be identified in standard radiographs. Arthroscopic examination revealed a transient cranial displacement of the posterior osteochondral layer at 30-40 degrees knee flexion that replaced beyond 40 degrees knee flexion with a visible jump. Surgical management in this case of DLP comprised open debridement of the soft tissue interface and fusion of the two osseous layers using double-threaded screws leading to an excellent outcome at 12 months follow-up. PMID- 18677494 TI - Parecoxib has non-significant long-term effects on bone healing in rats when administered for a short period after fracture. AB - INTRODUCTION: Selective and non-selective cyclo-oxygenase (COX) inhibitors impair bone healing by inhibiting prostaglandin synthesis. The purpose of this study was to evaluate the long-term effect of parecoxib, a selective COX-2 inhibitor, on bone healing in rats, when it is applied in a pattern similar to clinical treatment patterns, that is, in a high dose and for a short period after bone fracture. METHOD: Closed non-displaced mid-diaphyseal fractures in the middle of the left femoral shaft were generated in each animal. In the study group, parecoxib sodium (1.06 mg/kg) was administered intra-peritoneally every day for 7 days. In the control group, normal saline was administered intra-peritoneally every day for 7 days. In both groups fracture healing (bone union and callus formation) was evaluated with X-rays 28 and 42 days after surgery. RESULTS: Bone healing was lower in the study group (60 vs. 80% in the control group 28 days after fracture and 80 vs. 90% 42 days after fracture) but this difference was not statistically significant (P > 0.05). CONCLUSION: Parecoxib does not have a significant long-term effect on bone healing in rats, when it is administered in a high dose and for a short period after bone fracture. PMID- 18677497 TI - Fibula and tibia fusion with cancellous allograft vitalised with autologous bone marrow: first results for infected tibial non-union. AB - BACKGROUND AND AIMS: Autogenous bone grafting has been used in reconstructing bone defects and in stimulating fracture healing, producing high healing rates in the treatment of infected tibial non-unions. A novel therapeutic alternative is now available known as "vitalised allograft", a cancellous bone graft procured from femoral heads from living human donors and "vitalised" through the injection of autologous bone marrow. The aim of this study is to summarise the initial results of the fibula and tibia fusion using vitalised cancellous allograft in the treatment of infected tibial non-unions. PATIENTS AND METHODS: We initiated a follow-up of 15 prospective non-randomized patients who received a vitalised allograft in the treatment of infected tibial non-unions in order to produce bony union. The patients included 13 men and 2 women with an average age of 48 years. All patients received a multi-stage surgical approach. After establishing an infection-free environment, allogenic cancellous bone grafting was performed, intended as the final surgical procedure in fibula and tibia fusion. Our follow up included a clinical and radiographic investigation of the calf in four planes. We analysed union-rate and time required for bony consolidation, as well as recurrent infections, re-fractures, potential graft-resorption, and time needed for graft and bone remodelling. RESULTS: With an average follow-up of 17.1 months, infection control was obtained in 14 of 15 patients, producing an infection arrest rate of 93.3%. Radiographs indicated consolidation in 11 out of 15 cases, with a union rate of 73.3%. Bone union was achieved on average in 17.1 weeks. CONCLUSIONS: Fibula and tibia fusion with allogenic cancellous bone grafting, vitalised through autogenic bone marrow, could well become an innovative treatment option for infected tibial non-unions. We need, however, to analyse a higher number of cases over a longer follow-up period in order to assess more accurately recurrent infections and re-fractures. PMID- 18677498 TI - Two-stage revision of infected hip arthroplasty using a shortened post-operative course of antibiotics. AB - INTRODUCTION: We present a series of 30 consecutive patients with 31 infected total hip arthroplasties treated by a single surgeon over a 4-year period in whom a shortened post-operative course of antimicrobial chemotherapy was used. METHODS: The treatment protocol consisted of a two-stage exchange with removal of infected components, insertion of an interim antibiotic eluting cement spacer and re-implantation of an extensively coated uncemented prosthesis on the femoral side. Systemic antibiotic treatment following each stage consisted of an abridged course of 5 days post-operative intra-venous administration followed by complete cessation of anti-microbial therapy. RESULTS: At a mean follow-up of 35 months (minimum 24 months), there were no cases of recurrent prosthetic infection and no patient had required revision for aseptic loosening or mechanical instability on the femoral side. The combination of effective-staged surgical joint debridement, a shortened post-operative course of systemic antibiotic treatment and an adequate latent period before re-implantation has led to encourage early results in this series of revised chronic hip joint prosthetic infections. PMID- 18677499 TI - Changes in Na+, K+ concentrations in perspiration and perspiration volume with alternating current iontophoresis in palmoplantar hyperhidrosis patients. AB - Various treatments are currently available for palmoplantar hyperhidrosis. We have treated palmoplantar hyperhidrosis patients effectively with the use of alternating current (AC) iontophoresis. However, much remains unknown about the physiological changes that occur with AC iontophoresis, and its mechanism of action. We measured the changes in Na(+), K(+) concentration in perspiration and perspiration volume with AC iontophoresis in palmoplantar hyperhidrosis patients. We found that hyperhidrosis patients have significantly higher perspiration volume and Na(+) concentration in perspiration than healthy controls. Looking at the temporal changes with AC iontophoresis, we found a significant decrease in perspiration volume and Na(+) concentration in perspiration after six iontophoresis treatments. This result is further evidence that Na(+) concentration in perspiration is closely involved with perspiration volume. However, looking at the changes in perspiration volume and Na(+) concentration in perspiration before and after a single AC iontophoresis treatment, we found that while perspiration volume did not decrease in hyperhidrosis patients after a single treatment, there was a significant decrease in Na(+) concentration. In healthy controls as well, Na(+) concentration in perspiration decreased significantly after a single treatment. These findings suggest that the effect of AC iontophoresis may be due to a complex mechanism involving changes in reabsorption of ductal Na(+). PMID- 18677500 TI - Lessons learned from the outcome of bloodless emergency laparotomies on Jehovah's Witness women presenting in the extremis with ruptured uterus. AB - OBJECTIVE: We compared the outcomes of bloodless emergency laparotomies for uterine rupture in twenty Jehovah's Witness (JW) women who were in a state of haemorrhagic shock with 45 other women who received blood transfusion. METHODS: The case records of twenty Jehovah's Witness patients, who underwent bloodless emergency laparotomies for ruptured uterus complicated by haemorrhagic shock between 1 January 2000 and 31 December 2006 were reviewed. We also reviewed the case records of 45 other patients who underwent similar surgical procedures for similar indications and in whom blood was transfused. The outcome variables compared were maternal deaths, infectious morbidity, acute respiratory distress syndrome, disseminated intravascular coagulation, length of post laparotomy ventilatory support, and length of postpartum hospitalization. RESULTS: Demographic and obstetric characteristics were similar in both groups. Seventeen out of the twenty Jehovah's Witness patients who refused blood transfusion survived the laparotomies and were discharged home in good condition. Three died of peritonitis. Six out of the 45 patients who received blood transfusion also died of peritonitis and one died of disseminated intravascular coagulopathy. The average duration of post partum hospitalization was 8 days in the group of patients transfused and 7 days in the group not transfused. Disseminated intravascular coagulation occurred post partum in two of the patients transfused. No case of acute respiratory distress syndrome or amniotic fluid embolism was identified in either group. CONCLUSION: Patients who are in haemorrhagic shock from ruptured uterus and refuse blood transfusion can still be salvaged in a low resource setting. The study adds evidence that major operative procedures can be carried out on Jehovah's Witness patients without blood transfusions or blood products. PMID- 18677501 TI - Interobserver variability of modified Ferriman-Gallwey hirsutism score in a Turkish population. AB - OBJECTIVE: The aim of the study was to evaluate the interobserver variability of modified Ferriman-Gallwey (mFG) hirsutism scores on each body area in a Turkish population. DESIGN: A cross-sectional study of simultaneous mFG scoring design was used. Observers did not make any interview with the subjects and were masked to the previous score results. Analyses included percentage of agreement, kappa coefficients, the Bland and Altman plot, confidence intervals, minimum and maximum kappa coefficients. SETTING: The study was performed at a teaching and research hospital. PATIENTS: Hundred and twenty-one Turkish women without any complaints of excessive body hair were studied. INTERVENTIONS: Interventions included two special trained physicians, simultaneous and independent mFG scoring. MAIN OUTCOME MEASURES: The main outcome measures were mFG scores in each body area. RESULTS: Agreement analysis demonstrated that the scores of the two physicians were quite concordant. The mean kappa value for nine body area was 0.744 and the highest kappa values from the upper back and the lowest kappa values from the upper lip revealed kappa=0.847, kappa=0.585, respectively. The highest (upper lip) and the lowest (arm) mean range scores for the two researchers among the 9 areas were 1.46-1.55 and 0.17-0.12, respectively. Only 68.6 and 67.8% of the mFG scores observed by each of the two observers were equal or below 8. CONCLUSION: The mFG scoring system was found to be clinically useful. The upper lip was observed to have the highest score of androgen sensitive area of the body as well as the highest interobserver variability. The cut-off value to establish the diagnosis of hirsutism should be population-specific. PMID- 18677502 TI - Bi-orienting chromosomes: acrobatics on the mitotic spindle. AB - To maintain their genetic integrity, eukaryotic cells must segregate their chromosomes properly to opposite poles during mitosis. This process mainly depends on the forces generated by microtubules that attach to kinetochores. During prometaphase, kinetochores initially interact with a single microtubule that extends from a spindle pole and then move towards a spindle pole. Subsequently, microtubules that extend from the other spindle pole also interact with kinetochores and, eventually, each sister kinetochore attaches to microtubules that extend from opposite poles (sister kinetochore bi-orientation). If sister kinetochores interact with microtubules in wrong orientation, this must be corrected before the onset of anaphase. Here, I discuss the processes leading to bi-orientation and the mechanisms ensuring this pivotal state that is required for proper chromosome segregation. PMID- 18677503 TI - Besides Purkinje cells and granule neurons: an appraisal of the cell biology of the interneurons of the cerebellar cortex. AB - Ever since the groundbreaking work of Ramon y Cajal, the cerebellar cortex has been recognized as one of the most regularly structured and wired parts of the brain formed by a rather limited set of distinct cells. Its rather protracted course of development, which persists well into postnatal life, the availability of multiple natural mutants, and, more recently, the availability of distinct molecular genetic tools to identify and manipulate discrete cell types have suggested the cerebellar cortex as an excellent model to understand the formation and working of the central nervous system. However, the formulation of a unifying model of cerebellar function has so far proven to be a most cantankerous problem, not least because our understanding of the internal cerebellar cortical circuitry is clearly spotty. Recent research has highlighted the fact that cerebellar cortical interneurons are a quite more diverse and heterogeneous class of cells than generally appreciated, and have provided novel insights into the mechanisms that underpin the development and histogenetic integration of these cells. Here, we provide a short overview of cerebellar cortical interneuron diversity, and we summarize some recent results that are hoped to provide a primer on current understanding of cerebellar biology. PMID- 18677506 TI - Sex-related differences in evaporative heat loss: the importance of metabolic heat production. AB - We evaluated the hypothesis that different rates of metabolic heat production between sexes, during exercise at the same percentage of maximum oxygen consumption [VO2 max] give proportional differences in evaporative heat loss. Seven males and seven females, exercised at 41.3 +/- 2.7% VO2 Max for 60-min at 40 degrees C and 30% relative humidity. Whole-body direct air calorimetry measured rate of whole-body evaporative heat loss (H e) while metabolic heat production (M - W) was measured by indirect calorimetry. M -W was greater in males (243 +/- 18 W m(-2)) relative to females (201 +/- 4 W m(-2)) (P 0.1 in over 93%, >0.25 in over 86%, and >0.5 in over 82% of patients with Legionella-CAP. Patients with adverse medical outcomes (59%, n = 17) including need for ICU admission (55%, n = 16) and/or inhospital mortality (14%, n = 4) had significantly higher median PCT values on admission (4.27 [IQR 2.46-9.48] vs 0.97 [IQR 0.29-2.44], p = 0.01), while the PSI (124 [IQR 81-147] vs 94 [IQR 75-116], p = 0.19), the CURB65 (2 [IQR 1-2] vs 1 [1-3], p = 0.47), CRP values (282 [IQR 218 343], p = 0.28 vs 201 [IQR 147-279], p = 0.28), and leukocyte counts (12 [IQR 10 21] vs 12 [IQR 9-15], p = 0.58) were similar. In receiver operating curves, PCT concentrations on admission had a higher prognostic accuracy to predict adverse outcomes (AUC 0.78 [95%CI 0.61-96]) as compared to the PSI (0.64 [95%CI 0.43 0.86], p = 0.23), the CURB65 (0.58 [95%CI 0.36-0.79], p = 0.21), CRP (0.61 [95%CI 0.39-0.84], p = 0.19), and leukocyte count (0.57 [95%CI 0.35-0.78], p = 0.12). Kaplan-Meier curves demonstrated that patients with initial PCT values above the optimal cut-off of 1.5 had a significantly higher risk of death and/or ICU admission (log rank p = 0.003) during the hospital stay. In patients with CAP due to Legionella, PCT levels on admission might be an interesting predictor for adverse medical outcomes. PMID- 18677520 TI - Atypical abdominal hernias in the emergency department: acute and non-acute. AB - The objective of this pictorial essay is to review uncommon abdominal hernias, many of which present to the Emergency Department with abdominal pain. These hernias may be congenital, post-traumatic, or iatrogenic in origin. They may present as an acute (surgical) abdomen without localizing signs or symptoms. They may present with an obvious antecedent event such as motor vehicle trauma or simply present as an incidental finding. Multi-detector computed tomography is currently the study of choice to diagnose abdominal hernia and to evaluate the possible complications such as small bowel obstruction and/or strangulation. This modality can delineate a "zone of transition" (abnormally dilated bowel transitioning to normal or decreased bowel caliber) or identify the involved anatomy. It can also suggest compromised blood supply. PMID- 18677521 TI - Diversity of promoter elements in a Geobacter sulfurreducens mutant adapted to disruption in electron transfer. AB - The delta-proteobacterium, Geobacter sulfurreducens, can obtain energy by coupling the oxidation of organic matter to the reduction of insoluble Fe(III) or the anode of a microbial fuel cell. Because Fe(III) oxide or the anode surface, in contrast to oxygen, nitrate, or sulfate, is not soluble nor can it be reduced readily, Geobacter species have developed mechanisms which allow electrons to be delivered across outer membrane to the cell surface. OmcB is an outer-membrane c type cytochrome important for G. sulfurreducens Fe(III) respiration. In the absence of OmcB, cells lost the ability to reduce soluble or insoluble Fe(III). However, the omcB deletion mutant can slowly adapt to growth on soluble Fe(III) over prolonged incubation in the medium with acetate as the electron donor. We discuss available information about predicted or experimentally validated promoters and transcription regulatory sites identified upstream of operons with transcriptional expression significantly changed in the adapted omcB mutant. DNA sequences of upstream regions of coregulated operons in the adapted mutant are divergent, suggesting the presence of recognition sites for different transcriptional regulators and indicating that adaptation of the omcB mutant to growth on soluble Fe(III) has shifted the relevant expression networks involved to a more diverse molecular basis. PMID- 18677522 TI - Non-LTR retrotransposons in fungi. AB - Non-long terminal repeat (non-LTR) retrotransposons have contributed to shaping the structure and function of genomes. Fungi have small genomes, usually with limited amounts of repetitive DNA. In silico approach has been used to survey the non-LTR elements in 57 fungal genomes. More than 100 novel non-LTR retrotransposons were found, which belonged to five diverse clades. The present survey identified two novel clades of fungal non-LTR retrotransposons. The copy number of non-LTR retroelements varied widely. Some of the studied species contained a single copy of non-LTR retrotransposon, whereas others possessed a great number of non-LTR retrotransposon copies per genome. Although evolutionary relationships of most elements are congruent with phylogeny of host species, a new case of possible horizontal transfer was found between Eurotiomycetes and Sordariomycetes. PMID- 18677523 TI - The foramen spinosum: a landmark in middle fossa surgery. AB - The foramen spinosum is an easily identifiable landmark in microsurgery of the middle cranial fossa, and knowledge of the variations in its relationship to the surrounding neurovascular structures is important when operating in this area. We studied the anatomical relationship of the foramen spinosum to the foramen ovale, the mandibular branch of the trigeminal nerve, the greater superficial petrosal nerve, and the petrous part of the internal carotid artery in 12 cadaver heads. We also tried to define an external landmark for early identification of the location of the foramen spinosum in ten dry skulls. We found considerable variations in the anatomy around the foramen spinosum. This knowledge may improve the identification and preservation of the neurovascular structures when using approaches to the middle cranial fossa. PMID- 18677524 TI - Aneurysm clipping after endovascular treatment with coils: a report of 13 cases. AB - In 1996, Civit et al. (Neurosurgery, 38:955-961, 1996) reported a series of eight patients whose aneurysms were clipped after previous embolization with coils. This paper highlighted the safety of this surgery in second line, with a low complication rate and a favorable outcome. The two major surgical indications were either after deliberate partial occlusion of the aneurysm (N=3) or partial occlusion after endovascular treatment (N=3). Reviewing 13 additional patients from 1996 to June 2005, the authors compared the surgical indications and focused on the technical problems of clipping after coiling. Thirteen patients (men=6, women=7) with aneurysm clipping following one or more endovascular embolizations have been operated on since 1996. The patients' files were reviewed retrospectively by both a senior consultant neurosurgeon and a neuroradiologist. Demographic data included sex, age at admission, relevant medical history, initial endosaccular treatment and its quality (partial or complete effectiveness), the rationale for surgery, and the complications arising from the different treatments. In addition to the patient's clinical follow-up, angiograms were performed soon after the surgical procedure, 3 months, 1 year, and 5 years after the coiling, respectively. None of the initial endovascular treatments was complete. Surgical indication was related firstly to anatomical particularities of the aneurysm (width of the neck, N=5; arterial branches from the aneurysm, N=4; no individualized neck in a small aneurysm, N=1); secondly to a shift of the coils with delayed aneurysm regrowth and repermeabilization, N=4; and thirdly to rebleeding, N=3. All the patients who were operated on underwent complete surgical exclusion of their aneurysm (controlled by angiogram). Twelve out of 13 patients recovered satisfactorily (92.3%), attaining the same neurological state they presented prior to surgery. One patient died after the operation. He had already been in a serious condition because of severe rebleeding following the embolization. Aneurysm clipping following a previous endovascular embolization procedure is a rare, although not so exceptional, indication. It is a safe and effective procedure, probably under-used. Nowadays, "hemostatic" and incomplete embolization of an aneurysm increases the risk of future growth and rebleeding of the residual pouch. An additional aneurysm clipping may therefore be required rapidly after embolization. PMID- 18677525 TI - Thermal denaturation of wild type and mutant recombinant acetylcholinesterase from amphioxus: effects of the temperature of in vitro expression and of reversible inhibitors. AB - We have studied the thermal inactivation at 37 degrees C of wild type and mutant ChE2 (C310A, F312I, C466A, C310A/F312I, and C310A/C466A) from amphioxus (Branchiostoma floridae) expressed in vitro in COS-7 monkey cells under three sets of conditions: 30 degrees C for 48 h, 30 degrees C for 24 h and 37 degrees C for 24 h, and 37 degrees C for 48 h. We found biphasic denaturation curves for all enzymes and conditions, except wild type and C310A ChE2 expressed at 30 degrees C for 48 h. Generally, single mutants are more unstable than wild type, and the double mutants are even more unstable. We propose a model involving stable and unstable conformations of the enzymes to explain these results, and we discuss the implications of the model. We also found a correlation between the melting temperature of the ChEs and the rates at which they denature at 37 degrees C, with the denaturation of the unstable conformation dominating the relationship. Reversible cholinergic inhibitors protect the ChEs from thermal denaturation, and in some cases produce monophasic denaturation curves; we also propose a model to explain this stabilization. PMID- 18677526 TI - Laparoscopic renal cooling device. AB - A new medical device has been designed for inducing hypothermia in kidneys, via laparoscopic deployment, without leaving residual fluid in the abdomen. It was tested in laboratory and animal trials to ascertain its suitability for achieving a target renal temperature of 15 degrees C for tissue preservation within a 30 min period. A simple theoretical model was developed to calculate cooling rates under ideal circumstances. In the laboratory, a potato was used as the model for the kidney and an average minimum temperature of 15.8 degrees C was reached in 30 min. The theoretical model calculated this as 9.9 degrees C. In the animal trials, an average minimum temperature of 11.4 degrees C was reached in 30 min, compared to the calculated temperature of 9.8 degrees C from the theoretical model. In conclusion, the new device can cool as affectively as other techniques trialled, with the added advantage that it does not release cold fluid or ice into the abdomen. PMID- 18677527 TI - The effect of copper on the structure of the ammonia-oxidizing microbial community in an activated sludge wastewater treatment plant. AB - Molecular approaches based on both whole-cell and extracted DNA were applied to assess chronic and acute effects of copper on the ammonia oxidizing bacteria (AOB) community in an activated sludge system. The ammonia monooxygenase amoA gene was chosen as the functional marker to evaluate changes in the AOB community. Using in situ polymerase chain reaction, we were able to visualize the peripheric distribution of the amoA gene-possessing bacteria in activated sludge flocs. The AOB biomass content was constant in both chronic and acute toxicity experiments, but the ammonia oxidizing activity, measured as ammonia uptake rate, was different. The AOB community structural changes due to the copper presence were evaluated by multivariate analysis of the DGGE bands profiles. The chronic contamination caused a change in the AOB community compared to the control. In contrast, acute inputs led to a temporary change in the AOB community, after which the community was similar to the control. Recovery after acute intoxication was achieved after 72 h. The present study reports on the effects of chronic and acute copper contamination on the ammonia uptake ability of the AO microorganisms and the structure of the AOB community in a wastewater system and, as a consequence, gives indications on the response of wastewater plants under similar conditions. PMID- 18677528 TI - Antibiotic resistance in bacteria isolated from the deep terrestrial subsurface. AB - Various natural environments have been examined for the presence of antibiotic resistant bacteria and/or novel resistance mechanisms, but little is known about resistance in the terrestrial deep subsurface. This study examined two deep environments that differ in their known period of isolation from surface environments and the bacteria therein. One hundred fifty-four strains of bacteria were isolated from sediments located 170-259 m below land surface at the US Department of Energy Savannah River Site (SRS) in South Carolina and Hanford Site (HS) in Washington. Analyses of 16S rRNA gene sequences showed that both sets of strains were phylogenetically diverse and could be assigned to several genera in three to four phyla. All of the strains were screened for resistance to 13 antibiotics by plating on selective media and 90% were resistant to at least one antibiotic. Eighty-six percent of the SRS and 62% of the HS strains were resistant to more than one antibiotic. Resistance to nalidixic acid, mupirocin, or ampicillin was noted most frequently. The results indicate that antibiotic resistance is common among subsurface bacteria. The somewhat higher frequencies of resistance and multiple resistance at the SRS may, in part, be due to recent surface influence, such as exposure to antibiotics used in agriculture. However, the HS strains have never been exposed to anthropogenic antibiotics but still had a reasonably high frequency of resistance. Given their long period of isolation from surface influences, it is possible that they possess some novel antibiotic resistance genes and/or resistance mechanisms. PMID- 18677530 TI - An unusual left ventricular outflow pseudoaneurysm: usefulness of echocardiography and multidetector computed tomography for surgical repair. AB - Left ventricular outflow tract (LVOT) pseudoaneurysm is a rare but potentially lethal complication, mainly after aortic root endocarditis or surgery. Usually it originates from a dehiscence in the mitral-aortic intervalvular fibrosa and it arises posteriorly to the aortic root. Due to these anatomical features, its imaging assessment is challenging and surgical repair requires complex procedures. An unusual case of LVOT pseudoaneurysm is described. It was detected by transthoracic ecocardiography 7 months after aortic root replacement for acute endocarditis. Multidetector computed tomography (MDCT) confirmed the presence of a pouch located between the aortic root and the right atrium. Computed tomography also detected the origin of the pseudoaneurysm from the muscular interventricular septum of the LVOT, rather below the aortic valve plane. It was repaired with an extracardiac surgical approach, sparing the aortic root bioprosthesis previously implanted. The high-resolution three-dimensional details provided by the preoperative MDCT allowed us to plan a simple and effective surgical strategy. PMID- 18677531 TI - Endovascular treatment of a superior mesenteric artery aneurysm secondary to Behcet's disease with Onyx (ethylene vinyl alcohol copolymer). AB - Behcet's disease is a complex multisystemic chronic inflammatory disease that is characterized by oral and genital aphtous ulcers and vasculitis. Aneurysms of major arteries are the most important cause of mortality in Behcet's disease. Four patients with superior mesenteric artery (SMA) aneurysms related to Behcet's disease have been reported in the literature. We report here the first successful endovascular treatment of a giant, wide-necked SMA aneurysm secondary to Behcet's disease. We performed a balloon-assisted embolization technique using ethylene vinyl alcohol copolymer (Onyx, ev3, Irvine, CA, USA). There were no signs of recurrence during 2-year follow-up. PMID- 18677532 TI - Failed retrieval of an inferior vena cava filter during pregnancy because of filter tilt: report of two cases. AB - Thromboembolic disease during pregnancy is an important cause of obstetric morbidity and mortality. Pregnant patients with venous thromboembolism are usually managed by conventional anticoagulation. However, this must be discontinued during vaginal or caesarian delivery to avoid haemorrhage and to reduce the risk of possible epidural haematoma. Retrievable inferior vena cava filters (IVCFs) offer protection against pulmonary embolism during this high-risk period, when anticoagulation is discontinued, while avoiding potential long-term sequelae of a permanent IVCF. Here we report two patients who presented in the third trimester of pregnancy with floating ileofemoral deep vein thrombosis. Both patients were initially treated with standard anticoagulation; however, shortly before delivery both patients had a retrievable IVCF placed in a suprarenal position. In both patients, retrieval failed at 28 days after insertion because of filter tilt. The timing and mechanism of filter tilt remains uncertain. We believe that a number of factors could have been involved, including change in the anatomic configuration with lateral displacement of the IVCF as a result of the gravid uterus as well as forceful uterine contractions during labour, which modified the shape and diameter of the IVC. We showed that failure to retrieve the IVCF has had considerable implications for the two young patients regarding long-term anticoagulation and have highlighted the need for further clinical trials regarding the safe use of retrievable IVCFs during pregnancy. PMID- 18677534 TI - Image texture characterization using the discrete orthonormal S-transform. AB - We present a new efficient approach for characterizing image texture based on a recently published discrete, orthonormal space-frequency transform known as the DOST. We develop a frequency-domain implementation of the DOST in two dimensions for the case of dyadic frequency sampling. Then, we describe a rapid and efficient approach to obtain local spatial frequency information for an image and show that this information can be used to characterize the horizontal and vertical frequency patterns in synthetic images. Finally, we demonstrate that DOST components can be combined to obtain a rotationally invariant set of texture features that can accurately classify a series of texture patterns. The DOST provides the computational efficiency and multi-scale information of wavelet transforms, while providing texture features in terms of Fourier frequencies. It outperforms leading wavelet-based texture analysis methods. PMID- 18677535 TI - Naturally induced humoral immunity to West Nile virus infection in raptors. AB - West Nile virus (WNV) infection can be fatal to many bird species, including numerous raptors, though population- and ecosystem-level impacts following introduction of the virus to North America have been difficult to document. Raptors occupy a diverse array of habitats worldwide and are important to ecosystems for their role as opportunistic predators. We documented initial (primary) WNV infection and then regularly measured WNV-specific neutralizing antibody titers in 16 resident raptors of seven species, plus one turkey vulture. Most individuals were initially infected and seroconverted between July and September of 2003, though three birds remained seronegative until summer 2006. Many of these birds became clinically ill upon primary infection, with clinical signs ranging from loss of appetite to moderate neurological disease. Naturally induced WNV neutralizing antibody titers remained essentially unchanged in some birds, while eight individuals experienced secondary rises in titer presumably due to additional exposures at 1, 2, or 3 years following primary infection. No birds experienced clinical signs surrounding or following the time of secondary exposure, and therefore antibodies were considered protective. Results of this study have implications for transmission dynamics of WNV and health of raptor populations, as well as the interpretation of serologic data from free-ranging and captive birds. Antibodies in raptors surviving WNV may persist for multiple years and protect against potential adverse effects of subsequent exposures. PMID- 18677536 TI - The cost-effectiveness of three strategies for the surgical treatment of symptomatic primary hyperparathyroidism. AB - INTRODUCTION: Modern surgical approaches to the treatment of primary hyperparathyroidism [unilateral neck exploration (UNE) and minimally invasive parathyroidectomy (MIP)] have become commonplace in recent years. However, the cost-effectiveness of these strategies has been questioned since the effectiveness of the gold standard, bilateral neck exploration (BNE), is well established. The objective of our study was to determine the incremental cost effectiveness of UNE and MIP compared with BNE for treatment of primary hyperparathyroidism (HPT). METHODS: Patients presenting to a tertiary endocrine surgical center for treatment of HPT over a 38-month period were included in the study. The primary measure of effectiveness was the rate of postoperative complications (hypocalcemia and paresthesias) observed in our cohort. A decision analytic model was constructed to determine the incremental cost-effectiveness ratios (ICERs) of the UNE and MIP strategies compared with the BNE strategy. Deterministic and probabilistic sensitivity analyses were conducted to evaluate uncertainty around model-based estimates of costs and effectiveness. RESULTS: A total of 94 patients (56 BNEs, 19 UNEs, and 19 MIPs) provided estimates of mean costs (BNE = $4524, UNE = $4784, MIP = $4961) and success rates (BNE = 0.91, UNE = 0.86, MIP = 0.93) for each treatment arm. The gold standard BNE strategy dominated the UNE strategy (lower cost, higher effectiveness) under most model formulations. The MIP strategy had an ICER of $28,439 per complication avoided, which is likely to be above societal willingness to pay to avoid primarily minor postoperative complications. CONCLUSION: Our results suggest that within our institution, and in several different model formulations, bilateral neck exploration remains the cost-effective strategy for the treatment of primary hyperparathyroidism. PMID- 18677537 TI - The azygos vein: to resect or not? PMID- 18677538 TI - The integrity of esophagogastric junction anatomy in patients with isolated laryngopharyngeal reflux symptoms. AB - BACKGROUND: Distortion of esophagogastric junction anatomy in patients with gastroesophageal reflux disease produces permanent dilation of the gastric cardia proportional to disease severity, but it remains unclear whether this mechanism underlies reflux in patients with isolated laryngopharyngeal reflux symptoms. METHOD: In a prospective study, 113 patients were stratified into three populations based on symptom complex: laryngopharyngeal reflux symptoms, typical reflux symptoms, and both laryngopharyngeal and typical symptoms. Subjects underwent small-caliber upper endoscopy in the upright position. Outcome measures included gastric cardia circumference, presence and size of hiatal hernia, and prevalence of esophagitis and Barrett's esophagus within each group. RESULTS: There were no differences in gastric cardia circumference between patient groups. The prevalence of Barrett's esophagus was 20.4% overall and 15.6% in pure laryngopharyngeal reflux patients. Barrett's esophagus patients had a greater cardia circumference compared to those without it. In the upright position, patients with isolated laryngopharyngeal reflux display the same degree of esophagogastric junction distortion as those with typical reflux symptoms, suggesting a similar pathophysiology. CONCLUSION: This indicates that, although these patients may sense reflux differently, they have similar risks as patients with typical symptoms. Further, the identification of Barrett's esophagus in the absence of typical reflux symptoms suggests the potential for occult disease progression and late discovery of cancer. PMID- 18677539 TI - Distal pancreatectomy is not associated with increased perioperative morbidity when performed as part of a multivisceral resection. AB - PURPOSE: To evaluate the indications for and the outcomes from distal pancreatectomy. METHODS: Retrospective chart review of 171 patients who underwent distal pancreatectomy at Brigham and Women's Hospital between January 1996 and August 2005. RESULTS: Nearly one-third of distal pancreatectomies were performed as part of an en bloc resection for a contiguous or metastatic tumor. Fifty-six percent of the patients underwent a standard distal pancreatectomy +/- splenectomy (group 1), whereas 44% of distal pancreatic resections included additional organs or contiguous intraperitoneal or retroperitoneal tumor (group 2). The overall post-operative complication rate was 37%; the most common complication was pancreatic duct leak (23%). When compared to patients undergoing standard distal pancreatectomy, those with a more extensive resection including multiple viscera and/or metastatic or contiguous tumor resection had no significant difference in overall complication rate (35% v. 39%, p = 0.75), leak rate (25% v. 20%, p = 0.47), new-onset insulin-dependent diabetes mellitus (3% v. 4%, p = 1.0), and mortality (2% v. 4%, p = 0.656). CONCLUSION: This series includes a large number of patients in whom distal pancreatectomy was performed as part of a multivisceral resection or with en bloc resection of contiguous tumor. Complications were no different in these patients when compared to patients undergoing straightforward distal pancreatectomy. PMID- 18677540 TI - Clinicopathological feature and surgical outcome of choledochal cyst in different age groups: the implication of surgical timing. AB - BACKGROUND/AIMS: Surgical resection of choledochal cysts (CC) has become standard treatment. However, surgery is not universally recommended in early infancy and/or asymptomatic patients. In order to investigate the optimal timing of CC excision, we analyzed clinicopathological data and surgical results from different age groups. MATERIAL AND METHODS: This retrospective review included 107 patients (77 females, 30 males) who underwent CC resection at the National Taiwan University Hospital between January 1988 and December 2005. Patient demographic, clinical, and surgical data were collected and analyzed. RESULTS: The patients were divided into three groups according to age at the time of surgery: <1 year old (group I, n = 26), 1-16 years old (group II, n = 48), and >16 years old (group III, n = 33). About two thirds of the patients in group I had jaundice, while abdominal pain related to inflammation was the commonest symptom in groups II and III. Group I suffered significantly fewer surgical complications and less severe liver fibrosis than groups II or III. CONCLUSION: CC surgery in infancy and in asymptomatic patients is safe and may prevent the complications of this condition. The results support a recommendation for early excision. PMID- 18677541 TI - Esophageal extension encountered during transhiatal resection of gastric or gastroesophageal tumors: attaining a negative margin. AB - INTRODUCTION: Over the last several decades, the incidence of gastroesophageal junction tumors has been increasing. Often, patients present late in the course of their disease. However, if the disease is localized, then complete surgical resection remains the standard of cure and the best chance for cure. On occasion, these tumors involve a significant portion of both the distal esophagus and proximal stomach. MATERIALS AND METHODS: In order to completely remove these tumors with an adequate surgical margin and lymph node dissection, a total gastrectomy and total esophagectomy with colonic interposition may be required. We have utilized this approach on six patients with excellent clinical results. In this manuscript, we discuss the technical considerations involved in this approach and present our results. PMID- 18677542 TI - Loss of heterozygosity predicts poor survival after resection of pancreatic adenocarcinoma. AB - BACKGROUND: American Joint Committee on Cancer (AJCC) staging for pancreatic adenocarcinoma is a validated predictor of prognosis but insufficiently discriminates postresection survival. We hypothesized that genetic analysis of resected cancers would correlate with tumor biology and postoperative survival. METHODS: Resected pancreatic ductal and ampullary adenocarcinomas (n = 50) were analyzed for loss of heterozygosity (LOH) at 15 markers including 5q(APC), 6q(TBSP2), 9p(p16), 10q(PTEN), 12q(MDM2), 17p(TP53), and 18q(DCC/SMAD4). KRAS exon 1 mutations were detected by sequencing. The primary endpoint of this interim data analysis was survival at 18 month median follow-up. RESULTS: Negative margins were achieved in 43 (86%) cases. AJCC stage was: Ia/b (3), IIa (16), IIb (31). KRAS mutations were detected in 31 cases (62%) and LOH in 26 (52%) with mean fractional allelic loss score 23 +/- 16%. Median survival was significantly shorter with LOH (15.2 months versus not reached; p = 0.021) and KRAS mutations (19.6 months versus not reached; p = 0.038). Combining KRAS mutation with LOH was a powerful negative predictor in Cox regression (HR = 10.6, p = 0.006). Stage, nodal and margin status were not predictive of survival. CONCLUSION: LOH and KRAS mutations indicate aggressive tumor biology and correlate strongly with survival in resected pancreatic ductal and ampullary carcinomas. Genetic analysis may improve risk stratification in future clinical trials. PMID- 18677543 TI - No effects of olive oils with different phenolic content compared to corn oil on 1,2-dimethylhydrazine-induced colon carcinogenesis in rats. AB - BACKGROUND: Some epidemiological and experimental studies suggest that olive oil, despite its elevated caloric content, may have protective activity against colon cancer, partially due to its phenolic content. However, little experimental evidence exists to support this claim in vivo. AIM OF THE STUDY: To test the effect of olive oils with different phenolic content in a well-characterized model of colon carcinogenesis, comparing them with corn oil (CO). METHODS: F344 rats were fed AIN-76 based diets for the entire experimental period; the diets contained 23% (w/w) of lipids from three different sources: extra-virgin olive oil rich in phenolic compounds (EV), rectified olive oil (ROO) with the same fatty acid composition but devoid of phenolic compounds and CO as a control diet. One week later, rats were induced with 1,2-dimethylhydrazine (DMH) (150 mg/kg b.w. x 2 times) to measure preneoplastic lesions (aberrant crypt foci (ACF) and mucin depleted foci (MDF)) and tumours in the intestine. RESULTS: Thirteen weeks after DMH, the numbers of ACF and MDF were similar in the different groups (ACF/colon were 344.9 +/- 27.0, 288.6 +/- 28.5 and 289.8 +/- 21.4 in CO, EV and ROO groups, respectively, means +/- SE; MDF/colon were 8.83 +/- 1.2, 8.41 +/- 1.5 and 8.75 +/- 1.6 in CO, EV and ROO groups, respectively, means +/- SE). Thirty two weeks after DMH, the incidence of tumours (rats with tumours/rats in the group) did not differ among the different groups (20/21, 18/19 and 20/20 in the CO, EV, and ROO groups, respectively). Similarly, the number of tumours/ rat in the colorectum (both adenomas and cancers) was not different in the three different groups (2.33 +/- 0.26, 2.42 +/- 0.41 and 2.25 +/- 0.40 in CO, EV and ROO groups, respectively, means +/- SE). CONCLUSIONS: Olive oil, irrespective of its phenolic content, does not affect DMH-induced colon carcinogenesis in F344 rats compared with CO. PMID- 18677544 TI - Overweight and obesity and their relation to dietary habits and socio-demographic characteristics among male primary school children in Al-Hassa, Kingdom of Saudi Arabia. AB - BACKGROUND: Several studies were carried out to study the prevalence of overweight and obesity among Saudi children, but those assessed the association between eating habits, socio-demographic differentials and obesity in these children are scarce. OBJECTIVES: To assess the magnitude of obesity and overweight among male primary schoolchildren and to find the possible association between obesity/overweight with dietary habits and socio-demographic differentials among them. STUDY DESIGN AND METHODS: A cross-sectional descriptive study including 1,139 Saudi male enrolled in the fifth and sixth grades in public primary schools in Al Hassa, KSA, through a multistage random sampling technique, submitted to interview using Youth and Adolescent Food Frequency Questionnaire, gathering data regarding dietary intake, some dietary habits, followed by anthropometric measurements with calculation of body mass index, the interpretation of which was based on using Cole's tables for standard definition of overweight and obesity. Socio-demographics data were collected through parental questionnaire form. Data analysis was carried out using SPSS 12 (SPSS Inc. Chicago, IL, USA), univariate as well as multivariate analyses were conducted. RESULTS: The age ranged from 10 to 14 years. The prevalence of overweight among the included subjects was 14.2% while obesity was 9.7%, more in urban, older age students, mothers of obese and overweight were less educated, more working. Missing and or infrequent intake of breakfast at home, frequent consumption of fast foods, low servings of fruits, vegetables, milk and dairy product per day, with frequent consumption of sweets/candy and carbonated drinks were all predictors of obesity and overweight among the included male schoolchildren. CONCLUSION: The prevalence of childhood obesity is escalating and approaching figures reported in the developed countries. Less healthy dietary habits and poor food choices may be responsible for this high prevalence. PMID- 18677545 TI - Analyzing deforestation rates, spatial forest cover changes and identifying critical areas of forest cover changes in North-East India during 1972-1999. AB - Deforestation is recognized as one of the most significant component in LULC and global changes scenario. It is imperative to assess its trend and the rates at which it is occurring. The changes will have long-lasting impact on regional climate and in turn on biodiversity. In North-East India, one of the recognized global biodiversity hotspots, approximately 30% of total forest cover is under pressure of rapid land use changes. This region harbors variety of rare and endemic species of flora and fauna. It also has a strong bearing on regional climatic conditions. Extensive shifting cultivation, compounded by increasing population pressure and demands for agriculture land are the prime drivers in addition to other proximate drivers of deforestation. It is therefore of prime concern to analyse forest cover changes in the region, assess rate of change and extent and to identify the areas, which show repetitive changes. We analyzed forest cover maps from six temporal datasets based on satellite data interpretation, converted to geospatial database since 1972 till 1999. The states of Meghalaya, Nagaland and Tripura show highest changes in forest cover. Arunachal Pradesh shows least dynamic areas and maintains a good forest cover owing to its topographical inaccessibility in some areas. The present study reports the forest cover changes in the region using geospatial analysis and analyse them to devise proper management strategies. PMID- 18677546 TI - Using the Sediment Quality Triad to characterize baseline conditions in the Anacostia River, Washington, DC, USA. AB - The Sediment Quality Triad (SQT) consists of complementary measures of sediment chemistry, benthic community structure, and sediment toxicity. We applied the SQT at 20 stations in the tidal portion of the Anacostia River from Bladensburg, MD to Washington, DC to establish a baseline of conditions to evaluate the effects of management actions. Sediment toxicity was assessed using 10-day survival and growth tests with the freshwater amphipod, Hyalella azteca and the midge, Chironomus dilutus. Triplicate grabs were taken at each station for benthic community analysis and the Benthic Index of Biotic Integrity (B-IBI) was used to interpret the data. Only one station, #92, exhibited toxicity related to sediment contamination. Sediments from this station significantly inhibited growth of both test species, had the highest concentrations of contaminants, and had a degraded benthic community, indicated by a B-IBI of less than 3. Additional sediment from this station was tested and sediment toxicity identification evaluation (TIE) procedures tentatively characterized organic compounds as the cause of toxicity. Overall, forty percent of the stations were classified as degraded by the B-IBI. However, qualitative and quantitative comparisons with sediment quality benchmarks indicated no clear relationship between benthic community health and contaminant concentrations. This study provides a baseline for assessing the effectiveness of management actions in the Anacostia River. PMID- 18677548 TI - Biological responses to sediment remediation based on thin layer placement near a former pulp mill in Ward Cove, AK (USA). AB - In 2001, 28 acres of the bottom of Ward Cove, AK (USA) were remediated using thin layer placement, to enhance the natural recovery of contaminated sediments. The remediated areas were part of an 80-acre area of concern identified offshore from the site of a former sulfite pulp mill. The primary chemicals of concern were those commonly associated with organic enrichment near pulp mills: ammonia and 4 methylphenol. The primary remedial objectives were to reduce the toxicity of the contaminated sediments and to stimulate colonization of the remediated areas by benthic macroinvertebrates. In 2004, the initial monitoring event for the remediated areas was conducted, and included evaluations of physical/chemical sediment variables (i.e., total organic carbon, grain size distribution, ammonia, and 4-methylphenol), sediment toxicity (i.e., using the 10-day amphipod survival test with Eohaustorius estuarius), and in situ benthic macroinvertebrate communities. Results of the monitoring event showed that conditions in the remediated areas had improved considerably in the 3 years since thin layer placement had occurred. At most stations, concentrations of both ammonia and 4 methylphenol were very low, and amphipod survival was >or=90%. In addition, benthic macroinvertebrates appeared to be rapidly colonizing the remediated areas, based on evaluations of several community metrics (i.e., taxa richness, diversity, and dominance), as well as key indicator species of organic enrichment and associated transitional areas (i.e., primarily the polychaetes Capitella capitata and Nephtys cornuta, and the bivalve molluscs Axinopsida serricata and Parvilucina tenuisculpta). In general, colonization was consistent with the patterns identified for areas recovering from organic enrichment on the continental shelf of southern California and elsewhere. Based on these results, thin layer placement was considered successful in enhancing the natural recovery of the remediated sediments in Ward Cove. PMID- 18677547 TI - The occurrence of glyphosate, atrazine, and other pesticides in vernal pools and adjacent streams in Washington, DC, Maryland, Iowa, and Wyoming, 2005-2006. AB - Vernal pools are sensitive environments that provide critical habitat for many species, including amphibians. These small water bodies are not always protected by pesticide label requirements for no-spray buffer zones, and the occurrence of pesticides in them is poorly documented. In this study, we investigated the occurrence of glyphosate, its primary degradation product aminomethylphosphonic acid, and additional pesticides in vernal pools and adjacent flowing waters. Most sampling sites were chosen to be in areas where glyphosate was being used either in production agriculture or for nonindigenous plant control. The four site locations were in otherwise protected areas (e.g., in a National Park). When possible, water samples were collected both before and after glyphosate application in 2005 and 2006. Twenty-eight pesticides or pesticide degradation products were detected in the study, and as many as 11 were identified in individual samples. Atrazine was detected most frequently and concentrations exceeded the freshwater aquatic life standard of 1.8 micrograms per liter (microg/l) in samples from Rands Ditch and Browns Ditch in DeSoto National Wildlife Refuge. Glyphosate was measured at the highest concentration (328 microg/l) in a sample from Riley Spring Pond in Rock Creek National Park. This concentration exceeded the freshwater aquatic life standard for glyphosate of 65 microg/l. Aminomethylphosphonic acid, triclopyr, and nicosulfuron also were detected at concentrations greater than 3.0 microg/l. PMID- 18677549 TI - Ecology and abundance of zooplankton in Karanja reservoir. AB - The present work aims to study the Zooplankton seasonal distribution and diversity in Karanja reservoir, Bidar district for the period of two years from October 2001 to September 2003. We have recorded 36 species of which, 15 species belongs to rotifera, 11 species belongs to cladocera, nine species belongs to copepoda and ostracoda three species. Among zooplankton, particularly rotifera was the dominant group throughout the study period and highest count was recorded in the month of March 2003. Cluster analysis was also revealed by dendrogram and Shannon diversity index is also presented. Zooplankton community is also correlated with physicochemical parameters. PMID- 18677550 TI - Effectiveness, efficiency, and e-learning. PMID- 18677551 TI - An integrated method for improving the dairy production sector in developing countries: the case of Cameroon. AB - During the past six years, ten research topics were carried out with the aim of developing an integrated method to improve production and sustainability of dairy systems in Cameroon. This involved reviewing dairy research done in the country, carrying out a participatory rural appraisal and an economic opportunity survey in selected dairy farms, setting up on-farm interventions, investigating cow reproduction, evaluating milk quality and the impact of integrated interventions. Guidelines for improvement of the dairy sector were set up. It was found that the developed integrated method had a positive impact on dairy farms. Farmers who adopted interventions had nearly 200% higher economic returns. In order to boost the Cameroonian dairy sector, it is suggested that the government acts as a motivating force by organizing the market, ensuring the monitoring of epizootic diseases and providing artificial insemination services and organizing breeding societies. It is also suggested that the integrated method becomes a discipline in dairy science and be applied in other developing countries. PMID- 18677552 TI - Abstracts of the Annual Symposium of the Society for the Study of Inborn Errors of Metabolism. Lisboa, Portugal. September, 2-5, 2008. PMID- 18677553 TI - High-throughput crystallization-to-structure pipeline at RIKEN SPring-8 Center. AB - A high-throughput crystallization-to-structure pipeline for structural genomics was recently developed at the Advanced Protein Crystallography Research Group of the RIKEN SPring-8 Center in Japan. The structure determination pipeline includes three newly developed technologies for automating X-ray protein crystallography: the automated crystallization and observation robot system "TERA", the SPring-8 Precise Automatic Cryosample Exchanger "SPACE" for automated data collection, and the Package of Expert Researcher's Operation Network "PERON" for automated crystallographic computation from phasing to model checking. During the 5 years following April, 2002, this pipeline was used by seven researchers to determine 138 independent crystal structures (resulting from 437 purified proteins, 234 cryoloop-mountable crystals, and 175 diffraction data sets). The protocols used in the high-throughput pipeline are described in this paper. PMID- 18677554 TI - Synchronization of Na/K pump molecules by an oscillating electric field. AB - Synchronization of the Na/K pump molecules in a cell membrane was studied in frog skeletal muscle fibers using double Vaseline-gap voltage-clamp techniques. We found that the pumping rate of naturally random-paced pump molecules can be artificially synchronized by a pulsed, symmetric, oscillating membrane potential with a frequency comparable to the physiological turnover rate. The synchronized pump currents show separated outward and inward components, where the magnitude of the outward component is about three times the randomly-paced pump currents, and the magnitude-ratio of the outward to inward pump currents is close to 3:2, which reflects the stoichiometric ratio of the pump molecules. Once synchronized, the pumping rate is restricted to the field frequency, and the pump currents are mainly dependent on the field frequency, but not the field strength. In contrast to previous work, which by restraining the pumps at a presteady state succeeded in triggering the steps of the pump cycle only individually and between interruptions, here we synchronize the pumps running continuously and in a normal running mode. PMID- 18677555 TI - Mitochondrial membrane cholesterol, the voltage dependent anion channel (VDAC), and the Warburg effect. AB - Normal cells of aerobic organisms synthesize the energy they require in the form of ATP via the process of oxidative phosphorylation. This complex system resides in the mitochondria of cells and utilizes oxygen to produce a majority of cellular ATP. However, in most tumors, especially those with elevated cholesterogenesis, there is an increased reliance on glycolysis for energy, even in conditions where oxygen is available. This aerobic glycolysis (the Warburg effect) has far reaching ramifications on the tumor itself and the cells that surround it. In this brief review, we will discuss how abnormally high membrane cholesterol levels can result in a subsequent deficiency of oxidative energy production in mitochondria from cultured Morris hepatoma cells (MH-7777). We have identified the voltage dependent anion channel (VDAC) as a necessary component of a protein complex involved in mitochondrial membrane cholesterol distribution and transport. Work in our laboratory demonstrates that the ability of VDAC to influence mitochondrial membrane cholesterol distribution may have implications on mitochondrial characteristics such as oxidative phosphorylation and induction of apoptosis, as well as the propensity of cancer cells to exhibit a glycolytic phenotype. PMID- 18677556 TI - Tapping healthcare resource by older Mexicans with diabetes: how migration to the United States facilitates access. AB - This study employs semi-structured interviews conducted in two small migrant towns in Michoacan, Mexico to determine how older Mexicans with diabetes access healthcare and treatment. Barriers in access to healthcare services are identified as well as how family members and migration are utilized to offset these limitations. Results indicate that former migrants who were economically successful in the USA have greater options in the type of services they are able to use. In addition, residents with diabetes who have family members living in the USA receive remittances in the form of money and medical supplies that aid in the treatment of their illness. Implications for policy are discussed. PMID- 18677557 TI - Preimplantation HLA haplotyping using tri-, tetra-, and pentanucleotide short tandem repeats for HLA matching. AB - PURPOSE: To aid couples wishing to conceive children who are HLA matched to a sibling in need of a hematopoietic progenitor cell transplant, we developed a preimplantation HLA haplotype analysis of embryos that utilizes tri-, tetra-, and pentanucleotide STR markers. METHODS: For preimplantation HLA genotyping, we use polymorphic STR markers located across the HLA and flanking regions, selecting exclusively tri-, tetra-, and pentanucleotide repeats. These markers can be resolved using either capillary electrophoresis (CE) or polyacrylamide gels. RESULTS: We have developed 43 reliable STR markers for preimplantation HLA matching. Selected STR markers enabled unambiguous identification of embryos whose HLA haplotypes were matched with the affected patient using polyacrylamide gel or capillary electrophoresis. CONCLUSIONS: The use of tri-, tetra-, and pentanucleotide repeat markers and polyacrylamide gels for STR genotyping in HLA matching is a simple and cost effective approach to clinical testing. PMID- 18677558 TI - Characterization of the weak estrogen receptor alpha agonistic activity of exemestane. AB - Third generation aromatase inhibitors (AI) have shown good clinical efficacy in comparison to the anti-estrogen tamoxifen. The steroidal AI, exemestane (EXE) has previously been shown to act as an androgen, but this report demonstrates the estrogen-like activity of EXE. Based on genome-wide microarray analysis, high correlation was seen between EXE-Only (EXE O, hormone-free) and hormone containing AI-resistant lines. In addition, the top regulated genes in the EXE O lines were mostly estrogen-responsive genes. This estrogen-like activity of EXE was further validated using estrogen receptor (ER) activity assays, where in comparison to 17beta-estradiol (E2), EXE was able to induce ER activity, though at a higher concentration. Also, this EXE-mediated ER activity was blocked by the ER antagonist ICI as well as the ERalpha-specific antagonist methyl-piperidino pyrazole (MPP). Similarly, EXE was able to induce proliferation of breast cancer cell lines, MCF-7 and MCF-7aro, as well as activate transcription of known estrogen-responsive genes, i.e., PGR, pS2 and AREG. These results suggest that EXE does have weak estrogen-like activity. PMID- 18677559 TI - Reversal of antiprogestin resistance and progesterone receptor isoform ratio in acquired resistant mammary carcinomas. AB - To explore mechanisms related to hormone resistance, three resistant variants of the MPA mouse breast cancer tumor model with low levels of progesterone receptor (PR) isoform A (PR-A)/high PR-B expression were developed by prolonged selective pressure with antiprogestins. The resistant phenotype of one tumor line was reversed spontaneously after several consecutive passages in syngeneic BALB/c mice or by 17-beta-estradiol or tamoxifen treatment, and this reversion was significantly associated with an increase in PR-A expression. The responsive parental tumors disclosed low activation of ERK and high activation of AKT; resistant tumors on the other hand, showed the opposite, and this was associated with a higher metastatic potential, that did not revert. This study shows for the first time in vivo a relationship between PR isoform expression and antiprogestin responsiveness, demonstrating that, whereas acquired resistance may be reversed, changes in kinase activation and metastatic potential are unidirectional associated with tumor progression. PMID- 18677560 TI - The prognostic and predictive values of circulating HER-2/neu extracellular domain in patients with metastatic breast cancer. PMID- 18677561 TI - Elevated beta1,4-galactosyltransferase-I induced by the intraspinal injection of lipopolysaccharide. AB - Beta1,4-galactosyltransferase-I (beta1,4-GalT-I) is one of the best studied glycosyltransferases. Previous studies demonstrated that beta1,4-GalT-I was a major galactosyltransferase responsible for selectin-ligand biosynthesis and that inflammatory responses of beta1,4-GalT-I deficient mice were impaired. In this study, we investigate the expression of beta1,4-GalT-I in lipopolysaccharide (LPS)-induced neuroinflammatory processes. The results of this study demonstrated that beta1,4-GalT-I was strongly induced by intraspinal administration of LPS. More than 90% galactose-containing glycans and beta1,4-GalT-I were expressed in immune cells. The ELISA assay shows focal injection LPS also induces TNF-alpha alteration. Double staining indicated beta1,4-GalT-I overlapped with TNF-alpha. Moreover, RT-PCR for beta1,4-GalT-I mRNA showed that beta1,4-GalT-I mRNA in microglia in vitro was affected in a dose- and time dependent manner in response to LPS or TNF-alpha stimulation. All these results indicated that the increase of beta1,4-GalT-I might attribute to the effect of TNF-alpha excreting during inflammation. E-selectin, which ligand was modified by beta1,4-GalT-I, was correlated with galactose-containing glycans following injecting LPS into spinal cord. We therefore suggest that beta1,4-GalT-I may play an important role in regulating immune cell migration into the inflammatory site. PMID- 18677562 TI - Corticosterone in relation to tissue cadmium, mercury and selenium concentrations and social status of male lesser scaup (Aythya affinis). AB - Combined lesser scaup (Aythya affinis) and greater scaup (A. marila) populations have declined steadily from the 1970s. Accompanying the population decline have been two shifts in lesser scaup demographics: a decrease in the proportion of young birds and an increase in male to female ratio. In addition, there are concerns about potential effects of contaminants and trace elements. These metals may influence the stress response and corticosterone secretion. We examined impacts of cadmium, selenium and mercury on the stress response in relation to social status in male lesser scaup near Yellowknife, NWT May to June 2004 and 2005. Kidney cadmium and liver selenium and mercury ranged 0.78-93.6, 2.12-9.64, and 0.56-3.71 microg/g, dry weight, respectively. Results suggest that corticosterone release may be influenced by complex contaminant interactions in relation to body condition and body size. When cadmium was high and birds were in good body condition, there was a negative relationship between liver selenium and corticosterone (R(2) = 0.60, n = 10, P = 0.008) but not in birds with poor body condition (R(2) = 0.07, n = 9, P = 0.50). Unfortunately we were unable to draw any conclusions about metals and social status in relation to corticosterone or glucose and T(4). This study emphasizes the complex nature of biological systems and the importance of considering interactions to characterize effects of metals. PMID- 18677563 TI - Dp71f modulates GSK3-beta recruitment to the beta1-integrin adhesion complex. AB - Previously, it was shown that Dp71f binds to the beta1-integrin adhesion complex to modulate PC12 cell adhesion. The absence of Dp71f led to a failure in the beta1-integrin adhesion complex formation. One of the structural proteins which links the beta1-integrin cytoplasmic domain to the actin cytoskeleton is ILK. GSK3-beta is an ILK substrate and the carboxi-terminal region of dystrophin 427 is a substrate for hierarchical phosphorylation by GSK3-beta. Dp71f contains the carboxi-terminal domain present in dystrophin 427. By using co immunoprecipitation assays, in the present work it is demonstrated that in the neuronal PC12 cell line an interaction between Dp71f and GSK3-beta occurs. This interaction was corroborated by in vitro pulldown assays. We show that GSK3-beta is recruited to the beta1-integrin complex and that a reduced expression of Dp71f induces a reduced GSK3-beta recruitment to the beta1-integrin complex. In addition, the present work establishes that adhesion of PC12 cells to laminin does not influence the phosphorylation status of Dp71f. PMID- 18677564 TI - Inhibition of P-glycoprotein over-expression by shRNA-mdr1b in rat astrocytes. AB - The development of multiple drug resistance (MDR) is a significant problem in epilepsy therapy. The primary factor responsible for antiepileptic drug (AEDs) resistance is the over-expression of the MDR gene product, P-glycoprotein (Pgp). To model a therapeutic approach for decreasing drug resistance in patients with intractable epilepsy, we established a model of coriaria lactone (CL) induced Pgp overexpression in rat astrocytes and administered a recombinant adenovirus Ad5 EGFP-shRNA1-U6 to deliver an anti-mdr1b short hairpin RNA (shRNA) for 5 days. We then investigated the gene-silencing effects of shRNA by quantitative real-time RT-PCR, Western-blot, and Rho123 accumulation assay. The results showed that over expression of mdr1b and Pgp was successfully suppressed, the ability of intracellular Rho123 retention was increased, and drug efflux was decreased in the adenovirus treated astrocytes. In conclusion, MDR was reversed in rat astrocyte model. These findings may be favorable for developing new therapeutic strategies for treating intractable epilepsy. PMID- 18677565 TI - Towards an implantable biochip for glucose and lactate monitoring using microdisc electrode arrays (MDEAs). AB - A complete electrochemical cell-on-a-chip that uses the MicroDisc Electrode Array (MDEA) working electrode (WE) design was evaluated for eventual intramuscular implantation for the continuous amperometric monitoring of glucose and lactate in an animal trauma model. The microfabricated ECC MDEA5037 comprises two discrete electrochemical cells-on-a-chip (ECCs), each with a reference, counter, and MDEA working electrode. Each MDEA comprises 37 microdiscs (diameter = 50 microm) arranged in a Hexagonal Closed Packed (HCP) arrangement with a center to center distance (d) of 100 microm. Cyclic Voltammetry (CV) and Electrical Impendence Spectroscopy (EIS) reveals that this device scales in its interfacial properties with a corresponding MDEA 050 device that comprises 5,184 microdiscs. Parallel development of miniaturized mixed-signal integrated electronics for wireless reprogramming, data acquisition and communication addresses the key issues involved in developing measurement electronics, AD/DA processing, power management and telemetry for implantable amperometric biosensors. A generalized electronics platform based on the Texas Instruments TI NC01101 chip has been developed that may be readily applied to many types of biotransducers with minor modifications. PMID- 18677567 TI - Best interests: a reappraisal. PMID- 18677566 TI - eNpHR: a Natronomonas halorhodopsin enhanced for optogenetic applications. AB - Temporally precise inhibition of distinct cell types in the intact nervous system has been enabled by the microbial halorhodopsin NpHR, a fast light-activated electrogenic Cl(-) pump. While neurons can be optically hyperpolarized and inhibited from firing action potentials at moderate NpHR expression levels, we have encountered challenges with pushing expression to extremely high levels, including apparent intracellular accumulations. We therefore sought to molecularly engineer NpHR to achieve strong expression without these cellular side effects. We found that high expression correlated with endoplasmic reticulum (ER) accumulation, and that under these conditions NpHR colocalized with ER proteins containing the KDEL ER retention sequence. We screened a number of different putative modulators of membrane trafficking and identified a combination of two motifs, an N-terminal signal peptide and a C-terminal ER export sequence, that markedly promoted membrane localization and ER export defined by confocal microscopy and whole-cell patch clamp. The modified NpHR displayed increased peak photocurrent in the absence of aggregations or toxicity, and potent optical inhibition was observed not only in vitro but also in vivo with thalamic single-unit recording. The new enhanced NpHR (eNpHR) allows safe, high-level expression in mammalian neurons, without toxicity and with augmented inhibitory function, in vitro and in vivo. PMID- 18677568 TI - Best interests: what problems in family law should health care law avoid? AB - This article comments briefly on three specific issues in Shazia Choudhry's paper "'Best Interests' What can healthcare law learn from family law?" The three issues are: (1) the implications of 'best interests' and 'welfare science' for women within the family law and the health care law context, (2) the risk of capture by the 'welfare science' industry, and (3) the proposal that a committee of medical experts and medical ethicists should be set up to provide reports to the Court of Protection on cases brought under the Mental Capacity Act 2005 (MCA). I argue that the risk of capture by 'welfare science' is equally large in health care law and that a committee of the kind envisaged by Choudhry is unlikely to contribute significantly to conflict resolution under the MCA. PMID- 18677569 TI - Best interests and treatment for mental disorder. AB - This paper considers the role of the concept of best interests in the treatment of mental disorder. It considers the Mental Capacity Act 2005 where treatment of an incapacitated person's mental disorder is authorized if treatment is in the patient's own best interests. It also examines the Mental Health Act 1983 as amended by the Mental Health Act 2007 where treatment without consent of a detained patient is allowed where necessary for the patient's health or safety or for the protection of others. Under both statutory regimes treatment must be in the best interests of the patient. This paper argues that 'best interests' is open to interpretation to include treatment interventions carried out primarily to protect other people. PMID- 18677570 TI - Best interests in the MCA 2005--what can healthcare law learn from family law? AB - The 'best interests' standard is a highly seductive standard in English law. Not only does it appear to be fairly uncontroversial but it also presents as the most sensible, objective and 'fair' method of dealing with decision making on behalf of those who are perceived to be the most vulnerable within society. This article aims to provide a critical appraisal of how the standard has been applied within family law, to outline how the standard is to be applied within healthcare law and, finally, to assess the relevance of the family law experience of the best interests standard to the operation of the standards as envisaged by the MCA. PMID- 18677571 TI - Effect of pluronic p85 on amino acid transport in bovine brain microvessel endothelial cells. AB - A synthetic amphiphilic block copolymer Pluronic P85 (P85) was shown to be among the most potent inhibitors of Pgp efflux system in the blood-brain barrier (BBB) and capable of enhancing delivery of Pgp substrates to the brain. The purpose of this work is to evaluate the effects of P85 on amino acid transport in BBB. Primary bovine brain microvessel endothelial cells (BBMEC) grown on membrane inserts were used as an in vitro BBB model. Expression of amino acid transporters, like large neutral amino acid transporter 1, cationic amino acid transporter 1, and small neutral amino acid transporter 1, were confirmed by reverse transcriptase polymerase chain reaction. Effects of P85 on amino acid transporters were examined using their substrates: (3)H-phenylalanine, (3)H lysine, and (3)H-methylaminoisobutyric acid, respectively. BBMEC permeability studies were carried out in apical (AP) to basolateral (BL) and BL to AP directions. P85 added at the AP side had little, if any, effect on AP to BL ("blood to brain") transport for all examined amino acids in BBMEC monolayers. However, 0.1% P85 added at the BL side significantly increased the BL to AP transport of these substrates. Furthermore, the effective concentrations of P85 were also shown to induce plasma membrane depolarization and increase intracellular sodium concentration in BBMEC, which can contribute to the effects of the copolymer on the energy-dependent transport systems. All together, despite profound effects on transport system(s) at the brain side of cell monolayers, P85 had no effect on AP to BL transport of amino acids in brain microvessel endothelial cell model. PMID- 18677572 TI - The inhibitory effect of an RGD-human chitin-binding domain fusion protein on the adhesion of fibroblasts to reacetylated chitosan films. AB - Biomaterials used for tissue engineering applications must provide a structural support for the tissue development and also actively interact with cells, promoting adhesion, proliferation, and differentiation. To achieve this goal, adhesion molecules may be used, such as the tripeptide Arg-Gly-Asp (RGD). A method based on the use of a carbohydrate-binding module, with affinity for chitin, was tested as an alternative approach to the chemical grafting of bioactive peptides. This approach would simultaneously allow the production of recombinant peptides (alternatively to peptide synthesis) and provide a simple way for the specific and strong adsorption of the peptides to the biomaterial.A fusion recombinant protein, containing the RGD sequence fused to a human chitin binding module (ChBM), was expressed in E. coli. The adhesion of fibroblasts to reacetylated chitosan (RC) films was the model system selected to analyze the properties of the obtained proteins. Thus, the evaluation of cell attachment and proliferation on polystyrene surfaces and reacetylated chitosan films, coated with the recombinant proteins, was performed using mouse embryo fibroblasts 3T3. The results show that the recombinant proteins affect negatively fibroblasts anchorage to the materials surface, inhibiting its adhesion and proliferation. We also conclude that this negative effect is fundamentally due to the human chitin binding domain. PMID- 18677573 TI - Twelve-month treatment of psychiatric disorders in the South African Stress and Health Study (World Mental Health Survey Initiative). AB - BACKGROUND: The proportion of people with mental disorders in treatment is relatively small in low and middle income countries. However, little is known about patterns of recent service use in a country like South Africa. METHODS: A nationally representative household survey of 4,351 adult South Africans was carried out. Twelve-month DSM-IV disorders were determined using the WHO composite international diagnostic interview (CIDI). Prevalence and correlates of treatment were assessed among respondents with anxiety, mood and substance use disorders. RESULTS: One-fourth (25.5%) of respondents with a 12-month disorder had received treatment in the past 12 months either from a psychiatrist (3.8%), nonpsychiatrist mental health specialist (2.9%), general medical provider (16.6%), human services provider (6.6%), or complementary-alternative medical (CAM) provider (5.9%). Only 27.6% of severe cases had received any treatment. In addition, 13.4% of respondents with no disorder had accessed services in the past year. Blacks were significantly more likely than other racial groups to access the CAM sector while Whites were more likely to have seen a psychiatrist. CONCLUSIONS: The majority of South Africans with a 12-month mental disorder have unmet treatment needs. In addition to a greater allocation of resources to mental health services, more community outreach and awareness initiatives are needed. PMID- 18677574 TI - Equine herpesvirus infections in yearlings in South-East Queensland. AB - Twelve nasal swabs were collected from yearling horses with respiratory distress and tested for equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4) by real-time PCR targeting the glycoprotein B gene. All samples were negative for EHV-1; however, 3 were positive for EHV-4. When these samples were tested for EHV 2 and EHV-5 by PCR, all samples were negative for EHV-2 and 11 were positive for EHV-5. All three samples that were positive for EHV-4 were also positive for EHV 5. These three samples gave a limited CPE in ED cells reminiscent of EHV-4 CPE. EHV-4 CPE was obvious after 3 days and was characterised by syncytia. None of the samples produced cytopathic effect (CPE) on African green monkey kidney (Vero) cells or hamster kidney (BSR) cells. Four of the samples, which were positive in the EHV-5 PCR, produced CPE on rabbit kidney (RK13) cells and equine dermis (ED) cells. EHV-5 CPE on both cell lines was slow and was apparent after four 7-day passages. On RK13 cells, the CPE was characteristic of equid herpesvirus, with the formation of syncytia. However, in ED cells, the CPE was characterised by ring-shaped syncytia. For the first time, a case of equine respiratory disease involving dual infection with EHV-4 and EHV-5 has been reported in Queensland (Australia). This was shown by simultaneously isolating EHV-4 and EHV-5 from clinical samples. EHV5 was recovered from all samples except one, suggesting that EHV5 was more prevalent in young horses than EHV2. PMID- 18677575 TI - Characterization of Santa Catarina (Brazil) coal with respect to human health and environmental concerns. AB - The current paper presents the concentration, distribution, and modes of occurrence of trace elements of 13 coals from south Brazil. The samples were collected in the state of Santa Catarina. Chemical analyses and the high ash yields indicate that all studied coals are rich in mineral matter, with SiO(2) and Al(2)O(3) dominating as determined by inductively coupled plasma-atomic emission spectrometry (ICP-AES). Quartz is the main mineral species and is associated with minor levels of feldspars, kaolinite, hematite, and iron-rich carbonates. The contents of trace elements, including As, Pb, Cd, Ni, Cr, Mn, Be, V, U, Zn, Li, Cu, Tl, and Ni, in coals were determined. A comparison of ranges and means of elemental concentrations in Santa Catarina, Brazil, and world coals shows that the ranges of most elements in Santa Catarina coal are very close to the usual worldwide concentration ranges in coal. PMID- 18677576 TI - Left ventricular volumes and mass in marathon runners and their association with cardiovascular risk factors. AB - BACKGROUND: To assess left ventricular volumes and mass by cardiac magnetic resonance imaging in relation to conventional cardiovascular risk factors and coronary atherosclerotic plaque burden in master marathon runners aged > or =50 years. METHODS: Cardiac MRI was performed in 105 clinically healthy male marathon runners (mean age 57.3 +/- 5.7 years, range 50-71 years) on a 1.5 T MR system (Avanto, Siemens, Germany). Cine steady state free precession images in standard long and short axes views were acquired to assess left ventricular volumes and mass. Cardiovascular risk factors (blood pressure, HDL/LDL cholesterol, smoking, body mass index) were assessed and coronary artery calcification (CAC) was quantified by electron beam computed tomography. RESULTS: Left ventricular muscle mass (mean LVMM = 140 +/- 27 g; 73 +/- 13 g/m(2)) increased with increasing left ventricular end-diastolic volume (mean LVEDV = 137 +/- 32 ml; 72 +/- 15 ml/m(2)) (r = 0.41, P < 0.0001) and with systolic (r = 0.33, P = 0.005) and diastolic (r = 0.28, P = 0.005) blood pressures. Left ventricular EDV increased up to the age of 55 years, but decreased thereafter. Runners with LVMM > or =150 g had significantly higher CAC scores than runners with LVMM <150 g (median CAC score 110 vs. 25, P = 0.04). CONCLUSIONS: Increases in LVMM and LVEDV may not only represent a response to exercise but are dependent on age and blood pressure, also. In addition, a left ventricular hypertrophy without an increase in volume may be an indicator for early subclinical cardiac alterations in response to risk factor exposure. PMID- 18677577 TI - Ghrelin and obestatin levels in rheumatoid arthritis. AB - BACKGROUND: Ghrelin is a powerful, endogenous orexigenic peptide. In addition, ghrelin has anti-inflammatory effects, and it has been reported that ghrelin down regulates pro-inflammatory cytokines, including interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha. Obestatin appears to decrease food intake and appetite, and its potential role in inflammation is not yet clear. The aims of this study were to assess total and acylated (active) ghrelin and obestatin serum levels and their relations with inflammatory status in rheumatoid arthritis (RA) patients. DESIGN: Fasting blood samples were obtained from 37 patients with RA, 29 patients with Behcet's disease (BD) and 28 healthy controls (HC). Total ghrelin and obestatin levels were measured by radioimmunoassay and acylated ghrelin was quantified by enzyme-linked immunosorbent assay. RESULTS: Patients with RA had lower total ghrelin, but higher obestatin levels than patients with BD (p<0.05 for both), but when compared with HC group differences were not significant. There was no difference across groups in terms of acylated ghrelin. Total ghrelin level was not correlated with any study parameters in the all groups. Obestatin level correlated with erythrocyte sedimentation rate and DAS-28 in the RA group, the level of IL-6 in the BD group, and with the level of TNF alpha in the HC group (r=0.400, p<0.05; r=0.412, p<0.05, r=0.543, p<0.01 and r=0.528, p<0.05, respectively). CONCLUSIONS: Our results did not show a significant correlation between circulating ghrelin and clinical or laboratory markers of disease activity in RA. Surprisingly, obestatin correlated with some inflammatory markers. So, obestatin seems to be more valuable than ghrelin in the pathogenesis of RA. PMID- 18677578 TI - Rhamnus alpinus leaf extract suppresses lipopolysaccharide-induced, monocyte derived macrophage chemokine secretion. AB - Periodontitis is a chronic inflammatory disease of bacterial etiology that affects tooth-supporting tissues. The aim of the present study was to investigate the effect of Rhamnus alpinus extracts on lipopolysaccharide (LPS)-induced chemokine secretion by human macrophage-like cells. Phorbol myristic acid differentiated macrophages were stimulated with Aggregatibacter actinomycetemcomitans LPS in the absence and presence of various concentrations of the extracts. The secretion of interleukin-8 (IL-8), regulated on activation normal T cell expressed and secreted (RANTES), and monocyte chemotactic protein (MCP)-1 was measured using enzyme-linked immunosorbent assays (ELISA). Activation of NF-kappaB p65 was evaluated with an ELISA-based kit containing immobilized oligonucleotides with an NF-kappaB consensus binding site. A. actinomycetemcomitans LPS (1 microg/ml) induced a marked increase in the secretion of IL-8 and RANTES by monocyte-derived macrophages. At non-cytotoxic concentrations, the R. alpinus leaf extract, which contains polyphenols, inhibited the secretion of RANTES and, to a lesser extent, IL-8 in a dose dependent manner. The extract also decreased the basal levels of MCP-1 secreted by monocyte-derived macrophages. The extract appeared to exert its anti inflammatory effect by inhibiting NF-kappaB p65 activation. Our results suggest that the leaf extract of R. alpinus possesses a therapeutic potential through its capacity to limit the infiltration of immune cells into periodontal sites. This may impede the progression and aggravation of inflammation given that the migration of immune cells plays an important role in the outcome of periodontitis. PMID- 18677579 TI - Sequential expression of TLR4 and its effects on the myocardium of rats with myocardial ischemia-reperfusion injury. AB - This study was designed to explore the role of toll like receptor 4 (TLR4) in myocardial ischemia reperfusion injury (MI/RI). Male Sprague-Dawley rats were randomly divided into sham and IR groups (36/group). The rats were sacrificed at various times following reperfusion (0, 1/2, 1, 2, 4 and 8 h). The histopathological and ultrastructural changes in the myocardium were examined under a light microscope and a transmission electron microscope. The TLR4 protein and mRNA expression were detected by immunohistochemistry and real-time reverse transcription polymerase chain reaction, respectively. The levels of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 in the myocardium were measured by enzyme-linked immunosorbent assays. The injury to the myocardium was severe in the IR group, and there were no significant improvements in histopathology and ultrastructure of the myocardium during the first 8 h following reperfusion. Positive TLR4 protein staining was observed in both sham and IR groups. The TLR4 protein levels were significantly increased in the IR group, peaking at 1 h post reperfusion. Additionally, the TLR4 mRNA levels were also up-regulated in the IR group. At all time points, IR rats had significantly higher TNF-alpha and IL-6 levels than the sham rats (P<0.05). The TLR4 mRNA expression positively correlated with the levels of TNF-alpha and IL-6 (r=0.728 and 0.676, P<0.01). Myocardial TLR4 expression was elevated at the early stage of myocardial ischemia reperfusion. Activated TLR4 may play a role in MI/RI by increasing TNF-alpha and IL-6 expression. PMID- 18677580 TI - High level of sialate-O-acetyltransferase activity in lymphoblasts of childhood acute lymphoblastic leukaemia (ALL): enzyme characterization and correlation with disease status. AB - Previous studies had established an over-expression of 9-O-acetylated sialoglycoproteins (Neu5,9Ac(2)-GPs) on lymphoblasts of childhood acute lymphoblastic leukaemia (ALL). Here, we report the discovery and characterization of sialate-O-acetyltransferase enzyme in ALL-cell lines and lymphoblasts from bone marrow of children diagnosed with B- and T-ALL. We observed a positive correlation between the enhanced sialate-O-acetyltransferase activity and the enhanced expression of Neu5,9Ac(2)-GPs in these lymphoblasts. Sialate-O acetyltransferase activity in cell lysates or microsomal fractions of lymphoblasts of patients was always higher than that in healthy donors reaching up to 22-fold in microsomes. Additionally, the V (max) of this enzymatic reaction with AcCoA was over threefold higher in microsomal fractions of lymphoblasts. The enzyme bound to the microsomal fractions showed high activity with CMP-N acetylneuraminic acid, ganglioside GD3 and endogenous sialic acid as substrates. N-acetyl-7-O-acetylneuraminic acid was the main reaction product, as detected by radio-thin-layer chromatography and fluorimetrically coupled radio-high performance liquid chromatography. CMP and coenzyme A inhibited the microsomal enzyme. Sialate-O-acetyltransferase activity increased at the diagnosis of leukaemia, decreased with clinical remission and sharply increased again in relapsed patients as determined by radiometric-assay. A newly-developed non radioactive ELISA can quickly detect sialate-O-acetyltransferase, and thus, may become a suitable tool for ALL-monitoring in larger scale. This is the first report on sialate-O-acetyltransferase in ALL being one of the few descriptions of an enzyme of this type in human. PMID- 18677581 TI - Glycosylation at the fetomaternal interface: does the glycocode play a critical role in implantation? AB - During pregnancy, the heavily glycosylated surfaces of the implanting blastocyst and maternal uterine epithelium interact in a highly controlled and specific manner. Examination of this interface in species that show interdigitation of embryonic and maternal surfaces (epitheliochorial placentation) shows that each has its own particular pattern of glycosylation or glycotype, and that closely related and/or interbreeding species e.g. horse and donkey or llama and guanaco, have very similar glycotypes. Implantation of interspecies hybrids is facilitated, when the blastocyst has an outer cell layer bearing glycans that are compatible with the maternal host. We refer to this mutual compatibility as a glycocode. The probability that hybrid embryo glycotypes differ from those normally associated with the host species may account for the high pregnancy failure rates seen in interspecies breeding. We suggest the maternal host selects between genotypically distinct embryos, and this selection depends partly on cell surface glycosylation. We infer that the glycocode plays a critical role in implantation, for if the survival of modified genotypes results in fitter offspring with altered placental glycosylation, selection pressure downstream may in turn act to drive adaptations in the maternal surface glycotype to produce a complementary glycocode, thus leading eventually to the creation of new species. We speculate that glycan microheterogeneity plays a specific role in this process. PMID- 18677582 TI - Plasma levels of zinc, copper, copper/zinc ratio, and activity of carbonic anhydrase in equine piroplasmosis. AB - We have determined the plasma concentrations of copper, zinc, copper/zinc ratio, and carbonic anhydrase activity in horses infected with Babesia equi. The study was conducted in 14 horses with the disease and 10 healthy animals that served as controls. The infection was confirmed by the clinical manifestations of the disease and by Giemsa staining of thin blood smears showing the parasites inside red blood cells. The horses with piroplasmosis had lower plasma levels of zinc, elevated copper, and increased activity of carbonic anhydrase. Consequently, the copper/zinc ratio was also higher than in the healthy controls. PMID- 18677583 TI - Evidence for involvement of ERK, PI3K, and RSK in induction of Bcl-2 by valproate. AB - Valproate, an anticonvulsant and mood stabilizer, up-regulates Bcl-2, a neurotrophic/neuroprotective protein. In this study, we investigated the molecular mechanism through which Bcl-2 is up-regulated by valproate using cultured human neuron-like cells. Valproate, within therapeutically relevant ranges, induced time- and concentration-dependent up-regulations of both Bcl-2 messenger RNA and protein implicating an underlying gene transcriptional-mediated mechanism. Bcl-2 up-regulations were associated with ERK1/2 and PI3K pathway activations and elevated levels of activated phospho-RSK and phospho-CREB, convergent targets of the ERK1/2 and PI3K pathways. Valproate increased transcriptional activity of a human bcl-2 promoter-reporter gene construct. This effect was attenuated, but not blocked, by mutation of a CREB DNA binding site, a CRE site in the human bcl-2 promoter sequence. ERK and/or PI3K pathway inhibitors and RSK1 small hairpin RNA knockdown reduced, but did not abolish, baseline and valproate-induced promoter activities and lowered Bcl-2 protein levels. These data collectively suggest that valproate induces Bcl-2 regulation partially through activations of the ERK and PI3K cascades and their convergent kinase, RSK, although other unknown mechanism(s) are likely involved. Given the known roles of Bcl-2 in the central nervous system, the current findings offer a partial yet complex molecular mechanistic explanation for the known neurobiological effects of valproate including neurite growth, neuronal survival, and neurogenesis. PMID- 18677584 TI - [Editorial: hypertensiology]. PMID- 18677585 TI - [Hypertension and the kidney]. AB - Hypertension and proteinuria are common risk factors for cardiovascular morbidity and mortality, as well as for the progression of renal disease. Renal functional impairment represents an independent cardiovascular risk factor by itself. Strict antihypertensive therapy and measures to maximally reduce proteinuria can substantially improve the prognosis of renal patients. This review summarizes current evidence for the role of hypertension and proteinuria with regard to renal and cardiovascular outcomes and for the importance of intensive antihypertensive and anti-proteinuric measures. PMID- 18677586 TI - An update on the relationship between the kidney, salt and hypertension. AB - The relationship between salt intake and blood pressure has been noteworthy for a long time. Nevertheless, the complex hemodynamic alterations associated with the salt-induced blood pressure changes have become clear only quite recently. Despite this knowledge, the concept of Guyton, which postulates that any increase in blood pressure should lead to a pressure natriuresis normalising blood pressure over the long term is still valid. As a conclusion, we have to remember that an elevation of arterial pressure can only be maintained if renal function as indicated by pressure natriuresis is impaired. PMID- 18677587 TI - [Hypertension and overweight]. AB - Obesity shows a strong association with hypertension, primarily due to secretion of an abundance of para- and endocrine peptides by the visceral adipose tissue that play a key role in the pathogenesis of high blood pressure and are responsible for the accelerated atherothrombosis encountered in overweight individuals. Increased activity of the RAAS and the sympathetic nerve system are the main pathophysiologic factors; hypertension in obese subjects is characterized by increased peripheral arterial resistance, high cardiac output, hypervolemia and salt sensitivity. Medical therapy sees blockade of the RAAS in front. Priority in treatment should be given to improvements in life style, primarily in increasing physical activity and decreasing caloric intake, and to medical treatment of associated risk factors according to the modern concept of global risk management. PMID- 18677588 TI - [Arteriell wall-stiffness: a new important cardiovascular risk factor]. AB - In addition to blood pressure (BP), Augmentation index (AIx) and pulse wave velocity (PWV) are two further parameters to assess arterial function. Even so these functions are partly interdependent, each of them is a strong and independent predictor of vascular risk and end organ damage. In consideration of the high general interest in cardiovascular disease, the diagnostic improvement in assessing abnormal vascular function by these new techniques should be promptly put into practice. The earlier the recognition of vascular dysfunction, the better the chance to prevent vascular or end organ damage by treatment. As dysfunction is directly derived from the vessels where damage should be prevented, AIx and PWV are closer to the object of interest than risk scores, containing predominantly variables which are independent of the vascular tree. Furthermore, they should become abnormal prior to anatomic alteration such as plaques and thus are preferable in early risk diagnostics. As long as BP is the only vascular function diagnosed, its value is similar to that of high cholesterol or waist circumference. But in joining AIx and PWV, these parameters present information on vessel wall function, such as elasticity and on unfavourable hemodynamic flow conditions. An advantage of AIx and PWV over BP recording is their lower variability as shown in a comparison in 97 hypertensive patients. The standard deviation (SD) of three consecutive blood pressure recordings was systolic/diastolic: 8.7/6.0 mmHg, the SD of three consecutive HR recording 3.2 beats/min, the SD of three AIx-recordings: 3.9% (TensioClinicgerat) and SD of three PWV-recordings: 1.6 m/sec (TensioClinicgerat). PMID- 18677589 TI - The benefit of strength training on arterial blood pressure in patients with type 2 diabetes mellitus measured with ambulatory 24-hour blood pressure systems. AB - BACKGROUND: An ambulatory 24-hour BP-monitoring (ABPM) is of paramount importance, while patients are engaged in their usual activities, for a better representation of blood pressure (BP). ABPM provides not only automated measurements of brachial-artery pressure over a 24-hour period but also a highly reproducible circadian profile. The purpose of this investigation was to evaluate the effect of strength training (ST) on BP in patients with type 2 diabetes mellitus (T2D) and to obtain new and important information on BP profiles over 24 hour by using an ABPM. MATERIAL AND METHODS: We recruited ten patients (mean age: 59.7 +/- 7.3) from our Diabetes Department who participated in a 4-month systematic ST program on three non-consecutive days of the week. The ST program consisted of exercises for all major muscle groups. The numbers of sets for each muscle group were systematically increased from 3 at the beginning of the program to 4, 5 and finally 6 sets per week at the end of the program. The ABPM equipment (oscillometric Model Mobil-O-Graph CE 0434) was applied before and after 4-month training period. Routine HbA1C levels were measured using standard techniques. All subjects took a cycling test to measure maximum oxygen uptake (VO2peak) and maximum workload (Wmax) before and after the training period. Maximal strength was determined by one repetition maximum (1RM) in kp for the bench press, bench pull and leg press exercises, using the Concept 2 Dyno. RESULTS: Analysis of the pooled daytime and night-time data showed a significant reduction of mean arterial BP (from 93.8 +/- 19.2 to 90.6 +/- 14.3 mmHg; p > 0.01) after a 4-month ST (-3.4% mmHg). VO2peak (p < 0.05), Wmax (p < 0.05), 1RM for all muscle groups (p < 0.01), lean body mass (p < 0.05) and percent body fat (p < 0.05) improved significantly after a 4-month of ST. HbA1C showed a significant reduction by 14.5% (from 8.3 +/- 1.7 to 7.1 +/- 0.9%; p < 0.01). CONCLUSION: We found a significant reduction of mean arterial BP after a 4-month ST, measured by the ABPM system. These results demonstrate that ST may not only increase muscle strength but also decrease BP and perhaps the risk of future CVD development. PMID- 18677591 TI - [Don't leave alone! Or maybe do? What it means to acknowledge vegetative patients as human subjects]. AB - Acknowledging vegetative patients as subjects within the therapeutic and nursing practice requires a clarification of the terms of subject, person and autonomy. There are good reasons for using the terms person and human being synonymously, instead of making the person status of human beings dependent on their state of consciousness. Furthermore, the understanding that the body is the bearer of human personality and the medium of communication is of crucial importance. Like the human countenance, the human body as a whole has got a response structure, too. Hence, it can be argued that the mere presence of a comatose human alone, but also many nonverbal signals and reactions should be understood as a request for communication. How far signals and responses coming from the patient in individual cases can be interpreted as personal messages or as pure body reactions depends, however, very much on the interpretation. PMID- 18677590 TI - The neonatal coagulation system and the vitamin K deficiency bleeding - a mini review. AB - Coagulation factors do not cross the placental barrier but are synthesized independently by the conceptus. At birth, activities of the vitamin K dependent factors II, VII, IX, and X and the concentrations of the contact factors XI and XII are reduced to about 50% of normal adult values. The levels of the factors V, VIII, XIII, and fibrinogen are similar to adult values. Plasma concentrations of the naturally occurring anticoagulant proteins (antithrombin, protein C, and protein S) are significantly lower at birth than during the adult years. Plasminogen is reduced by approximately 50%. Platelet counts are within the normal range, regarding function, however, neonatal platelets seem to be hyporeactive. The von Willebrand factor contains large multimers and its concentration is increased. Properties and functions of vitamin K as well as requirement and plasma concentrations in newborns are reviewed. Regarding vitamin K deficiency bleeding (VKDB), the classical nomenclature is used: "early" (presenting within the first 24 h of life), "classical" (day 1-7 after birth), and "late" (8 days to 6 months). After the presentation of the history of vitamin K prophylaxis, vitamin K levels are described as can be expected after the administration of prophylactic doses at various routes. Subsequently, the actual schedule of vitamin K prophylaxis as recommended by the "Osterreichische Gesellschaft fur Kinder- und Jugendheilkunde" is given as follows: i) the oral treatment of healthy full-term babies and orally fed preterm babies, ii) the parenteral treatment of small preterm and sick full-term babies, and iii) the treatment of mothers under medication with enzyme-inducing drugs with vitamin K during the last 15-30 days of pregnancy. The regimes of prophylactic vitamin K treatment of different countries are also given. Finally, the therapeutic use of vitamin K is addressed; the potential use of fresh-frozen plasma, prothrombin complex preparations, and recombinant factor VIIa is discussed. PMID- 18677592 TI - [Klippel-Trenaunay-Weber syndrome]. AB - Klippel-Trenaunay syndrome is an inherited disorder that is known to affect the venous system. Besides venous pathologies, arterial stenosis and aneurysmatic dilatations are extremely rare. In this report, we present a 7-year-old boy with Klippel-Trenaunay-(Weber) syndrome who developed iliac and femoral artery aneurysms and stenosis at different levels in his lower limb. PMID- 18677593 TI - [Books & organizations]. PMID- 18677594 TI - A novel cell subset: interferon-producing killer dendritic cells. AB - Recent reports introduce a novel cell subset of DCs with antigenic phenotypes shared by both NK cells and B cells, but without surface markers of pDCs and T cells, appearing to be a chimera of NK cells and DCs, namely interferon-producing killer dendritic cells (IKDCs). IKDCs not only secret type I and type II interferons to recognize and kill tumor cells effectively, but also express MHC II molecules to present antigens. Thus, IKDCs are considered as important immunosurveilance cells for tumors, providing a link between innate and adaptive immunity. PMID- 18677595 TI - Signal interaction between nitric oxide and hydrogen peroxide in heat shock induced hypericin production of Hypericum perforatum suspension cells. AB - Heat shock (HS, 40 degrees C, 10 min) induces hypericin production, nitric oxide (NO) generation, and hydrogen peroxide (H(2)O(2)) accumulation of Hypericum perforatum suspension cells. Catalase (CAT) and NO specific scavenger 2-4 carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) suppress not only the HS-induced H(2)O(2) generation and NO burst, but also the HS-triggered hypericin production. Hypericin contents of the cells treated with both NO and H(2)O(2) are significantly higher than those of the cells treated with NO alone, although H(2)O(2) per se has no effects on hypericin production of the cells, which suggests the synergistic action between H(2)O(2) and NO on hypericin production. NO treatment enhances H(2)O(2) levels of H. perforatum cells, while external application of H(2)O(2) induces NO generation of cells. Thus, the results reveal a mutually amplifying action between H(2)O(2) and NO in H. perforatum cells. CAT treatment inhibits both HS-induced H(2)O(2) accumulation and NO generation, while cPTIO can also suppress H(2)O(2) levels of the heat shocked cells. The results imply that H(2)O(2) and NO may enhance each other's levels by their mutually amplifying action in the heat shocked cells. Membrane NAD(P)H oxidase inhibitor diphenylene iodonium (DPI) and nitric oxide synthase (NOS) inhibitor S,S'-1,3-phenylene-bis(1,2-ethanediyl)-bis-isothiourea (PBITU) not only inhibit the mutually amplifying action between H(2)O(2) and NO but also abolish the synergistic effects of H(2)O(2) and NO on hypericin production, showing that the synergism of H(2)O(2) and NO on secondary metabolite biosynthesis might be dependent on their mutual amplification. Taken together, data of the present work demonstrate that both H(2)O(2) and NO are essential for HS-induced hypericin production of H. perforatum suspension cells. Furthermore, the results reveal a special interaction between the two signal molecules in mediating HS-triggered secondary metabolite biosynthesis of the cells. PMID- 18677596 TI - Novel alleles among soybean Bowman-Birk proteinase inhibitor gene families. AB - Trypsin inhibitors have been found in various animals, plants and microorganisms. There were two types of trypsin inhibitors in soybean including Bowman-Birk protease inhibitors (BBI) and Kunitz inhibitors (KTI). The different BBI genes from wild soybean (G. soja) and cultivated soybean (G. max) formed a multigene family. We constructed a cDNA library of cultivar 'SuiNong 14' seed at the R7 growth stage using the SMART Kit. Seventeen contigs or singletons were highly homologous to soybean protease inhibitors. Contigs of 5, 35, 8 and 9 were highly homologous to BBI family members BBI-A1, BBI-A2, BBI-C and BBI-D, respectively. Sequence analyses showed there were novel allelic variations among the 4 BBI members in SuiNong 14. Based on the comparison of soybean seed cDNA libraries from different developmental stages, it was apparent that the expression of trypsin inhibitors increased during seed development in soybean. Phylogenetic analysis of BBI gene sequences among dicotyledonous and monocotyledonous plants demonstrated that these genes shared a common progenitor. PMID- 18677597 TI - Characterization of mitochondrial genome of Chinese wild mulberry silkworm, Bomyx mandarina (Lepidoptera: Bombycidae). AB - The complete mitochondrial genome of Chinese Bombyx mandarina (ChBm) was determined. The circular genome is 15682 bp long, and contains a typical gene complement, order, and arrangement identical to that of Bombyx mori (B. mori) and Japanese Bombyx mandarina (JaBm) except for two additional tRNA-like structures: tRNA( Ser(TGA))-like and tRN( AIle(TAT))-like. All protein-coding sequences are initiated with a typical ATN codon except for the COI gene, which has a 4-bp TTAG putative initiator codon. Eleven of 13 protein-coding genes (PCGs) have a complete termination codon (all TAA), but the remaining two genes terminate with incomplete codons. All tRNAs have the typical clover-leaf structures of mitochondrial tRNAs, with the exception of tRNA( Ser(TGA))-like, with a four stem and-loop structure. The length of the A+T-rich region of ChBm is 484 bp, shorter than those of JaBm (747 bp) and B. mori (494-499 bp). Phylogenetic analysis among B. mori, ChBm, JaBm, and Antheraea pernyi (Anpe) showed that B. mori is more closely related to ChBm than JaBm. The earliest divergence time estimate for B. mori-ChBm and B. mori-JaBm is about 1.08+/-0.18-1.41+/-0.24 and 1.53+/-0.20 2.01+/-0.26 Mya, respectively. ChBm and JaBm diverged around 1.11+/-0.16-1.45+/ 0.21 Mya. PMID- 18677598 TI - Reduction of drosopterin content caused by a 45-nt insertion in Henna pre-mRNA of Drosophila melanogaster. AB - Phenylalanine hydroxylase is assumed to be a key enzyme in drosopterin metabolism, but direct in vivo evidence to support this hypothesis is still absent. In the present study, we found a new natural recessive purple eye mutant of Drosophila melanogaster, Hn ( bp ), which was a 45-nt insertion mutant in the second exon of Henna. The insertion resulted in a predicted protein with 15 additional amino acids as compared to the wild-type protein. Further analysis of protein structure showed that the predicted mutant protein probably had two more beta-sheets, which may cause instability of two alpha-helices near the catalytic centre of the enzyme in the Biopterin-Hydroxyl binding domain. Hn ( bp ) mutant showed eye color defect with decrease of mRNA level, as well as drosopterin content reduction. The drosopterin defect could be fully rescued by expression of wild type Henna in the Hn ( bp ) background by GMR-GAL4 UAS-Henna/UAS-Henna:Hn ( bp )/Hn ( bp ) transgenic line. All taken together, it can be concluded that the mutation in Henna is responsible for drosopterin reduction in mutant Hn ( bp ), which provides key in vivo evidence to support the hypothesis that Henna is involved in drosopterin synthesis. PMID- 18677599 TI - Effect of mammogenic hormones on the expression of FGF7, FGF10 and their receptor in mouse mammary gland. AB - To investigate the regulation of estrogen, progesterone and prolactin stimulating the development of mammary gland, the Kunming mice were used as experimental animals in this study. Through the experiment in vitro, the effect of mammogenic hormones were systematically investigated on expression of FGF7 and FGF10 and their receptor in different periods. The results are as follows: in mammary glands of mice, 17 beta-estradiol increased the expression of FGF7; progesterone did not affect the expression of FGF7; prolactin up-regulated the expression of FGF7 significantly in pregnancy and lactation. 17 beta-estradiol increased the expression of FGF10; progesterone and prolactin reduced the expression of FGF10 significantly in virgin; prolactin significantly increased the expression of FGF10 in pregnancy. When 17 beta-estradiol in the body was in relatively high proportion, it would lower the expression of KGFR; while 17 beta-estradiol in the body was in relatively low proportion, it would increase the expression of KGFR. Low concentration of progesterone increased the expression of KGFR and high progesterone did not affect the expression of KGFR. Prolactin increased the expression of KGFR significantly in pregnancy and lactation. PMID- 18677600 TI - Expression profiling analysis for genes related to meat quality and carcass traits during postnatal development of backfat in two pig breeds. AB - The competitive equilibrium of fatty acid biosynthesis and oxidation in vivo determines porcine subcutaneous fat thickness (SFT) and intramuscular fat (IMF) content. Obese and lean-type pig breeds show obvious differences in adipose deposition; however, the molecular mechanism underlying this phenotypic variation remains unclear. We used pathway-focused oligo microarray studies to examine the expression changes of 140 genes associated with meat quality and carcass traits in backfat at five growth stages (1-5 months) of Landrace (a leaner, Western breed) and Taihu pigs (a fatty, indigenous, Chinese breed). Variance analysis (ANOVA) revealed that differences in the expression of 25 genes in Landrace pigs were significant (FDR adjusted permutation, P<0.05) among 5 growth stages. Gene class test (GCT) indicated that a gene-group was very significant between 2 pig breeds across 5 growth stages (P (ErmineJ)<0.01), which consisted of 23 genes encoding enzymes and regulatory proteins associated with lipid and steroid metabolism. These findings suggest that the distinct differences in fat deposition ability between Landrace and Taihu pigs may closely correlate with the expression changes of these genes. Clustering analysis revealed a very high level of significance (FDR adjusted, P<0.01) for 2 gene expression patterns in Landrace pigs and a high level of significance (FDR adjusted, P<0.05) for 2 gene expression patterns in Taihu pigs. Also, expression patterns of genes were more diversified in Taihu pigs than those in Landrace pigs, which suggests that the regulatory mechanism of micro-effect polygenes in adipocytes may be more complex in Taihu pigs than in Landrace pigs. Based on a dynamic Bayesian network (DBN) model, gene regulatory networks (GRNs) were reconstructed from time-series data for each pig breed. These two GRNs initially revealed the distinct differences in physiological and biochemical aspects of adipose metabolism between the two pig breeds; from these results, some potential key genes could be identified. Quantitative, real-time RT-PCR (QRT-PCR) was used to verify the microarray data for five modulated genes, and a good correlation between the two measures of expression was observed for both 2 pig breeds at different growth stages (R=0.874+/-0.071). These results highlight some possible candidate genes for porcine fat characteristics and provide some data on which to base further study of the molecular basis of adipose metabolism. PMID- 18677601 TI - Efficient generation of transgenic chickens using the spermatogonial stem cells in vivo and ex vivo transfection. AB - The highly efficient novel methods to produce transgenic chickens were established by directly injecting the recombinant plasmid containing green fluorescent protein (GFP) gene into the cock's testis termed as testis-medianted gene transfer (TMGT), and transplanting transfected spermatogonial stem cells (TTSSCs). For the TMGT approach, four dosages of pEGFP-N1 DNA/cationic polymer complex were injected intratesticularly. The results showed: (1) 48 h after the injection, the percentages of testis cells expressing GFP were 4.0%, 8.7%, 10.2% and 13.6% in the 50, 100, 150 and 200 microg/mL group, respectively. The difference from the four dosage groups was significant (P<0.05). On day 25 after the injection, a dosage-dependent and time-dependent increase in the number of transgenic sperm was observed. The percentages of gene expression reached the summit and became stable from day 70 to 160, being 12.7%, 12.8%, 15.9% and 19.1%, respectively. The difference from the four dosage groups was also significant (P<0.05). (2) 70 d after the injection, strong green fluorescent could be observed in the seminiferous tubules by whole-mount in-situ hybridization. (3) 70 d after the injection, the semen was collected and used to artificially inseminate wild-type females. The blastoderms of F(1) and F(2) transgenic chicken expressed GFP were 56.2% (254/452) and 53.2% (275/517), respectively. The detection of polymerase chain reaction (PCR) of F(1) and F(2) transgenic chicken blood genomic DNA showed that 56.5% (3/23) of F(1) and 52.9% (9/17) of F(2) were positive. Southern blot showed GFP DNA was inserted in their genomic DNAs. (4) Frozen whole mount tissue sections of F(1) and F(2) transgenic chicken liver, heart, kidney and muscle showed that the rates of green fluorescent positive were between 50.0% and 66.7%. (5) With the TTSSCs method, SSCs ex vivo transfected with GFP were transplanted into recipient roosters whose endogenic SSCs had been resoluted. The donor SSCs settled and GFP expression became readily detectable in the frozen whole mount tissue sections of recepient testes. Moreover, sperms carrying GFP could be produced normally. The results of artificially inseminating wild-type females with these sperms showed 12.5% (8/64) of offspring embryo expressed GFP and 11.1% (2/18) hatched chicks were tested transgenic. Our data therefore suggest TMGT and TTSSCs are the feasible methods for the generation of transgenic chickens. PMID- 18677602 TI - Prevention of pathological change and cognitive degeneration of Tg2576 mice by inoculating Abeta(1-15) vaccine. AB - This study aims to discuss the effect of preventing pathological changes and cognitive degeneration of Tg2576 mice by inoculating the subunit fragment of Abeta vaccine. Thirty-two Tg2576 mice were randomly divided into four groups, each having eight mice: Group I, the control group, inoculated with adjuvants; Group II, the Abeta(42) group, inoculated with Abeta(42) vaccine; Group III, the Abeta(1-15) group, inoculated with Abeta(1-15) vaccine; and Group IV, the Abeta(36-42) group, inoculated with Abeta(36-42) vaccine. The titer of the serum antibody against Abeta(42) (Group II) was significantly higher than that of the control group (Group I), and a low level of antibodies could be detected in the brain homogenate in the three vaccine-inoculated groups. Morris water maze test showed that the Abeta(42) group, Abeta(1-15) group and Abeta(36-42) group were obviously improved compared with the control group. The cultured splenocytes sampled from each group were induced by Con A or their respective antigens, and the cell proliferation of the three vaccine-inoculated groups was significantly higher than that of the control group. In the Abeta(42) group, IL2 and IFN-gamma were relatively low and IL4 and IL10 were relatively high. By contrast, IL4 and IL10 were much higher in the Abeta(1-15) group and IL2 and IFN-gamma were much higher in the Abeta(36-42) group. The immunohistochemical test showed a large number of senile plaques in the brain cortex and hippocampus of the mice in the control group, no senile plaque in the brain of the Abeta(1-15) group and Abeta(42) group mice, and a small number of senile plaques in the brain of the Abeta(36-42) group mice. The results suggest that the subunit fragment of Abeta(1 15) vaccine could prevent not only cognitive and behavioral degeneration but also Abeta deposition and formation of senile plaques in Tg2576 mice. PMID- 18677603 TI - Diversity and ecological distribution of endophytic fungi associated with medicinal plants. AB - A total of 973 isolates of endophytic fungi were recovered from 1,144 tissue fragments of the six medicinal plant species belonging to 4 families collected in the Beijing Botanical Garden. Of these isolates 778 sporulated and were identified into 21 taxa by morphological characteristics. Among the taxa 11 belonged to Coelomycetes, 6 to Ascomycetes, and 4 to Hyphomycetes. Various numbers of endophytic fungi (5-8 taxa) were obtained from each plant. Alternaria alternata was the dominant species in the 6 plants, and Microsphaeropsis conielloides was also dominant in Eucommia ulmoides. There were high colonization rates (47.9%-63.1%) and isolation rates (0.7-0.93) of endophytic fungi, and they were conspicuously higher in twigs than those in leaves in the 6 plants examined. The colonization and isolation rates of endophytic fungi increased with the twig age. The results based on the analyses of cluster and Sorenson's similarity coefficients indicated that some endophytic fungi showed a certain degree of host and tissue preference. PMID- 18677604 TI - Effects of pH on the peripheral light-harvesting antenna complex for Rhodopseudomonas palustris. AB - In this work steady-state absorption spectroscopy, circular dichroism spectroscopy and sub-microsecond time-resolved absorption spectroscopy were used to investigate the effect of pH on the structures and functions of LH2 complex for Rhodopseudomonas palustris. The results revealed that: (1) B800 Bchla was gradually transformed to free pigments absorbing around 760 nm on the minutes timescale upon the induction of strong acidic pH, and subsequently there disappeared the CD signal for Q(y) band of B800 in the absence of B800. In addition, Carotenoids changed with the similar tendency to B850 BChl. (2) The introduction of strong basic pH gave rise to no significant changes for B800 Bchla, while B850 BChla experienced remarkable spectral blue-shift from 852 to 837 nm. Similar phenomenon was seen for the CD signal for Q(y) band of B850. Carotenoids displayed strong and pH-independent CD signals in the visible range. (3) In the case of both physiological and basic pH, broad and asymmetrical positive T( n ) <-- T(1) transient absorption appeared following the pulsed photo excitation of Car at 532 nm. By contrast, the featureless and weak positive signal was observed on the sub-microsecond timescale in the acidic pH environment. The aforementioned experimental results indicated that acidic pH induced removal of B800 Bchla prevented the generation of the carotenoid triplet state ((3)Car), which is known to be essential for the photo-protection function. Nevertheless, carotenoids can still perform this important physiological role under the basic pH condition, where the spectral blue shift of B850 exerts little effect on the overall structure of the cyclic aggregate, therefore favoring the formation of carotenoid triplet state. PMID- 18677605 TI - Cytogenetic biomonitoring of oral mucosa cells from adults exposed to dental X rays. AB - PURPOSE: Although it has been clearly demonstrated that X-rays play a key role in diagnosing medical and dental problems, this type of ionizing radiation is also able to induce noxious activities, such as genetic damage. The aim of the present study was to evaluate DNA damage (micronucleus) and cellular death in exfoliated buccal mucosa cells from healthy individuals (smokers and nonsmokers) following dental X-ray exposure. MATERIAL AND METHODS: A total of 39 healthy people who had submitted to panoramic dental radiography were included in the study: 9 smokers and 30 nonsmokers. RESULTS: The results indicated no significant statistically differences (P>0.05) in micronucleated oral mucosa cells before and after dental X-ray exposure. On the other hand, X-ray exposure did increase other nuclear alterations closely related to cytotoxicity, such as karyorrhexis, pyknosis, and karyolysis. It seems that cigarette smoke did not affect X-ray outcomes induced in buccal cells. CONCLUSION: These data indicate that dental panoramic radiography may not induce chromosomal damage, but it is able to promote cytotoxicity. Because cellular death is considered a prime mechanism in nongenotoxic mechanisms of carcinogenesis, dental X-ray should be used only when necessary. PMID- 18677606 TI - Monte Carlo study on a flattening filter-free 18-MV photon beam of a medical linear accelerator. AB - PURPOSE: Several studies on the dosimetric properties of unflattened photon beams have shown some advantages for radiotherapy. In this study, the effect of removing the flattening filter from an 18-MV photon beam was investigated using the Monte Carlo method. MATERIALS AND METHODS: The 18-MV photon beam of an Elekta SL25 linear accelerator was simulated using the MCNP4C Monte Carlo (MC) code. Beam dosimetric features, including central axis absorbed doses, beam profiles, and photon energy spectra, were calculated for flattened and unflattened 18-MV photon beams. RESULTS: A 4.24-fold increase in the dose rate was seen for the unflattened beam with a field size of 10 x 10 cm(2). A decrease in the out-of field dose up to 30% was seen for the unflattened beam. For the unflattened beam, photon energy spectra were softer, and the mean energies of the spectra were higher for a smaller field size. CONCLUSION: Our study showed that the increase in dose rate and lower out-of-field dose can be possible advantages for an unflattened 18-MV beam. PMID- 18677607 TI - Comparison of 16-multidetector-row computed tomography and angiocardiography for evaluating the central pulmonary artery diameter and pulmonary artery index in children with congenital heart disease. AB - PURPOSE: The pulmonary artery (PA) is involved in most congenital heart diseases; and in these patients it is necessary to evaluate precisely the PA configuration and development. The accuracy of 16 multidetector row computed tomography (16 MDCT) in evaluating the central PA was evaluated. MATERIALS AND METHODS: 16-MDCT and angiocardiography (ACG) were performed in 26 patients with various congenital heart diseases aged 7 days to 9 years (median 1.2 years). We reconstructed coronal oblique images along the long axis of the right and left PAs and measured the PA diameter and Nakata's PA index, which were compared with those obtained by ACG. RESULTS: Correlations between PA diameters [R (2) = 0.80, standard error of the estimate (SEE)=1.3, n=52] and PA indices (R(2)=0.81, SEE=42, n=26) obtained from coronal oblique images and ACG were excellent. Bland-Altman plots showed a mean +/- SD difference of -0.3+/-1.3 mm for the PA diameter and +/-15.1 +/-41.5 for the PA index. CONCLUSION: 16-MDCT might be useful for evaluating the central PA in patients with congenital heart disease. PMID- 18677608 TI - Infectious mediastinitis after cardiovascular surgery: role of computed tomography. AB - PURPOSE: The aim of this study was to evaluate the effectiveness of computed tomography (CT) findings in the diagnosis of mediastinitis after cardiovascular surgery with median sternotomy. MATERIAL AND METHODS: A total of 122 patients were divided into two groups: the early group (21 days after surgery). Among them, six patients were ultimately diagnosed with infectious mediastinitis. CT findings in each patient were evaluated. Mediastinal fluid collections or free gas bubbles were regarded as the primary findings of mediastinitis. RESULTS: In the early group, sensitivity and specificity of the primary CT findings were 100% and 39%, respectively. In the late group, the sensitivity was 100% and the specificity 85%. Mediastinal fluid collections were observed in all six patients with mediastinitis. CONCLUSION: Mediastinal fluid collections or free gas bubbles are not specific during the early postoperative period. However, after 2 21 days, these observations could be indicative of mediastinitis. PMID- 18677609 TI - Initial experience of using the filter protection device during carotid artery stenting in Japan. AB - PURPOSE: No filter protection devices for carotid artery stenting (CAS) have been formally approved for use in Japan; however, as of April 2008, the Angioguard XP (AGXP) was approved. This article describes our initial results using the AGXP during CAS for the treatment of carotid artery stenosis. MATERIAL AND METHODS: A group of 15 patients (14 men) with a mean age of 72.3 years (range 53-81 years) were treated by CAS using the AGXP. Among them, 10 were symptomatic with >50% stenosis of the common or internal carotid artery (ICA), and 5 were asymptomatic with >70% stenosis. The rates of technical success, periprocedural stroke, ICA flow impairment, filter movement, and development of new ischemic lesions on diffusion-weighted imaging (DWI) were assessed. RESULTS: CAS using the AGXP was successful in all cases. There was one minor stroke, and flow impairment occurred in six patients. Filter movement averaged 1.9 vertebral bodies. DWI showed new ipsilateral ischemic lesions in eight of the patients. CONCLUSION: Initial clinical experience using the AGXP for CAS has been generally sufficient. However, attention must be paid to three problems when using the AGXP: the filter may move after placement; the filter may disturb blood flow in the ICA; and debris may pass around the filter. PMID- 18677611 TI - Detection of incidental breast tumors by noncontrast spiral computed tomography of the chest. AB - PURPOSE: The aim of this study was to investigate the frequency of breast tumors and breast cancers with noncontrast spiral chest computed tomography (CT). MATERIAL AND METHODS: A clinical study was conducted to evaluate findings in the mammary region of 1008 consecutive patients with no mammary symptoms or signs who underwent noncontrast spiral CT of the chest from April 2003 to March 2006. RESULTS: Six cases of breast abnormality were detected among the 1008 women. Three were primary breast cancers, one was metastatic breast cancer, and two were benign tumors. All four breast cancer patients were over 70 years old. The characteristics of the tumor margins on CT scans corresponded to the mammography and ultrasonography findings. The mammographic background density ranged from inhomogeneous high-density breast to fatty breast. The detection rate of primary breast cancer by noncontrast spiral CT was 0.30%. CONCLUSION: Noncontrast spiral chest CT occasionally detects nonsymptomatic breast cancers, especially in elderly patients. PMID- 18677610 TI - Pancreatic adenocarcinoma: analysis of the effect of various concentrations of contrast material. AB - PURPOSE: The aim of this study was to compare the efficacy of two contrast materials with moderate and high iodine concentrations for the depiction of pancreatic adenocarcinoma. MATERIALS AND METHODS: A series of 107 patients with histologically proven pancreatic adenocarcinoma underwent helical computed tomography. A fixed dose of 100 ml of iopamidol 300 (mg I/ml) was administered to 50 patients (group A) and iopamidol 370 (mg I/ml) to 57 patients (group B) at the same injection rate (3 ml/s). Unenhanced helical scans and contrast-enhanced scans for three phases (30, 70, and 300 s after starting the infusion of contrast material) were obtained. We evaluated enhancement of the aorta, portal vein, hepatic parenchyma, pancreatic parenchyma, and pancreatic adenocarcinoma during each phase. RESULTS: During all phases, both aortic and pancreatic enhancement were significantly greater in group B than in group A (P<0.01). Enhancement of the portal vein and hepatic parenchyma was significantly greater at 70 and 300 s in group B than in group A (both P<0.01). Tumor-to-pancreas contrast was significantly greater in group B than in group A at both 30 s (P<0.01) and 70 s (P<0.05). CONCLUSION: Administration of contrast material with a high iodine concentration is more effective for depicting pancreatic adenocarcinomas. PMID- 18677612 TI - Computed tomography and magnetic resonance imaging findings of soft tissue perineurioma. AB - Soft tissue perineurioma is an uncommon benign peripheral nerve sheath tumor, although it is the most common subtype of perineuriomas. We present a case of soft tissue perineurioma in the left groin of a 48-year-old man. Precontrast computed tomography showed a homogeneous hypodense mass that showed faint enhancement. The mass appeared with hypointensity on T1-weighted magnetic resonance (MR) images and heterogeneous hyperintensity on T2-weighted MR images. Slight contrast uptake was noted on enhanced T1-weighted MR images with fat suppression. Although these CT and MR imaging findings were nonspecific, the overall imaging features are similar to those of schwannomas. PMID- 18677613 TI - Endovascular treatment of ureteroarterial fistulas with stent-grafts. AB - Ureteroarterial fistula is a rare condition with life-threatening hematuria that should be diagnosed and treated immediately. We reported two patients of ureteroarterial fistula who underwent successful endovascular treatment by stent grafts. They had undergone pelvic surgery followed by placement of an indwelling ureteral stent for stricture of the ureter before sudden hematuria occurred. Routine contrast-enhanced computed tomography did not reveal extravasation or a pseudo-aneurysm. Angiograms with multiple oblique views demonstrated small pseudoaneurysms of the iliac artery overriding the ipsilateral ureter. The endovascular treatment of ureteroarterial fistula using stent-grafts was successful, and the hematuria disappeared immediately after stent-graft deployment. The hematuria did not recur up to the last follow-up visits of 3 and 19 months, respectively. PMID- 18677614 TI - Magnetic resonance imaging findings of vulvar epithelioid sarcoma. AB - We describe here a case of vulvar epithelioid sarcoma. Epithelioid sarcoma is a malignant appendicular skin tumor that is rarely of vulvar origin. Magnetic resonance imaging (MRI) revealed a solid mass with an ulcer-like lesion and multilobulated contour. To our knowledge, this is the first case of vulvar epithelioid sarcoma described in the MRI literature. PMID- 18677615 TI - Cerebral air embolism treated with hyperbaric oxygen therapy following percutaneous transthoracic computed tomography-guided needle biopsy of the lung. AB - A 71-year-old man presented with cough and sputum for 12 months. Chest radiography showed a homogeneous opacity in the right lower lobe. Computed tomography (CT) showed a nodular opacity, 2 cm in diameter, in the posterior segment of the right lower lobe. Mild emphysematous changes were also seen. With the patient in a prone position, a 19-gauge 7.8-cm introducer was placed in the lesion during a single inspiratory breath-hold. A coaxial 20-gauge automated needle was inserted through the introducer using a biopsy gun. Although the patient did not complain of any symptoms, postbiopsy CT showed air in the left ventricle and ascending aorta. After 5 h of bed rest, we found weakness in his left lower extremity. He was transferred to a hyperbaric oxygen chamber and recovered the next day. Air embolism is a rare, potentially fatal complication of percutaneous lung biopsy. Although the true effect of hyperbaric oxygen therapy is controversial, knowledge regarding the prompt management of such cases may help radiologists who perform this procedure. PMID- 18677616 TI - Unruptured pseudoaneurysm of the cystic artery with acute calculous cholecystitis incidentally detected by computed tomography. AB - Pseudoaneurysm of the cystic artery is rare; to our knowledge, fewer than 30 cases have been reported worldwide. We report the first case of an unruptured pseudoaneurysm of the cystic artery with concurrent acute calculous cholecystitis. We incidentally detected the aneurysm by contrast-enhanced computed tomography (CT) in the edematous, thickened wall of the gallbladder neck in a 71-year-old man, whereas in most of the reported cases the disease presented as hemobilia. Because of the high risk of aneurysm rupture in this location, we avoided such interventions as percutaneous cholecystostomy and laparoscopic cholecystectomy. The aneurysm was localized pathologically in the undisrupted gallbladder wall, and elective open cholecystectomy with ligation of the cystic artery was performed successfully. Our case highlights the usefulness of CT for both diagnosis and patient management. Open cholecystectomy with ligation of the cystic artery is demonstrated as a reasonable first line of treatment for this unusual condition. PMID- 18677618 TI - Polymer MS-MS by MALDI: some advances and some challenges. PMID- 18677617 TI - Endocrine and gene expression changes following forced swim stress exposure during cocaine abstinence in mice. AB - RATIONALE: Stress can reinstate previous cocaine-seeking long after drug is no longer present. However, little is known regarding the effect of chronic drug exposure and subsequent drug abstinence on responsivity to stress. OBJECTIVE: To determine the effect of acute (24-h) and prolonged (14-day) drug-free periods in cocaine-experienced mice on behavioral, endocrine, and molecular outputs following stress exposure. MATERIALS AND METHODS: Mice were administered a cocaine binge (15 mg/kg, every hour for 3h) for 2 weeks. Following a 24-h or 14 day drug-free period, stress responsivity, along with levels of anxiety, were measured using the forced swim test and elevated zero maze, respectively. In addition, alterations in the levels of plasma corticosterone, corticotrophin releasing factor (CRF) mRNA, brain-derived neurotrophic factor (BDNF) mRNA, and histone acetylation at their respective promoters were examined following stress exposure. RESULTS: At both acute and prolonged abstinence time points, behavioral measures were essentially unaltered; however, cocaine-experienced mice exhibited an augmented corticosterone response to the forced swim stress compared to saline treated mice. Stress exposure increased BDNF mRNA levels in the ventral tegmental area (VTA) and nucleus accumbens (NAc) only in cocaine-experienced mice following a prolonged, but not acute, drug-free period. Increased BDNF mRNA in the NAc was associated with an increase in acetylated histone 3 (AcH3) at the BDNF I promoter. CRF mRNA levels were increased in the amygdala (AMYG); however, this was not associated with alterations in histone acetylation at the promoter. CONCLUSION: These results demonstrate that drug history and prolonged abstinence can alter the endocrine and molecular responses to stress, which may facilitate the reinstatement of drug-seeking behaviors. PMID- 18677619 TI - Development and validation of an efficient HPLC method for quantification of voriconazole in plasma and microdialysate reflecting an important target site. AB - Voriconazole is a very potent antifungal agent used to treat serious fungal infections (candidiasis); it is also the therapy of choice for aspergillosis. After standard dosing, several factors affect exposure of voriconazole, resulting in large variability and demanding further elucidation of drug distribution. For measurements at the site of action, microdialysis is considered to be an outstanding minimally invasive method. For determination of voriconazole in microdialysate and human plasma a new, efficient, reliable, and robust HPLC assay using UV detection at 254 nm has been developed and validated. After simple sample preparation using acetonitrile for plasma and for microdialysate, 20 microL were injected and separated on an RP-18 column. The chromatographic run time was less than 4 min. Overall, the assay showed high precision (CV 93.9 to 99.5%) and accuracy (RE -96.7 to +107%) for both matrices. Of the 36 drug products typically co-administered with voriconazole, none except ambroxol interfered with its peak signal, and this interference was successfully managed. In summary, the method is highly suitable for application in (pre)clinical microdialysis studies, e.g., of critically ill patients with invasive mycoses. PMID- 18677621 TI - Screening of humic and fulvic acids in estuarine sediments by near-infrared spectrometry. AB - Diffuse reflectance near-infrared spectroscopy (NIR) combined with partial least squares (PLS) data treatment has been employed for the rapid and nondestructive determination of sedimentary humic substances. Forty one samples of surface estuarine sediments, taken during distinct seasonal periods from different locations across Ria de Arousa (northwest of Spain), were scanned at wavelengths from 833 to 2,976 nm (12,000 to 3,360 cm(-1)). Twenty four samples were randomly selected, from previous hierarchical cluster analysis of their NIR spectra, for the calibration set, and the 17 remaining samples were assigned to the validation set. NIR spectra of calibration samples were correlated to measured values of humic acids (HAs) and fulvic acids (FAs), which ranged from 1.53 to 28.17 mg/g and from 0.37 to 2.45 mg/g, respectively, using PLS regression and multiplicative scattering correction on the raw and first-derivative NIR spectra, respectively. Low root mean square error of prediction values of 4.3 mg HA/g sediment and 0.25 mg FA/g sediment were obtained. Good residual prediction deviation values of 1.16 and 1.2 were obtained for HA and FA, respectively, allowing the PLS models built to be considered as appropriate tools for screening purposes. PMID- 18677622 TI - Serum concentrations of sertraline and N-desmethyl sertraline in relation to CYP2C19 genotype in psychiatric patients. AB - OBJECTIVE: To investigate the impact of CYP2C19 genotype on serum concentrations of sertraline and N-desmethyl sertraline in psychiatric patients. METHODS: Patients treated with sertraline (n = 121) were divided into six subgroups according to CYP2C19 genotype: CYP2C19*17/*17, CYP2C19*1/*17, CYP2C19*1/*1, CYP2C19*17/def, CYP2C19*1/def and CYP2C19def/def (def = allele encoding defective CYP2C19 metabolism, i.e. *2 and *3). Dose-adjusted serum concentrations were compared by linear mixed model analyses using the CYP2C19*1/*1 subgroup as reference. RESULTS: Subgroups carrying one or two alleles encoding defective CYP2C19 metabolism achieved significantly higher mean dose-adjusted serum concentrations of sertraline and N-desmethyl sertraline compared to the CYP2C19*1/*1 subgroup (P < 0.05). The effect of CYP2C19 genotype was expressed as 3.2-fold (sertraline) and 4.5-fold (N-desmethyl sertraline) higher dose-adjusted serum concentrations in the CYP2C19def/def subgroup compared to the CYP2C19*1/*1 subgroup (P < 0.01). The CYP2C19*17 allele had no influence on the dose-adjusted serum concentrations of sertraline and N-desmethyl sertraline. CONCLUSION: The significantly higher serum concentrations associated with alleles encoding defective CYP2C19 metabolism might be of relevance for the clinical outcome of sertraline treatment. PMID- 18677623 TI - The effectors responsible for gastrointestinal nematode parasites, Trichinella spiralis, expulsion in rats. AB - Helper T (Th2) cells type 2 have a central role in host protective responses to gastrointestinal nematode parasite, Trichinella spiralis infection, but the actual effector mechanisms involved in parasite expulsion are still uncertain. Recent evidences suggest that mast cell recruitment and activation may associate with parasite elimination from host intestines in mice. On the other hand, IgE production may induce defensive responses to primary infection with the helminth in rats. The differences of host effector mechanisms to the same experimental infections might disturb our understanding of the host protective mechanisms to gastrointestinal nematode parasite infection. In order to redefine these differences, we investigated in detail the relationship between intestinal immune responses and worm expulsion following T. spiralis infection among several rat strains including mutants. As a result, there were significant correlations of parasite expulsion with mast cell hyperplasia in addition to serum IgE level. Moreover, mast cell-deficient and dysfunction rats showed delayed worm elimination from their gut. Therefore, the present study suggests that mast cells should also be one of the prominent effector cells involved in T. spiralis parasite expulsion in rats as well as mice. PMID- 18677624 TI - Recombinant Cryptosporidium parvum p23 as a target for the detection of Cryptosporidium-specific antibody in calf sera. AB - Cryptosporidium parvum is a widely distributed coccidian parasite and causes enteric disease in humans and animals. In addition to being a cause of life threatening disease in immunodeficient people, mostly AIDS patients, C. parvum has been reported as a common serious primary cause of outbreaks of diarrhea in newborn calves, especially newborn ruminants (De Graaf et al. in Int J Parasitol 29:1269-1287, 1999). To obtain the recombinant P23 protein, we isolated the mRNA from oocyst of C. parvum and amplified the cDNA of P23 gene by reverse transcriptase PCR. Sequencing of cDNA showed 100% homology to the known P23 sequences. The double strand P23-cDNA was then cloned in pGEX-5X-2 expression vector. Western blot analysis of recombinant P23 showed that it could be recognized by the positive C. parvum serum. Since P23 is an immunodominant surface glycoprotein expressed in the early phase of infection (Jakobi and Petry in Microbes Infect 8:2186-2194, 2006) and the immunogenic epitopes are also found in the residual chain of amino acid sequence of this glycoprotein, the recombinant P23 was used for the screening of 437 serum samples collected from calves (#264) and cattle (#173). The dot blot analysis showed that from 264 calf and 173 cattle sera, 33% and 37% sera were positive, respectively. Due to the simple handling and equipment, dot blot analysis with P23 could be recommended for calves screening against cryptosporidiosis. PMID- 18677626 TI - Dynamic compression inhibits fibronectin fragment induced iNOS and COX-2 expression in chondrocyte/agarose constructs. AB - Mechanical loading and the fibronectin fragments (FN-fs) are known to stimulate the anabolic and catabolic processes in articular cartilage, possible through pathways mediated by *NO. This study examined the combined effects of dynamic compression and the NH(2)-hep I or COOH-hep II FN-fs on the expression levels of iNOS and COX-2 and production of *NO and PGE(2) release. Both types of fragments induced iNOS and COX-2 expression and stimulated the production of *NO release. This response was inhibited by dynamic compression. Inhibitor experiments indicated that both dynamic compression and the iNOS inhibitor were important in restoring cell proliferation and proteoglycan synthesis in the presence of the FN fs. This is the first study which demonstrates a downregulation of the FN-f induced iNOS and COX-2 expression by dynamic compression. The combination of mechanical and pharmacological interventions makes this study a powerful tool to examine further the interactions of biomechanics and cell signalling in osteoarthritis. PMID- 18677625 TI - Diversity and structure of ectomycorrhizal and co-associated fungal communities in a serpentine soil. AB - The community of ectomycorrhizal (ECM) and co-associated fungi from a serpentine site forested with Pinus sylvestris and Quercus petraea was explored, to improve the understanding of ECM diversity in naturally metalliferous soils. ECM fungi were identified by a combination of morphotyping and direct sequencing of the nuclear ribosomal internal transcribed spacer region 2 and of a part of the large subunit region. Co-associated fungi from selected ECM were identified by restriction fragment length polymorphism and sequencing of representative clones from libraries. Polymerase chain reaction with species-specific primers was applied to assess patterns of association of ECM and co-associated fungi. Twenty ECM species were differentiated. Aphyllophoralean fungi representing several basidiomycete orders and Russulaceae were dominant. Phialocephala fortinii was the most frequently encountered taxon from the diverse assemblage of ECM co associated fungi. A ribotype representing a deeply branching ascomycete lineage known from ribosomal deoxyribonucleic acid sequences only was detected in some ECM samples. A broad taxonomic range of fungi have the potential to successfully colonise tree roots under the extreme edaphic conditions of serpentine soils. Distribution patterns of ECM-co-associated fungi hint at the importance of specific inter-fungal interactions, which are hypothesised to be a relevant factor for the maintenance of ECM diversity. PMID- 18677627 TI - [New-onset urinary incontinence in the first six month after admission into a nursing home: prevalence, incidence and remission, risk and protective factors]. AB - BACKGROUND: Urinary incontinence is a common health problem, particularly in nursing homes. However, neither its prevalence at the time of entry nor its incidence in the first six months of residence has been assessed in the German nursing home population. Few studies have been performed to identify risk factors for new-onset urinary incontinence. Possible protective factors have not yet been analyzed for Germany. AIM: The study assesses the entry and overall prevalence of urinary incontinence in German nursing homes as well as its incidence during the first six months after entry. It also identifies factors that increase the risk of new-onset urinary incontinence after entry as well as those that promote preservation/restoration of the continence capacity. METHOD: A secondary analysis (cross-section and longitudinal section) was conducted to examine resident- related data recorded by the electronic health care documentation system of a provider with 47 nursing homes. The random sample comprised 2466 (cross-section) and 2000 persons (longitudinal section). RESULTS: Urinary incontinence was seen in 79.5% of residents at the time of entry. Its prevalence rose to 83% after six months. The overall prevalence was 87.3% (2153 of 2466). Bivariate analysis showed a disproportionately high incidence of new-onset urinary incontinence among residents with adequate physical and mental resources and low care needs at the time of entry. Constructing course variables in the bivariate analysis revealed that new-onset urinary incontinence correlated highly with deterioration of the overall physical constitution, the ability to walk and move, and the mental condition, whereas restoration of the continence capacity was clearly associated with improvement in these areas. CONCLUSIONS: The results show that preserving and promoting the mental and physical capacities of residents are the most important preventive and rehabilitating measures for counteracting the development of incontinence after nursing home entry. PMID- 18677628 TI - Urinary incontinence - the family caregivers' perspective. AB - Urinary incontinence can be expected to present an issue in domestic care with various conditions and ways of dealing with the situation. This pilot study investigates the kinds of coping strategies developed by family caregivers of urinary incontinent persons. Therefore, it seeks to explore how family caregivers experience and shape everyday life with an incontinent family member.Based on a qualitative design, problem-centered interviews were carried out with 10 family caregivers (4 men, 6 women) and analyzed according to Grounded Theory.The results show that family caregivers place major emphasis on maintaining continence, which is conceived as an expression of concern for the care-dependent person. The data facilitated the development of four major concepts: incontinence as a cause of dependency, feelings of shame, striving for continence and coping with incontinence. These concepts indicate that family caregivers look for strategies to provide the incontinent family member with the opportunity of (self-) controlled excretion. Yet this kind of support is linked to feelings of shame on both parts. PMID- 18677629 TI - [Stereotypes of ageing and sporting activity in the over-50s]. AB - Stereotypes of ageing affect activity in older adults. However, no representative studies on the impact of "images of ageing" on sporting activity existed until now. In a telephone survey, 2002 over-50s were asked about sporting activity and their opinions about ageing. A factor analysis of the queried attitudes disclosed five dimensions of ageing stereotypes. A subsequent cluster analysis identified three categories of attitudes towards ageing: "fit and performance-hungry", "conventional", and "easy-going and committed". Physical activity was described as the key criterion for a healthy old age in all clusters. Analysis also revealed a significant correlation between cluster affiliation and amount of sporting activity. The percentage of non-exercisers is lowest in Cluster 1, the "fit and performance-hungry" category (29.4%), and highest in Cluster 3, the "worryfree committed" (51.6%). Sports for health and trendy sports are the sports of choice, while conventional sports attract little interest. PMID- 18677631 TI - Severe neuropsychiatric symptoms in two siblings with intermediate type of Niemann-Pick disease. PMID- 18677630 TI - The G79A polymorphism of protein Z gene is an independent risk factor for cerebral venous thrombosis. AB - BACKGROUND AND PURPOSE: Protein Z (PZ), a vitamin Kdependent protein, plays a role in inhibiting coagulation. Its plasma level or PZ gene polymorphisms have been discussed as risk factors for stroke with conflicting results reported between various studies. Only one of these polymorphisms was studied in a cohort of patients suffering from cerebral venous thrombosis (CVT). METHODS: We performed a retrospective genetic study comparing 100 healthy controls to 54 patients referred to our hemostasis unit after CVT occurrence. We compared the distribution of three PZ gene polymorphisms that may influence PZ plasma levels: A-13G in the promoter and G79A in intron F were tested using previously described techniques, and we developed a technique to evaluate the G-103A in intron A. RESULTS: The G79A polymorphism was significantly more frequent in patients than in controls (p = 0.012): the presence of at least one A allele led to an odds ratio of 2.57 with a 95 % confidence interval of 1.23-5.34. The A-13G polymorphism also showed a nonsignificant trend towards a higher prevalence in patients. CONCLUSION: The G79A polymorphism of the PZ gene was shown to be a new independent risk factor for cerebral venous thrombosis. Nevertheless, these results have to be confirmed by a prospective study including plasma PZ evaluation. PMID- 18677632 TI - Anterior cysts of the spine: a difficult differential diagnosis to amyotrophic lateral sclerosis. AB - We describe three patients referred to our ALS/MND clinic with suspected diagnosis of amyotrophic lateral sclerosis (ALS). The patients were all male, middle aged, and their initial symptoms were weakness and fasciculations in upper limb muscles. Results of clinical and electrophysiological examination in all cases were in accordance with possible ALS according to the revised El Escorial criteria. Other conditions mimicking ALS appeared to be excluded by extensive technical examinations and laboratory tests. Only repeated MRI examinations revealed anterior spinal cysts several years after symptom onset. This report intends to highlight this rare and difficult differential diagnosis of ALS and underlines the value of the revised El Escorial criteria in conjunction with electrophysiology to asses the certainty of the diagnosis ALS. PMID- 18677633 TI - Improvement of motor functions by noisy vestibular stimulation in central neurodegenerative disorders. AB - Through the cerebellar vermis, the vestibular nerves are known to influence the basal ganglia and the limbic system. By means of noisy galvanic vestibular stimulation (GVS), it may be possible to ameliorate movement disorders, particularly akinesic symptoms, in patients with central neurodegenerative disorders. We evaluated the effect of 24-hour noisy GVS on a power-law temporal autocorrelation exponent of daytime wrist activity, separately for higher (local maxima) and lower (local minima) levels of activity, in 14 hospitalized patients. The power-law exponent for the local maxima was significantly (p < 0.002) lower with the noisy GVS than with sham stimulation, suggestive of more frequent switching behavior from low to high levels of activity or less severe akinesia. The noisy GVS may thus potentially improve certain motor dysfunctions in patients with distinct central neurodegenerative diseases. PMID- 18677634 TI - Age-dependent differences in demographics, risk factors, co-morbidity, etiology, management, and clinical outcome of acute ischemic stroke. AB - BACKGROUND: Comparisons between younger and older stroke patients including comorbidities are limited. METHODS: Prospective data of consecutive patients with first ever acute ischemic stroke were compared between younger (< or = 45 years) and older patients (> 45 years). RESULTS: Among 1004 patients, 137 (14 %) were < or = 45 years. Younger patients were more commonly female (57 % versus 34 %; p < 0.0001), had a lower frequency of diabetes (1 % versus 15 %; p < 0.0001), hypercholesterolemia (26 % versus 56 %; p < 0.0001), hypertension (19 % versus 65 %; p < 0.0001), coronary heart disease (14 % versus 40 %; p < 0.0001), and a lower mean Charlson co-morbidity index (CCI), (0.18 versus 0.84; p < 0.0001). Tobacco use was more prevalent in the young (39 % versus 26 %; P < 0.0001). Large artery disease (2 % versus 21 %; p < 0.0001), small artery disease (3 % versus 12 %; p = 0.0019) and atrial fibrillation (1 % versus 17 %; p = 0.001) were less common in young patients, while other etiologies (31 % versus 9 %; p < 0.0001), patent foramen ovale or atrial septal defect (44 % versus 26 %; p < 0.0001), and cervical artery dissection (26 % versus 7 %; p < 0.0001) were more frequent. A favorable outcome (mRS 0 or 1) was more common (57.4 % versus 46.9 %; p = 0.023), and mortality (5.1 % versus 12 %; p = 0.009) was lower in the young. After regression analysis, there was no independent association between age and outcome (p = 0.206) or mortality (p = 0.073). Baseline NIHSS score (p < 0.0001), diabetes (p = 0.041), and CCI (p = 0.002) independently predicted an unfavorable outcome. CONCLUSIONS: Younger patients were more likely to be female, had different risk factors and etiologies and fewer co-morbidities. There was no independent association between age and clinical outcome or mortality. PMID- 18677635 TI - Surgical resection plus stereotactic 125I brachytherapy in adult patients with eloquently located supratentorial WHO grade II glioma - feasibility and outcome of a combined local treatment concept. AB - OBJECTIVE: The current pilot study analyzed feasibility, risk and effectiveness of 1) microsurgery plus stereotactic iodine-125 ((125)I) brachytherapy (SBT) for large (diameter > 4 cm), circumscribed, and complex located WHO grade II glioma and 2) SBT alone for small (diameter < 4 cm), and complex located recurrences. METHODS: Lowactivity temporary (125)I seeds were used. The applied reference dose was 54 Gy and the dose rate was low (median, 10 cGy/h). Time to progression and time to additional external beam radiation (EBR) and/or chemotherapy were estimated with the Kaplan-Meier method. Any adverse sequel potentially attributable to treatment was classified as morbidity. Treatment effects of SBT were estimated according to the modified MacDonald criteria. RESULTS: Thirtyone patients (de novo group: n = 18, recurrence group: n = 13) were included. The median tumor volume before surgery was 66 ml. A planned partial tumor resection achieved eligibility for SBT in all patients. Transient morbidity of microsurgery and SBT was 27.8 % and 6.4 %, respectively. There was no permanent morbidity. Radiogenic complications did not occur. Complete response, partial response, and stable disease were seen in 8, 9, and 14 patients, respectively. Ten patients exhibited tumor progression (overall 5-year progression- free survival > 60 %). The 5-year probability to receive chemotherapy and/or EBR was 18 %. CONCLUSION: A planned partial tumor resection of large and complex located WHO grade II glioma is safe. SBT of small and complex located residual of recurrent tumors is safe and minimally invasive. Combined treatment may provide the possibility to withhold EBR and/or chemotherapy for a considerable number of patients and deserves further prospective evaluation. PMID- 18677636 TI - Increased serum ferritin levels in amyotrophic lateral sclerosis (ALS) patients. AB - Iron misregulation promotes oxidative stress, a proposed pathological mechanism in neurodegenerative disease. The aim of this study was to evaluate serum iron metabolism indicators in 60 amyotrophic lateral sclerosis (ALS) patients and 44 age matched controls. Serum ferritin levels were significantly increased in ALS patients compared to controls (p < 0.001), while no differences in the levels of serum iron, transferrin, iron saturation or total iron binding capacity were found. Likewise no differences in C reactive protein (CRP) or caeruloplasmin were detected, suggesting that the elevated ferritin levels in ALS did not merely indicate an acute phase response. The increased ferritin level may reflect a general increase in stored iron or be a consequence of ongoing muscle degeneration. PMID- 18677637 TI - Small vessel versus large vessel vascular dementia: risk factors and MRI findings. AB - OBJECTIVE: The aim of this study was a cross-sectional comparison of clinical and MRI characteristics and risk factor profiles between patients with small vessel disease (lacunae and white matter hyperintensities) and large vessel disease (large territorial or strategical infarcts) in a large cohort of VaD patients. METHODS: Patients with VaD (NINDS-AIREN) were included in a large multicenter treatment trial (the VantagE study). All patients were examined by a neurologist and interviewed about their medical history. Based on MRI, patients were classified as having large vessel VaD, small vessel VaD, or a combination. Other MRI characteristics included white matter hyperintensities (WMH), medial temporal lobe atrophy (MTA) and general cortical atrophy. RESULTS: Of the 706 patients, 522 (74 %) had small vessel disease, 126 (18 %) had large vessel disease and 58 (8 %) had both. Patients with small vessel disease were older and less educated, and showed more cortical and medial temporal lobe atrophy than patients with large vessel disease. The most prevalent vascular risk factors (hypertension, diabetes and smoking) were equally distributed between the different types of VaD. However, patients with large vessel disease had more hypercholesterolemia and cardiac risk factors compared to patients with small vessel disease. CONCLUSION: Cerebrovascular disease underlying VaD consists in the majority of small vessel disease and in about one fifth of large vessel disease. This study demonstrates heterogeneity between these two groups with regard to risk factor profile and atrophy scores on MRI. PMID- 18677638 TI - Development of classification models for early identification of persons at risk for persistent cognitive decline. AB - OBJECTIVE: To develop two classification models for use in primary care to aid early identification of persons at risk for persistent cognitive decline. METHODS: Data were used from the Longitudinal Aging Study Amsterdam (LASA), an ongoing populationbased study. The study sample consisted of 2,021 non-demented men and women aged 58-88 years. Data on relevant predictors of persistent cognitive decline were collected at baseline. RESULTS: The incidence of persistent cognitive decline after three years of follow-up was 4.0 %. In the first model, in which predictors already known or otherwise easily assessed (first set) were included, age was the strongest predictor of persistent cognitive decline, with an increased risk for persons > 75. In addition, having memory problems, low education, and a MMSE score of < or = 24, resulted in a predictive value for persistent cognitive decline of 43.5 %. In the second classification model, in addition to the first set, predictors requiring additional measurement (e.g. markers determined in blood) were included in the analyses. Age was again the strongest predictor of persistent cognitive decline. In persons > 75 years, having a low total cholesterol level (< 5.0 mmol/L) and a MMSE score of < or = 24 resulted in a predictive value of 30.0 %. CONCLUSIONS: Both models lead to a substantial increase of the predictive value for persistent cognitive decline, that is from 4.0 % to 43.5 % and 30.0 %, and may identify to a large extent a different subsample of persons who are at risk for persistent cognitive decline. The developed classification trees could be useful for case finding of persons at risk for future persistent cognitive decline who are therefore at risk for dementia, in a feasible and cost-effective manner. PMID- 18677639 TI - Employment in multiple sclerosis. Exiting and re-entering the work force. AB - Multiple sclerosis (MS) is associated with significant economic burden and high rates of unemployment. This investigation evaluated patient and disease characteristics associated with work loss and work initiation using the NARCOMS patient registry. Patient and disease characteristics associated with transitions to unemployment or employment were evaluated cross-sectionally and prospectively over the course of two assessment periods (mean interval of 1.56 +/- 0.93 years). Eligible participants included 8,867 patients for the cross-sectional component, and 8,122 for longitudinal analyses. At Time 1 and Time 2 56-58 % of MS patients were not employed. At Time 1, unemployed participants more likely to have a progressive disease course, had a longer symptom duration, greater levels of disability as measured by the PDDS, and greater functional limitations across all domains of the performance scales (p < 0.0001 for all). At Time 2, increasing MS symptoms in the past 6 months increased the odds of becoming unemployed. In addition, specific problems in mobility, hand function, fatigue, and cognitive performance domains were associated with increased odds of becoming unemployed. Less severe problems in similar areas, including mobility, hand function, and cognitive functioning were also predictive of work initiation among patients not employed. MS is associated with high rates of unemployment. Specific physical and mental health limitations confer risk of employment cessation over time, as well as the likelihood of employment initiation. This study has implications for rehabilitation interventions to target specific MS related limitations that place patients at greatest risk for work status changes. PMID- 18677640 TI - Is in utero early-exposure to interferon beta a risk factor for pregnancy outcomes in multiple sclerosis? AB - OBJECTIVE: There exist controversial and discrepant results on the risk of spontaneous abortions and teratogenesis induced by interferon treatment in people with MS.Aim of this study is to evaluate risks of the administration of INFbeta related not only to the foetus, but also to children development up to 12-months developmental milestones. METHODS: The study design is retrospective with a follow-up of babies until 18-months of their life. Thirty-eight women out of 240 with MS followed-up at Clinic MS Center of the University Hospital of Catania, Italy became pregnant in the period june 1997-may 2006. Patients were grouped into three arms: in utero exposed to INFbeta, never treated and patients who discontinued INFbeta before starting conception. Pregnancy outcomes, birth weight, 12-month developmental milestones were collected with an ad hoc questionnaire. RESULTS: Newborns of in utero exposed to INFbeta patients were little smaller for birth weight (3079.6 +/- 313.3 g), but not statistically significant, if compared with the other groups. Developmental milestones appeared within the normal range in all groups. CONCLUSIONS: Our results were particularly favourable on pregnancy outcomes, because we observed only a smaller birth weight which was not detrimental for the further development of children. We believe that INFbeta therapy might not be considered to be a reason for interruption of an intact pregnancy once the drug has been discontinued until delivery. PMID- 18677641 TI - Common variable immunodeficiency-associated myelitis: report of treatment with infliximab. PMID- 18677642 TI - Pain and motor function in carpal tunnel syndrome: a clinical, neurophysiological and psychophysical study. AB - OBJECTIVE: Patients with carpal tunnel syndrome (CTS) complain of motor symptoms. The study is aimed to understand which features are associated with the presence of motor symptoms in CTS. METHODS: We recruited 282 consecutive CTS patients. After selection, 129 patients (203 hands) were included. Patients were asked about the presence and severity of hand weakness (HW) and hand clumsiness (HC). They underwent a self-administered questionnaire on symptoms, clinical evaluation and neurographic study. Quantitative sensory testing (QST) was performed on the patients with unilateral right CTS. RESULTS: HW and HC may be found in 56 % and 48 % of CTS hands, respectively. HW was related to the severity of sensory symptoms (pain, numbness and tingling) but not to clinical-neurographic measures of median nerve involvement. HC was related to the severity of sensory symptoms and to the clinical-neurographic signs of motor but not sensory nerve damage. Motor symptoms were significantly more frequent in right hands. QST showed a relationship between the presence and severity of HW and HC and the warm threshold. CONCLUSIONS: Motor symptoms may be found in approximately half of CTS hands. Clinical and neurographic signs of median nerve motor damage appear to be poorly correlated to motor symptoms. The factor that can help reconcile the discrepancy between motor symptoms and motor signs is pain. Pain modulation on motor function may take place at various anatomical levels in CTS. Nociceptive C fibers may be involved in pain-motor interactions finally leading to motor symptoms. PMID- 18677643 TI - JNCL patients show marked brain volume alterations on longitudinal MRI in adolescence. AB - Juvenile neuronal ceroid lipofuscinosis (JNCL, CLN3) is an inherited lysosomal disease. We used longitudinal MRI, for the first time, to evaluate the rate of brain volume alterations in JNCL. Six patients (mean ages of 12.4 years and 17.3 years) and 12 healthy controls were studied twice with 1.5 T MRI. White matter (WM), gray matter (GM) and CSF volumes were measured from the sets of T1-weighted 3-dimensional MR images using a fully automated image-processing procedure. The brain volume alterations were calculated as percentage change per year. The GM and whole brain volumes decreased and the CSF volume increased significantly more in the patients than in controls (p-values for the null hypothesis of equal means were 0.001, 0.004, and 0.005, respectively). We found no difference in the WM volume change between the populations. In patients, the GM volume decreased 2.4 % (SD 0.5 %, p 0.0001 for the null hypothesis of zero mean change between observations), the whole brain volume decreased 1.1 % (SD 0.5 %, p = 0.003), and the CSF volume increased 2.7 % (SD 1.8 %, p = 0.01) per year. In normal controls, only the mean white matter volume was significantly altered (0.8 % increase, SD 0.7 %, and p = 0.001). CONCLUSION: We demonstrated by longitudinal MRI that the annual rate of the gray matter loss in adolescent JNCL patients is as high as 2.4 %. PMID- 18677644 TI - Fulminant tumefactive multiple sclerosis: therapeutic implications of histopathology. PMID- 18677646 TI - Hans Steinert (1875-1911). PMID- 18677645 TI - The association between smoking and smell and taste impairment in the general population. AB - BACKGROUND: The aim of this study was to evaluate the effect of smoking on taste and smell impairment in a large population- based study. STUDY DESIGN: Cross sectional survey in Dortmund, Germany. METHODS: The population sample was randomly drawn from the city's central registration office. Following a standardized interview, validated taste and smell tests were performed. Descriptive statistics and logistic regression was used in the analysis. RESULTS: Among the 1312 study participants, 3.6 % were functionally anosmic, and 18 % had olfactory dysfunction. Approximately 20 % recognized only three or less of the four tastes when presented at suprathreshold concentrations, indicating signs of taste impairment. Current smoking in general increased the risk for impairment of olfactory function (odds ratio 1.71, 95 % CI 1.19-2.47), but not the risk for taste impairment. Heavy smokers of 20 or more cigarettes/day had significant increased risks for impairment in both senses. CONCLUSIONS: Our results reveal that both olfactory and gustatory function are compromised in a significant proportion of the general population. Smoking increases significantly the risk of impairment of olfactory function. Our findings add an important detail to the large body of evidence that describes adverse health effects of smoking. PMID- 18677647 TI - Spontaneous abortion and the prophylactic effect of folic acid supplementation in epileptic women undergoing antiepileptic therapy. AB - BACKGROUND: Antiepileptic drugs (AEDs) like phenytoin (PHE), carbamazepine (CBZ), barbiturates and valproic acid (VPA) interfere with folic acid absorption and metabolism, which in turn can be the cause of adverse pregnancy outcome. OBJECTIVE: To study the prophylactic effect of folic acid supplementation with regard to spontaneous abortion and preterm delivery (fetal demise after week 20 of gestational age) in pregnant women receiving AED therapy, as well as benefits of most common dosage and preconceptional commencement. METHODS: Prospective examination of 104 patients, registered in EURAP from 1999-2004 at a single center and a retrospective analysis of data from our epilepsy databank completed with medical records and patients interviews of the Department of Neurology of Innsbruck University Hospital from 1971 to 1999. RESULTS: 388 pregnancies in 244 patients were analyzed. Pregnancies with folic acid supplementation showed significant reduction of spontaneous abortion. With regard to monotherapies, in the group of women taking VPA, supplementation of folic acid had significant benefit. Other examined monotherapies (CBZ, PHE, and PB) known to interfere with folic acid showed no significant results. CONCLUSIONS: This study confirms the prophylactic effect of folic acid supplementation on spontaneous abortion. For AED therapy, folic acid supplementation should be part of the therapy of every pregnant epileptic woman, especially for those treated with VPA. PMID- 18677648 TI - Intracranial arachnoid cysts--do they impair mental functions? AB - OBJECT: Arachnoid cysts are benign, congenital malformations of the arachnoid with a predilection for the temporal fossa. Radiologically, most arachnoid cysts appear to be expansive lesions, and functional imaging has shown that arachnoid cysts may cause a reorganization of cortical functions. METHODS: The present article is based on a literature search for articles reporting dyscognition or psychiatric symptoms in arachnoid cyst patients. Emphasis is placed on systematic, prospective studies reporting results from larger series of patients. A total of five articles were identified that had investigated cognition in larger series of patients. A variety of cognitive functions were found to be impaired, including verbal perception and memory, more complex verbal tasks, visuospatial functions, and visual attention. In addition, numerous case reports were identified that link arachnoid cysts to a wide variety of neuropsychiatric and cognitive symptoms. CONCLUSIONS: This survey indicates that arachnoid cysts may affect cognition and also that they do so in a reversible manner, as the patients' cognitive impairment seems to improve after surgical decompression of the cyst. PMID- 18677650 TI - How does acculturation affect the use of complementary and alternative medicine providers among Mexican- and Asian-Americans? AB - Researchers have found that immigrants in the United States gradually relinquish cultural practices and adopt health behaviors similar to native born individuals as they acculturate. Few studies have looked at acculturation and Complementary and Alternative Medicine (CAM) use, particularly ethnic forms of CAM. This study uses data from the 2001 California Health Interview Survey-Complementary and Alternative Medicine (CHIS-CAM) supplement to estimate the prevalence of CAM provider use among Mexican- and Asian- Americans and examine the relationship of acculturation on use. Multinomial logistic regression models were used to predict the probability of provider use based on socio-demographic variables, health status and acculturation. Mexican- and Asian- Americans who have spent more time in the US were more likely to use chiropractors or massage therapists compared to no CAM provider. Both groups were less likely to use ethnic-specific CAM providers with more time in the US compared to chiropractors or massage therapists. PMID- 18677651 TI - Immunomagnetic bead-based cell concentration microdevice for dilute pathogen detection. AB - A cell concentration microdevice for immunomagnetic pathogen isolation from a dilute sample is presented. Cells are driven by integrated on-chip pumps through a fluidized bed of immobilized immunomagnetic beads. Off-chip polymerase chain reaction and capillary electrophoretic analysis are used to determine capture efficiencies of E. coli and to optimize the system. Beads are immobilized after each split in a bifurcated channel system to ensure a balanced distribution of beads in all the capture channels. The addition of a pumping flutter step to repeatedly drive sample through the bead bed was found to enhance capture. Capture efficiencies of 70% and a limit of detection of 2 cfu/microL were achieved; specific capture of E. coli at a concentration of 100 cfu/microL in a 100-fold background of S. aureus is shown. This capture/concentration system is an important step in overcoming the macro-to-micro interface challenge in the development of microdevices for pathogen detection. PMID- 18677653 TI - Gene cloning, overexpression, and characterization of the nitrilase from Rhodococcus rhodochrous tg1-A6 in E. coli. AB - A DNA fragment containing the entire coding sequence of nitrilase gene was amplified from Rhodococcus rhodochrous tg1-A6 with high nitrilase activity using PCR and sequenced. The open reading frame of the nitrilase gene contains 1,101 base pairs, which encodes a putative polypeptide of 366 amino acid residues. The nitrilase gene was cloned into an expression vector pET-28a and expressed in an Escherichia coli strain BL21(DE3). The enzymatic activity of nitrilase, which converts various nitriles to the corresponding carboxylic acids, was detected to reach 24.5 U/ml at 9 h in the recombinant bacteria. PMID- 18677652 TI - Differential expression of dysadherin in papillary thyroid carcinoma and microcarcinoma: correlation with E-cadherin. AB - Dysadherin is a novel glycoprotein, with an anti-cell-cell adhesion function. The aim of the present study was to examine the expression of dysadherin in thyroid papillary microcarcinoma (PMC), to associate it with the expression of E-cadherin and to investigate whether there are differences with papillary carcinoma (PC). A statistically significant difference in dysadherin and E-cadherin expression between PC and PMC and a negative correlation between E-cadherin and dysadherin expression regardless of tumor size were noted. Based on these findings it is hypothesized that retained cell-cell adhesion, through maintenance of the E cadherin adhesion system, in PMC prevents neoplastic cells from dissociating easily from each other and metastasizing. Increased dysadherin expression is possibly one of the post-transcriptional mechanisms responsible for E-cadherin downregulation in thyroid papillary neoplasia. PMID- 18677654 TI - CD137-guided isolation and expansion of antigen-specific CD8 cells for potential use in adoptive immunotherapy. AB - The efficient isolation and ex vivo expansion of antigen-specific T cells are crucial for successful adoptive immunotherapy against uncontrollable infections and cancers. Several methods have been reported for this purpose, for example, employing MHC-multimeric complexes, interferon-gamma secretion, and antibodies specific for molecules expressed on T-cell surfaces, including CD25, CD69, CD107a, CD137, and CD154. Of the latter, CD137 has been shown to be one of the most promising targets since it is only expressed on CD8(+) T cells early after encountering antigen, while being almost undetectable on resting cells. However, detailed comparisons between CD137-based and other methods have not yet been conducted. In this study, we therefore compared three approaches (with CD137, CD107a, and tetramers) using HLA-A24-restricted CMV pp65 and EBV BRLF1 epitopes as model antigens. We found that the CD137-based isolation of antigen-stimulated CD8(+) T cells was comparable to tetramer-based sorting in terms of purity and superior to the other two methods in terms of subsequent cell expansion. The method was less applicable to CD4(+) T cells since their CD137 upregulation is not sufficiently high. Collectively, this approach is most likely to be optimal among the methods tested for the isolation and expansion of antigen-specific CD8(+) cells. PMID- 18677655 TI - Effect of single to triple dose DEC on microfilaremics up to 5 years. AB - A 5-year study was conducted with 200 selected microfilaremic subjects bearing Mean microfilarial density (MMD) of 11.6 from 16 small foothill villages around Susunia Hill, Bankura, West Bengal during June 2001 to May 2002. All of them were treated with a single dose (6 mg/kg body weight) to triple dose diethylcarbamazine (DEC). On day 180, 365, 730, 1,095, 1,460, and 1,825, recurrence of microfilaremia were 0.0%, 6.0%, 15.5% (6 + 9.5), 27% (15.5 + 11.5), 42% (27 + 15), and 49% (42 + 7), respectively, and corresponding MMD were 0.0, 1.2, 1.4, 2.7, 6.2, and 7.8, respectively. Culex quinquefasciatus was incriminated as the vector and their infection and infectivity rates reduced to 4.2% and 0.81% from the initial values of 6.40% and 1.33%, respectively, after DEC treatment to selected mff carriers. Single-dose DEC treatment was found effective for reduction of microfilaremia and MMD but it was not always sufficient to eliminate all the microfilariae, especially when MMD was high (>40). For microfilaremics with high MMD, a single course (6 mg/kg body weight for 3 days) DEC treatment may be safe for the first instance. PMID- 18677656 TI - Trans-Sylvian selective amygdalohippocampectomy for medically intractable temporal lobe epilepsy: a single-centre experience. AB - Selective amygdalohippocampectomy (SelAH) is now widely used in the surgical treatment of patients with medically intractable medial temporal lobe epilepsy (MTLE). We present our single-centre experience of a uniform series of drug resistant MTLE patients treated exclusively with SelAH. A retrospective study of 50 adult patients (25 males, 25 females, mean age 34 years) suffering from medically intractable MTLE was conducted. All patients has been investigated under the same protocol and operated by SelAH performed by one surgeon. Immediate and long-term follow-up was also available and is presented. Forty-two patients (84%) became seizure free (Engel's I); eight of them (16%) were classified as Engel's II; four (8%) as Engel's III; and one as Engel's IV (2%). In 42 of them the final result has been achieved after a single operation, while in the rest eight after a second or third surgical procedure. The mean follow-up period was 6.2 years. Two permanent complications (4%) with no treatment-related mortality have been detected. SelAH performed in well selected cases, where concordance between electrophysiological and MR findings exists, can be a safe and effective surgical treatment method for medically intractable MTLE. PMID- 18677657 TI - An open, randomized, prospective, comparative study of topical pimecrolimus 1% cream and topical ketoconazole 2% cream in the treatment of seborrheic dermatitis. AB - BACKGROUND: Seborrheic dermatitis (SD) is a chronic inflammatory disorder that mainly affects the seborrheic region. While ketoconazole is often used, pimecrolimus has been used successfully in SD. OBJECTIVE: To compare the efficacy and tolerability of pimecrolimus in comparison with ketoconazole in the treatment of SD. METHODS: A total of 48 patients with SD were included in the study. Patients were randomized into two groups: 23 and 25 patients in the pimecrolimus and ketoconazole groups, respectively. Clinical measures were assessed by erythema, scaling and infiltration, which were evaluated using a four-point scale (0 to 3) at 2, 6, and 12 weeks. RESULTS: Of these 48 patients, 38 completed the study (18 and 20 patients in the pimecrolimus and ketoconazole groups, respectively). The mean percentage decrease in clinical severity scores from baseline to the last follow-up period was 86.2% and 86.1% in the pimecrolimus and ketoconazole groups, respectively. Both pimecrolimus and ketoconazole were effective in SD. Differences between the two groups were not statistically significant. Side effects were observed more frequently with pimecrolimus than with ketoconazole and this difference was statistically significant. CONCLUSION: Our study showed that pimecrolimus had a comparable efficacy profile with that of ketoconazole, but side effects appeared more frequently in the pimecrolimus group than in the ketoconazole group. PMID- 18677658 TI - Neuroanatomy: a single institution study of knowledge loss. AB - BACKGROUND: Most of the literature on neuroanatomy education has focused on its instructional method. Little is known about the retention of acquired knowledge in the basic neurosciences upon graduation from medical school. METHODS: Twenty four graduating medical students at the University of Saskatchewan, Canada answered 20 multiple-choice questions from the original first year neuroanatomy midterm examination, 33 months after the original exam date. The course involved 58 instructional hours in the dissecting lab and classroom during the first year of medical school. RESULTS: Relative knowledge loss in this cohort was 60%, and the mean multiple-choice exam score dropped from 82% to 33%. Two students received passing grades on the retest (50% and 55%) and the rest failed. CONCLUSIONS: Most graduating medical students were unable to pass a first year exam in the basic neurosciences. Lack of knowledge reinforcement and poor applicability to the clinical setting may be to blame, and suggests that teaching foundational concepts, useful for general practice, are more worthwhile. PMID- 18677659 TI - Faculty development: yesterday, today and tomorrow. AB - Medical education has evolved to become a discipline in its own right. With demands on medical faculties to be socially responsible and accountable, there is now increasing pressure for the professionalisation of teaching practice. Developing a cadre of professional and competent teachers, educators, researchers and leaders for their new roles and responsibilities in medical education requires faculty development. Faculty development is, however, not an easy task. It requires supportive institutional leadership, appropriate resource allocation and recognition for teaching excellence. This guide is designed to assist those charged with preparing faculty for their many new roles in teaching and education in both medical and allied health science education. It provides a historical perspective of faculty development and draws on the medical, health science and higher education literature to provide a number of frameworks that may be useful for designing tailored faculty development programmes. These frameworks can be used by faculty developers to systematically plan, implement and evaluate their staff development programmes. This guide concludes with some of the major trends and driving forces in medical education that we believe will shape future faculty development. PMID- 18677660 TI - Medical education in India. AB - Medical schools in India produce the largest number of doctors than anywhere else in the world (30,408 from 271 medical schools), corresponding to the rapid proliferation of medical colleges in the last two decades, especially within the private sector. The Medical Council of India (MCI), the regulatory body, is required to approve any significant reforms in medical curricula. The accreditation process for medical schools focuses largely on the infrastructure and human resources required and little on the process and quality of education or outcomes. The implementation of the recommendations of MCI regarding recognition or de-recognition of a medical college is governed by the Ministry of Health and Family Welfare, whilst individual universities also have variable sets of regulations for their affiliated medical schools. As a result, there is no uniformity in the standard of medical education across the country. PMID- 18677661 TI - Implementation of the Bologna two-cycle system in medical education: where do we stand in 2007? - results of an AMEE-MEDINE survey. AB - BACKGROUND: The Bologna Declaration aims to harmonize European higher education. At workshops held at AMEE Conferences (2001/2007), it was observed that medical educators seem unaware of Bologna Declaration policies in their own countries. Specifically the objective to structure higher education in two cycles evokes strong opinions, but an overview on the implementation progress is lacking. METHOD: To determine the present state of implementation of the Bologna two-cycle system in medical education, an AMEE-MEDINE survey was sent to all forty-six signatory countries, inquiring about legislative decisions. RESULTS: Not all answers were unequivocal, but it appears that only seven countries decided for adoption and nineteen decided not to adopt it. The remainder fifteen have not decided or leaves the decision to their medical schools. Non-European countries seem to reject the system more often than European countries. DISCUSSION: We found that very few persons are well informed about national policies and harmonization of medical education does not seem to be enhanced by the Bologna Declaration. Our findings point in the direction of a diversification regarding curricula structure. There is a need for clarity and dialogue on many aspects of Medical Education. The Bologna process could serve as a vehicle to reach this goal. PMID- 18677662 TI - From music to macromolecules: using rich media/podcast lecture recordings to enhance the preclinical educational experience. AB - BACKGROUND: Little is known regarding the impact of lecture recordings on medical education. AIM: This study was designed to assess the impact of lecture recordings on the educational experience of first-year medical students. METHODS: Students were provided with digital lecture recordings in Molecular Foundations of Medicine, an integrated preclinical science block. Students could access recordings as audio/visual rich media, in which lectures were linked to slide presentations, and as audio-only podcasts. Student reactions were assessed by a mandatory questionnaire on the use of recordings and by a voluntary follow-up questionnaire on the effects of recordings on learning, stress and anxiety. RESULTS: Student response to lecture recordings was universally positive. A high proportion of the class accessed recordings, with rich media being the preferred format. Students felt that the lecture recordings helped them learn course material and reduced stress and anxiety. Finally, the availability of lecture recordings had no apparent adverse effect on classroom attendance. CONCLUSIONS: The availability of lecture recordings aided medical students in their studies and reduced stress and anxiety. Student response to the recordings was universally positive, and no negative outcomes were noted by students or faculty. PMID- 18677663 TI - Healing the healer: a tool to encourage student reflection towards health. AB - BACKGROUND: Research has demonstrated that students' health falters while in medical school with healthy behaviors continuing to deteriorate during residency. Medical education can be focused toward helping students find health for themselves. Physicians who are most likely to practice healthy lifestyles are more likely to encourage their patients to do so. AIMS: Create a tool that encourages self-reflection, education and self-care for physicians-in-training. METHOD: Users completed a web-based tool that helped them create a personal health plan focusing on the themes of nutrition, lifestyle choices, family history, mind-body influences and spiritual connection. A six-question survey was completed by 500 users. RESULTS: The results support the main objectives of the tool, which were to encourage self-reflection, positive lifestyle habits and education towards key aspects of health and well-being. CONCLUSION: Having medical students and residents develop their own health plans can be an efficient method towards encouraging self-care, understanding foundational health concepts and instilling skills to teach health promotion to their patients. PMID- 18677665 TI - Power of powerpoint and role of chalkboard. PMID- 18677666 TI - Comparison of factual and action knowledge in year 5 and year 6 medical students. PMID- 18677667 TI - Do you know? PMID- 18677669 TI - eMedical teacher. PMID- 18677671 TI - Body schema and body awareness of amputees. AB - AIM: The phantom phenomenon is a well-known example of the difference between body awareness and body schema. The present study is aimed at showing how body changes and prosthesis use are reflected in body schema and body awareness-the latter relating to the image that various amputees have of their bodies. SUBJECT AND METHODS: (i) Examining the configuration of body schema: A trial examining the spatial location of the phantom limb (50 people with lower or upper limb loss); (ii) examining the functional aspect of body schema: The distribution of weight power between intact and prosthetic limbs (34 people with tibial amputation); (iii) examining body awareness: Body Focus Questionnaire by Fisher (44 people with lower limb amputation, 33 intact people); and (iv) Questionnaire on anamnesis- and prosthesis-wearing habits (people participating in research methods [i] and [iii] mentioned above). RESULTS: We found that when the amputees wore their prostheses, the configuration of body schema did not change, however, the people who had not used their prosthesis for a long period of time (in our study, at least for six years), the phantom limb shortened, a phenomenon known as telescoping. The functional adaptation of the prosthesis to the body schema starts in a short time (within two weeks) after wearing it, and it becomes close to normal in carrying body weight after a longer period of time (two years). In the beginning phase of rehabilitation, the awareness of legs is similar to that of the control group, while later on it this awareness decreases. Over time, however, the lost limb, regardless of having a prosthesis or not, loses its importance. People with a more serious or vascular amputation of the upper limbs have a clearer image of them. Limb parts having a greater cortical representation appear more intensively in phantom sensations, while the strength of the cortical representation in body schema has no significance. CONCLUSION: From both configuration and functional aspects, wearing a prosthesis helps maintain a body schema in which the phantom limb remains similar to the intact one, which can be explained by the connectional schema model. This is needed for movements to be carried out properly. Although the amputee can see the prosthesis and senses the phantom limb, they do not consider it their own since they are aware of the loss. Therefore, the fact that a prosthesis is worn will not be represented in body awareness as the highest level of mental structure. PMID- 18677672 TI - Comparison of plantar pressure distribution patterns between foot orthoses provided by the CAD-CAM and foam impression methods. AB - Foot orthotic treatment is one of the major conservative methods used to handle foot problems. Total plantar contact foot orthoses are used to reduce and redistribute peak pressures. For the fabrication of a total plantar contact foot orthosis, the computer-aided design and computer-aided manufacturing (CAD-CAM) method has been applied. In this study, the plantar foot-orthosis interface pressure data during walking were collected by the Novel Pedar-mobile in-shoe plantar pressure measuring system. The data were collected under three conditions: (i) Flat insole, (ii) foot orthosis provided by the CAD-CAM method, and (iii) foot orthosis provided by the foam impression method. The Swiss Comfort CAD-CAM foot orthotics system was used in this study. For conditions (ii) and (iii), foot shapes were collected in partial weight bearing and subtalar neutral conditions. Thirty normal subjects were recruited for this study. The plantar foot surface was divided into eight plantar foot regions and then was investigated. These regions included the heel, the medial and lateral arches, the medial, mid and lateral forefoot, the hallux, and the lateral toes. The results showed that the orthoses provided by both the CAD-CAM and foam impression methods could decrease the peak pressure and the maximum force in the heel region, and increase the peak pressure and the maximum force in the medial arch region. Both orthoses redistributed the peak pressure and the maximum force from the heel to the medial arch region. The peak pressure in the mid forefoot region was different between the orthoses provided by the CAD-CAM and foam impression methods. PMID- 18677673 TI - General effect on high-risk persons when general practitioners are trained in intensive treatment of type 2 diabetes. AB - OBJECTIVE: Within the frame of a randomized clinical trial to examine whether training of general practitioners (the intervention group) in intensive lifestyle modification and pharmacological treatment of patients with type 2 diabetes has a spillover effect on individuals with impaired fasting glycaemia (IFG) or impaired glucose tolerance (IGT). DESIGN: A high-risk screening study for type 2 diabetes with an intervention programme, where general practices were randomized to provide standard treatment versus intensive lifestyle modification and pharmacological treatment to newly diagnosed diabetic patients. SETTING: General practices in Denmark. SUBJECTS: Of 1821 individuals identified with IFG or IGT, results from oral glucose tolerance tests after one and three years were available in 1510 individuals. MAIN OUTCOME MEASURES: Progression rates from IFG and IGT to diabetes and effect of intervention were estimated in a regression model using interval censoring. RESULTS: A total of 442 persons developed diabetes. There was no significant overall effect of intervention on progression rates. For risk factors, no difference in rate of change was found between randomization groups, but a difference was found between general practices within the same randomization groups. CONCLUSION: General practitioners identify a high number of incident diabetes cases in individuals with IFG or IGT found by high risk screening. Intervention at the general practitioner's level in intensive treatment type 2 diabetes does not have a significant spillover effect reducing the risk of diabetes from pre-diabetic conditions. This could indicate that intervention strategies should be specifically targeted at individuals with IFG or IGT, either by training general practitioners or directly at the individual level. PMID- 18677674 TI - Slow progression of colorectal symptoms in patients with systemic sclerosis: an 8 year follow-up. PMID- 18677675 TI - [The differential diagnosis of dementia--a remaining challenge]. PMID- 18677676 TI - [Semantic dementia. A case report in the context of an independent medical examination]. AB - Semantic dementia (SD) is a relatively rare primary degenerative brain disease, often with onset before the age of 65, which belongs to the group of frontotemporal lobar degenerations. The central characteristic of SD is a progressive loss of semantic knowledge with manifestation in aphasia as well as impaired face and object recognition. A reliable discrimination of SD from other neurodegenerative conditions, in particular from Alzheimer's disease, may be a challenge for neurologists as well as neuropsychologists. In the first place, a sound knowledge base is expected from the expert in order to minimize false diagnoses. To illustrate this, a detailed case history of a 55-year old patient is presented who was referred for an independent medical and neuropsychological examination for disability benefits. The referral question was Pick's disease. However, the clinical manifestation as well as the results of a comprehensive neuropsychological evaluation clearly indicated the diagnosis of SD with neuroimaging and neuropsychological evidence of marked right temporal lobe atrophy. The case history highlights a number of problems inherent in current practice of dementia assessment. For differential diagnosis of dementing conditions, a thorough neuropsychological assessment appears to be indispensable. PMID- 18677677 TI - [E-learning about dementia: a randomized study]. AB - BACKGROUND: Curricular medical training on dementia at the Department of Psychiatry and Psychotherapy at the University of Erlangen comprises of a traditional lecture. This setting was compared with two different E-Learning formats using a randomized study design. METHODS: 104 students (average age 26.3 +/- 3.6 years) were randomized into 3 groups: Interactive e-Learning; virtual lecture (slides and audio) and standard lecture (control group; 90 minutes). RESULTS: Overall, the response rate was 40.4 %. Assessment of formal knowledge using a multiple-choice test yielded no differences between the three groups. In the students' evaluation, the interactive e-learning showed the best results (1.86 +/- 0.69), followed by the standard lecture (2.0 +/- 0.71) and the virtual lecture (2.6 +/- 0.8). Nevertheless, the students would not prefer e-learning to the standard lecture when both methods are directly compared. CONCLUSION: Our findings suggest that e-learning is equivalent to a standard lecture when formal knowledge is assessed. Evaluation results are best for interactive e-learning formats. The detailed reasons for the preference of different learning styles should be further investigated. PMID- 18677678 TI - [Forensic risk calculation: basic methodological aspects for the evaluation of the applicability and validity of diverse methods]. AB - Risk assessment instruments have been the subject of a number of validation studies which have mainly examined the psychometric properties known primarily from psychological test development (objectivity, reliability and validity). Hardly any attention was paid to the fact that validation of forensic risk assessment instruments is confronted with a whole row of methodical challenges. Risk assessments include a quantitative and a qualitative component in that they state the probability (quantitative) of a particular offense (qualitative) to occur. To disregard the probabilistic nature of risk calculations leads to methodically faulty assumptions on the predictive validity of an instrument and what represents a suitable statistical method to test it. For example, ROC analyses are considered to be state of the art in the validation of risk assessment instruments. This method does however not take into account the probabilistic nature of prognoses and its results can be interpreted only to a limited degree. ROC analyses for example disregard certain aspects of an instrument's calibration which might lead in an instrument's validation to high ROC values while demonstrating only low validity. Further shortcomings of validation studies are that they ignore changes of risk dispositions or that they don't differentiate between offense specific risks (e. g. any recidivism vs. violent or sexual recidivism). The paper discusses and reviews different quality criteria of risk assessment instruments in view of methodological as well as practical issues. Many of these criteria have been ignored so far in the scientific discourse even though they are essential to the evaluation of the validity and the scope of indication of an instrument. PMID- 18677679 TI - [Treatment of compulsive buying]. AB - Although compulsive buying is receiving increasing attention in research, it is largely ignored in clinical practice. Compulsive buying disorder (CBD) is defined as excessive and mostly senseless spending or excessive shopping impulses that cause marked distress, interfere with social or occupational functioning, and often results in financial problems. It is currently conceptualized as an "impulse control disorder not otherwise specified" (ICD-10 F63.9). CBD is associated with significant psychiatric co-morbidity particularly mood and anxiety disorders, obsessive-compulsive disorders, binge eating disorder, substance use disorders, personality disorders, and other impulse control disorders. Previous research indicates that many compulsive buyers also suffer from compulsive hoarding. There is no evidence-based treatment approach for CBD and treatment research on this topic is limited. Open label trials suggest that antidepressants could improve compulsive buying. However, small randomized controlled trials failed to demonstrate significant improvement over Placebo and the high placebo-response rate prevents any definitive statement on the efficacy of antidepressants. Two controlled cognitive-behavioral therapy (CBT) trials reported that group CBT is significantly more effective compared to waiting list control groups in the treatment of compulsive buying. Clinical and therapeutic implications are discussed. PMID- 18677680 TI - [Autism: exploring historical psychiatric and psychological concepts]. AB - Autism today is a widely used term, yet what is understood by autism has changed considerably since first being introduced in scientific discourse almost 100 years ago. Autism is one example for the influence of the psychoanalytic school of Sigmund Freud on scientific psychiatry at the beginning of the 20th century. In particular psychoanalysis had an impact on Eugen Bleuler's concept of schizophrenia. The Swiss psychiatrist did not only acknowledge and follow a biological, but also a psychological approach to psychiatry and thus opened up his subject to psychoanalytic thoughts. This paper provides insights into the term's conceptual history--or, more specifically and precisely--sheds light on the expansion of the term's scope, which has gotten to be used for more and more symptoms and phenomena. When Bleuler first presented the term autism, he used it to refer to a classical schizophrenic symptom. Since, however, Bleuler was not very specific and exclusive in his definition, the term was soon used for other phenomena as well, such as to describe a schizoid symptom in the sense of today's schizoid personality disorder (schizoid autism). The concepts of autistic hebephrenia and depressive autism are further examples how the term was used and give insight into how the contents behind the term changed, got less and less specific and widened its scope. Due to its growing vagueness its suitability and usability as a psychopathological term decreased. This process further was strengthened when the word autism got more and more widely used in colloquial language for different aspects of day-to-day routine and thinking. Thus in psychiatry today, autism is exclusively used in connection with the so-called autism spectrum disorders, but has, as other formerly exclusively technical terms, different and rather unspecific meanings in everyday communication. PMID- 18677681 TI - [Diagnostic pathways for polyneuropathy. Part 2: Neurophysiology, laboratory diagnostics, biopsy]. PMID- 18677682 TI - Evaluation of tissue oxygen measurements for flap monitoring in an animal model. AB - Tissue oxygen tension (p(ti)O(2)) measurements are common in neurosurgery but uncommon in plastic surgery. We examined this technique as a monitoring method with probe placement in the subcutaneous tissue and addressed the importance of probe placement. Myocutaneous flaps were raised in an animal model and p(ti)O(2) measurements performed at different levels in the subcutaneous fat. Flap artery and vein were occluded until a 50% p(ti)O(2) reduction had occurred (T(1/2)). We found no significant effect of depth (P>0.10) on the level of p(ti)O(2). T(1/2)(arterial) was 7.2 minutes and T(1/2)(venous) was 18 minutes. We found no significant relation between initial levels of p(ti)O(2) and T(1/2). Location of the probe and absolute p(ti)O(2) value is of little relevance for flap monitoring. It is the relative change in p(ti)O(2) that is important. The p(ti)O(2) technique is well suited for monitoring in the subcutaneous tissue and is highly sensitive to changes in both arterial and venous blood flow. PMID- 18677683 TI - Vertical mattress suture technique: an alternative vascular anastomosis. AB - The objective of this study was to introduce the vertical mattress (VM) suture technique and compare its patency and postoperative healing with other commonly used suture techniques. Thirty-two Sprague-Dawley rats were randomly assigned to one of four suture techniques to be applied to both femoral arteries: continuous (C), interrupted (I), VM, and VM with flap (VMF) sutures. The operating time and the degree of postanastomotic leakage were recorded. The milking test was employed 5 and 15 minutes and 14 days after the operation to attain patency. Blood flow velocity was determined using ultrasonography at 1 hour and on days 1 and 14 postoperation. Rats then were sacrificed, and anastomotic lining tissues were harvested for histopathologic examination. Anastomoses by C and I were executed in 12.06 and 18.31 minutes. Modified suture techniques took longer to execute anastomosis (23.09 minutes for VM and 32.44 minutes for VMF). VM suture technique was superior to other suture techniques in terms of anastomotic leakage, quality of filling, and blood flow velocity. Also, absence of the luminal closure or constriction, intimal injury and hyperplasia, foreign body inflammation, and inversion were noted in histopathology. In conclusion, the VM suture technique could be considered for microvascular surgery. PMID- 18677684 TI - Magnetic resonance imaging of iron oxide labelled stem cells: applications to tissue engineering based regeneration of the intervertebral disc. AB - Minimally-invasive monitoring of regeneration in diseased tissue is an important aspect of stem cell therapy. Magnetic resonance imaging (MRI) based tracking of cells labelled with ferumoxides has the potential for non-invasive in vivo detection and longitudinal assessment of implanted cells. Cells labelled with ferumoxides appear as hypointense regions on MR images and thus can be distinguished from the surroundings. Application of this methodology to intervertebral disc degeneration (IVD), and detection of labelled cells implanted into the disc for tissue regeneration was examined. Mesenchymal stem cells labelled with a ferumoxide contrast agent were imaged in vitro to quantitatively characterize the signal intensity loss using MRI relaxation parameters (T1, T2, and T2*). To determine whether labelled cells could be detected within scaffolds suitable for implantation, labelled cells were seeded within both natural and synthetic polymers and imaged using MRI. Labelled cells were loaded within poly(ethylene glycol) hydrogels and imaged in vitro using both MRI and confocal microscopy. Labelled cells were also loaded into fibrin gels, and detected ex vivo within rat IVDs using MRI. Lastly, the effect of ferumoxide labelling on cell viability was investigated. Quantitatively, labelled cells demonstrate the greatest signal intensity loss and contrast on T2*-weighted images. Labelled cells can be detected in both synthetic and natural polymers, and can be distinguished from the native tissue environment of the rat IVD. Finally, labelling does not significantly impair cell viability. Consequently, this technique shows promise as a potential method for in vivo longitudinal tracking of stem cell based regeneration of the IVD. PMID- 18677685 TI - [The assessment of nursing care needs in cancer patients]. AB - In order to explore the status quo of the assessment of nursing care needs in newly admitted inpatients on oncology wards a survey in a German university hospital was carried out. Research questions included: Do the nurses collect data regarding nursing care needs of admitted cancer patients? When are the data collected? What data are collected? What is documented, when is it documented, and is the patient involved in the documentation? What, if any, factors on the part of nurses have an impact on the way in which these data are collected? Two methods were used to collect research data: nursing documents of hospitalised cancer patients were analysed (n = 68), followed by an interview with the nurses (n = 81). Beforehand, an instrument was developed and tested for both investigative methods. The main result was: the nurses' perception of the needs' assessment was incongruent with data in the patients' documentation files. The study showed that nurses consider their documentation of nursing care needs to be more comprehensive with regard to the time and content/areas of documentation than it actually is. With regard to the psychological, social, and spiritual care needs, for example, this means: in the nurses' own opinion, they had taken into account these needs far more strongly than they actually had. According to the analysis of the care documents, patients were involved in the documentation in 15% of the cases. According to the nurses, general communication skills, the use of assessment instruments, and the standardisation of the documentation need to be improved. The review of international literature confirms the deficiency recorded by this survey. The oncology patients' perspective should be included more strongly in the assessment of nursing care needs. In this way, individual nursing care needs are assumed to be understood more appropriately. Self assessment instruments could be a suitable resource in this context. The instruments have to be introduced by education, and their validity, reliability, practical relevancy, and applicability need to be checked continuously. PMID- 18677686 TI - [A nurse-led multidisciplinary malnutrition program to assess and treat patients with malnutrition or those at risk of malnutrition]. AB - Malnutrition is a world-wide problem that does not only affect developing countries. People in industrialised countries, in particular the elderly, the poor and the sick, are at risk. In Switzerland, it is estimated that between 15 and 20% of patients (depending on patient population) show signs of malnutrition upon hospital admission. In response and based on a systematic review of the literature, a working group of the internal medicine department at the University Hospital Basel has developed a malnutrition programme comprising a screening instrument, a multidisciplinary care guideline, an information brochure, and a training programme for nursing staff. It is the aim of this article to report how the programme to recognise and treat malnutrition was developed, implemented, and evaluated. PMID- 18677687 TI - [Assessment instruments for the measurement of the health-related self-care of patients with heart failure]. AB - Heart Failure is a frequent disease that is accompanied by much suffering for the patients and high costs for the health care system. An improvement of the health related self-care is supposed to improve the outcomes of patients with heart failure. In order to evaluate the efficiency of interventions valid and reliable instruments measuring the health-related self-care are needed in practice and research. The aim of the systematic literature review is to create a synopsis of the present nursing assessment instruments for the measurement of the health related self-care of patients with heart failure and to evaluate these instruments' reliability and validity. With a systematic literature search in three electronic databases and hand-search studies using instruments that measure the health-related self-care of patients with heart failure have been identified. The instruments are described and their reliability and validity are discussed. RESULTS: Five Instruments were identified: European Heart Failure Self-care Behaviour Scale (EHFScBS); Revised Heart Failure Self-care Behaviour Scale (rHFScBS); Heart Failure Self-Care Inventory (HFS-CI), Self-Management of Heart Failure Scale (SMHF); Self-Care of Heart Failure Index (SCHFI). As far as the psychometric properties are concerned, each of these scales has its strengths and weaknesses. The EHFScBS, the HFS-CI, and the SCHFI can be recommended with restriction for use in practice and research. However, further testing in order to evaluate reliability and validity is necessary for all instruments. PMID- 18677688 TI - [Swiss Research Agenda for Nursing (SRAN): the development of an agenda for clinical nursing research in Switzerland]. AB - In many Anglo-Saxon and North European countries nursing research agendas have been developed to address priorities in nursing research in accordance with a nationally defined health policy. In Switzerland, due to lack of a nationwide governmental health policy, co-ordination of nursing research so far was scarce. The "Swiss Research Agenda for Nursing (SRAN)" project developed an agenda for clinical nursing research between 2005 and 2007. Based on literature reviews, expert panels and a national survey a project team formulated an agenda which passed a consensus conference. The agenda recommends aspects that should lead research and defines seven research priorities for nursing in Switzerland for the time between 2007 and 2017. Nursing research should prioritize to investigate 1) the effectiveness of nursing interventions; 2) the influences of service adaptations in a changing health care system; 3) the phenomena in patients requiring nursing care; 4) the influence of the work environment on the quality of nursing care; 5) the functioning of family and social systems; 6) varieties of life circumstances and their integration; and 7) the implementation of ethical principles in nursing. Written in German and French, the Swiss Research Agenda for Nursing for the first time formulates priorities for nursing research in Switzerland and can be used for strategic discussions. As a next step, the development of an action plan to enhance nursing research will take place in Switzerland. PMID- 18677689 TI - [Living with a chronic abdominal wound--the patients' perspective]. AB - Chronic abdominal wounds lead to prolonged hospital stays. However, no data exist that describe the experience of persons living with a chronic abdominal wound. The aim of this qualitative study was to explore the experience of persons living with chronic abdominal wounds and elicit their expectations in health professionals. Narrative interviews were conducted with five women and four men. Using content analysis techniques, five categories with one to four subcategories emerged from the data. "Returning to everyday life without a wound" represents the main goal of the participants and is driven by their hope to achieve this outcome. "Everyday life with the wound" illustrates the reality of participants' lives, which is affected by suffering. "The patients' work" demonstrates the component that patients contribute to managing their wound. "The work of primary support persons" shows the importance of the work of people close to the patient. "The work of professionals" includes the expectations that patients have in their caregivers. Teamwork between professionals, patients and primary support persons seems to be an essential condition for the successful healing of an abdominal wound. PMID- 18677690 TI - [Mobility and balance]. AB - Quality of life is strongly associated with the mobility of elderly people. Falls often cause restricted mobility, a decline in activities of daily living and an increased risk of institutionalisation. Frailty, commonly associated with aging, is a biologic syndrome of decreased resistance to stressors, resulting from declines across multiple physiological systems. Changes in mobility and gait constitute part of the frailty syndrome. Since more than one third of persons over the age of 65 fall each year, prevention of falls is very important. Already while taking the patients' history special emphasis should be laid on matters associated with an increased risk of falling, such as the use of more than four medications. To assess mobility several brief tests exist (i.e. Timed up & go [17], Walking while Talking [20]) which immediately yield information regarding mobility and falling risk. Patients with poor performance on such tests or those with a history of several falls should undergo a spatio-temporal gait analysis in order to determine a possible cause as well as suitable interventions. Additionally, the objective measurement of temporo-spatial gait parameters under dual task conditions may detect deficits in cognitive function. Several interventions have been shown to have favourable effects on gait stability and the occurrence of falls. Proprioceptive problems can be partially compensated for by wearing special shoes. Also, different movement exercises such as Tai Chi Chuan, Jaques-Dalcroze eurhythmics and social dancing are associated with better balance and gait safety, and a reduction of falls. PMID- 18677691 TI - [Falls and osteoporoses]. AB - Falls are common in each period of life. They become more important in old age as they may lead to impairment and handicap, as well as to an increased need for social and financial support, or to admission to nursing homes. Although geriatric syndromes are well known, the entity of osteoporoses, gait disturbances and falls is not established in community based health services. There are many reasons for the negotiation--lack of awareness in professionals as well in the people themselves, lack of time and lack of financing by health insurance, fear of consequences, as admission to a nursing home. Therefore it would be of interest to raise the awareness in medical professions, especially in general practitioners. Cochrane data analysis proves the efficiency of fall prevention programmes, mainly of exercise and environmental adaptations. PMID- 18677692 TI - [Laboratory tests in the elderly and their relationship with frailty]. AB - Frailty is a geriatric syndrome that has been characterized by an increased vulnerability against external stressors. The aim of laboratory investigations in the context of frailty is to screen for conditions that are pathophysiologically related to the development of this syndrome. Successful therapy of these conditions may help to stabilize the elderly individual who is at risk for functional decline and increasing frailty. In this review laboratory parameters for the detection of anemia, impaired vitamin D intake, thyroid and gonadal function are described in their relationship with developing frailty. PMID- 18677693 TI - [Diabetes mellitus]. AB - Diabetes mellitus has a prevalence of up to 25% in the age group above 75 years and is thus a major disease of the elderly. Geriatric syndromes (e.g. immobility or falls, incontinence or intellectual decline) occur more often in the diabetic elderly. Moreover there is a mutual interaction between the syndromes and the control and therapy of diabetes. This fact can be explained by the molecular mechanisms of frailty. Advanced glycation end products, inflammatorial cytokines like IL-6 or TNF-alpha are elevated in subjects with metabolic syndrome as well as in frail elderly. Insulin has anabolic effects inhibiting protein catabolism. The situation of frail elderly with diabetes sometimes can be improved by starting an insulin therapy because of its anabolic effects beyond the action of normalizing blood glucose. PMID- 18677694 TI - [Cerebrovascular disease]. AB - The present article presents correlations of frailty associated functional changes in older patients with the development and progression of cerebrovascular disease (CVD). Special focus is drawn on the impact of well established risk factors for the development of CVD such as diabetes, hypertension and smoking on frailty markers especially in the light of functional recovery following an acute cerebrovascular event. PMID- 18677695 TI - [Chronic heart failure]. AB - As a consequence of the increasing life expectancy the number of patients suffering from chronic heart failure has been growing continuously in the past few decades, especially in the group of the old and oldest. Frailty is a clinical syndrome that geriatricians attach great importance to. Like many other diseases chronic heart failure can cause frailty. Based on the experience that we see only a small correlation between the functional capacity of patients with heart failure and the results of cardiological findings, the model of peripheral myopathy in chronic heart failure was developed. Different pathophysiological changes may cause the increasing exercise intolerance in patients with chronic heart failure. We can already consider different experimental approaches to the therapy of frailty caused by chronic heart failure. At the moment we have to focus our efforts on an optimal therapy of heart failure, especially with angiotensin-converting-enzyme inhibitors and beta-blockers, and on individual endurance and strength training. PMID- 18677696 TI - [Urinary incontinence]. AB - Urinary incontinence is a common health problem of older adults and affects more than 50% of nursing home residents. Different types of urinary incontinence can be distinguished: urgency, stress, overflow (urinary retention) and extra urethral incontinence. The pathogenesis of urinary incontinence in older patients is multifactorial. Age-related physiologic changes, urological or gynaecological ailments, neurological diseases, behavioural patterns and functional decline frequently contribute to incontinence to some extent. These pathogenetic factors may appear in close relation to frailty. As incontinence on the one hand may substantially contribute to decompensation of health reserves, and on the other hand is an indicator for frailty, early assessment and treatment are important. Clinical diagnostics focus on history, physical examination, bladder diary and measurement of postvoid residual volume. Diagnostic and therapeutic approaches to urinary incontinence should focus on finding and treating reversible causes. Behavioural interventions such as toileting procedures may reduce the level of incontinence even in severe cases. Frail older adults with functional decline and urgency incontinence often do not respond well to drug treatment. Paramount for the adequate care for incontinent seniors is the instruction of the affected person and--if necessary--of their caregiver in handling incontinence aids. PMID- 18677697 TI - [Modern intensive care]. AB - Frailty associated functional changes in the elderly lead to different problems treating geriatric patients in an intensive care unit. Patients suffering from frailty have less physiological reserves in acute illness, complications are more frequent and recovery is clearly delayed. Frailty is also an established and independent predictor for mortality. Therefore understanding of physiological ageing and knowledge of geriatric syndromes must be part of modern intensive care medicine. The importance of Frailty for intensive care is shown by means of selected clinical pattern. PMID- 18677698 TI - [Physical therapy and rehabilitation]. AB - The characteristic of increasing frailty in the elderly is amyotrphia, the so called sarcopenia. At this physiological ageing is as important as lifestyle and environmental influences. With specific physical activities, older people have the ability to gain positive effects in strength, balance and quality of life. This fits as well in prevention as in rehabilitation after sickness. PMID- 18677699 TI - [A health economics approach]. AB - Frailty is a complex term and has a broad influence on the elderly patients. For evaluating the monetary impact of frailty it is necessary to approach in a differentiated way. It is of great interest to discover the relevance of multidisciplinary assessment and accordant preventive actions. In this article the costs of osteoporosis and (hip) fractures are briefly presented. PMID- 18677700 TI - Hemodynamics of the maternal venous compartment: a new area to explore in obstetric ultrasound imaging. PMID- 18677701 TI - Cardiac output following fetoscopic coagulation of major placental vessels in fetal sheep. AB - OBJECTIVES: To measure changes in cardiac output (CO) after partial cord occlusion in fetal sheep in order to investigate pathophysiological fetal adaptation mechanisms in a simulated acute placental insufficiency model under standardized conditions, with the aim of finding relevant methods for monitoring human fetuses during stress situations. METHODS: We used minimally invasive, percutaneous endoscopic techniques to close umbilical vessels in mid-gestational fetal sheep. Placental blood flow was reduced by preferentially closing first arterial and then the concomitant venous umbilical vessels within a short time interval. The investigations were carried out on 11 pregnant ewes at a median gestational age of 86 (range, 73-95) days. Major placental arteries and veins were occluded permanently by coagulation with bipolar forceps under percutaneous fetoscopic control. The fetal CO and Doppler parameters in the ductus venosus (DV), umbilical artery (UA) and umbilical vein (UV) were measured. RESULTS: In spite of heart rate reduction, the CO was not significantly affected by vessel occlusion (mean +/- SD, 500 +/- 194 mL/min before and 457 +/- 219 mL/min after coagulation) because stroke volume slightly increased from 2.65 +/- 1.16 mL/beat to 3.1 +/- 1.5 mL/beat. The right to left CO ratio remained at 1.4. The UV flow/combined CO ratio decreased from 34 +/- 14% to 25 +/- 10% after vessel coagulation. The pulsatility index in the DV increased from 0.4 +/- 0.1 to 0.7 +/ 0.4. The DV blood flow volume remained relatively constant after the intervention. The DV shunting rate, given as DV/UV flow ratio, increased significantly from 30.8 +/- 4.7% to 59.3 +/- 25.0%. CONCLUSIONS: The nearly simultaneous closure of arterial and venous umbilical vessels changed the flow pattern in the UA and significantly reduced placental blood perfusion. It did not distinctly change the blood flow volume rate through the DV, and consequently the DV shunting rate was increased. The combined CO was not significantly affected by the vascular obstruction, whereas the fraction of combined CO directed to the placenta was reduced. PMID- 18677702 TI - Reliability of transvaginal ultrasonography at 7-9 weeks' gestation in the determination of chorionicity and amnionicity in twin pregnancies. AB - OBJECTIVES: To assess the agreement in the diagnosis of chorionicity and amnionicity between transvaginal ultrasound examination at 7-9 weeks' gestation and at the 11-14-week transabdominal scan. METHODS: This was a retrospective analysis of prospectively collected data of twin pregnancies over a 3-year period. Chorionicity and amnionicity were assessed in women with viable twin pregnancies who underwent a transvaginal ultrasound scan at between 7 and 9 weeks' gestation. These findings were compared with the diagnoses of chorionicity and amnionicity at the subsequent 11-14-week scan in these women. RESULTS: Chorionicity and amnionicity were documented in 67 viable twin pregnancies at both 7-9 and 11-14 weeks' gestation. There was agreement in the chorionicity and amnionicity reported at each of the two scans in 65 out of 67 (97%) cases. Of the dichorionic-diamniotic (DCDA) pregnancies reported at 7-9 weeks, 53 out of 54 (98%) were confirmed at the 11-14-week scan and one (2%) was found to be monochorionic-diamniotic (MCDA). However, at birth these twins were of different sex, confirming DCDA twins as initially diagnosed at 7-9 weeks. Of the 12 pregnancies diagnosed as MCDA at 7-9 weeks, all were found to be MCDA at the 11 14-week scan. There was one monochorionic-monoamniotic (MCMA) pregnancy diagnosed at 7-9 weeks that was subsequently found to be MCDA at the 11-14-week scan. CONCLUSION: Transvaginal ultrasound examination at 7-9 weeks' gestation shows very high agreement with the 11-14-week scan in the diagnosis of chorionicity and amnionicity in twin pregnancies, suggesting that it provides a similar level of accuracy. Accuracy may be higher for DC twins than MC twins, which may relate to the gestational age at which the sonographic appearance of the amniotic sac develops. PMID- 18677703 TI - Prenatal sonographic and MRI findings in a pregnancy complicated by uterine sacculation: case report and review of the literature. AB - Approximately 15% of women have a retroverted uterus prior to pregnancy, and retroversion occurs in 11% of women in the first trimester of pregnancy. However, the uterus usually moves to an upward position before 14 weeks' gestation. Incarceration and sacculation of a retroverted uterus occur in 1 in 3000 pregnancies and are difficult to diagnose. They have often been missed until shortly before delivery and can lead to serious obstetric emergencies such as labor dystocia, uterine rupture, retained placenta and uncontrollable postpartum hemorrhage. Performing a Cesarean section without correct diagnosis may cause difficulties in identifying the bladder and the cervix, and therefore in opening the lower uterine segment. This leads to bladder injuries, vaginal transsection and trans- or supracervical hysterectomy. Early diagnosis and detailed scanning are crucial for the obstetric management and operative approach.We report a case of an incarcerated uterus in a patient presenting at 24 weeks' gestation with severe bilateral flank and lower abdominal pain. The symptoms were misdiagnosed as appendicitis. Digital examination revealed a ventralized vaginal axis. The cervix was not palpable. The clinical course, and two- and three-dimensional ultrasound and magnetic resonance imaging findings, are presented. The delivery was performed by midline laparotomy Cesarean section. The management for different gestational ages and a review of the literature are discussed. PMID- 18677704 TI - Ultrastructure of the eggshell and its formation in Planipapillus mundus (Onychophora: Peripatopsidae). AB - Although the majority of onychophorans are viviparous or ovoviviparous, oviparity has been described in a number of species found exclusively in Australia and New Zealand. Light microscopy and scanning and transmission electron microscopy were used to examine developing eggs and the reproductive tract of the oviparous Planipapillus mundus. Deposited eggs and fully developed eggs dissected from the terminal end of the uteri have an outer thick, slightly opaque chorion, and an inner thin, transparent vitelline membrane. The chorion comprises an outermost extrachorion, sculptured with domes equally spaced over the surface; a middle exochorion, with pores occurring in a pattern of distribution equivalent to that of the domes of the extrachorion above; and an innermost, thick endochorion consisting of a spongelike reticulum of cavities comparable to the respiratory network found in insect eggs. The vitelline membrane lies beneath the chorion, from which it is separated by a fluid-filled space. The vitelline membrane tightly invests the developing egg. Examination of oocytes in the ovary and developing eggs at various stages of passage through the uterus indicate that the majority of chorion deposition occurs in the midregion of the uterus, where vast networks of endoplasmic reticulum are present in the columnar epithelium. The vitelline membrane, however, is believed to begin its development as a primary egg membrane, surrounding the developing oocytes in the ovary. The vitelline membrane is transformed after fertilization, presumably by secretions from the anterior region of the uterus; hence, it should be more accurately referred to as a fertilization membrane. Aspects of the reproductive biology of P. mundus are also included. PMID- 18677705 TI - The utility of ultra-performance liquid chromatography/electrospray ionisation time-of-flight mass spectrometry for multi-residue determination of pesticides in strawberry. AB - The utility of ultra-performance liquid chromatography/orthogonal-acceleration time-of flight mass spectrometry (UPLC/TOFMS) for the rapid qualitative and quantitative analysis of 100 pesticides targeted in strawberry was assessed by comparing results with those obtained using a validated in-house UPLC tandem mass spectrometry (MS/MS) multi-residue method. Crude extracts from retail strawberry samples received as part of the 2007 annual UK pesticide residues in food surveillance programme were screened for the presence of pesticide residues using UPLC/TOFMS. Accurate mass measurement of positive and negative ions allowed their extraction following 'full mass range data acquisition' with negligible interference from background or co-eluting species observed during UPLC gradient separation (in a cycle time of just 6.5 min per run). Extracted ion data was used to construct calibration curves and to detect and identify any incurred residues (i.e. pesticides incorporated in or on the test material following application during cultivation, harvest and storage). Calibration using matrix-matched standards was performed over a narrow concentration range of 0.005-0.04 mg kg(-1) with determination coefficients (r2) > or =0.99 for all analytes with the exception of malathion/fenarimol/fludioxanil (r2 = 0.98), quassia/pymetrazine (r2 = 0.97) and fenthion sulfone (r2 = 0.95). Residues found in selected samples ranged from 0.025-0.28 mg kg(-1) and were in excellent agreement with results obtained using UPLC/MS/MS. Mass measurement accuracies of < or =5 ppm were achieved consistently throughout the separation, mass range and concentration range of interest thus providing the opportunity to obtain discrete elemental compositions of target ions. PMID- 18677706 TI - Structural evaluation of the glucuronides of morphine and formoterol using chemical derivatization with 1,2-dimethylimidazole-4-sulfonyl chloride and liquid chromatography/ion trap mass spectrometry. AB - For the first time chemical derivatization of isomeric drug glucuronides with 1,2 dimethylimidazole-4-sulfonyl chloride (DMISC) has been successfully applied as a tool for determining the site of conjugation. This provides a way to differentiate between glucuronide isomers containing aliphatic and phenolic hydroxyl groups. The analyses were performed with liquid chromatography/electrospray ion trap mass spectrometry (LC/ESI-MSn). DMISC has previously been shown to react selectively with phenols in estrogens, thus improving sensitivity in ESI-MS. The model compounds selected for this study were commercially available standards of formoterol, morphine, morphine-3-glucuronide (M3G), and morphine-6-glucuronide (M6G). Formoterol glucuronides were produced with an enzymatic method in house. Both formoterol and morphine possess one phenolic and one aliphatic hydroxyl group where glucuronidation could take place. The product ion mass spectra of the native morphine glucuronides were indistinguishable due to the initial neutral loss of monodehydrated glucuronic acid (176 u). However, a significant difference between the isomers was observed with DMISC derivatization, as only the form with a free phenol, M6G, gave a detectable reaction product. Formoterol formed two detectable glucuronide isomers in the enzymatic reaction. Their respective sites of conjugation could not be directly determined from the product ion spectra. Reaction with DMISC, however, gave a detectable product with only one of the isomers. Based on previous experience of the preferred DMISC reactions with phenols, and interpretation of the fragmentation pattern of the derivative, it was concluded that the reactive isomer had a free phenol, and was thus conjugated on the aliphatic chain. PMID- 18677707 TI - A molecular mechanics force field for lignin. AB - A CHARMM molecular mechanics force field for lignin is derived. Parameterization is based on reproducing quantum mechanical data of model compounds. Partial atomic charges are derived using the RESP electrostatic potential fitting method supplemented by the examination of methoxybenzene:water interactions. Dihedral parameters are optimized by fitting to critical rotational potentials and bonded parameters are obtained by optimizing vibrational frequencies and normal modes. Finally, the force field is validated by performing a molecular dynamics simulation of a crystal of a lignin fragment molecule and comparing simulation derived structural features with experimental results. Together with the existing force field for polysaccharides, this lignin force field will enable full simulations of lignocellulose. PMID- 18677708 TI - Accuracy and convergence of free energy differences calculated from nonequilibrium switching processes. AB - The molecular-dynamics-based calculation of accurate free energy differences for biomolecular systems is a challenging task. Accordingly, convergence and accuracy of established equilibrium methods has been subject of many studies, often focusing at small test systems. In contrast, the potential of more recently proposed nonequilibrium methods, derived from the Jarzynski and Crooks equalities, has not yet fully been explored. Here, we compare the performance of these methods by calculating free energy differences for test systems at different levels of complexity and varying extent of the involved perturbations. We consider the interconversion of ethane into methanol, the switching of a tryptophane-sidechain in a tripeptide, and the binding of two different ligands to the globular protein snurportin 1. On the basis of our results, we suggest and assess a new nonequilibrium free energy method, Crooks Gaussian Intersection (CGI), which combines the advantages of existing methods. CGI is highly parallelizable and, for the test systems considered here, is shown to outperform the other studied equilibrium and nonequilibrium methods. PMID- 18677709 TI - Evaluation of role of angiotensin III and aminopeptidases in prostate cancer cells. AB - BACKGROUND: The aim of this study was to perform a comprehensive evaluation of angiotensin III (Ang-III) and related converting enzymes, aminopeptidase A (APA) and N (APN), in prostate cancer. METHODS: We investigated the effects of Ang-III on the in vitro growth of human prostate cancer cells and the expression of APA and APN in cells treated with Ang-III or hormonal agents. Furthermore, we performed real-time quantitative PCR to investigate the expression pattern of APA and APN in 86 prostate tissue samples including normal prostate, untreated and hormone refractory prostate cancer (HRPC). RESULTS: Ang-III stimulated cell proliferation, and the proliferative effect was inhibited by olmesartan, an AT(1) receptor blocker (ARB). Western blot analysis showed that phosphorylation of mitogen-activated protein kinase (MAPK) was enhanced by Ang-III and inhibited by olmesartan. APN mRNA level in HRPC was significantly lower than that in normal prostate and untreated prostate cancer tissue. In LNCaP cells, APN expression was augmented by Ang-III, whereas APA expression was not modulated. Hormonal agents, such as estradiol (E2) and dexamethasone (Dex), enhanced APA expression, but did not modulate APN expression in LNCaP cells. CONCLUSIONS: Our results suggest that Ang-III and related converting enzymes contribute to cell proliferation of prostate cancer, and may be implicated in cancer progression. PMID- 18677710 TI - Breaking bad news to cancer patients: survey and analysis. AB - PURPOSE: To find out how patients perceived the disclosure of news about their cancer as regards the physician counselling and how they perceived the flow of information between hospital-based and family physicians. METHODS: 272 cancer patients were polled with a 16-item questionnaire. RESULTS: 252 cancer patients, 92.6% of those asked, completed the questionnaire. 37.7% (f:35.4%, m:41.8%) stated that the fact that they had cancer was presented to them 'very empathically' or 'empathically'. 62.3% (f:64.7%, m:58.3%) stated that it was presented to them 'not so empathically' or ' not at all empathically'. When patients had been counselled by family physicians they were more likely to state that it had been done 'very empathically' or 'empathically', in contrast to when they had been counselled by hospital-oncologists or self-employed specialists (81.8% vs. 41.2% vs. 41.2%; p=0.001). Significantly more patients thought that they had been given adequate opportunity to ask the questions they considered important when counselled by a family physician (81.8%) as compared to counselling by a hospital-oncologist (43.5%; p=0.002) or a self-employed specialist (44.3%; p=0.001). 56.8% preferred to discuss the suggested cancer therapies with an oncologist. 87.5% of patients considered the exchange of information between the hospital-based specialists and their family physician 'very important' or 'important'; more than half of all patients stated that this exchange of information was 'rather poor' or 'poor'. CONCLUSIONS: Oncologists should involve family physicians in disclosing bad news to patients. There are considerable deficiencies regarding information-exchange in cancer care in Austria. PMID- 18677711 TI - Improved performance of first-trimester combined screening for trisomy 21 with the double test taken before a gestational age of 10 weeks. AB - OBJECTIVE: To evaluate if there is a performance difference in the combined screening for trisomy 21 between the double tests performed before and after 10 + 0 weeks of gestation. METHODS: The study included all 97 trisomy 21 cases from January 2004 to December 2007, in a screening program where the double test was measured in week 8 + 0 to 13 + 6 and the nuchal translucency in week 11 + 3 to 13 + 6. RESULTS: As many as 87 of the 97 cases were diagnosed in the screening program (detection rate = 90%). Among the 53 cases having the double test taken before 10 + 0 weeks, no cases were missed [detection rate (DR) = 100%, CI = 0.94 1.00] and among the 44 having the double test taken after 10 + 0 weeks, 10 cases were missed (DR = 77%, CI = 0.65-0.90). This difference in DR was highly significant (P = 0.0009). No difference was observed between false positive rates or median maternal age of the two groups. CONCLUSION: A significantly higher DR was obtained when the double test was taken early in pregnancy (<10 + 0 weeks), than when it was taken later (>or=10 + 0 weeks). The results could not be explained by a difference in either the false positive rate or the maternal age between the two groups. PMID- 18677712 TI - False positive FISH diagnosis of monosomy X in uncultured amniotic fluid cells due to a chromosome Y deletion. PMID- 18677713 TI - Serum levels of pigment epithelium-derived factor (PEDF) are positively associated with visceral adiposity in Japanese patients with type 2 diabetes. AB - BACKGROUND: Pigment epithelium-derived factor (PEDF) inhibits endothelial cell injury. Further, serum levels of PEDF are elevated in the metabolic syndrome. These observations suggest that PEDF may be elevated as a counter-system against vascular cell damage in the metabolic syndrome. However, little is known about the regulation of PEDF in patients with diabetes. In order to clarify the determinants of serum PEDF, here, we examined the relationship between the 1-year changes in PEDF levels and those in anthropometric and metabolic variables in type 2 diabetic patients. METHODS: Eighty-six consecutive outpatients with type 2 diabetes underwent a complete history and physical examination, determination of blood chemistries, and serum levels of PEDF at baseline and 1 year after. PEDF gene expression in cultured subcutaneous or omental adipocytes were analysed by quantitative real-time reverse transcription-polymerase chain reactions. RESULTS: Multiple regression analyses revealed that waist circumference, triglycerides, creatinine, and TNF-alpha were independently associated with PEDF. Further, the percent changes in serum levels of PEDF during 1-year observational periods were positively correlated with those of BMI. In addition, PEDF mRNA levels in cultured adipocytes were increased in parallel to the BMI values of subjects from whom adipocytes were derived, especially in omental adipocytes. CONCLUSION: These results demonstrated that serum levels of PEDF were positively associated with metabolic components and TNF-alpha in Japanese patients with type 2 diabetes. Our present study suggests that PEDF may be generated from adipose tissues and play some role in visceral obesity in type 2 diabetic patients. PMID- 18677714 TI - Validation of an item bank for detecting and assessing psychological distress in cancer patients. AB - OBJECTIVE: To validate an item bank for assessing and detecting psychological distress in cancer patients by (1) identifying whether additional items are required in the full item bank; (2) identifying any item bias in the existing item bank; (3) linking levels of distress against thresholds derived from gold standard psychiatric interviews (PSE/SCAN/SCID). METHOD: A Rasch analysis was conducted on a heterogeneous sample of cancer patients (n=4919) who had completed a combination of eight psychological distress screening instruments. A subset of patients had completed a psychiatric interview along with the HADS (n=381) or PHQ 9 (n=440). Item thresholds were plotted along the latent trait. Furthermore, items were assessed for differential item functioning (DIF) by age and gender. Finally, optimum thresholds were derived for the HADS and PHQ-9 and plotted along the latent trait distribution for the entire item bank. RESULT: Item thresholds exceeded the range of person measures, although a gap was still present along the latent trait. No DIF was observed for either age or gender. Putative cut-offs were derived for the item bank detecting moderate to severe levels of psychological distress. CONCLUSION: The item bank covers the majority of levels of emotional distress reported by cancer patients. Additionally, initial thresholds have been derived on the item bank, which correspond to a formal psychiatric assessment. Further work is required to ascertain the stability of the item bank over time and by diagnosis and stage of disease, as well as to determine additional thresholds for levels of distress. PMID- 18677715 TI - Patient-physician communication during oncology consultations. AB - OBJECTIVE: The aim of this study was to characterize the content of patient physician communication in standard oncology care. METHODS: The sample consisted of 19 patients with gastrointestinal cancer. The consultations were audio recorded, transcribed verbatim, and analyzed according to qualitative content analysis. RESULTS: The analysis resulted in seven main categories: Disease and treatment, Healthcare planning, Everyday living, Psychological well-being, Coping with disease, Expressions of concerns and feelings, and Other aspects of communication. The main focus during the consultations was on disease and treatment. Physicians tended to concentrate on response to treatment and types and severity of side effects and how to treat them. More patient-centered subjects of psychosocial character like coping and psychological well-being were discussed only briefly, if at all. CONCLUSIONS: This study adds to the information given by the existing communication analysis systems, and hence we suggest a development of the psychosocial content categories of those systems to make them more valid. PMID- 18677716 TI - Pre-treatment symptom cluster in breast cancer patients is associated with worse sleep, fatigue and depression during chemotherapy. AB - OBJECTIVE: The concept of symptom clusters is relatively new in cancer patients' symptom management. This study, which spanned four cycles of chemotherapy, combined three commonly seen pre-treatment symptoms in cancer patients (i.e. sleep disturbances, fatigue and depression) into one symptom cluster, to explore the associations between pre-treatment cluster categories and longitudinal profiles of these same symptoms during chemotherapy. METHODS: This was a prospective study. Seventy-six women with newly diagnosed stage I-III breast cancer, scheduled to receive at least four cycles of adjuvant or neoadjuvant anthracycline-based chemotherapy participated. Data were collected at seven time points before and during treatment. Sleep quality was measured with the Pittsburgh Sleep Quality Index. Fatigue was measured with the Multidimensional Fatigue Symptom Inventory--Short Form. Depressive symptoms were measured with the Center of Epidemiological Studies--Depression. Patients were divided into three groups based on the number of symptoms they experienced before the start of chemotherapy (i.e. no symptoms, 1-2 symptoms or all three symptoms) and a symptom cluster index (SCI) was computed. RESULTS: All women reported worse sleep, more fatigue and more depressive symptoms during treatment compared with baseline (all p's<0.01); however, those women with a higher SCI (i.e. more symptoms pre treatment) continued to experience worse symptoms during treatment compared with those who began with fewer symptoms (all p's<0.01). CONCLUSIONS: A higher clinically relevant-based pre-treatment symptom cluster was associated with more sleep disturbances, greater fatigue and more depressive symptoms during chemotherapy. Specific interventions for these pre-treatment symptoms may improve the frequency and severity of these same symptoms during chemotherapy, when they are most severe and most disruptive to quality of life. PMID- 18677718 TI - Imaging mass spectrometry using peptide isoelectric focusing. AB - Imaging Mass Spectrometry (IMS) has emerged as a powerful technique in the field of proteomics. The use of Immobilized pH Gradient-IsoElectric Focusing (IPG-IEF) is also a new trend, as the first dimension of separation, in shotgun proteomics. We report a combination of these two outstanding technologies. This approach is based on the separation of shotgun-produced peptides by IPG-IEF. The peptides are then transferred by capillarity to a capture membrane, which is then scanned by the mass spectrometer to generate MS images. This high-throughput methodology allows a preview of the sample to be obtained in a single day. We report the application of this new pipeline for differential comparison of the membrane proteome of two different strains of Staphylococcus aureus bacteria in a proof-of principle experiment. PMID- 18677719 TI - Software for the calculation of isotope patterns in tandem mass spectrometry. AB - Software, available at no cost on the Internet, is described which uses polynomial expansion algorithms to calculate the isotope patterns for precursor ion, neutral loss, and MSn product ion tandem mass spectra. Such information is useful for determining product ion and neutral loss composition, identification of analytes in complex samples, deconvolution of overlapping spectra, and correction of peak heights or areas in quantitative analysis. The effect of less than unit mass resolution on the isotope patterns is described and experimental examples of the use of the software are presented. PMID- 18677717 TI - Physical, psychological, and social sequelae following hematopoietic stem cell transplantation: a review of the literature. AB - OBJECTIVE: This article reviews recent literature on adults' quality of life following hematopoietic stem cell transplantation (HSCT). METHODS: We identified 22 prospective reports with at least 20 participants at baseline through a search of databases (Medline and PsycInfo) and handsearching of articles published from 2002 to October 2007. If longitudinal data were not available or were scarce for a particular topic or time point, cross-sectional studies were reviewed. RESULTS: Although physical, psychological, and social aspects of quality of life tend to improve during the years following transplantation, a significant proportion of HSCT survivors experience persistent anxiety and depressive symptoms, fatigue, sexual dysfunction, and fertility concerns. Despite ongoing treatment side effects, the majority of HSCT survivors resume their work, school, or household activities. CONCLUSION: We conclude that theory-driven research with larger samples is needed to identify subgroups of HSCT survivors with adjustment difficulties. Such research would examine survivors' evolving standards and definitions of quality of life to improve the accuracy and meaningfulness of assessment and incorporate biological, psychological, and contextual factors that may contribute to positive adjustment. PMID- 18677720 TI - Oligosaccharide relative quantitation using isotope tagging and normal-phase liquid chromatography/mass spectrometry. AB - The growing interest in the conversion of plant biomass into biofuels has recently highlighted the lack of analytical techniques that are able to profile the fine structures of plant cell-wall polysaccharides. Here we present a new liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) platform called Oligosaccharide Quantitation using Isotope Tagging (OliQuIT) developed for profiling the oligosaccharides derived from glycosyl hydrolase digestion of polysaccharides. The method is demonstrated using different arabinoxylan-derived oligosaccharide samples, which are reductively aminated with either the light (12C6) or heavy (13C6) form of aniline. The complex oligosaccharide mixtures are analysed by capillary normal-phase (NP)-LC and ESI MS. Importantly, arabinoxylan oligosaccharide isomers are separated by NP-LC and their relative abundance in different samples can be determined from the intensities of ions labeled with the different isotopes. OliQuIT will be of use in multiple applications, including screening for plant varieties with improved saccharification properties, characterizing glycosyl hydrolase specificities and analysing plant glycosyl transferase mutants. PMID- 18677721 TI - Comparing welfare estimates from payment card contingent valuation and discrete choice experiments. AB - This study presents the first comparison of willingness to pay estimates derived from the payment card (PC) contingent valuation and discrete choice experiment (DCE) methods. A within-sample experiment was used to elicit women's preferences for Chlamydia screening. The willingness to pay estimate derived from the DCE was larger than that derived from the PC. To investigate why the willingness to pay estimates were different, a range of validity tests were conducted. Both methods produced theoretically valid results, and there was no difference in the reported difficulty of completing the tasks. Evidence of a prominence effect was found in the PC responses. Responses to the DCE satisfied tests of non-satiation. Responses to both methods were compared with revealed preference data. There were significant differences between stated screening intention in both methods and actual screening uptake. Future work should address the external validity of stated preference methods. PMID- 18677722 TI - Pecuniary compensation increases participation in screening for colorectal cancer. AB - The participation rate in medical screening programmes is typically below 100%, which means that not all potential health benefits are fully exploited. In this paper, the prospect of pecuniary compensation is tested as a method of increasing the participation rate. We propose a model explaining the individual's probability of participating in screening for colorectal cancer when he is offered pecuniary compensation, given that he did not participate when first invited. The participant's decision is based on both known and uncertain factors. The estimation is conducted in two steps, where a binary probit model is used in each. We find that pecuniary compensation increases the probability of participation, and that an individual's participation probability systematically varies with variables such as travel expenses, income, age, education level, expected benefit from the screening, use of health-care services, genetic predisposition and subjective health status. Using the results from the estimation, we predict changes in the participation rate for different levels of compensation and estimate the cost per additional individual screened. The cost per additional individual screened is 808, including 25 in compensation; this cost increases with the level of compensation. PMID- 18677723 TI - Weight and wages: fat versus lean paychecks. AB - Past empirical work has shown a negative relationship between the body mass index (BMI) and wages in most cases. We improve on this work by allowing the marginal effect of non-linear BMI groups to vary by gender, age, and type of interpersonal relationships required in each occupation. We use the National Longitudinal Survey of Youth 1979 (1982-1998). We find that the often-reported negative relationship between the BMI and wages is larger in occupations requiring interpersonal skills with presumably more social interactions. Also, the wage penalty increases as the respondents get older beyond their mid-twenties. We show that being overweight and obese penalizes the probability of employment across all race-gender subgroups except black women and men. Our results for the obesity wage association can be explained by either consumers or employers having distaste for obese workers. PMID- 18677724 TI - Transferability indices for health economic evaluations: methods and applications. AB - In this paper, we have elaborated an index in two phases to measure the degree of transferability of the results of the economic evaluation of health technologies. In the first phase, we have considered the objective factors (critical and non critical) to derive a general transferability index, which can be used to measure this internal property of the studies of economic evaluation applied to health technologies. In the second phase, with a more specific index, we have measured the degree of applicability of the results of a given study to a different setting. Both indices have been combined (arithmetic and geometric mean) to obtain a global transferability index. We have applied the global index to a sample of 27 Spanish studies on infectious diseases. We have obtained an average value for the index of 0.54, quite far from the maximum theoretical value of 1. We also found that 11 studies lacked some critical factor and were directly deemed as not transferable. PMID- 18677725 TI - Effects of the German reference drug program on ex-factory prices of prescription drugs: a panel data approach. AB - This paper examines effects of the German social health insurance system's reference drug program (RDP) for prescription drugs on ex-factory prices. Moreover, we analyze whether manufacturers adapt prices of their products that are not subject to reference pricing as a consequence of changes in reference prices of their products that are subject to reference pricing. We use econometric panel data methods based on a large panel data set of nearly all German prescription drugs on a monthly basis between October 1994 and July 2005. They provide information on ex-factory prices, reference prices, manufacturers, type of prescription drug, and market entries and exits. Our results show that there is no full price adjustment: A 1%-change in reference prices leads to a 0.3%-change in market prices. Price adjustment, however, is fast - it mostly happens in the first month. Furthermore, the first introduction of a reference price reduces market prices of the affected products by approximately 7%. Finally, we observe a significant time effect that is positive in the market without reference prices and negative in the market with reference prices. PMID- 18677726 TI - Sustained water oxidation photocatalysis by a bioinspired manganese cluster. PMID- 18677727 TI - Target-catalyzed transfer reactions for the amplified detection of RNA. PMID- 18677728 TI - Directed assembly of sub-nanometer thin organic materials with programmed-size nanopores. PMID- 18677729 TI - Anion-directed self-assembly of flexible ligands into anion-specific and highly symmetrical organic solids. PMID- 18677730 TI - Highly enantioselective direct Michael addition of nitroalkanes to nitroolefins catalyzed by La(OTf)3/N,N'-dioxide complexes. PMID- 18677731 TI - The role of bent acyclic allene gold complexes in axis-to-center chirality transfers. PMID- 18677732 TI - Unmasking representative structures of TMP-active Hauser and turbo-Hauser bases. PMID- 18677733 TI - Solution-state NMR spectroscopy of a seven-helix transmembrane protein receptor: backbone assignment, secondary structure, and dynamics. PMID- 18677734 TI - Gold-catalyzed waste-free generation and reaction of azomethine ylides: internal redox/dipolar cycloaddition cascade. PMID- 18677735 TI - Mechanism of binding of fluoroquinolones to the quinolone resistance-determining region of DNA gyrase: towards an understanding of the molecular basis of quinolone resistance. AB - We have studied the bacterial resistance to fluoroquinolones that arises as a result of mutations in the DNA gyrase target protein. Although it is known that DNA gyrase is a target of quinolone antibacterial agents, the molecular details of the quinolone-gyrase interaction remain unclear. The mode of binding of ciprofloxacin, levofloxacin, and moxifloxacin to DNA gyrase was analyzed by means of docking calculations over the surface of the QRDR of GyrA. The analysis of these binding models allows study of the resistance mechanism associated with gyrA mutations more commonly found in E. coli fluoroquinolone-resistant strains at the atomic level. Asp87 was found to be critical in the binding of these fluoroquinolones because it interacts with the positively charged nitrogens in these bactericidal drugs. The role of the other most common mutations at amino acid codon Ser83 can be explained through the contacts that the side chain of this residue establishes with fluoroquinolone molecules. Finally, our results strongly suggest that, although Arg121 has never been found to be associated with fluoroquinolone resistance, this residue makes a pivotal contribution to the binding of the antibiotic to GyrA and to defining its position in the QRDR of the enzyme. PMID- 18677736 TI - Split-ubiquitin and the split-protein sensors: chessman for the endgame. PMID- 18677737 TI - Enhanced glycosylation with mutants of endohexosaminidase A (endo A). PMID- 18677738 TI - Two lysine residues are responsible for the enzymatic activities of indole prenyltransferases from fungi. PMID- 18677739 TI - Peptide-grafted nanodiamonds: preparation, cytotoxicity and uptake in cells. AB - Nanodiamonds that were prepared by high pressure/high temperature were functionalized with biomolecules for biological applications. Nanodiamonds (NDs, < or =35 nm) that were coated by silanization or with polyelectrolyte layers were grafted with a fluorescent thiolated peptide via a maleimido function; this led to an aqueous colloidal suspension that was stable for months. These substituted NDs were not cytotoxic for CHO cells. Their capacity to enter mammalian cells, and their localisation inside were ascertained after labelling the nucleus and actin, by examining the cells by confocal, reflected light and fluorescence microscopy. PMID- 18677740 TI - Signal transducers and activators of transcription as targets for small organic molecules. AB - Signal transducers and activators of transcription (STATs) are a family of transcription factors that are of central importance for cellular signaling and have therefore emerged as attractive target proteins for cell-permeable small molecules. This review outlines the basic concept of STAT signaling, the relevance of individual members of the STAT family for cellular signaling and human disease, and generally applicable approaches taken to the identification of small-molecule inhibitors of STATs. PMID- 18677741 TI - Investigation into the mechanism of phenolic couplings during the biosynthesis of glycopeptide antibiotics. PMID- 18677745 TI - Patterns of structural complexity in Alzheimer's disease and frontotemporal dementia. AB - The goal of this project was to utilize an information theoretic formalism for medical image analysis initially proposed in [Young et al. (2005): Phys Rev Lett 94:098701-1] to detect and quantify subtle global and regional differences in spatial patterns in patients suffering from Alzheimer's disease (AD) and frontotemporal dementia (FTD) by estimating the structural complexity of anatomical brain MRI. The sensitivity and specificity of the results are compared with those of a recent analysis, currently considered state of the art for MR studies of neurodegeneration. The previous study used regional estimates of cortical thinning and/or volume loss to differentiate between normal aging, AD, and FTD. The analysis illustrates that the structural complexity estimation method, a general multivariate approach to the study of variation in brain structure which does not depend on highly specialized volumetric and thickness estimates, is capable of providing sensitive and interpretable diagnostic information. PMID- 18677746 TI - Neural networks involved in artistic creativity. AB - Creativity has been proposed to be either the result of solely right hemisphere processes or of interhemispheric interactions. Little information is available, however, concerning the neuronal foundations of creativity. In this study, we introduced a new artistic task, designing a new tool (a pen), which let us quantitatively evaluate creativity by three indices of originality. These scores were analyzed in combination with brain activities measured by functional magnetic resonance imaging (fMRI). The results were compared between subjects who had been formally trained in design (experts) and novice subjects. In the experts, creativity was quantitatively correlated with the degree of dominance of the right prefrontal cortex over that of the left, but not with that of the right or left prefrontal cortex alone. In contrast, in novice subjects, only a negative correlation with creativity was observed in the bilateral inferior parietal cortex. We introduced structure equation modeling to analyze the interactions among these four brain areas and originality indices. The results predicted that training exerts a direct effect on the left parietal cortex. Additionally, as a result of the indirect effects, the activity of the right prefrontal cortex was facilitated, and the left prefrontal and right parietal cortices were suppressed. Our results supported the hypothesis that training increases creativity via reorganized intercortical interactions. PMID- 18677747 TI - FISH analysis in cell touch preparations and cytological specimens from formalin fixed fetal autopsies. AB - Postmortem studies on still-borns and miscarriages are important to define the sex and eventually the morphologic anomalies correlated to chromosomal aberrations. When the conditions for carrying out a cytogenetic study do not exist, these chromosomal alterations can be investigated by nucleic acid fluorescent in situ hybridization (FISH), which can be performed on interphase nuclei, usually on formalin-fixed paraffin embedded tissues or on fresh cytological specimens. The objective of the present study is to prove whether this technique can be successfully applied to formalin-fixed cell touch preparations and cytological specimens obtained from foetal autopsies. The study was carried out 12 abortions some of which were spontaneous and some of which were therapeutic. The materials were formalin-fixed. Cell touch preparations and cytological specimens were obtained. The FISH was performed using X/Y probes (Vysis) and the Aneuvysion Kit (05J38-030, Vysis), the probes being for chromosomes 13/21 and X/Y/18. To verify the reliability of the technique, the same reactions were also performed on fresh analogous materials. The slides were evaluable, and the probes hybridized to interphase nuclei showed distinct signals. All the samples were adequate for FISH analysis without any notable difference in the results. Moreover, it is technically possible to perform this analysis not only on fresh but particularly on formalin-fixed cytological specimens. On the other hand, the use of this type of cytological samples, as compared to formalin-fixed and paraffin embedded tissue sections, has the advantage of presenting intact, noncut nuclei with preserved cytomorphology, avoiding the problems of overlapping nuclei and making the identification of the real chromosomal arrangement easier. PMID- 18677748 TI - Chromophobe renal cell carcinoma with calcification: Report of a case with rare finding on aspiration smears. AB - Chromophobe renal cell carcinoma (ChRCC) is a distinct variant of renal cell carcinoma. It is important to identify this entity because it has a significantly better prognosis than clear cell (conventional) renal cell carcinoma. We report the case of a 56-year-old male who presented with 10 days history of painless hematuria. There was no palpable swelling or systemic symptoms. On radiological examination the possibility of a right renal neoplasm was considered. Ultrasound guided aspiration was done and cytological features of ChRCC were noted. In addition, numerous foci of calcification were seen, which was an unusual cytological feature for ChRCC. The characteristic cytomorphology of ChRCC makes it possible to render a preoperative diagnosis on fine needle aspiration cytology (FNAC). We report the case to emphasize the importance and possibility of diagnosing of ChRCC on cytology. In addition, the extensive calcification as seen in our case is an unusual cytological finding not previously reported in cytology smears. PMID- 18677749 TI - Lymphoepithelial cysts of the pancreas: endosonography-guided fine needle aspiration. AB - Lymphoepithelial cyst (LEC) of the pancreas is a rare lesion. Here, we report three cases that we encountered during 2007 in our institution, which were evaluated by endosonography (EUS) and fine needle aspiration (FNA). All three patients were male with ages ranging from 47 to 77 years. Each patient had a single, cystic mass of the body/tail of the pancreas that was found incidentally on CT scan. The size of the masses ranged from 3 to 6.5 cm radiologically. For each patient, EUS showed a well-circumscribed, lobulated mass with smooth contours; the remainder of the pancreas appeared normal without ductal dilation. FNA of all three masses demonstrated predominantly anucleated squamous cells, amorphous keratinous debris, and lymphocytes. Cholesterol crystals were present in one case. One patient underwent surgical resection and the diagnosis of LEC was confirmed histologically. In conclusion, pancreatic LECs are extremely rare, benign cystic lesions. FNA study may help to render a correct diagnosis and reduce unnecessary surgery. PMID- 18677750 TI - Primary pulmonary Hodgkin's lymphoma: a rare pitfall in transthoracic fine needle aspiration cytology. AB - Primary pulmonary Hodgkin's lymphoma (PPHL) is extremely rare. At an extranodal location such as the lung this lymphoma is likely to be confused with the more commonly occurring carcinomas at this site. We report the fine needle aspiration cytology (FNAC) findings of a PPHL in a 36-year-old male with a view to discuss the pitfalls and clues to the accurate cytologic diagnosis. This patient presented with a large, heterogeneously enhancing mass involving the anterior segment of right upper lobe without any evidence of nodal involvement. A CT guided transthoracic FNAC of this mass revealed large connective tissue fragments with entrapped voluminous cells amidst a polymorphous population of eosinophils, polymorphs, and lymphocytes. The large cells showed abundant often stripped off cytoplasm, an irregular nucleus with nucleolus and were initially diagnosed as non-small cell carcinoma of the lung. In view of the locally advanced stage, patient received a carboplatin and gemcite-based chemotherapy with complete response but postchemotherapy patient refused local surgery. Two years later, the patient developed enlarged nodes which were diagnosed as Hodgkin's lymphoma, and a review of prior lung tumor confirmed the diagnosis of PPHL. Hence the rare diagnosis of PPHL should be kept in mind when a cytopathologist observes large cells embedded in collagenous tissue fragments with dominant cell dispersal amidst an inflammatory infiltrate in an aspirate from a primary lung tumor. PMID- 18677751 TI - Cryptococcal autofluorescence on fine needle aspiration cytology of lymph node. PMID- 18677752 TI - Significance of intercellular spaces (windows) in effusion fluid cytology: a study of 46 samples. AB - The presence and nature of intercellular windows were studied on 46 body cavity fluid samples chosen on the basis of an unequivocal diagnosis on May-Grunwald Giemsa (MGG)/Papanicolaou-stained smears and cell blocks. Of these, 24 cases had adenocarcinoma (AC) and seven had reactive mesothelium (RM) with 15 having distinct populations of both. Mucicarmine and PAS stains were used wherever indicated. The specificity of windows for predicting reactive mesothelium was evaluated. Intercellular windows were found in all cases of reactive mesothelium coinciding with the presence of the fuzzy peripheral microvillous borders. Surprisingly, as many as 17/39 (44%) of the adenocarcinomas also exhibited this feature, of which 13 had a distinctly visible evenly distributed ciliated cell membrane. In addition, 30/39 (77%) cases of AC exhibited a "window-like" appearance caused by cytoplasmic vacuolation. None of the adenocarcinoma clusters with true window formation showed positivity for the mucin stains, whereas all the clusters with pseudowindows caused by vacuolation were stained. Thus the specificity of intercellular windows for RM was merely 56%, though the sensitivity was 100%. On the other hand, the absence of windows was 100% specific for adenocarcinoma. Intercellular windows, though a feature of reactive mesothelial cell populations, can also be found in cases of ciliated adenocarcinomas and may not have a significant predictive value. PMID- 18677753 TI - Fine-needle aspiration cytology of a rheumatoid arthritis-associated interstitial lung disease. PMID- 18677754 TI - Pineal germinoma morphological features in a liquid-based cerebrospinal fluid sample. PMID- 18677755 TI - High-grade neuroendocrine carcinoma presenting as an abscess: diagnosis by fine needle aspiration and review of the literature. AB - Neuroendocrine carcinomas (NEC) comprise about 2% of all carcinomas. Review of the literature indicates only a few cases of NEC presenting as an abscess, all having been diagnosed by tissue biopsy. Here, we report the FNA diagnosis of a high-grade neuroendocrine carcinoma presenting as an abscess of the axilla/chest wall in a 68-year-old man and review the pertinent literature. PMID- 18677756 TI - Myxoid liposarcoma of the breast in a 25-year-old female as a diagnostic pitfall in fine needle aspiration cytology: report of a rare case. AB - Primary sarcomas of the breast are extremely rare comprising less than 1.0% of all malignant tumors of the breast. It is even rarer to be reported in a 25-year old female. This can cause a diagnostic dilemma not only for the clinician but also for the cytopathologist. A 25-year-old woman presented with a well defined firm, mobile lump in her right breast. With fine needle aspiration cytology (FNAC) and ultrasonography a diagnosis of fibroadenoma was made. The patient underwent lumpectomy. Histopathologically it was diagnosed as myxoid liposarcoma. This case is reported to highlight the fact that, even though rare in young females but the possibility of a breast lump being a myxoid liposarcoma does exist. This report discusses a primary myxoid liposarcoma of female breast, considers cytologic differential diagnoses with review of the relevant literature. PMID- 18677757 TI - Fine needle biopsy and genetics, two allied weapons in the diagnosis, prognosis, and target therapeutics of solid pediatric tumors. AB - The recognition that genetic defects identify some pediatric solid tumors and may represent prognostic markers has provided cytologists with an extra tool for dealing with such tumors. Using some entities as archetypes, we discuss the importance of the association of fine needle biopsy and genetics, in the diagnosis, prognosis, and therapy selection of solid pediatric tumors. Immunocytochemistry is important to differentiate neuroblastoma, PNET/Ewing sarcoma, alveolar rhabdomyosarcoma, lymphoma, and desmoplastic small round cell tumor. Despite its usefulness in many cases, it is not conclusive and some of the aforementioned tumors even share the expression of some antibodies. The detection of specific diagnostic translocations will thus provide additional information and allows a precise cytologic diagnosis. Kidney tumors are also frequent in children. Although no genetic abnormalities have been identified so far in nephroblastoma, other kidney tumors, such as mesoblastic nephroma, whose cytology pattern can masquerade nephroblastoma, are also characterized by specific translocations. Kidney tumors in children have also been associated recently with typical genetic alterations such as Xp11.2RCC. Concerning prognosis and therapy selection, neuroblastoma is a sort of paradigm. The identification of MYCN oncogene status as an independent prognostic factor is determinant, not only in the assessment of clinical evolution, but also in the identification of risk groups, and consequently in the appropriate therapy selection. Cytopathologists should be aware of the genetic alterations characterizing pediatric tumors in order to collect extra material to perform cytogenetics, FISH, PCR, and Southern blotting, to achieve the correct identification of such genetic changes. PMID- 18677758 TI - Differential diagnostic features of small cell carcinoma in the uterine cervix. AB - Small cell carcinoma (SMCC) of the uterine cervix is rare and known to be an aggressive tumor, but there are only few reports on the cytologic features of cervical SMCC. This rare small cell lesion should be distinguished from malignant lymphoma (ML), squamous cell carcinoma in situ (SCIS), and chronic lymphocytic cervicitis (CLC). By clarifying cytologic features and reevaluating the significance of cervical cytologic smears to reveal these cervical lesions, we can improve the diagnostic specificity and patient's outcome. The clinical record and available cervical smears from 13 cases of SMCC, four cases of malignant lymphoma, 20 cases of SCIS, and five cases of CLC were analyzed. The cytologic differential diagnostic points of SMCC were nuclear molding and smearing (100%), salt and pepper chromatin (100%), exudative and necrotic background (91.7%), various architectures including individual cells (83.3%), tight clusters (75%) and feathering and strip (50%), and inconspicuous nucleoli (75%). Early diagnosis of the cervical SMCC by cytology and treatment is important for better outcome of patients. PMID- 18677759 TI - Aspiration biopsy cytology of extraabdominal desmoid tumor concurrently occurring in a patient with tumoral calcinosis. AB - Extraabdominal fibromatosis or desmoid tumor (DT) is a slow growing locally aggressive soft tissue tumor that can occur anywhere in the body. We report the aspiration biopsy cytology features of a case of DT of the right neck area in a 35-year-old man who had a long standing history of tumoral calcinosis. The aspirate was interpreted as "benign spindle cell lesion" and confirmed as DT on histologic examination of the resected mass. We discuss the possible differential diagnoses of other benign or malignant lesions on fine-needle aspiration (FNA) biopsy and especially discuss the aspiration cytology features of DT compared with those of tumoral calcinosis. We also discuss the value of immunohistochemical markers that help in differentiating DT from other entities. PMID- 18677760 TI - Ovarian carcinoma cells in effusions show increased S-phase fraction compared to corresponding primary tumors. AB - The objective of this study was to analyze large-scale genomic patterns during disease progression from primary tumor to effusion in ovarian carcinoma, and to study the association between DNA ploidy parameters in effusions, proliferation/survival markers, and clinicopathologic characteristics. DNA ploidy status, DNA index (DI), and S-phase fraction (SPF) were compared in 22 matched primary carcinomas (all prechemotherapy specimens) and effusions (14 prechemotherapy and 8 postchemotherapy specimens) using image analysis. The association between these parameters and previously studied cell survival/proliferation biomarkers, previous administration of chemotherapy, chemotherapy response and survival was analyzed in a larger series of 54 effusions. The majority of specimens were aneuploid irrespective of anatomic site, with no significant differences in DI. SPF was significantly higher in effusions compared to matched primary tumors (P = 0.007 for all 22 pairs, P = 0.011 for 14 matched prechemotherapy specimens). Higher SPF was related to higher Ki-67 score (P = 0.045), and both SPF and DI were directly associated with higher level of Survivin (P < 0.001 for both). DI and SPF in effusions showed no association with histological grade, FIGO stage, residual disease volume, previous chemotherapy, response to chemotherapy at primary disease, recurrence or survival. Ovarian carcinoma cells in effusions have increased proliferation compared to corresponding primary tumors, as evidence of disease progression. DNA ploidy parameters in cancer cells in effusions are unaltered by chemotherapy and appear to be unrelated to chemotherapy response and to survival, suggesting that large-scale genomic patterns at this anatomic site are not useful in segregating patients into prognostic groups. PMID- 18677761 TI - Use of the ThinPrep method in bile duct brushings: analysis of morphologic parameters associated with malignancy and determination of interobserver reliability. AB - Recent work suggests the ThinPrep method can improve diagnostic sensitivity and accuracy in bile duct brushings. However, the proportion of atypical and suspicious diagnoses remains high. The aim of this study was to identify the most useful morphologic features in ThinPrep bile duct cytology and evaluate interobserver reliability. We evaluated 100 bile duct brushings prepared by ThinPrep, all with either histology or long term clinical follow-up (55 malignant, 45 benign). Morphologic features were evaluated by four experienced cytopathologists blind to clinical information and follow-up diagnoses. These features included cellularity, blood or diathesis, mitoses, inflammation, three dimensional groups, discohesive atypical cells, macronucleoli, well-defined cytoplasmic borders, and nuclear features of malignancy (nuclear membrance irregularity, chromatin clumping). The data were analyzed by intraclass correlation (ICC) and stepwise multiple logistic regression. Reviewers showed unanimous agreement in 29% of cases, one degree of disagreement in 58% of cases, and full disagreement in 13% of cases. Of benign cases, 38% were thought to be diagnostic of malignancy by at least one of the four reviewers. Sensitivity for the morphologic parameters varied from 18 to 67%; the highest sensitivity was for discohesive atypical cells, well-defined cytoplasmic borders, nuclear features of malignancy, and cellularity (67, 62, 51 and 46%, respectively). Specificity of parameters varied from 16 to 100%; the highest specificity was for mitoses, three dimensional groups, nuclear features of malignancy, and macronucleoli (100, 98, 93, and 93%, respectively). Interobserver reliability (ICC) was very good for specimen cellularity (0.72) and nuclear features of malignancy (0.60). In logistic regression analysis, only nuclear features of malignancy and increasing patient age separated benign from malignant. On ThinPrep bile duct brushings, nuclear features of malignancy are most useful in distinguishing benign from malignant, and interobserver reliability for this parameter is very good. Discohesive atypical cells show moderate sensitivity and specificity, while three dimensional clusters and macronucleoli are specific but not sensitive for malignancy, and are not significant in multivariate logistic regression models. The relatively high proportion of benign cases thought to be diagnostic of malignancy by at least one reviewer argues for a consensus approach to this diagnosis. PMID- 18677762 TI - Pleomorphic lobular carcinoma in pleural fluid: diagnostic pitfall for atypical mesothelial cells. AB - Pleomorphic lobular carcinoma (PLC) is a subtype of infiltrating lobular carcinoma because of its dyscohesiveness, linear infiltration pattern, and lack of membranous E-cadherin staining. However, it differs from classic lobular carcinoma because of its high-grade cytology and more aggressive clinical behavior. In breast fine-needle aspiration biopsies, PLC can be confused with invasive ductal carcinoma, particularly the apocrine variant. In this report, we illustrate how metastatic PLC in body fluid specimens shows many of the same cytomorphologic changes that occur in reactive/atypical mesothelial cells. Fortunately, the immunohistochemical staining pattern of PLC can help to distinguish it from other possible diagnoses in the differential, such as reactive/atypical mesothelial cells and other metastatic neoplasms. However, the frequent apocrine features seen in this variant of breast carcinoma can cause nonspecific immunohistochemical positivity that may make the interpretation difficult. This is the first report illustrating the cytopathology and immunohistochemical findings of pleomorphic lobular carcinoma in body cavity fluid cytology. Our case highlights the important issues and pitfalls to be aware of when making this diagnosis. PMID- 18677763 TI - Retraction halos: nature's dotting pen in effusion cytology processed by ultrafast Papanicolaou stain. AB - Effusion cytology is challenging when solitary adenocarcinoma cells resemble histiocytes or mesothelial cells, and when adenocarcinomas look like histiocytic hyperplasia. It is also challenging to locate rare tumor cells in markedly inflammatory background. Over a period of 9 years, 9,474 cases of body cavity fluids including 3,642 cases of pleural, 1,900 cases of peritoneal and 345 cases of pericardial effusions and 3,587 cases of pelvic washes were processed with Ultrafast Papanicolaou stain. Retraction halos around spherical solitary cells and tissue fragments were observed whenever the background was inflammatory, whereas absent around histiocytes or histiocytic aggregates, and inconspicuous around solitary mesothelial cells. Retraction halos were helpful when the tumor fragments are obscured by marked inflammation, the quantity of tumor fragments is sparse, or the tumor cells resemble histiocytes or mesothelial cells. In short, retraction halos are nature's dotting pen for effusion cytology and are particularly useful in challenging cases. PMID- 18677764 TI - Renal late effects in patients treated for cancer in childhood: a report from the Children's Oncology Group. AB - Improvements in childhood cancer therapy have led to increasing numbers of long term survivors. These survivors are at risk for a variety of late effects due to the disease itself, treatment exposures (surgery, chemotherapy, and radiotherapy), underlying medical problems, and health behaviors. The COG LTFU Guidelines are risk-based, exposure-related recommendations for the identification and management of late effects due to therapies utilized in the treatment of childhood cancer, and are designed for asymptomatic survivors presenting for routine medical follow-up 2 or more years after completion of cancer therapy. The COG Guidelines Task Force on Urinary Tract Complications conducted an extensive review of the medical literature via MEDLINE. Specific treatment exposures which were reviewed include nephrectomy, chemotherapy regimens known to be nephrotoxic (cisplatin, carboplatin, ifosfamide, and methotrexate), and renal irradiation. Literature sources were ranked according to the strength of evidence and are cited in the review. This review summarizes the literature that supported the recommendations for cancer survivors at risk for nephrotoxicity previously outlined in the Children's Oncology Group Long-Term Follow-Up Guidelines for Survivors of Childhood, Adolescent and Young Adult Cancers (COG LTFU Guidelines). PMID- 18677766 TI - Mass spectrometric characterization of naphthenic acids in environmental samples: a review. AB - There is a growing need to develop mass spectrometric methods for the characterization of oil sands naphthenic acids (structural formulae described by C(n)H(2n+z)O(2) where n is the number of carbon atoms and "z" is referred to as the "hydrogen deficiency" and is equal to zero, or is a negative, even integer) present in environmental samples. This interest stems from the need to better understand their contribution to the total acid number of oil sands acids; along with assessing their toxicity in aquatic environments. Negative-ion electrospray ionization has emerged as the analytical technique of choice. For infusion samples, matrix effects are particularly evident for quantification in the presence of salts and co-elutants. However, such effects can be minimized for methods that employ chromatographic separation prior to mass spectrometry (MS) detection. There have been several advances for accurate identification of classes of naphthenic acid components that employ a range of MS hyphenated techniques. General trends measured for degradation of the NAs in the environment appear to be similar to those obtained with either low- or high-resolution MS. Future MS research will likely focus on (i) development of more reliable quantitative methods that use chromatography and internal standards, (ii) the utility of representative model naphthenic acids as surrogates for the complex NA mixtures, and (iii) development of congener-specific analysis of the principal toxic components. PMID- 18677765 TI - A novel G6PD mutation leading to chronic hemolytic anemia. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an important cause of hemolytic anemia worldwide. Severely affected patients have chronic hemolysis with exacerbations following oxidative stress. Mutations causing severe chronic non-spherocytic hemolytic anemia (CNSHA) commonly cluster in Exon 10, a region important for protein dimerization. An African-American male presented at age 2 weeks with pallor and jaundice, and was found to have hemolytic anemia with G6PD deficiency. His severe clinical course was inconsistent with the expected G6PD A( ) variant. DNA sequencing revealed two common mutations (A(-)) and a third novel Exon 10 mutation. This inherited haplotype represents a novel triple G6PD coding mutation causing chronic hemolysis. PMID- 18677767 TI - Bioengineering and Imaging Research Opportunities Workshop V: a summary on Imaging and Characterizing Structure and Function in Native and Engineered Tissues. AB - The Fifth Bioengineering and Imaging Research Opportunities Workshop (BIROW V) was held on January 18-19, 2008. As with previous BIROW meetings, the purpose of BIROW V was to identify and characterize research and engineering opportunities in biomedical engineering and imaging. The topic of this BIROW meeting was Imaging and Characterizing Structure and Function in Native and Engineered Tissues. Under this topic, four areas were explored in depth:1) Heterogeneous single-cell measurements and their integration into tissue and organism models;2) Functional, molecular, and structural imaging of engineered tissue in vitro and in vivo;3) New technologies for characterizing cells and tissues in situ;4) Imaging for targeted cell, gene, and drug delivery. PMID- 18677768 TI - In vivo multispectral, multiparameter, photoacoustic lymph flow cytometry with natural cell focusing, label-free detection and multicolor nanoparticle probes. AB - Compared with blood tests, cell assessment in lymphatics is not well-established. The goal of this work was to develop in vivo lymph tests using the principles of flow cytometry. Cells in living animals were counted by laser (420-2,300 nm) generation of photoacoustic (PA) signals in individual cells hydrodynamically focused by lymph valves into a single file flow, and using endogenous absorption as intrinsic cell-specific markers, or gold nanorods, nanoshells, and carbon nanotubes as multicolor probes. PA data were verified by high-speed transmission, photothermal, and fluorescent imaging. Counting of melanoma and immune-related cells in normal, apoptotic, and necrotic states in lymphatics in vivo was demonstrated to have the unprecedented sensitivity as one metastatic cell among millions of white blood cells. The time-resolved PA spectral identification of flowing cells was achieved using multicolor labels and laser pulses of different wavelengths and time delays. Multiparameter, noninvasive, portable flow cytometer can be used for preclinical studies on animals with the potential of translation to humans for in vivo PA mapping of colorless lymph vessels and sentinel nodes with simultaneous single cell detection and metastasis assessment without labeling or use of contrast dyes and/or novel low-toxic multicolor probes with different absorption spectra. PMID- 18677769 TI - Electrostatic interactions between hyaluronan and proteins at pH 4: how do they modulate hyaluronidase activity. AB - Hyaluronan (HA) hydrolysis catalyzed by hyaluronidase (HAase) is inhibited at low HAase over HA ratio and low ionic strength, because HA forms electrostatic complexes with HAase, which is unable to catalyze hydrolysis. Bovine serum albumin (BSA) was used as a model to study the HA-protein electrostatic complexes at pH 4. At low ionic strength, there is formation of (i) neutral insoluble complexes at the phase separation and (ii) small positively-charged or large negatively-charged soluble complexes whether BSA or HA is in excess. According to the ionic strength, different types of complex are formed. Assays for HA and BSA led to the determination of the stoichiometry of these complexes. HAase was also shown to form the various types of complex with HA at low ionic strength. Finally, we showed that at 0 and 150 mmol L(-1) NaCl, BSA competes with HAase in forming complexes with HA and thus induces HAase release resulting in a large increase in the hydrolysis rate. These results, in addition to data in the literature, show that HA-protein complexes, which can exist under numerous and varied conditions of pH, ionic strength and protein over HA ratio, might control the in vivo HAase activity. PMID- 18677770 TI - Identification of ARHGEF17, DENND2D, FGFR3, and RB1 mutations in melanoma by inhibition of nonsense-mediated mRNA decay. AB - Gene identification by nonsense-mediated mRNA decay inhibition (GINI) has proven to be a strategy for genome-wide discovery of genes containing inactivating mutations in colon and prostate cancers. Here, we present the first study of inhibition of the nonsense-mediated mRNA decay (NMD) pathway in melanoma. We used a combination of emetine and actinomycin D treatment to stabilize mRNAs containing premature termination codons (PTCs), followed by microarray analysis and sequencing to identify novel tumor suppressor genes (TSGs) in a panel of 12 melanoma cell lines. Stringent analysis of the array data was used to select 35 candidate genes for sequencing. Of these, 4 (11%) were found to carry PTCs, including ARHGEF17, DENND2D, FGFR3, and RB1. While RB1 mutations have previously been described in melanoma, the other three genes represent potentially novel melanoma; TSGs. ARHGEF17 showed a G1865A mutation leading to W622X in a cell line derived from a mucosal melanoma; in RB1 a C1411T base change resulting in Q471X was discovered in a cell line derived from an acral melanoma; and the FGFR3 and DENND2D genes had intronic insertions leading to PTCs in cell lines derived from superficially spreading melanomas. We conclude that although the false positive rate is high, most likely due to the lack of DNA mismatch repair gene defects, the GINI protocol is one approach to discover novel TSGs in melanoma. PMID- 18677772 TI - Ectopic expression of LIM-nebulette (LASP2) reveals roles in cell migration and spreading. AB - LIM-nebulette (LASP2) is a small focal adhesion protein and a member of the nebulin family of actin binding proteins. This recently identified splice variant of the nebulette locus is widely expressed and highly enriched in neuronal tissue. Other than that LIM-nebulette is a focal adhesion protein and interacts with zyxin, nothing is known about its function. Given that LIM-nebulette has an identical modular organization and overlapping tissue distributions to that of LASP1, we have analyzed the role of LIM-nebulette in comparison with that of LASP1. We find that LIM-nebulette is a dynamic focal adhesion protein that increases the rate of attachment and spreading of fibroblasts on fibronectin coated surfaces. Additionally, LIM-nebulette is recruited from the cortical cytoskeleton in non-motile cells to focal adhesions at the leading edge of stimulated cells. In confluent cultures of HeLa and NIH3T3 cells, LIM-nebulette co-localizes with alpha-catenin in putative adherens junctions, whereas LASP1 is devoid of these areas. Interestingly, overexpression of LIM-nebulette in PC6 cells inhibits neurite outgrowth in response to growth factors. Collectively, our data indicate that LIM-nebulette and LASP1 have distinct roles in the actin cytoskeleton. PMID- 18677773 TI - Subcellular curvature at the perimeter of micropatterned cells influences lamellipodial distribution and cell polarity. AB - This paper employs substrates that are patterned with shapes having well-defined geometric cues to characterize the influence of curvature on the polarization of highly metastatic B16F10 rat melanoma cells. Substrates were patterned using microcontact printing to define adhesive islands of defined shape and size on a background that otherwise prevents cell adhesion. Cells adherent to these surfaces responded to local curvature at the perimeter of the adhesive islands; convex features promoted the assembly of lamellipodia and concave features promoted the assembly of stress filaments. Cells adherent to rectangular shapes displayed a polarized cytoskeleton that increased with the aspect ratio of the shapes. Shapes that combined local geometric cues, by way of concave or convex edges, with aspect ratio were used to understand the additive effects of shape on polarization. The dependence of cell polarity on shape was determined in the presence of small molecules that alter actomyosin contractility and revealed a stronger dependence on contractility for shapes having straight edges, in contrast to those having curved edges. This study demonstrates that the cytoskeleton modulates cell polarity in response to multiple geometric cues in the extracellular environment. PMID- 18677774 TI - Somatostatin analogues for acute bleeding oesophageal varices. AB - BACKGROUND: Somatostatin and its derivatives are sometimes used for emergency treatment of bleeding oesophageal varices in patients with cirrhosis of the liver. OBJECTIVES: To study whether somatostatin or its analogues improve survival or reduce the need for blood transfusions in patients with bleeding oesophageal varices. SEARCH STRATEGY: PubMed and The Cochrane Library were searched (November 2007). Reference lists of publications, contacts with authors. SELECTION CRITERIA: All randomised trials comparing somatostatin or analogues with placebo or no treatment in patients suspected of acute or recent bleeding from oesophageal varices. DATA COLLECTION AND ANALYSIS: The outcome measures extracted were: mortality, blood transfusions, use of balloon tamponade, initial haemostasis and rebleeding. Intention-to-treat analyses including all randomised patients were conducted if possible; a random-effects analysis was preferred if there was significant heterogeneity between the trials (P < 0.10). The trials were divided in two groups: trials with a low risk of bias, which had concealed allocation of patients and were double-blind, and other trials. MAIN RESULTS: We included 21 trials (2588 patients). The drugs did not reduce mortality significantly (relative risk 0.97, 95% confidence interval (CI) 0.75 to 1.25, for the trials with a low risk of bias, and 0.80, 95% CI 0.63 to 1.01, for the other trials). Units of blood transfused were 0.7 (0.2 to 1.1) less with drugs in the trials with a low risk of bias and 1.5 (0.9 to 2.0) less in the other trials. Number of patients failing initial haemostasis was reduced, relative risk 0.68 (0.54 to 0.87). Number of patients with rebleeding was not significantly reduced for the trials with a low risk of bias, relative risk 0.84 (0.52 to 1.37) while it was substantially reduced in the other trials, relative risk 0.36 (0.19 to 0.68). Use of balloon tamponade was rarely reported. AUTHORS' CONCLUSIONS: The need for blood transfusions corresponded to one half unit of blood saved per patient. It is doubtful whether this effect is worthwhile. The findings do not suggest a need for further placebo-controlled trials of the type reviewed here. A large placebo controlled trial enrolling thousands of patients is needed if one wishes to rule out the possibility that a worthwhile effect on mortality might have been overlooked. PMID- 18677775 TI - Ursodeoxycholic acid for primary biliary cirrhosis. AB - BACKGROUND: Primary biliary cirrhosis is an uncommon autoimmune liver disease with unknown aetiology. Ursodeoxycholic acid (UDCA) has been used for primary biliary cirrhosis, but the effects remain controversial. OBJECTIVES: To evaluate the benefits and harms of UDCA on patients with primary biliary cirrhosis against placebo or no intervention. SEARCH STRATEGY: We searched The Cochrane Hepato Biliary Group Controlled Trials Register, The Cochrane Central Register of Controlled Trials on The Cochrane Library, MEDLINE, EMBASE, SCI-EXPANDED, The Chinese Biomedical CD Database, LILACS, and the references of identified studies. The last search was performed in January 2007. SELECTION CRITERIA: Randomised clinical trials evaluating UDCA versus placebo or no intervention in patients with primary biliary cirrhosis. DATA COLLECTION AND ANALYSIS: The primary outcomes were mortality and mortality or liver transplantation. Binary outcomes were reported as odds ratio (OR) or relative risk (RR) and continuous outcomes as weighted mean difference, all with 95% confidence intervals (CI). Meta-regression was used to investigate the associations between UDCA effects and quality of the trial, UDCA dose, trial duration, and patient's severity of primary biliary cirrhosis. We also used Bayesian meta-analytic approach to estimate the UDCA effect as sensitivity analysis. MAIN RESULTS: Sixteen randomised clinical trials evaluating UDCA against placebo or no intervention were identified. Data from three trials have been updated. Nearly half of the trials had high risk of bias. The combined results demonstrated no significant effects favouring UDCA on mortality (OR 0.97, 95% CI 0.67 to 1.42) and mortality or liver transplantation (RR 0.92, 95% CI 0.71 to 1.21). The findings were supported by the Bayesian meta analyses. UDCA did not improve pruritus, fatigue, autoimmune conditions, liver histology, or portal pressure. UDCA seemed to improve biochemical variables, like serum bilirubin, ascites, and jaundice, but the findings were based on few trials with sparse data. The use of UDCA is significantly associated with adverse events, mainly weight gain. AUTHORS' CONCLUSIONS: This systematic review did not demonstrate any benefit of UDCA on mortality and mortality or liver transplantation of patients with primary biliary cirrhosis. The few beneficial effects could not be due to random errors or outcome reporting bias. PMID- 18677776 TI - Behavioural and cognitive-behavioural interventions for outwardly-directed aggressive behaviour in people with learning disabilities. AB - BACKGROUND: Outwardly directed aggressive behaviour is a significant part of problem behaviours presented by people with learning disabilities. Prevalence rates between 3.3% to 36% have been reported in the literature. Such behaviours often run a long term course and are a major cause of social exclusion. OBJECTIVES: To determine the efficacy of behavioural and cognitive behavioural interventions for outwardly-directed aggressive behaviour for people with learning disabilities. SEARCH STRATEGY: The Cochrane Library (CENTRAL) 2007 (Issue 1), MEDLINE 1966 to February 2007, EMBASE 1980 to February 2007, PsycINFO 1872 to February 2007 and Dissertation Abstracts late 1960s to February 2007 were searched. Where appropriate, research filters were used. SELECTION CRITERIA: Studies were selected if more than four participants, children or adults, were allocated by random or quasi-random methods to either intervention or standard treatment/wait list. DATA COLLECTION AND ANALYSIS: References identified by electronic searches, examinations of bibliography and personal contacts were screened against inclusion criteria by two independent reviewers. Two reviewers independently extracted and entered data into RevMan (Cochrane Collaboration software). MAIN RESULTS: Four studies based on adult populations with learning disabilities were deemed to be suitable for inclusion in the current version of this review. Data were only available in a form suitable for meta-analysis in three studies, but due to heterogeneity of populations and interventions, meta analysis was not performed. Direct interventions based on cognitive-behavioural methods (modified relaxation, assertiveness training with problem solving, and anger management) appear to have some impact on reduction of aggressive behaviour at the end of treatment and in some studies also at follow up (up to six months). AUTHORS' CONCLUSIONS: The existing evidence on the efficacy of cognitive behavioural and behavioural interventions on outwardly directed aggression in children and adults with learning disabilities is scant. There is a paucity of methodologically sound clinical trials. Given the impact of such behaviours on the affected individual, his or her carers and on service providers, effective interventions are essential. It is also important to investigate cost efficacy of treatment models against existing treatments. We recommend that randomised controlled trials of sufficient power are carried out using primary outcomes of reduction in outward directed aggression, improvement in quality of life and cost efficacy as measured by standardised scales. PMID- 18677777 TI - Antioxidant supplements for preventing gastrointestinal cancers. AB - BACKGROUND: Oxidative stress may cause gastrointestinal cancers. The evidence on whether antioxidant supplements are effective in preventing gastrointestinal cancers is contradictory. OBJECTIVES: To assess the beneficial and harmful effects of antioxidant supplements in preventing gastrointestinal cancers. SEARCH STRATEGY: We identified trials through the trials registers of the four Cochrane Review Groups on gastrointestinal diseases, The Cochrane Central Register of Controlled Trials in The Cochrane Library (Issue 2, 2007), MEDLINE, EMBASE, LILACS, SCI-EXPANDED, and The Chinese Biomedical Database from inception to October 2007. We scanned reference lists and contacted pharmaceutical companies. SELECTION CRITERIA: Randomised trials comparing antioxidant supplements to placebo/no intervention examining occurrence of gastrointestinal cancers. DATA COLLECTION AND ANALYSIS: Two authors (GB and DN) independently selected trials for inclusion and extracted data. Outcome measures were gastrointestinal cancers, overall mortality, and adverse effects. Outcomes were reported as relative risks (RR) with 95% confidence interval (CI) based on random-effects and fixed-effect model meta-analysis. Meta-regression assessed the effect of covariates across the trials. MAIN RESULTS: We identified 20 randomised trials (211,818 participants), assessing beta-carotene (12 trials), vitamin A (4 trials), vitamin C (8 trials), vitamin E (10 trials), and selenium (9 trials). Trials quality was generally high. Heterogeneity was low to moderate. Antioxidant supplements were without significant effects on gastrointestinal cancers (RR 0.94, 95% CI 0.83 to 1.06). However, there was significant heterogeneity (I(2) = 54.0%, P = 0.003). The heterogeneity may have been explained by bias risk (low-bias risk trials RR 1.04, 95% CI 0.96 to 1.13 compared to high-bias risk trials RR 0.59, 95% CI 0.43 to 0.80; test of interaction P < 0.0005), and type of antioxidant supplement (beta carotene potentially increasing and selenium potentially decreasing cancer risk). The antioxidant supplements had no significant effects on mortality in a random effects model meta-analysis (RR 1.02, 95% CI 0.97 to 1.07, I(2) = 53.5%), but significantly increased mortality in a fixed-effect model meta-analysis (RR 1.04, 95% CI 1.02 to 1.07). Beta-carotene in combination with vitamin A (RR 1.16, 95% CI 1.09 to 1.23) and vitamin E (RR 1.06, 95% CI 1.02 to 1.11) significantly increased mortality. Increased yellowing of the skin and belching were non serious adverse effects of beta-carotene. In five trials (four with high risk of bias), selenium seemed to show significant beneficial effect on gastrointestinal cancer occurrence (RR 0.59, 95% CI 0.46 to 0.75, I(2) = 0%). AUTHORS' CONCLUSIONS: We could not find convincing evidence that antioxidant supplements prevent gastrointestinal cancers. On the contrary, antioxidant supplements seem to increase overall mortality. The potential cancer preventive effect of selenium should be tested in adequately conducted randomised trials. PMID- 18677778 TI - Vitamin K for upper gastrointestinal bleeding in patients with liver diseases. AB - BACKGROUND: Upper gastrointestinal bleeding is one of the most frequent causes of morbidity and mortality in the course of liver cirrhosis. Several treatments are used for upper gastrointestinal bleeding in patients with liver diseases. One of them is vitamin K administration, but it is unknown whether it benefits or harms patients with liver disease and upper gastrointestinal bleeding. OBJECTIVES: To assess the beneficial and harmful effects of vitamin K for patients with liver disease and upper gastrointestinal bleeding. SEARCH STRATEGY: We searched The Cochrane Hepato-Biliary Group Controlled Trials Register (September 2007), which comprises references identified from comprehensive electronic database searches and handsearching of relevant journals and abstract books of conference proceedings, The Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library (Issue 3, 2007), MEDLINE (1950 to September 2007), EMBASE (1980 to September 2007), Science Citation Index Expanded (1945 to September 2007), and LILACS (1982 to November 2007). Additional randomised trials were sought from two registries of clinical trials, ClinicalTrials.gov and Sistema de Informacion Esencial en Terapeutica y Salud, the reference lists of the trials found, and reviews identified by the electronic searches. SELECTION CRITERIA: Randomised clinical trials. DATA COLLECTION AND ANALYSIS: Data from randomised clinical trials were to be summarised by standard Cochrane Collaboration methodologies. MAIN RESULTS: We could not find any randomised trials on vitamin K for upper gastrointestinal bleeding in patients with liver diseases. AUTHORS' CONCLUSIONS: This updated review found no randomised clinical trials on the safety and efficacy of vitamin K for upper gastrointestinal bleeding in patients with liver diseases. The effects of vitamin K need to be tested in randomised clinical trials. Unless randomised clinical trials provide evidence of a treatment effect and the trade off between potential benefits and harms are established, policy makers, clinicians, and academics should not use vitamin K for upper gastrointestinal bleeding in patients with liver diseases. PMID- 18677779 TI - Preoperative biliary drainage for obstructive jaundice. AB - BACKGROUND: Obstruction in the biliary drainage system causes a rise in serum bilirubin levels (obstructive jaundice). Studies have shown that surgery for severe obstructive jaundice is associated with high peri-operative mortality and morbidity. Jaundice has been considered as a potential risk factor for poor outcome, and pre-operative biliary drainage has been proposed as a method of reversing the pathophysiologic disturbance seen in patients with obstructive jaundice. OBJECTIVES: To determine the benefits and harms of pre-operative biliary drainage (ie, endoscopic sphincterotomy with stent insertion or percutaneous transhepatic biliary drainage) in obstructive jaundice. SEARCH STRATEGY: We searched The Cochrane Hepato-Biliary Group Controlled Trials Register (CHBG), the Cochrane Central Register of Controlled Clinical Trials (CENTRAL) in The Cochrane Library, MEDLINE, EMBASE, the Chinese BioMedical Literature on disc (CBM disc), and the Chinese Medical Current Contents (CMCC). All databases were searched up to October 2006. SELECTION CRITERIA: We considered for inclusion randomised clinical trials comparing biliary drainage followed by surgery and direct surgery performed for obstructive jaundice. DATA COLLECTION AND ANALYSIS: We collected the available data on the characteristics of the trial, methodological quality of the trials, mortality, morbidity, and hospital stay as reported in each trial. We analysed the data with both the fixed-effect and the random-effects models, using RevMan Analysis. For each outcome, we calculated the odds ratio (OR) with 95% confidence intervals (CI) based on intention-to-treat analysis. MAIN RESULTS: Five trials with 320 patients (160 in each group) were included. Four trials (n = 235) compared percutaneous transhepatic biliary drainage with direct surgery, and one trial (n = 85) compared pre-operative endoscopic drainage with direct surgery. All trials were of low methodological quality. There was no significant difference in mortality (OR 1.14, 95% CI 0.60 to 2.10) between the pre-operative biliary drainage group and the direct surgery group. No significant difference was found in mortality (OR 1.16, 95% CI 0.56 to 2.41), overall morbidity (OR 1.35, 95%CI 0.48 to 3.83), and in different complications between the percutaneous transhepatic biliary drainage group and the direct surgery group. The trial comparing pre-operative endoscopic drainage and direct surgery showed no significant difference in mortality (OR 1.09, 95% CI 0.32 to 3.68), but found higher morbidity in the endoscopic drainage group. The overall hospital stay was 8 to 17 days shorter in the direct surgery group. AUTHORS' CONCLUSIONS: Our analyses neither supports nor refutes pre-operative biliary drainage for patients with obstructive jaundice needing surgery. In some specific lesion site it may cause more complications. Pre-operative biliary drainage also prolonged hospital stay and increased cost. However, the strength of evidence is low because of the poor quality of the included trials. More rigorously designed randomised clinical trials with larger sample size and advanced techniques and drugs are needed. PMID- 18677780 TI - Hepatitis B immunisation in persons not previously exposed to hepatitis B or with unknown exposure status. AB - BACKGROUND: The benefits and harms of hepatitis B vaccination in persons not previously exposed to hepatitis B infection or with unknown exposure status have not been established. OBJECTIVES: To assess the benefits and harms of hepatitis B vaccination in people not previously exposed to hepatitis B infection or with unknown exposure status. SEARCH STRATEGY: Trials were identified from The Cochrane Hepato-Biliary Group Controlled Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, LILACS,Science Citation Index Expanded (last search, March 2007). Additionally, we contacted experts and vaccine manufacturers, and read through reference lists for eligible trials. SELECTION CRITERIA: Randomised clinical trials comparing hepatitis B vaccine versus placebo, no intervention, or another vaccine in persons not previously exposed to hepatitis B (HBsAg negative) or with unknown exposure status. DATA COLLECTION AND ANALYSIS: The primary outcome was hepatitis B infection (detecting HBsAg, HBeAg, HBV DNA, or anti-HBc). Secondary outcomes were lack of sero protection, antibody titre, clinical complications, adverse events, lack of compliance, and cost-effectiveness. Dichotomous outcomes were reported as relative risk (RR) with 95% confidence interval (CI), using intention-to-treat analysis assuming an unfavourable event for missing data. Sensitivity analyses based on methodological quality (risk of bias), available data analysis, intention-to-treat analysis assuming a favourable event for missing data, best case scenario, and worst-case scenario were conducted. MAIN RESULTS: Twelve trials were eligible. All had high risk of bias and reporting was inconsistent. Hepatitis B vaccine did not show a clear effect on the risk of developing HBsAg (RR 0.96, 95% CI 0.89 to 1.03, 4 trials, 1230 participants) and anti-HBc (RR 0.81, 95% CI 0.61 to 1.07; 4 trials, 1230 participants, random-effects) when data were analysed using intention-to-treat analysis assuming an unfavourable event for missing data. Analysis based on data of available participants showed reduced risk of developing HBsAg (RR 0.12, 95% CI 0.03 to 0.44, 4 trials, 576 participants) and anti-HBc (RR 0.36, 95% CI 0.17 to 0.76, 4 trials, 576 participants, random-effects). Intention-to-treat analysis assuming favourable outcome for missing data showed similar reduction in risk. Hepatitis B vaccination had an unclear effect on the risk of lacking protective antibody levels (RR 0.57, 95% CI 0.26 to 1.27, 3 trials, 1210 participants, random effects). Development of adverse events was sparsely reported. AUTHORS' CONCLUSIONS: In people not previously exposed to hepatitis B, vaccination has unclear effect on the risk of developing infection, as compared to no vaccination. The risk of lacking protective antibody levels as well as serious and non-serious adverse events appear comparable among recipients and non recipients of hepatitis B vaccine. PMID- 18677781 TI - Day-case versus overnight stay for laparoscopic cholecystectomy. AB - BACKGROUND: Although day-case elective laparoscopic cholecystectomy can save bed costs, its safety remains to be established. OBJECTIVES: To assess the safety and benefits of day-case surgery compared to overnight stay in patients undergoing elective laparoscopic cholecystectomy. SEARCH STRATEGY: We searched The Cochrane Hepato-Biliary Group Controlled Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL) in The Cochrane Library, MEDLINE, EMBASE, and Science Citation Index Expanded until April 2008 for identifying randomised trials using search strategies. SELECTION CRITERIA: Only randomised clinical trials, irrespective of language, blinding, or publication status, comparing day-case and overnight stay in elective laparoscopic cholecystectomy were considered for the review. DATA COLLECTION AND ANALYSIS: We collected the data on the characteristics of the trial, methodological quality of the trials, morbidity, prolonged hospitalisation, re-admissions, pain and quality of life from each trial. We analysed the data with both the fixed-effect and the random-effects models using RevMan Analysis. For each outcome we calculated the risk ratio, weighted mean difference, or standardised mean difference with 95% confidence intervals (CI) based on available case-analysis. MAIN RESULTS: Five trials with 429 patients randomised to the day-case group (215) and overnight stay group (214) were included in the review. All the trials were of high risk of bias. The trials recruited 49% of patients undergoing cholecystectomy. The selection criteria varied, but most included only patients without other diseases. The patients were living in easy reach of the hospital and with a responsible adult to take care of them. On the day of surgery, 81% of day-case patients were discharged. The drop-out rate after randomisation varied from 6.5% to 12.7%. There was no significant difference between day-case and overnight stay group as regards to morbidity, prolongation of hospital stay, re-admission rates, pain, quality of life, patient satisfaction and return to normal activity and work. AUTHORS' CONCLUSIONS: Day-case elective laparoscopic cholecystectomy seems to be a safe and effective intervention in selected patients (with no or minimal systemic disease and within easy reach of the hospital) with symptomatic gallstones. Because of the decreased hospital stay, it is likely to save costs. PMID- 18677782 TI - Outcomes of patients who participate in randomized controlled trials compared to similar patients receiving similar interventions who do not participate. AB - BACKGROUND: Some people believe that patients who take part in randomised controlled trials (RCTs) face risks that they would not face if they opted for non-trial treatment. Others think that trial participation is beneficial and the best way to ensure access to the most up-to-date physicians and treatments. This is an updated version of the original Cochrane review published in Issue 1, 2005. OBJECTIVES: To assess the effects of patient participation in RCTs ('trial effects') independent both of the effects of the clinical treatments being compared ('treatment effects') and any differences between patients who participated in RCTs and those who did not. We aimed to compare similar patients receiving similar treatment inside and outside of RCTs. SEARCH STRATEGY: In March 2007, we searched The Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, The Cochrane Methodology Register, SciSearch and PsycINFO for potentially relevant studies. Our search yielded 7586 new references. In addition, we reviewed the reference lists of relevant articles. SELECTION CRITERIA: Randomized studies and cohort studies with data on clinical outcomes of RCT participants and similar patients who received similar treatment outside of RCTs. DATA COLLECTION AND ANALYSIS: At least two review authors independently assessed studies for inclusion, assessed study quality and extracted data. MAIN RESULTS: We identified 30 new non-randomized cohort studies (45 comparisons): no new RCTs were found. This update now includes five RCTs (yielding 6 comparisons) and 80 non-randomized cohort studies (130 comparisons), with 86,640 patients treated in RCTs and 57,205 patients treated outside RCTs. In the randomised studies, patients were invited to participate in an RCT or not; these comparisons provided limited information because of small sample sizes (a total of 412 patients) and the nature of the questions they addressed. When the results of RCTs and non-randomized cohorts that reported dichotomous outcomes were combined, there were 98 comparisons; there was also heterogeneity (P < 0.00001, I(2) = 42.2%) between studies. No statistical significant differences were found for 85 of the 98 comparisons. Eight comparisons reported statistically significant better outcomes for patients treated within RCTs, and five comparisons reported statistically significant worse outcomes for patients treated within RCTs. There was significant heterogeneity (P < 0.00001, I(2) = 58.2%) among the 38 continuous outcome comparisons. No statistically significant differences were found for 30 of the 38 comparisons. Three comparisons reported statistically significant better outcomes for patients treated within RCTs, and five comparisons reported statistically significant worse outcomes for patients treated within RCTs. AUTHORS' CONCLUSIONS: This review indicates that participation in RCTs is associated with similar outcomes to receiving the same treatment outside RCTs. These results challenge the assertion that the results of RCTs are not applicable to usual practice. PMID- 18677783 TI - Cluster-controlled photofragmentation: the case of the Xe-pyrrole cluster. PMID- 18677784 TI - Model studies of the interfacial ordering of oleanolic acid in the cuticula. PMID- 18677785 TI - Asymmetric catalysis with metal complexes in nanoreactors. AB - Recently, the field of heterogeneous asymmetric catalysis, generally using chiral solid catalysts, has attracted much attention in the production of single enantiomers. Among versatile chiral solid catalysts, chiral metal complexes confined in nanoreactors often exhibit very unique enantioselectivity and catalytic activity compared to homogeneous catalysts. In this Focus Review, we summarize the recent advances in asymmetric reactions on chiral metal complexes confined in nanoreactors with an emphasis on the confinement effect and cooperative activation effect of the nanoreactor and new strategies for the preparation of chiral solid catalysts, such as the encapsulation of chiral metal complexes in the nanocages of mesoporous silica and incorporation of chiral ligands in the network of mesoporous organosilicas. PMID- 18677786 TI - Utilization of N,N-dialkylcarbamic acid derived from secondary amines and supercritical carbon dioxide: stereoselective synthesis of Z alkenyl carbamates with a CO2-soluble ruthenium-P(OC2H5)3 catalyst. AB - Reversible transformation of diethylamine (1) and supercritical carbon dioxide (scCO(2)) into N,N-diethylcarbamic acid (2) was confirmed by direct acquisition of (1)H and (13)C{(1)H} NMR spectra. The equilibrium between 1+CO(2) and 2 is strongly influenced by conditions of the supercritical state. Low temperature favors formation of carbamic acid, whereas high temperature causes decarboxylation. On the basis of the spectroscopic results of carbamic acid formation under scCO(2) conditions, the ruthenium-catalyzed formation of alkenyl carbamates from terminal alkynes, 1, and carbon dioxide was investigated to demonstrate the useful transformation of elusive carbamic acids. Selectivity toward the CO(2)-fixation products over enynes obtained by dimerization of the alkyne substrates was improved by the use of scCO(2) as a reaction medium. In particular, a CO(2)-soluble ruthenium complex, trans [RuCl(2){P(OC(2)H(5))(3)}(4)], was found to be effective in affording Z alkenyl carbamates with high stereoselectivity. PMID- 18677787 TI - A contractile DNA machine. PMID- 18677788 TI - Functionalized nanocompartments (synthosomes) with a reduction-triggered release system. PMID- 18677789 TI - Super robust nanoparticles for biology and biomedicine. PMID- 18677790 TI - Proton- and reductant-assisted dioxygen activation by a nonheme iron(II) complex to form an oxoiron(IV) intermediate. PMID- 18677792 TI - Mood and anxiety disorders more likely in people with arthritis. PMID- 18677793 TI - Chromosome "hot spot" linked to autism. PMID- 18677791 TI - An engineered aryl azide ligase for site-specific mapping of protein-protein interactions through photo-cross-linking. PMID- 18677794 TI - [Profit or loss--can handsurgery be profitable?]. PMID- 18677795 TI - Simvastatin with or without ezetimibe in familial hypercholesterolemia. PMID- 18677796 TI - Simvastatin with or without ezetimibe in familial hypercholesterolemia. PMID- 18677797 TI - Simvastatin with or without ezetimibe in familial hypercholesterolemia. PMID- 18677798 TI - Simvastatin with or without ezetimibe in familial hypercholesterolemia. PMID- 18677799 TI - Simvastatin with or without ezetimibe in familial hypercholesterolemia. PMID- 18677800 TI - Simvastatin with or without ezetimibe in familial hypercholesterolemia. PMID- 18677801 TI - Home automated defibrillators after myocardial infarction. PMID- 18677802 TI - Do risks of hormone therapy persist after discontinuation? PMID- 18677803 TI - Herb study is not a practice changer. PMID- 18677804 TI - The future of pain. PMID- 18677805 TI - Isolation and marriage patterns in four South Tyrolean villages (Italy) during the nineteenth century. AB - No information is currently available on the marriage patterns of German-speaking communities of the South Tyrol area. The aim of this study is to investigate the reproductive isolation of four South Tyrolean mountain villages during the 19th century. Data about 3953 marriages were drawn from existing pedigrees and completed with data from the parish registers of the studied villages to calculate the following indicators: age at marriage, endogamy, inbreeding from dispensations and from isonymy and repeated pairs of surnames among couples. The results show high levels of endogamy (78-87%) and an elevated age at marriage in all the studied villages. The percentages of consanguineous marriages (10-33%) vary considerably but result overall in relatively low inbreeding values (alpha 0.0015-0.0036; Ft 0.0098-0.0138). Levels of endogamy are consistent with the geographic characteristics of the area, while inbreeding values are lower than those observed in previous studies on Alpine communities. This is due to a low frequency of marriages between close relatives, probably related to the peculiar demographic and cultural characteristics of the studied populations that differentiate them from neighbouring Italian-speaking villages. PMID- 18677806 TI - Interpretation of immunohistochemistry for mismatch repair proteins is only reliable in a specialized setting. AB - We examined the validity of immunohistochemistry for mismatch repair (MMR) proteins in colorectal cancer specimens to identify patients at risk for Lynch syndrome (hereditary nonpolyposis colorectal cancer) and patients with sporadic microsatellite instable colorectal cancer. This was assessed by observer agreement for and accuracy of interpretation of immunohistochemistry. Seven pathologists from 5 different pathology laboratories evaluated 100 molecularly defined colorectal cancers stained for MLH1, PMS2, MSH2, and MSH6. Two of the pathologists were experienced in interpretation of immunohistochemistry for MMR proteins. After evaluation of a subset of 20 cases, a discussion meeting was organized, after which pathologists evaluated all 100 cases. Staining patterns were interpreted as aberrant, normal, or indefinite. In 82% of tumors, 5 or more pathologists reached the same interpretation, which was considered the consensus diagnosis. Consensus was reached slightly less frequently in microsatellite instable than in stable tumors, and interobserver variation was moderate to substantial (kappa: 0.49-0.79). More microsatellite instable tumors showed an indefinite staining pattern compared with microsatellite stable tumors. Three out of 7 pathologists, including the 2 experienced pathologists, did not miss a microsatellite instable tumor. Each pathologist found at least 1 tumor with an aberrant staining pattern, whereas consensus was a normal staining pattern and the tumor was microsatellite stable. We conclude that, if restricted to experienced pathologists, immunohistochemistry is a valid tool to identify patients at risk for Lynch syndrome and patients with sporadic microsatellite instable colorectal cancer. An indefinite or aberrant staining result has to be followed by molecular microsatellite instability analysis to confirm the presence of a defective DNA MMR system. PMID- 18677807 TI - Editorial comment on: recurrence patterns of squamous cell carcinoma of the penis: recommendations for follow-up based on a two-centre analysis of 700 patients. PMID- 18677808 TI - A heterogeneity test for fine-scale genetic structure. AB - For organisms with limited vagility and/or occupying patchy habitats, we often encounter nonrandom patterns of genetic affinity over relatively small spatial scales, labelled fine-scale genetic structure. Both the extent and decay rate of that pattern can be expected to depend on numerous interesting demographic, ecological, historical, and mating system factors, and it would be useful to be able to compare different situations. There is, however, no heterogeneity test currently available for fine-scale genetic structure that would provide us with any guidance on whether the differences we encounter are statistically credible. Here, we develop a general nonparametric heterogeneity test, elaborating on standard autocorrelation methods for pairs of individuals. We first develop a 'pooled within-population' correlogram, where the distance classes (lags) can be defined as functions of distance. Using that pooled correlogram as our null hypothesis reference frame, we then develop a heterogeneity test of the autocorrelations among different populations, lag-by-lag. From these single-lag tests, we construct an analogous test of heterogeneity for multilag correlograms. We illustrate with a pair of biological examples, one involving the Australian bush rat, the other involving toadshade trillium. The Australian bush rat has limited vagility, and sometimes occupies patchy habitat. We show that the autocorrelation pattern diverges somewhat between continuous and patchy habitat types. For toadshade trillium, clonal replication in Piedmont populations substantially increases autocorrelation for short lags, but clonal replication is less pronounced in mountain populations. Removal of clonal replicates reduces the autocorrelation for short lags and reverses the sign of the difference between mountain and Piedmont correlograms. PMID- 18677809 TI - Sources of information used by nurses to inform practice: An integrative review. AB - BACKGROUND: We present an integrative review of the literature about sources of information nurses use to inform practice. The demand for access to more and better information has been fueled by the evidence-based healthcare movement. Although the expectations for evidence-based practice have never been higher, the demands on care environments have never been greater. The goals of professional nursing are served by using the best available information to inform practice. To influence such activity, we must understand what sources of information nurses rely on for guidance. DATA SOURCES: We examined studies of any research design published between 1985 and 2006, as well as research dissertations in the same time frame. Databases searched included the Cumulative Index of Nursing and Allied Health Literature (CINAHL), Medline, the Educational Resources Information Center, the Library and Information Science Abstracts and the Library Literature and Information Science databases. REVIEW METHODS: The review question: what information sources do registered nurses turn to, to support direct patient care? Analysis included an overview of study design and practice setting, and an examination of sources accessed most frequently by nurses to guide practice. RESULTS: We present, in ranked order, the sources nurses accessed in order to guide practice. We note the high reliance on informal, interactive sources. An unexpected finding of high reliance on journals is explored in greater detail and found to be equivocal at best. We conclude with a critical discussion of what we see as embedded assumptions and expectations about how information-seeking supports nursing practice. CONCLUSION: Expectations embedded in the scope and context of nursing practice have influenced knowledge development in the area of information-seeking to support practice. It is important that future research in this area takes into account the expectations and information needs arising in emerging roles for nurses within evolving healthcare systems. PMID- 18677810 TI - [Clinical evaluation on fengweicao granule in treating benign prostatic hyperplasia]. AB - OBJECTIVE: To report the effect of Fengweicao Granule (FWCCG), a Chinese herbal preparation made of herba Pteris multifidae, in treating benign prostatic hyperplasia (BPH). METHODS: One hundred and fifty-five patients were assigned to two groups, the 108 patients in the treated group were administered orally with FWCG twice a day, 5 g every time; the 47 patients in the control group were treated with Proscar 5 mg per day for 3 months. The effects were evaluated by the change of the related indexes before and after treatment, including scoring by international prostatic symptom scale (I-PSS), maximum flowing rate of urine (MFR), residue urine volume (RU) in urinary bladder determined by abdominal B ultrasonography and volume of prostate. RESULTS: After being treated for 3 months, the I-PSS, MFR and RU improved remarkably in both groups (P <0.05 or P <0.01), but with no significant change in the volume of prostate, neither with significant difference in comparison between the two groups (P >0.05). CONCLUSION: FWCG has a good effect with less adverse reaction in treating BPH. PMID- 18677811 TI - Bibliography. Current world literature. Bladder cancer. PMID- 18677812 TI - [Success of the corticosteroid treatment during a rare and grave inaugural complication of the lupus: the acute pancreatitis]. PMID- 18677813 TI - Bibliography. Current world literature. Prevention. PMID- 18677814 TI - [Evaluation of chemist knowledge in treatment of acne in Tunisia]. PMID- 18677815 TI - [Emergent mania with atypical antipsychotics: review of the literature about one case]. PMID- 18677816 TI - A case of anaphylactic reaction to ibuprofen in a child with Still's disease. PMID- 18677817 TI - [Leflunomide-induced toxic epidermal necrolysis]. PMID- 18677818 TI - Patient safety and quality improvement. Notice of proposed rulemaking. AB - This document proposes regulations to implement certain aspects of the Patient Safety and Quality Improvement Act of 2005 (Patient Safety Act). The proposed regulations establish a framework by which hospitals, doctors, and other health care providers may voluntarily report information to Patient Safety Organizations (PSOs), on a privileged and confidential basis, for analysis of patient safety events. The proposed regulations also outline the requirements that entities must meet to become PSOs and the processes for the Secretary to review and accept certifications and to list PSOs. In addition, the proposed regulation establishes the confidentiality protections for the information that is assembled and developed by providers and PSOs, termed "Patient safety work product" by the Patient Safety Act, and the procedures for the imposition of civil money penalties for the knowing or reckless impermissible disclosure of patient safety work product. PMID- 18677819 TI - International Conference on Harmonisation; Guidance on Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the International Conference on Harmonisation Regions; availability. Notice. AB - The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled "Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions." The guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The guidance describes a process for the evaluation and recommendation by the ICH Q4B Expert Working Group (EWG) of selected pharmacopeial texts to facilitate their recognition by regulatory authorities for use as interchangeable in the ICH regions. Following favorable evaluations, ICH will issue topic-specific annexes with information about these texts and their implementation (the Q4B Outcomes). Implementation of the Q4B annexes is intended to avoid redundant testing by industry in favor of a common testing strategy in each ICH regulatory region. PMID- 18677820 TI - International Conference on Harmonisation; Guidance on Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions; Annex on Residue on Ignition/Sulphated Ash General Chapter; Availability. Notice. AB - The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled "Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions; Annex 1: Residue on Ignition/Sulphated Ash General Chapter.'' The guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The guidance provides the results of the ICH Q4B evaluation of the Residue on Ignition/Sulphated Ash General Chapter harmonized text from each of the three pharmacopeias (United States, European, and Japanese) represented by the Pharmacopoeial Discussion Group (PDG). The guidance conveys recognition of the three pharmacopeial methods by the three ICH regulatory regions and provides specific information regarding the recognition. The guidance is intended to recognize the interchangeability among these texts from the local regional pharmacopeias, thus avoiding redundant testing in favor of a common testing strategy in each regulatory region. Elsewhere in this issue of the Federal Register, FDA is announcing the availability of a guidance on the Q4B process entitled "Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions." PMID- 18677821 TI - International Conference on Harmonisation; Guidance on E15 Pharmacogenomics Definitions and Sample Coding; Availability. Notice. AB - The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled "E15 Definitions for Genomic Biomarkers, Pharmacogenomics, Pharmacogenetics, Genomic Data and Sample Coding Categories." The guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The guidance contains definitions of key terms in the discipline of pharmacogenomics and pharmacogenetics, namely genomic biomarkers, pharmacogenomics, pharmacogenetics, and genomic data and sample coding categories. In the effort to develop harmonized approaches to drug regulation, it is important to ensure that consistent definitions of terminology are being applied across all constituents of the ICH. The guidance is intended to facilitate the integration of the discipline of pharmacogenomics and pharmacogenetics into global drug development and approval processes. PMID- 18677822 TI - Medical devices; hearing aids; technical data amendments. Direct final rule. AB - The Food and Drug Administration (FDA) is amending its regulations governing hearing aid labeling to reference the most recent version of the consensus standard used to determine the technical data to be included in labeling for hearing aids. We are amending the regulations to require that manufacturers may use state-of-the-art methods to provide technical data in hearing aid labeling. FDA is also amending the regulations to update an address and remove an outdated requirement. FDA is amending the regulations in accordance with its direct final rule procedures. Elsewhere in this issue of the Federal Register, we are publishing a companion proposed rule under FDA's usual procedures for notice and comment rulemaking to provide a procedural framework to finalize the rule in the event we receive a significant adverse comment and withdraw this direct final rule. PMID- 18677823 TI - Medicare and Medicaid programs: hospice conditions of participation. Final rule. AB - This final rule revises the existing conditions of participation that hospices must meet to participate in the Medicare and Medicaid programs. The final conditions address the comments that we received on the proposed rule published on May 27, 2005. This final rule focuses on the care delivered to patients and their families by hospices and the outcome of that care. The final requirements continue to reflect the unique interdisciplinary view of patient care and allow hospices flexibility in meeting quality standards. These changes are an integral part of the Administration's efforts to achieve broad based improvements in the quality of health care and our efforts to improve the quality of care furnished through the Medicare and Medicaid programs. PMID- 18677824 TI - Medical devices; medical device reporting; baseline reports. Direct final rule. AB - The Food and Drug Administration (FDA) is amending its medical device reporting regulations to remove a requirement for baseline reports that the agency deems no longer necessary. Currently, manufacturers provide baseline reports to FDA that include the FDA product code and the premarket approval or premarket notification number. Because most of the information in these baseline reports is also submitted to FDA in individual adverse event reports, FDA is removing the requirement for baseline reports. The removal of this requirement will eliminate unnecessary duplication and reduce the manufacturer's reporting burden. FDA is amending the regulation in accordance with its direct final rule procedures. Elsewhere in this issue of the Federal Register, we are publishing a companion proposed rule under FDA's usual procedures for notice and comment to provide a procedural framework to finalize the rule in the event we receive a significant adverse comment and withdraw this direct final rule. PMID- 18677825 TI - Procedures for transportation workplace drug and alcohol testing programs: state laws requiring drug and alcohol rule violation information. Interim final rule. AB - The Office of the Secretary (OST) is amending its drug and alcohol testing procedures to authorize employers to disclose to State commercial driver licensing (CDL) authorities the drug and alcohol violations of employees who hold CDLs and operate commercial motor vehicles (CMVs), when a State law requires such reporting. This rule also permits third-party administrators (TPAs) to provide the same information to State CDL licensing authorities where State law requires the TPAs to do so for owner-operator CMV drivers with CDLs. PMID- 18677826 TI - Procedures for transportation workplace drug and alcohol testing programs. Final rule. AB - The Department of Transportation is amending certain provisions of its drug and alcohol testing procedures to change instructions to collectors, laboratories, medical review officers, and employers regarding adulterated, substituted, diluted, and invalid urine specimen results. These changes are intended to create consistency with specimen validity requirements established by the U.S. Department of Health and Human Services and to clarify and integrate some measures taken in two of our own Interim Final Rules. This Final Rule makes specimen validity testing mandatory within the regulated transportation industries. PMID- 18677827 TI - Medicare program; use of repayment plans. Final rule. AB - This final rule modifies Medicare regulations to implement section 935(a) of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 pertaining to the use of repayment plans (also known as extended repayment schedules or "ERS'') for Medicare provider and supplier overpayments. Under this provision, we are granting a provider or a supplier an ERS under certain terms and conditions as defined in the statute. This final rule establishes criteria and procedures to apply this requirement and to define the concepts of "hardship'' and "extreme hardship." PMID- 18677828 TI - Medicare program; appeals of CMS or CMS contractor determinations when a provider or supplier fails to meet the requirements for Medicare billing privileges. Final rule. AB - This final rule implements a number of regulatory provisions that are applicable to all providers and suppliers, including durable medical equipment, prosthetics, orthotics, and supplies (DMEPOS) suppliers. This final rule establishes appeals processes for all providers and suppliers whose enrollment, reenrollment or revalidation application for Medicare billing privileges is denied and whose Medicare billing privileges are revoked. It also establishes timeframes for deciding enrollment appeals by an Administrative Law Judge (ALJ) within the Department of Health and Human Services (DHHS) or the Departmental Appeals Board (DAB), or Board, within the DHHS; and processing timeframes for CMS' Medicare fee for-service (FFS) contractors. In addition, this final rule allows Medicare FFS contractors to revoke Medicare billing privileges when a provider or supplier submits a claim or claims for services that could not have been furnished to a beneficiary. This final rule also specifies that a Medicare contractor may establish a Medicare enrollment bar for any provider or supplier whose billing privileges have been revoked. Lastly, the final rule requires that all providers and suppliers receive Medicare payments by electronic funds transfer (EFT) if the provider or supplier, is submitting an initial enrollment application to Medicare, changing their enrollment information, revalidating or re-enrolling in the Medicare program. PMID- 18677829 TI - Medicare program; special enrollment period and Medicare premium changes. Final rule. AB - This final rule provides a special enrollment period (SEP) for Medicare Part B and premium Part A for certain individuals who are sponsored by prescribed organizations as volunteers outside of the United States and who have health insurance that covers them while outside the United States. Under the SEP provision, qualifying volunteers can delay enrollment in Part B and premium Part A, or terminate such coverage, for the period of service outside of the United States and reenroll without incurring a premium surcharge for late enrollment or reenrollment. This final rule also codifies provisions that require certain beneficiaries to pay an income-related monthly adjustment amount (IRMAA) in addition to the standard Medicare Part B premium, plus any applicable increase for late enrollment or reenrollment. The income-related monthly adjustment amount is to be paid by beneficiaries who have a modified adjusted gross income that exceeds certain threshold amounts. It also represents the amount of decreases in the Medicare Part B premium subsidy, that is, the amount of the Federal government's contribution to the Federal Supplementary Medicare Insurance (SMI) Trust Fund. PMID- 18677830 TI - Medicare and Medicaid programs; hospital conditions of participation: laboratory services. Final rule. AB - This final rule finalizes the hospital conditions of participation requirements for hospitals that transfuse blood and blood components. It requires hospitals to: Prepare and follow written procedures for appropriate action when it is determined that blood and blood components the hospitals received and transfused are at increased risk for transmitting hepatitis C virus (HCV); quarantine prior collections from a donor who is at increased risk for transmitting HCV infection; notify transfusion recipients, as appropriate, of the need for HCV testing and counseling; and extend the records retention period for transfusion-related data to 10 years. The intent is to aid in the prevention of HCV infection and to create opportunities for disease prevention that, in most cases, can occur many years after recipient exposure to a donor. PMID- 18677831 TI - Beneficiary travel under 38 U.S.C. 111 within the Unites. Final rule. AB - This document amends the beneficiary travel regulations of the Department of Veterans Affairs (VA) that provide a mechanism for payment of travel expenses within the United States under 38 U.S.C. 111 to help veterans and other persons obtain care and services from VA's Veterans Health Administration (VHA). The amended regulations more fully implement the statutory provisions governing such payments. PMID- 18677832 TI - [Anastomotic approach at the proximal side and auxiliary means for Stanford B type dissociation]. PMID- 18677833 TI - Myths, "zombies" and "damned lies" plague Canadian healthcare systems: what's a researcher to do? PMID- 18677834 TI - Incalculability as a feature of sugar production during the eighteenth century. PMID- 18677836 TI - Do Ontario patients with heart failure have equal access to life-saving technology? PMID- 18677835 TI - Malthus, population, and the generational bargain. PMID- 18677837 TI - ["Good policy" and local society: experience as a category in 18th-century administrative practice]. PMID- 18677838 TI - Re: To evaluate the role of diclofenac sodium on renal parenchyma of young albino rats. PMID- 18677839 TI - In conversation with Bonnie Brossart by Ken Tremblay. PMID- 18677840 TI - Spanish naturalists in Cuba and the West Indies, 1795-1802. PMID- 18677841 TI - Some aspects of the work of Robert Brown and the investigator naturalists in Madeira during August 1801. PMID- 18677842 TI - Urban demographic stagnation in early modern Germany: a simulation. PMID- 18677843 TI - Women and gender in early modern England. PMID- 18677844 TI - Parenting was for life, not just for childhood: the role of parents in the married lives of their children in early modern England. PMID- 18677845 TI - Cholangiocellular hepatocarcinoma. PMID- 18677846 TI - Toward a model of successful electronic health record adoption. AB - The Canadian healthcare landscape abounds with pressures to address wait times, chronic disease management, aging at home, information and service integration, health human resource shortages, pandemic planning and most importantly health outcomes of individuals receiving care in our system. Investment in clinical information technologies is often touted as significant to the successful resolution of most if not all of these issues. For example, Baker and Norton (2001) uncovered an alarming rate of preventable adverse events occurring within Canadian hospitals. A particularly high error rate associated with the administration of fluids and medications suggests that there is a dire need to introduce processes and tools to reduce human error in healthcare facilities. The implementation of clinical applications such as computerized physician order entry (CPOE) with integrated electronic medication administration records (MAR) has been identified as a key step to safer care (Bates and Gawande 2003; Leape et al. 2002; Leatt et al. 2006). It has been suggested that the full value of electronic health records (EHR) will only be realized with the implementation of CPOE and that its use (by physicians) is a reasonable proxy for adoption (Ash and Bates 2005). Considering recent surveys of Canadian and American hospitals, those that have fully implemented CPOE remain in the minority (Ash et al. 2004; Davis 2007; Gudbranson 2007); most have yet to tackle the challenges of the change imperative and adoption issues associated with the use of a complete EHR PMID- 18677847 TI - New study finds one in four hospitalizations is drug related. AB - A recent study by Samoy et al. (2006) found that almost 25% of patients admitted to the internal medicine ward of British Columbia's largest hospital are there because of sickness brought on by adverse drug reactions. PMID- 18677848 TI - Computerized physician order entry usage in North America: the doctor is in. AB - While computerized physician order entry (CPOE) is still in its infancy as far as adoption goes (<10% of North American hospitals have implemented CPOE), most organizations are planning to utilize CPOE in the future. However, one of the major questions on most hospitals' minds is, "Will physicians accept and use CPOE?"Recently, KLAS set out to discover what progress has been made in CPOE penetration and physician usage. The 2008 CPOE study is the sixth annual report of its kind and details the state of CPOE in North American hospitals. Among the findings are the percentage and depth of CPOE usage, the methods hospitals now employ to increase CPOE use and what successes and difficulties organizations have encountered with their CPOE initiatives. According to providers, the doctor is in and CPOE usage is growing. PMID- 18677849 TI - Perioperative nursing care of the bariatric surgical patient. AB - APPROXIMATELY TWO-THIRDS of the US population is overweight or obese. The effects of the comorbidities that accompany obesity often are severe and can be life threatening over time. Currently, the most effective and sustainable method of substantial weight loss is bariatric surgery. Bariatric surgery also has been successful in reversing comorbidities. THE BENEFITS, RISKS, AND COMPLICATIONS of common weight-loss surgical procedures (eg, Roux-en-Y gastric bypass, laparoscopic adjustable gastric banding, vertical-banded gastroplasty, biliopancreatic diversion and duodenal switch) are discussed. THE HEALTH CARE FIELD OF BARIATRICS is growing rapidly. More information is needed to support and guide changes in current standards of practice to better meet the needs of this patient population. PMID- 18677850 TI - [Tracheobronchoplasty combined with intracavitary irradiation to circumvent left pneumonectomy]. PMID- 18677851 TI - [A comment of the editor]. PMID- 18677852 TI - [Dioxins and sarcomas in Veneto: let us open a debate on the role of epidemiology]. PMID- 18677853 TI - [Dioxins and sarcomas in Veneto: the context, the method and the peer review]. PMID- 18677854 TI - [Fighting against HPV with a vaccine in Italy and in the world]. PMID- 18677855 TI - [Evaluation of the impact of HPV vaccination]. PMID- 18677856 TI - [Epidemiology of HPV genital infection and of its complications and use of HPV testing for cervical cancer screening]. PMID- 18677857 TI - [Biological basis for HPV vaccines]. PMID- 18677858 TI - [Neonatal mortality in campania region: analysis of causes of death by current data]. AB - OBJECTIVE: to analyze neonatal mortality by cause using the P-Perinatal classification system of the Italian National Institute of Statistics (ISTAT). DESIGN: comparisons by birth cohort (1991-2001 vs 1981-1990). Estimates of the rate ratio (RR) and 95% confidence intervals (CI). The 56 ISTAT causes of perinatal death were regrouped into 7 categories. SETTING: the study was carried out in the Campania region, which has the largest number of births in southern Italy and high neonatal mortality. The analysis covered 1627641 live births and 12865 neonatal deaths. RESULTS: neonatal mortality declined significantly over time (RR= 0.58; 95% CI 0.56-0.60). The decrease was greater for early (first 6 days of lfe) (RR=0.54; 95% CI 0.51-0.56) than for late neonatal mortality (6th to 29th day of life) (RR=0.75; 95% CI 0.69-0.81). Rates declined for all causes, but RR were particularly low for prematurity (RR=0.24; 95% CI 0.22-0.28), pre-partum hypoxia (RR=0.28; 95% CI 0.22-0.35), intra-partum hypoxia (RR=0.42; 95% CI 0.38 0.47). More limited declines were observed for congenital anomalies (RR=-0.67; 95% CI 0.61-0.72) and for RDS (Respiratory Distress Syndrome) (RR=0.82; 95% CI 0.77-0.87). Declines inpre- and intra-partum hypoxia and for other causes were observed in both early and late neonatal mortality. Mortality due to congenital abnormalities declined more in the early neonatal period (RR=0.59; 95% CI 0.53 0.65) than in the late neonatal period (RR=0.86; 95% CI 0.75-0.99). RDS, which was the leading cause of death in both periods, was the only cause for which there was an increase in late neonatal mortality (RR= 1.21; 95% CI 1.05-1.40). CONCLUSION: Available current data allow to describe different determinants of neonatal mortality by causes of death. Currently, improvement of RDS care is necessary in order to reduce high neonatal mortality. PMID- 18677859 TI - [Determinants of occupational injuries in the construction of the "high speed train" Bologna-Florence]. AB - OBJECTIVE: to study determinants of occupational injuries in tunnel construction using data from the surveillance system which had been implemented in order to monitor accidents during the construction of the "high speed train tracks in the Italian Regions Emilia-Romagna and Tuscany. DESIGN: retrospective cohort study. SETTING: 16 sites for the construction of 14 tunnels of the high speed railway tract Bologna-Firenze, in Italy. PARTICIPANTS: 1,602 workers (of 3,000 employed in the underground tunnelling), aged 18 - 67 years, operating during excavation with traditional method in 1999-2002. A total of 549 injuries occurred among 385 workers. The number of worked hours were used as time at risk. MAIN OUTCOME MEASURES: incidence rate ratios (IRR) and 95% confidence intervals for all injuries, serious injuries and first injuries were considered in separate multiple regression analyses (Poisson). RESULTS: residence, task and working phase were taken into consideration. An increased risk was found for younger workers, for carpenters (IRR "all-events" = 2.33; 95% CI=1.85-2.94; IRR" first events" = 2.12; 95% CI 1.62-2.77) and miners (IRR "all-events" = 1.76; 95% CI 1.39-2.24; IRR"first-events" = 1.71; 95% CI 1.30-2.24) vs. machinery operators. Construction of inverted arch turns out to have an incidence rate ratio three times higher than digging out (IRR "all-events" = 2.79; 95% CI 2.27-3.43; IRR "firsts-event = 2.98; 95% CI 2.33-3.81). The probability of "serious" injuries (>30 days) is higher for miners (IRR=2.45; 95% CI 1.65-3.64) and for carpenters (IRR=2.31; 95% CI 1.53-3.49). CONCLUSION: this study pointed out to indicate some determinants (age, task and work phase) of injuries in tunneling about which little had been published previously. These results are useful for addressing preventive measures, for control and prevention activities and point to the need to explore the effect of experience and to study, through a case crossover design, transient working and individual risk factors for traumatic injury within these working sites. PMID- 18677860 TI - [Mortality among the elderly in Tuscany in the period 1987-2003]. AB - AIM: to assess cause-specific mortality and its temporal trend in Tuscan elderly residents. DESIGN: descriptive epidemiologic study based upon death certificates, collected and registered since 1987 by the Tuscan Regional Mortality Registry (RMR) according to standardized procedures. METHODS: Major groups of causes: temporal trends are described calculating annual truncated age-adjusted mortality rates (based on age-specfic rates subdivided in three classes: 75-79; 80-84 and > or =85; standard: European population) and estimating annual percent changes (EAPC, Estimated Annual Percent Change) using ]oinpoint regression models. Most frequent specific causes: number of deaths and truncated age-adjusted mortality rates (based on age-specific rates subdivided in three classes: 75-79; 80-84 and > or =85; standard: European population) are compared between the first and the last quinquennium (1987-1991 and 1999-2003) by percent change. RESULTS: during 1987-2003, an average of 26667 annual deaths (65% of total) occurred in persons aged 75 and over in Tuscany. In the same period the number of elderly residents increased (2003 vs 1987: men +43.6%; women +41.5%) with a consequent increase in number of deaths (2003 vs 1987: men +13.8%; women +15.9%). The truncated age adjusted rates for all mortality causes decreased (EAPC = -1.35% in males; EAPC = -1.41% in females) while an increase was registered for few specific causes as Alzheimer's disease, senile dementia, arterial hypertension and lung cancer. CONCLUSIONS: the decreasing mortality trend observed in persons aged 75 and over in Tuscany is consistent with similar trends in other developed countries. The opposing trends for few specific causes of death need to be further investigated. PMID- 18677861 TI - [Use of hospital discharge records to estimate the incidence of malignant mesotheliomas]. AB - SETTING: cancer registries usually adopt strategies for active case finding. Interest in using administrative sources of data is rising OBJECTIVE: to assess the usefullness of Hospital discharge records (HDR) to supplement the traditional methods of case finding of the malignant mesothelioma (MM) Registry of the Piedmont Region. METHODS: HDRs have been used since 1996. We assessed the number of cases identified only through HDRs and their influence on MM incidence. RESULTS: cases identified through HDRs were about 10% of those with histologic confirmation of the diagnosis, 34% of those with cytologic confirmation, and 72% of those without morphologic examination. Cases diagnosed in hospitals located outside the region would have been easily (50%) missed. The age-standardised (standard: Italian pop. at the 1981 census) incidence rate of pleural MM increases from 2.2 to 2.7 per 100,000 per year among men, and from 1.1 to 1.2 among women, when including all cases identified from HDRs, irrespective of their diagnostic confirmation. Peritoneal MM incidence estimates are unaffected. Overall without access to the hospital discharge files, 179 cases out of 954 would not have been registered between 1996 and 2001. In the same calendar period 59 cases identified by means of active search by the Registry have not been found in the hospital discharge files. CONCLUSIONS: HDRs are useful in addition, but not in substitution, to active search of MM cases. PMID- 18677862 TI - [Incidence of malignant mesothelioma in Piedmont, 1990-2001]. AB - SETTING: age-, period-, and cohort-models allowed the prediction ofan increase in mortality from malignant mesotheliomas (MM) in Western Europe, expected to peak between 2015 and 2025 in different Countries, and around 2020 in Italy. Recently, however, a deceleration in the increase has been reported. OBJECTIVE: we updated to 31 december 2001 previous estimates ofMM incidence among residents in the Piedmont region (Italy) and assessed time-trends in incidence. DESIGN: the Registry of Malignant Mesotheliomas actively searches cases in hospital wards and pathology units' records since 1990; since 1996 the database of hospital admissions and discharges is also used. Cases are registered when eligible for inclusion in the National Mesothelioma Registry. RESULTS: the average yearly incidence of pleural MM, computed across four consecutive three-year calendar periods, from 1990-1992 to 1999-2001, increased in both sexes, for histologically confirmed as well as for all cases. No changes were observed in peritoneal MM incidence. CONCLUSIONS: the rise in incidence might be due to increased diagnostic awareness for MM. An analysis by birth-cohort, however, revealed a cohort-effect, therefore it is unlikely that diagnostic bias alone could explain the observed trend. Among men residing at diagnosis in the province of Turin, but not among women nor in both sexes outside Turin province, the increase in incidence has levelled off in the most recent calendar period; for them exposures stopped on average seven years earlier than those of residents in other areas of Piedmont. PMID- 18677863 TI - [Mortality due to chronic obstructive pulmonary disease: indicators of impact, access to health services, and outcome of hospital treatment in an Italian region]. AB - OBJECTIVE: to estimate mortality due to chronic obstructive pulmonary disease (COPD), risk of dying from COPD without hospital admission and mortality within 30 days after hospital admission for COPD in an Italian region. METHODS: population registries of death causes and hospital discharge reports were used as data sources; COPD was identified through ICD-9 codes = 490.X, 491.X, 492.X, 494.Xe 496.X. Age-adjusted mortality rates were calculated by direct standardisation using the Italian population from 1996 to 2000; logistic regression was used to estimate predictors of dying from COPD in multivariable models. RESULTS: there were 4,292 deaths among men and 2,713 among women; the mean annual mortality was 3.68/10,000 inhabitants among men and 2.29/10,000 among women. CONCLUSION: no statistically significant trend was observed from 1996 to 2000, but the increase in mortality rate was higher for females (13.2%) than for males (2.0%), suggesting a possible inversion in smoking habit between sexes. The risk of dying without hospitalisation was higher for males who resided outside Rome (OR 1.65; CI 95% 1.04-2.62). Mortality within 30 days after hospitalisation was 4.2% among 25,046 patients. Patients who died were more likely to be over 54 and male, to have comorbidities or complications, hospitalised in general wards rather than pneumology or intensive care units, and have been on ventilation. PMID- 18677864 TI - [Characteristics and effectiveness of smoking cessation programs in Italy. Results of a multicentric longitudinal study]. AB - AIM: to describe the characteristics and effectiveness of various smoking cessation programs offered by Italian treatment services operating within the National Health Service. DESIGN: prospective longitudinal multicentre study involving 41 smoking cessation services in 16 Italian regions. STUDY POPULATION: the study population includes patients entering smoking cessation programs between April 2003 and June 2004. The "study population" includes 1226 patients (54.2% males and 45.4% females), mean age 47 years. Patients have a middle/high level of education and a long history of smoking; most are highly dependent on nicotine and report previous attempts to quit smoking. METHODS: treatment effectiveness in smoking cessation is assessed six months after entering treatment service. Logistic Regression Model was used to determine the predictors of successfiul cessation, independent of treatment typology. The predictors were included as confounding variables in the logistic regression model that was used to evaluate the effectiveness of treatments. Besides the effect of treatment completion on smoking cessation was estimated. RESULTS: predictors of successful smoking cessation are: being male, presence of a partner, strong motivation to quit, previous attempts to give up smoking, mild nicotine dependence, and not suffering from mood disturbances. All treatments are effective in helping people to stop smoking: cessation rate ranges between 25.00% for patients receiving a single session of motivational counselling and 65.3% for those receiving nicotine replacement therapy combined to group therapy. Compared to a single session of motivational counseling, nicotine replacement therapy combined to group therapy is the most effective therapeutic program (OR 5.4; 95%CI 12.5-12.0). Treatment completion is a strong determinant ofsuccess (OR 4.8; 95%CI 3.5-6.4). CONCLUSION: enrolling people in any type of therapeutic program, in particular nicotine replacement therapy combined with group therapy increases the probability of successfully quitting smoking; moreover, patients that begin a smoking cessation program should be encouraged to complete the therapy PMID- 18677865 TI - [Prevalence of arterial hypertension treatment LHU 2 population in Umbria: a comparison between two data sources]. AB - OBJECTIVE: verify the consistency of two independent sources of data concerning the prevalence of arterial hypertension under pharmacological treatment. DESIGN: the pharmacological prescriptions database (A) and the Studio PASSI (Italian behavioural risk factor survey) (B) were assessed. Data selection was carried out according with the presence of at least one prescription of antihypertensive drug during the year 2004 (A), the statement of being under treatment with antihypertensive drugs (B). Prevalence rates were calculated. SETTING: resident population registered in the list of Umbrian Local Health Unit 2 (LHU 2); age range: 18-69 years. PARTICIPANTS: A) the pharmacological prescriptions database: 43,146 subjects selected from the 2004 archive, B) Studio PASSI: representative random sample of 200 persons drawn out from the resident population registered in the 2005 record of the LHU2 list. MAIN OUTCOME MEASURES: prevalence rate for arterial hypertension with pharmacological treatment. RESULTS: both studies showed similar prevalence rates: A) pharmacological prescriptions database = 17.2% (95% CI 17.1-17.4) and B) PASSI = 15.0% (95% CI10.4-20.7). Age and gender distribution were also comparable. Moreover, the PASSI survey produced additional socio-economical information. CONCLUSION: the hypothesis that the pharmacological prescription database can be used to monitor in a simple, timely and reproducible way major health-related events was validated. Small size surveys - such as the PASSI study - may add further information to the analysis of large administrative databases and they may serve to periodically verify the consistency of these databases when assessed for epidemiological purposes. PMID- 18677866 TI - [Work accidents notified to INAIL are not predictive of recognition]. PMID- 18677867 TI - Noel Plumridge on community tariff appeal. PMID- 18677868 TI - Clinical microsystems, part 1. The building blocks of health systems. AB - BACKGROUND: Wherever, however, and whenever health care is delivered-no matter the setting or population of patients-the body of knowledge on clinical microsystems can guide and support innovation and peak performance. Many health care leaders and staff at all levels of their organizations in many countries have adapted microsystem knowledge to their local settings. CLINICAL MICROSYSTEMS: A PANORAMIC VIEW: HOW DO CLINICAL MICROSYSTEMS FIT TOGETHER? As the patient's journey of care seeking and care delivery takes place over time, he or she will move into and out of an assortment of clinical microsystems, such as a family practitioner's office, an emergency department, and an intensive care unit. This assortment of clinical microsystems-combined with the patient's own actions to improve or maintain health--can be viewed as the patient's unique health system. This patient-centric view of a health system is the foundation of second-generation development for clinical microsystems. LESSONS FROM THE FIELD: These lessons, which are not comprehensive, can be organized under the familiar commands that are used to start a race: On Your Mark, Get Set, Go! ... with a fourth category added-Reflect: Reviewing the Race. These insights are intended as guidance to organizations ready to strategically transform themselves. CONCLUSION: Beginning to master and make use of microsystem principles and methods to attain macrosystem peak performance can help us knit together care in a fragmented health system, eschew archipelago building in favor of nation building strategies, achieve safe and efficient care with reliable handoffs, and provide the best possible care and attain the best possible health outcomes. PMID- 18677869 TI - Improving the translation of research into primary care practice: results of a national quality improvement demonstration project. AB - BACKGROUND: There is widespread evidence of inadequate translation of research findings into primary care practice. Theoretically sound demonstrations of how health care organizations can overcomes these deficiencies are needed. A demonstration project was conducted from January 1, 2003, through June 30, 2006, to evaluate the impact of a multicomponent intervention and improvement models intended to enhance adherence to clinical practice guidelines across eight broad clinical areas. METHODS: The demonstration project involving 530 clinicians and staff members from 99 primary practices in 36 states entailed practice performance reports (audit and feedback), practice site visits for academic detailing and participatory planning, and network meetings for sharing 4 of "best practice" approaches. Data from electronic medical records (EMRs) of 847,073 patients were abstracted to identify 31 process and 5 outcome quality measures for prevention and treatment of cardiovascular disease and diabetes, cancer screening, adult immunization, respiratory and infectious disease, mental health and substance abuse, obesity and nutrition, safe medication prescribing in the elderly, and a summary measure, the Summary Quality Index (SQUID). RESULTS: The yearly adjusted absolute improvement in the SQUID was 2.43% (95% confidence interval [C.I.], 2.24%-2.63%). Clinically and statistically significant improvements occurred for 29 of the 36 quality measures, including all 5 outcome measures. DISCUSSION: The findings suggest that a multicomponent quality improvement intervention involving audit and feedback, academic detailing and participatory planning activities, and sharing of "best practice" approaches in practices with EMRs can have a robust impact in quality of care for Americans seen in primary care practices. PMID- 18677870 TI - The effectiveness of root cause analysis: what does the literature tell us? AB - BACKGROUND: Root cause analysis (RCA) is an analysis framework used in health care to determine the systemic causes and prevent recurrences of adverse events. It is required by The Joint Commission for reported events and by the Department of Veterans Affairs (VA) National Center for Patient Safety for qualifying events in VA medical centers. The evidence on RCA effectiveness in improving patient safety was reviewed. METHODS: MEDLINE, Academic Search Premier, and the Cochrane Database were searched from database inception to September 2007. RCA case studies and articles that directly addressed the RCA framework were reviewed. RESULTS: Discussion of RCA did not emerge in the literature until the late 1990s, and there have been no controlled trials that test the RCA framework. Twenty three articles describe the RCA process, 38 articles present RCA case studies, and 12 articles analyze weaknesses of the RCA framework. Eleven of the case studies measure RCA effectiveness, 3 using clinical outcome measures and 8 using process measures. All 11 articles report improvement of safety following RCA. RCA participants report the difficulty in forming causal statements and in developing/implementing corrective actions. Criticisms of RCA include the uncontrolled study design and participant biases. DISCUSSION: Overall, the limited literature on RCA effectiveness provides anecdotal evidence that RCA improves safety. At the same time, it highlights the numerous theoretical problems with the analytical framework. Formal studies at the system level and cost-benefit analysis are needed to determine the effectiveness of RCA. Structured publication of case studies will support shared knowledge and will provide benchmarks for improvement. Enrichment of the RCA literature body will enable reproducibility of improvement work, optimization of analysis, and validation of the framework itself. PMID- 18677871 TI - Outcomes of an initial set of standardized performance measures for inpatient mental health. AB - BACKGROUND: In January 2006, the University HealthSystem Consortium (UHC) convened a committee of experts from academic health centers to identify an initial set of important standardized performance measures for inpatient psychiatric services and to evaluate the current state of performance in these measures at eight academic health centers. METHOD: The eight UHC academic medical centers completed a retrospective review of 20 inpatient psychiatric records on patients who were 18-65 years of age with a primary diagnosis of psychosis and a length of stay > or = 2 days. The performance measures, derived from practice standards and the consensus of an interdisciplinary committee of experts, focused on the processes of care, including screening, assessment, treatment, coordination, continuity, and safety. RESULTS: Although there was variability in organizational performance in a number of the psychiatric measures, some organizations demonstrated high levels of performance. Performance measures indicating the greatest improvement opportunities for organizations included notification of outpatient mental health provider of the psychiatric hospitalization within two days; collaboration with the outpatient mental health provider and/or primary care physician; and scheduling a follow-up appointment within seven days of discharge. DISCUSSION: This initial benchmarking project in mental health at academic health centers shows that there is a range of conformity to important processes of care in the inpatient mental health setting. The results of the notification, collaboration, and continuity measures in this study highlight national concerns regarding the lack of communication and collaboration between providers in the transition through the continuum of services. Future quality measurement projects in mental health services should integrate clinical process measures with outcome measures. PMID- 18677873 TI - A controlled trial of a rapid response system in an academic medical center. AB - A medical emergency team composed of house staff and existing float-pool nurses was successfully implemented on the general medical floor of an academic medical center without increasing personnel. The intervention had little noticeable impact, although the number of cardiac arrests and deaths were low both before and after the intervention. PMID- 18677872 TI - Measuring practice systems for chronic illness care: accuracy of self-reports from clinical personnel. AB - BACKGROUND: Purchasers, plans, and clinical practices involved in quality improvement initiatives are increasingly interested in measuring practice systems, particularly in relation to clinical quality and as part of pay-for quality initiatives. The validity of self-reports of the use of practice systems was examined. METHODS: In 11 medical groups in Minnesota, the Physician Practice Connections Readiness Survey, which was developed on the basis of the concepts and evidence base of the Chronic Care Model, was used to survey office practice personnel about practice systems. Participation rates by medical group ranged from 61% to 94%, with a mean of 76%, yielding surveys from 32 lead physicians and 241 other personnel. Survey results were compared with an on-site audit by trained surveyors. RESULTS: Overall agreement with the on-site audit ranged from 40.9% to 96.7% among lead physicians and from 33.9% to 81.9% among other personnel. Mean agreement was high for quality improvement (96.7% for lead physicians and 81.9% for other personnel), moderate for clinical information systems (71.2% for lead physicians and 66.0% for others), and low for the use of care management (less than 50% for both groups). Mean positive predictive value ranged from 55.2% to 100% among lead physicians and from 49.6% to 100% among other personnel. Both the presence of systems and the accuracy of reporting varied across medical groups. DISCUSSION: The accuracy of self-reports of practice systems varies by type of system being assessed and by type of respondent. Although self-assessment may be useful for quality improvement purposes, self-reported information on clinical practices systems should not be used for accountability purposes, including pay-for-quality efforts or public reporting unless additional documentation is required to ensure fair comparisons. PMID- 18677875 TI - Has health promotion reached the tipping point? PMID- 18677874 TI - Transparency in health care: leading the way. AB - A one-day conference sponsored by Dartmouth-Hitchcock Medical Center focused on external transparency (sharing data outside an organization) also featured discussion of internal transparency (sharing data inside an organization). PMID- 18677876 TI - Wellness in the workplace: a multi-stakeholder health-promoting initiative of the World Economic Forum. AB - The World Economic Forum and the World Health Organization, arguably the most powerful business and health organizations in the world, are collaborating on an effort to make workplace wellness a global priority. The authors discuss progress on this effort. PMID- 18677877 TI - Understanding barriers to participation in mammography by women with disabilities. AB - PURPOSE: The study purpose was to identify barriers to mammography screening among women with different disabilities and to suggest interventions to address barriers. METHODS: Forty-two women with self-reported disabilities, ages 40 to 69 years participated. They resided in 24 Connecticut towns, and most had a prior mammogram. Data were collected through six disability-specific focus groups from women with sensory, physical, psychiatric, and cognitive/intellectual impairments. Facilitator-conducted groups used a semistructured guide. Qualitative analysis applied an iterative coding process to generate themes and categories. RESULTS: We identified four themes (i.e., access, beliefs, social support, and comfort/ accommodations) and nine subthemes that characterized barriers. In all focus groups, women mentioned physical access and physical comfort/accommodations as types of barriers. Other major subthemes were communication and professional support. Women also described mammography facilitators. CONCLUSION: Despite frequent use of health care and personal strategies to facilitate mammography screening, women with disabilities reported barriers to getting mammograms. Findings suggest a multifaceted approach to address these barriers. PMID- 18677878 TI - Cigarette smoking and food insecurity among low-income families in the United States, 2001. AB - PURPOSE: To quantify the association between food insecurity and smoking among low-income families. DESIGN AND SETTING: A retrospective study using data from the 2001 Panel Study of Income Dynamics (PSID), a longitudinal study of a representative sample of U.S. men, women, and children and the family units in which they reside. SUBJECTS: Low-income families. MEASURES: Family income was linked with U.S. poverty thresholds to identify 2099 families living near or below 200% of the federal poverty level. Food insecurity (i.e., having insufficient funds to purchase enough food to maintain an active and healthy lifestyle) was calculated from the 18-core-item food security module of the U.S. Department of Agriculture. Current smoking status was determined. RESULTS: Smoking prevalence was higher among low-income families who were food insecure compared with low-income families who were food secure (43.6% vs. 31.9%; p < .01). Multivariate analysis revealed that smoking was associated with an increase in food insecurity of approximately six percentage points (p < .01). CONCLUSIONS: Given our finding that families near the federal poverty level spend a large share of their income on cigarettes, perhaps it would be prudent for food assistance and tobacco-control programs to work together to help low-income people quit smoking. PMID- 18677879 TI - Multi-ethnic adolescents' attitudes toward smoking: a focus group analysis. AB - PURPOSE: To explore young, multi-ethnic adolescents' attitudes and influences related to cigarette smoking for the purpose of developing and producing a youth led, tobacco prevention drama. DESIGN: Focus groups and demographic surveys. SETTING: Island of Oahu, Hawaii. PARTICIPANTS: Fifty-four multi-ethnic youth, ranging in age from 10 to 14 years, participated. The study was promoted in schools with a large representation of Hawaiian, Filipino, and Pacific Islander students. METHODS: Each of five focus groups was audio-recorded, and an observer recorded extensive notes throughout the sessions. Content analysis consisted of coding focus group notes for recurrent themes and using the audio recording as confirmation. RESULTS: Just more than one quarter (n = 15) of the youth had tried smoking, and two-thirds (n = 35) currently lived with someone who smoked. Participants expressed the feeling of being surrounded by smoking influences at home, in their communities, and at school. Youth were negatively affected by family members' tobacco use, and they desired skills that could enable them to help family members stop using tobacco. CONCLUSION: Family influences may play an important role in youth attitudes toward tobacco use, especially given the cultural significance of extended family and of filial piety that are reflected in many traditional, Asian and Pacific Islander families. Interventions targeting youth in Asian and Pacific Islander communities should incorporate key cultural references to the extended family and to a respect for elders to establish relevance in the life experiences of young people in these population groups. PMID- 18677880 TI - Credit card debt, stress and key health risk behaviors among college students. AB - PURPOSE: To examine cross-sectional associations between credit card debt, stress, and health risk behaviors among college students, focusing particularly on weight-related behaviors. DESIGN: Random-sample, mailed survey. SUBJECTS/SETTING: Undergraduate and graduate students (n = 3206) attending a large public university. MEASURES: Self-reported health indicators (e.g., weight, height, physical activity, diet, weight control, stress, credit card debt). RESULTS: More than 23% of students reported credit card debt > or = $1000. Using Poisson regression to predict relative risks (RR) of health behaviors, debt of at least $1000 was associated with nearly every risk indicator tested, including overweight/obesity, insufficient physical activity, excess television viewing, infrequent breakfast consumption, fast food consumption, unhealthy weight control, body dissatisfaction, binge drinking, substance use, and violence. For example, adjusted RR [ARR] ranged from 1.09 (95% Confidence interval [CI]: 1.02 1.17) for insufficient vigorous activity to 2.17 (CI: 0.68-2.82) for using drugs other than marijuana in the past 30 days. Poor stress management was also a robust indicator of health risk. CONCLUSION: University student lifestyles may be characterized by a variety of coexisting risk factors. These findings indicate that both debt and stress were associated with wide-ranging adverse health indicators. Intervention strategies targeting at-risk student populations need to be tailored to work within the context of the many challenges of college life, which may serve as barriers to healthy lifestyles. Increased health promotion efforts targeting stress, financial management, and weight-related health behaviors may be needed to enhance wellness among young adults. PMID- 18677881 TI - Worksite-based weight loss programs: a systematic review of recent literature. AB - OBJECTIVE: To update a previous systematic review on the effectiveness of worksite-based weight loss programs. DATA SOURCE: The following databases were searched: Medline, PsychlNFO, Embase, The Cochrane Library, and LexisNexis. STUDY INCLUSION AND EXCLUSION: Studies were limited to those published in English from 1995 to 2006 to which the following inclusion criteria were applied: (1) worksite intervention, (2) body weight assessed before and after intervention, and (3) study duration of at least 8 weeks. DATA EXTRACTION: Data were extracted on the following: study design; funding source; purpose of evaluation; participant and worksite characteristics; type, intensity and duration of intervention; primary and secondary outcomes; and methodological quality. DATA SYNTHESIS: Heterogeneity of study designs precluded quantitative data synthesis. Results. We identified 11 randomized controlled trials, most of which focused on education and counseling to improve diet and increase physical activity. Follow-up ranged from 2 to 18 months, with 56% to 100% of subjects completing the studies. The overall methodological quality of the studies was poor. Intervention groups lost significantly more weight than controls, with the mean difference in weight loss ranging from -0.2 to -6.4 kg. CONCLUSION: Worksite-based weight loss programs can result in modest short improvements in body weight; however, long-term data on health and economic outcomes are lacking. So What? There is a need for rigorous controlled studies of worksite-based interventions that integrate educational, behavioral, environmental, and economic supports. PMID- 18677882 TI - The effectiveness of tailored feedback and action plans in an intervention addressing multiple health behaviors. AB - PURPOSE: Examine the effects of three iterative tailored feedback letters addressing smoking; physical activity; and fruit, vegetable, and fat intake, and test the additional effects of providing feedback on action plans. DESIGN: A tailored, print-based intervention was developed and tested in a randomized control trial with a posttest after 9 months. SETTING: A total of 2827 respondents agreed to participate. They were recruited from a random sample of 35,000 addresses obtained through the Dutch national telephone company. SUBJECTS: The mean age was 49 years, and 55% were female. Intervention. The experimental group received three printed tailored letters, and the control group received three printed generic letters. Respondents from the experimental group randomly received either a third letter with tailored information or tailored information and action-planning feedback. MEASURES; The questionnaire assessed physical activity; smoking; consumption of fruit, vegetables and fat; motivational determinants; action plans; and demographics. RESULTS: Tailored information resulted in more improvement over time than generic information for the intake of fruit, vegetables, and fat and for physical activity. No differences between the conditions were found for smoking because of high cessation rates in all conditions. Action-planning feedback did not increase the effects. CONCLUSIONS: Tailored lifestyle information can be effective for adults in changing nutrition behavior and physical activity. PMID- 18677883 TI - Exploring obesogenic food environments in Edmonton, Canada: the association between socioeconomic factors and fast-food outlet access. AB - PURPOSE: To explore the relationship between the placement of fast-food outlets and neighborhood-level socioeconomic variables by determining if indicators of lower socioeconomic status were predictive of exposure to fast food. DESIGN: A descriptive analysis of the fast-food environment in a Canadian urban center, using secondary analysis of census data and Geographic Information Systems technology. SETTING: Edmonton, Alberta, Canada. MEASURES: Neighborhoods were classified as High, Medium, or Low Access based on the number of fast-food opportunities available to them. Neighborhood-level socioeconomic data (income, education, employment, immigration status, and housing tenure) from the 2001 Statistics Canada federal census were obtained. ANALYSIS: A discriminant function analysis was used to determine if any association existed between neighborhood demographic characteristics and accessibility of fast-food outlets. RESULTS: Significant differences were found between the three levels of fast-food accessibility across the socioeconomic variables, with successively greater percentages of unemployment, low income, and renters in neighborhoods with increasingly greater access to fast-food restaurants. A high score on several of these variables was predictive of greater access to fast-food restaurants. CONCLUSION: Although a causal inference is not possible, these results suggest that the distribution of fast-food outlets relative to neighborhood-level socioeconomic status requires further attention in the process of explaining the increased rates of obesity observed in relatively deprived populations. PMID- 18677884 TI - The effect of objectively measured crime on walking in minority adults. AB - PURPOSE: Evaluate the relationship between neighborhood crime and the amount of daily walking by minority adults. DESIGN: This was a cross-sectional study of minority adult walking behavior and crime. Setting. Oakland, California was chosen as the study area because of the substantial spatial variation in levels of criminal activity combined with detailed information on walking trips. PARTICIPANTS: The study was restricted to minority adults who responded to the 2000 Bay Area Travel Survey and lived in Oakland, California (n = 359). METHOD: Data on leisure and utilitarian walking were collected through the 2000 Bay Area Travel Survey and combined with crime data from the Oakland Police Department. A negative binomial model was used to test if violent, property, or quality of life crimes had significant associations with daily minutes walked, controlling for individual and neighborhood covariates. RESULTS: The model showed a significant negative association between violent crime and minutes walked per day (b = -.07; p = .016). Neither property nor quality of life crimes were correlated with amount of walking. CONCLUSIONS: Reductions in violent crime may increase opportunities for minority residents in urban areas to participate in physical activity such as walking, thereby providing another reason to pursue anticrime measures. Urban designers' efforts to increase physical activity by improving neighborhood walkability may consider violent crime prevention in their designs. PMID- 18677885 TI - The ecological approach in health promotion programs: a decade later. AB - PURPOSE: Present a 10-year update of the ecologic model of health promotion published by Richard et al. in the American Journal of Health Promotion in 1996. APPROACH: We adapted and simplified the model by leaving out settings, focusing on levels, and incorporating interpersonal and individual levels and agents that are in control of environmental conditions. SETTING: Health care facilities, schools, workplaces, living environments, and public environments were represented in program descriptions. PARTICIPANTS: Forty-three program coordinators responsible for 47 programs were interviewed in the United States and The Netherlands. METHODS: A systematic interview protocol elicited general program descriptions and relevant aspects of the ecologic approach. Program aspects from written reports were coded by the authors into levels, interventions, targets, and strategies. RESULTS: The programs had 234 strategies and 276 targets, with a mean of 2.15 levels. Twenty-seven distinct intervention strategies were identified, with the most common being modification of an organization to which the at-risk individuals belong, followed by policy and community changes. CONCLUSION: Our data fit the ecologic model and our adaptations and indicate that over the past decade health promotion practice may have changed to include more multilevel programs. Systematic analysis of program strategies within and across environmental levels allows better understanding of the social ecology of health-related behavior and potential leverage points for change. PMID- 18677886 TI - Planning worksite health promotion programs: models, methods, and design implications. AB - As the field of Worksite Health Promotion and Wellness matures, greater pressure is being place on programs that produce mitigation of health risks, improved employee productivity and increased levels of economic return. To achieve these expected results and to successfully contend with the challenges of today's workplace and work force it will likely become even more critical to carefully plan both new and existing programs each year. A model for the planning and development of worksite programs is presented and discussed along with a variety of design implications for different population segments. PMID- 18677887 TI - Don't let paperwork be a barrier. PMID- 18677888 TI - Pursuing a larger mission. PMID- 18677889 TI - An opportunity to improve lives and the bottom line. PMID- 18677890 TI - A city embraces change. PMID- 18677891 TI - Building a virtual bridge. PMID- 18677892 TI - EAST helps people with psychosis out west. PMID- 18677893 TI - Patient privacy in a digital world. PMID- 18677894 TI - [Possibilities of radical surgical treatment of patients with tumoral obstruction of extrahepatic biliary ducts]. AB - Experience of radical surgical treatment of 32 patients, suffering extrahepatic biliary ducts (EBD) tumor, was analyzed. The respectability index of a Clatskin tumor have constituted 12.5%, and of the EBD middle portion--25%. In 8 patients an open operative intervention was performed and in 1--laparoscopic procedure was applied. The respectability index of a distal EBD tumors have constituted 29.6%. Postoperative lethality was 15.6%. PMID- 18677895 TI - [Surgical treatment of patients with Leriche syndrome--immediate results]. AB - Comparative analysis of the various methods of surgical treatment efficacy was conducted in patients suffering Leriche syndrome, including bifurcational, one sided and multilevel reconstruction. Clinical indices of intra- and postoperative periods, the frequency of thrombotic and nonthrombotic complications occurrence, lethality for every kind of intervention were analyzed. It was established, that tactics of surgical treatment of Leriche syndrome ought to be individualized, but in presence of clinical and anatomical conditions the one-sided reconstruction must be preferred. PMID- 18677896 TI - [Syndrome of acute coronary insufficiency in ulcerative gastro-intestinal hemorrhage]. AB - In 2004- 2007 in the clinic 249 patients, in whom the ulcer disease gastro intestinal hemorrhage (GIH) occurred, were observed. Of 107 elderly patients 43 (40.1%) were hospitalized lately than 24 hours after GIH beginning. Gastroduodenofibroscopy was conducted for the hemorrhage origin investigation and the local hemostasis state determination. In a hospitalization day in 19 (17.7%) patients myocardial infarction on the GIH background was diagnosed. GIH and myocardial infarction were treated simultaneously. Timely elimination of the circulating blood volume deficiency under constant control of arterial and central venous pressure, the hourly diuresis and ECG dynamics was considered the main element of a treatment program. After infusion-transfusion therapy conduction the acute coronary insufficiency syndrome was eliminated on the 3-5th day. PMID- 18677897 TI - [Observation of primary esophageal melanoma]. PMID- 18677898 TI - [Era of medical imaging (II)]. PMID- 18677899 TI - [Recent advances in molecular pathogenesis and treatment of polycystic kidney disease]. AB - Polycystic kidney diseases (PKD) are common genetic disorders characterized by formation and progressive enlargement of cysts kidney, liver and other organs, leading to end stage renal disease. Regardless of the genetic defect underlying PKD, cystic epithelia seem to display common abnormalities: increased proliferation and apoptosis, loss of cellular differentiation and polarity, hypersecretion. The localization of multiples proteins, whose function are disrupted in PKD, in the primary cilium or at basal body at the base of the cilium highlight this neglected organelle as a common trigger of cystic diseases. Significant progresses have been made over the last few years towards a greater understanding of the molecular pathogenesis of cysts formation, particularly in the signaling pathways involved in cytogenesis: cAMP, mTOR, Wnt, Ras/MAPK. These advances have already brought several potential therapies targeting several key pathways of cystogenesis. PMID- 18677900 TI - [Arterial media calcification in patients with type 2 diabetes mellitus]. AB - Arterial calcification was previously viewed as an inevitable, passive, and degenerative process that occurred at the end stages of atherosclerosis. Recent studies, however, have demonstrated that calcification of arteries is a complex and regulated process. It may occur in conjunction with atherosclerosis or in an isolated form that is commonly associated with diabetes and renal failure. Higher artery calcium scores are associated with increased cardiovascular events, and some aspects of arterial calcification are similar to the biology of forming bone. Arterial calcification can thus be viewed as a distinct inflammatory arteriopathy, much like atherosclerosis and aneurysms, with its own contribution to cardiovascular morbidity and mortality. Current research involves efforts to define the complex interactions between cellular and molecular mediators of arterial calcification and, in particular, the role of endogenous calcification inhibitors. This review discusses the clinical relevance, cellular events, and suspected molecular pathways that control arterial calcification. PMID- 18677901 TI - [Nutrition of low birth weight infants]. AB - The low birth weight infant's (LBW) nutrition needs special attention. Energy requirements are increased due to a high metabolic rate and a low thermogenesis. Increased protein requirements are, easy to be absorbed (from lacto-serum). Lipids, as important energetic source requires an appropriate content in essential fatty acids and medium chain triglycerides (TGM) which are easy absorbed in absence of lipase and deficit of bile salt secretion. The carbohydrates should be represented by lactose or equivalents like di-, oligo or polysaccharides. The addition of maltodextrin avoid an exaggerate supply and incomplete digestion of lactose. Sodium requirement are increased due to exaggerated loss consequence of renal immaturity. The rapid rate of growth impose the addition of Calcium, Phosphorus, Iron, Copper, and vitamins, especially Vitamin D, E and Folic acid. The appropriate nutrition of LBW must cover his special needs to maintain the homeostasis and rapid growth. Precise nutrient requirements are, however, difficult to establish. PMID- 18677902 TI - Incubation of the coronary disease. Pediatric perspective. AB - The atherosclerosis modifications at the vascular level are evidentiated early by diminishing the blood flow mediated vasodilatation, thickening of the arterial wall and the appearance since childhood of the aortic fibrose plates. Their evolution is determined mainly by the atherosclerotic dislipidemia. The maximum risk for atherosclerosis and coronary disease appears in the subjects with low birth weight which in the fetal period and childhood can develop epigenetic modifications in the tissues and organ development. The study proposes to collect the clinical, epidemiological, experimental and epigenetic data in an unified explanation of the induction mechanism of atherosclerosis. PMID- 18677903 TI - [Triple-negative breast cancer--towards a new entity]. AB - Breast cancer is considered today as a heterogeneous group of diseases, with a wide spectrum of clinical, pathologic and molecular features. A new classification of breast carcinomas, based on gene expression profiles technologies (cDNA microarray), shows impact on the diagnosis and prognostic classification of tumors, prediction of response of individual patients to specific chemotherapeutic regimens, and identification of novel tumor targets. AIM: To identify the proportion of basal-like ("triple-negative") tumors in patients treated for breast carcinoma. OBJECTIVES: To evaluate the impact of molecular subtyping on treatment response, disease-free survival and overall survival of patients, and to establish and correlate particular biologic, clinical and diagnostic features of these cancers. METHODS: 96 patients with breast cancer, aged 28-78 years, were retrospectively selected using specific criteria from cases treated between 2001 and 2007 in the Oncology Department of the "Sf. Spiridon" Hospital Iasi. Tumors were divided into molecular subtypes using immunohistochemistry data. RESULTS: The most frequent pathology was ductal carcinoma (77 patients, 80.2%), and most tumors were intermediately or poorly differentiated (G2 43 patients, 44.8%; G3 37 patients, 38.5%) The proportion of basal-like tumors in the population sample analyzed was 15.62% (15 patients). Histological type (p=0.85), pTNM stage (p=0.618), and tumor differentiation grading (p=0.54) did not correlate with molecular subtype. Patients with basal like tumors had lower time-to-progression (mean 31.2 months, 95%CI 18.5-43.8, p=0.005), disease free survival (mean 24.46 months, 95%CI 12.6-36.3, p=0.016), and overall survival (mean 39.26 months, 95%CI 26.7-51.7, p=0.023). CONCLUSION: We have demonstrated, using the simple commonly available immunohistochemical markers ER, PR and HER2/neu, that patients with triple negative breast cancers have relatively poor prognosis, with significantly shorter survival and a poorer response to standard treatment strategies. PMID- 18677904 TI - [Colorectal polyps: clinical, endoscopic, and histopathologic features]. AB - AIM: To evaluate the prevalence of colorectal polyps in a population from North- Eastern Romania, which underwent colonoscopic examination and to identify their clinical, endoscopic and histopathologic characteristics. METHODS: A retrospective prevalence study of subjects aged 18-95 years, with no personal or familial history of familial adenomatous polyposis and inflammatory bowel disease, who underwent a colonoscopy. The number, size, gross endoscopic appearance, histopathological examination, and the anatomic location of colorectal polyps were analyzed, as well as the associated diseases. RESULTS: Enrolled were 1001 patients (647 men, 354 women), aged 18-95 years (mean: 62.8 +/ 11.4 years) who were diagnosed with colorectal polyps. Overall, there were 1.534 colorectal polyps, and the majority of them (83.0%) were present in patients over the age of 50 years, from urban area (73.9%) and most of the patients were men (63.8%), and had solitary polyps (61.1%). Three-quarters of polyps were distal (rectum, sigmoid, and descendent colon) in location, and most of them (75.8%) had a size less than 10 mm. Over half of all polyps (54.7%) were pedunculated. Based on the histological structural configuration, 76.5% of all polyps were adenomas, and of these 62.6% were tubular adenomas; malignant polyps have been diagnosed in 82 (5.4%) patients. Associated hemorrhoids, diverticulosis, colorectal cancer, and angiodysplasia were identified in 9.6%, 10% and 1.3%, respectively. CONCLUSION: The prevalence of colorectal polyps in our geographical area is in the range with that observed in other regions of our country. Most of polyps were identified in patients older than 50 years, predominantly men, located in distal colon, had a tubular adenoma histological structure, and frequently associated with hemorrhoids and diverticular disease. PMID- 18677905 TI - [Clinical characteristics and lipid profile of patients with life-threatening cardiovascular diseases assisted in different hospitals of Cluj-Napoca]. AB - The knowledge about lipid profile particularities in patients with atheromatous cardiovascular diseases may have therapeutic and prophylactic implications. MATERIAL AND METHOD: Study included patients with acute myocardial infarction (IMA) and stroke (AVC), cared in Cluj-Napoca's hospitals. RESULTS: The patients with IMA were from urban area in 76% of cases, 72% were males and 58% had arterial hypertension to. Cholesterolemia over 240 mg% level had 11.9% of patients (17.2% female, 9.8% male with IMA), HDL-cholesterolemia less then 35 mg% had 33.4% of male patients and less then 45 mg%, 64.1% of female IMA patients. Patients with AVC were only male who provided in 59% of cases from urban area, had age average of 63 years old, and 82% of them had arterial hypertension to. The average levels for cholesterol were higher in AVC patients (218.6 mg% from urban area, 211.3 mg% from rural area), for triglycerides were higher in patients with IMA and AVC from urban area. High average level of HDL-cholesterolemia had patient with antecedent alcohol consuming (49.5 mg%), women with IMA (45.09 mg %) and in contrast the smokers had the lowers levels (38.37 mg%). PMID- 18677906 TI - [The genetic polymorphism of MTHFR gene in schizophrenia]. AB - The 677C > T polymorphism of the MTHFR gene, resulting in hyperhomocysteinemia, has been shown to be implicated in the aetiology of schizophrenia. Previous studies showed that A1298 C polymorphism seems not to be related to schizophrenia. AIM OF THE STUDY: To analyze two genetic polymorphisms of the MTHFR gene, 677C > T and A1298 C in 44 patients with schizophrenia and evaluate its relationship with the risk of schizophrenia and with some clinical aspects. MATERIAL AND METHOD: We determined the presence of the 677C > T and A1298 C mutations of the MTHFR gene in 44 inpatients with schizophrenia and in 35 normal controls. The patients were assessed by psychiatric examination and scalar evaluation. RESULTS: 28 (66,7%) of the patient group had the T allele of the 677C > T genetic polymorphism, compared to 11 (34,3%) subjects of the control group. The intensity of the positive, negative and general symptoms was slightly higher in the patients presenting the T allele. The A1298C missense mutation was more frequent between control subjects (57,5%) compared to the patient group (39%). The intensity of the positive symptoms was slightly increased in the patients with the missense mutation in the position 1298, but the intensity of the negative and general symptoms did not differ. CONCLUSIONS: Our study confirms the role of the 677C > T genetic polymorphism in the susceptibility for schizophrenia. The relationship between A1298C genetic polymorphism and schizophrenia was not demonstrated in our study. PMID- 18677907 TI - [Neuropsychological disturbances in thalamic hemorrhage--clinical study]. AB - Neuropsychological disturbances are frequently associated with hemorrhagic stroke of the thalamus. MATERIAL AND METHOD: We present our personal findings on 117 patients admitted in the Ist Neurological Clinic, lasi with hemorrhagic stroke of the thalamus. RESULTS: Transcortical aphasia for the left-sided strokes and anosognosia or hemi-neglect were among the most prominent clinical features, especially for large thalamic hemorrhage. Visual or tactile extinction, memory disturbances, agitation and hallucinosis were also exhibited in various proportions. PMID- 18677908 TI - [Osteopenia in children with juvenile idiopathic arthritis]. AB - The aim of the study was to evaluate the presence and etiopathogenesis of osteopenia in 41 children with Juvenile Idiopathic Arthritis (JIA). METHODS: Bone status was evaluated by quantitative ultrasound using a Sunlight Omnisense 7000s Ultrasound Bone Sonometer. Measurements were performed at the distal radius and midshaft tibia. Results were obtained as Speed of sound (SOS) and Z-score. We used standardised clinical evaluation (modified Giannini's criteria, CHAQ). ESR, Fibrinogen, serum calcium, magnesium, alkaline phosphatase, protein electrophoresis, 25-OH vitamin D (RIA) and urinary Hydroxyproline were obtained in all patients. Osteopenia was present in 15 (36.5%) patients. Statistical analysis was performed with SPSS 13.0. RESULTS: Age, sex, age at onset, disease duration, life standards and duration of corticotherapy and methotrexate treatment were not related to osteopenia in our study. The disease activity, evaluated by clinical criteria, ESR and Fibrinogen, was strongly associated with osteopenia (p<0.001). Nutritional status was an independent risk factor for osteopenia (p<0.001). Low serum calcium (p=0.034), magnesium (p=0.010), 25-OH vitamin D (p=0.091) and alkaline phosphatase (p=0.31) were more frequent in patients with osteopenia. Hydroxyproline was increased in all patients with osteopenia (p<0.001). CONCLUSIONS: Osteopenia was a frequent (36.5%) complication of JIA in our study. The disease activity and nutritional status were the most important risk factors for osteopenia. The increase of bone reabsorption was the main pathogenic mechanism of osteopenia in our study. Calcium and magnesium deficits were related to osteopenia. Decrease of bone synthesis was not associated with osteopenia in the present study. PMID- 18677909 TI - [Factors influencing responsiveness to treatment in children with renal anemia in end stage renal disease]. AB - Human recombined erythropoietin had proven its efficiency in the treatment of anemia in chronic dialyzed patients and allows the elimination of complete necessity of transfusion. PURPOSE: We proposed ourselves to identify the factors that limit the answer to the combined treatment with iron and erythropoietin in a group study of 71 children diagnosed with ESRD in the IVth Pediatrics Clinic and included in the chronic dialysis program. MATERIAL AND METHOD: We divided the patients in three groups, in relation to the time in which the patients had reached and maintained the hemoglobin value of 10.5 g/dl under sustained iron treatment, combined with EPO. We analyzed the correlations established between responsively/non-responsively to treatment and the medium value of Hb, Ht, reticulocytes number, plasma value of iron and ferritin, the parameters of phospho-calcium metabolism, malnutrition as presence and degree, the age of entrance in dialysis and the duration of renal function support. RESULTS: We observed that Hb and Ht have low medium values (Hb-8.44 g/dl, Ht-25.65%) in the non-responsive group, by comparison to the responsive patients group (Hb-10.98 g/dl, Ht -32.31%). The most elevated values of iPTH around 420 pg/ml and the most frequent cases of protein-caloric malnutrition (26.8%) are seen in the non responsive group to EPO treatment. The age of 13 years old at the entrance in the chronic dialysis program and the time of support of renal function over 21 months are seen in the non-responder group. CONCLUSION: We consider the secondary hyperparathyroidism, the protein-caloric malnutrition, the older age in the moment of entrance in dialysis and the long time of support of renal function factors that are able to influence, in a negative manner, the response to this treatment. PMID- 18677910 TI - [Cryptococcus meningitis, five years of experience and literature review]. AB - The aim of this study was to determine the distribution and antifungal susceptibility profile of Cryptococcus spp. isolated from patients in northeast Romania. MATERIAL AND METHOD: Fungi isolated from blood and cerebrospinal fluid (CSF) cultures were identified by ID32C strips (bioMerieux, France). Susceptibility testing was performed using ATB FUNGUS2 strips (bioMerieux, France). RESULTS: A total of 20 significant strains have been isolated. Overall, Cryptococcus neoformans was the most frequent isolate (95%). We mention the first Cryptococcus albidus meningitis in Romania (strain with multiple resistance). All cryptococci tested were susceptible to amphotericin B with minimal inhibitory concentration (MIC) =1 microg/mL; 90% strains were susceptible to flucytosine. Resistance to fluconazole and itraconazole was observed in 2 and 1 case, respectively. Most of the patients had an associated infection, tuberculosis in 37.5% cases. Despite appropriate therapy 4 patients died (21%). CONCLUSION: First choice induction therapy with amphotericin B plus flucytosine followed by fluconazole is highly recommended since resistance to antifungal agents is still very low. PMID- 18677911 TI - [Persistent and recurrent skin infection with small-colony variant methicillin resistant Staphylococcus aureus]. AB - S. aureus is one of the problematic bacteria, capable to develop resistance mechanisms to all antibiotics that the bacteria are naturally susceptible. A particular phenotypic mechanism, especially against the antibiotics that repressed the synthesis of the cellular wall and aminoglycosides, was evidenced in subpopulations that grows in small-colonies and represents auxotrophic mutants for hemin, menadione or thymidine. This type of strains has been isolated most frequently from patients with osteomyelitis, septic arthritis or pulmonary infections after a long period of antibiotic treatment. The authors present the case of a patient with persistent and recurrent staphylococcal infection of the peritoneal dialysis exit site, treated with different antibiotics (ciprofloxacin, vancomycin, amoxicillin and clavulanic acid, cephalexin) from witch has been isolated a small-colony strain of methicillin-resistant S. aureus. Therapeutic failure can be explain by the slow multiplication of this strain in vivo, persistence into phagocytes and the protection offered by biofilm from the surface of the catheter. Bacteriologic diagnosis in these cases is difficult because of the culture, biochemical and susceptibility testing particularities of these strains. All these may lead failure to identification small colony variants of S. aureus and mis-evaluation of the frequency of infection with these strains in patients with long-term antibiotherapy. PMID- 18677912 TI - [Gamma interferon testing in blood and cerebrospinal fluid--rapid method for the diagnosis of tuberculous meningitis]. AB - AIM: To evaluate the benefit of this early method of diagnosis of tuberculous meningitis, gamma-interferon levels comparatively in blood and cerebrospinal fluid (CSF). MATERIAL AND METHOD: We have studied 50 patients with tuberculous meningitis admitted in the Clinic of Infectious Diseases Iasi between 2006-2007, from the epidemiological, clinical, diagnosis and therapeutical points of view. RESULTS: Tuberculous meningitis was more frequent in males, with a mean ration of 2.5. The age of patients varied between 2 and 78 years of age, with a mean value of 30.5 years. 35.6% of patients were children. In 32 cases the meningeal infection was secondary, disseminated from a primary site, most frequently from the lungs (68,7%). The bacteriologic confirmation was through culture in 13.3%, with an average of 23.4 days of incubation. In whole blood the sensitivity Quantiferon TB Gold assay (QTF) was 78.57%. In CSF the specificity was higher than in blood (96.1 vs. 88.4%), having a positive prediction value over 90%. Antituberculous therapy prior to QTF testing had a negative impact, 50 to 6 cases having negative QTF results. CONCLUSION: The performance indicators of TB Gold Quantiferon Test in serum or CSF are high, being more sensitive and rapid than the direct exam of CSF or BK culture. PMID- 18677913 TI - [Therapeutic algorithm of idiopathic scoliosis in children]. AB - Acquired deformations of spinal cord (scoliosis, kyphosis, lordosis) represent a frequent pathology in child; their treatment is complex, with variable results which depend on various parameters. Mild scoliosis, with an angle less than 30 degrees, is treated with physiotherapy and regular follow-up. If the angle is higher than 30 degrees, the orthopedic corset is required; the angle over 45 degrees impose surgically correction. The indications of every therapeutic method depend on many factors, the main target of the treatment is to prevent the aggravation of the curvature; concerning the surgery, the goal is to obtain a correction as normal as possible of the spinal axis. PMID- 18677914 TI - Outcome of incidental thyroid carcinoma in patients undergoing surgery for Graves' disease. AB - Thyroid malignancy detected incidentally in patients who are operated for Graves'disease (GD) has been reported at different rates. The aim of this study was to investigate the rate and outcome of incidental thyroid carcinoma in thyrotoxic GD patients managed by surgery in our institution. METHODS: A retrospective study was performed on a consecutive series of 92 patients with GD operated between January 2000 and December 2006 in the Department of Surgery of the Philipps University, Marburg. All patients were identified via the central electronic patient registry. Incidence and outcome of thyroid carcinoma incidentally discovered in patients undergoing surgery for GD were analysed. RESULTS: 82.6% were females and 17.4 were males. Mean age at time of surgery was 41.6 years (range: 12-72). Mean hospital stay was 5 days (range: 1-21). Two (2.2%) of all patients had a papillary thyroid microcarcinoma. Tumour size was 3 mm and 5 mm. Multifocality and lymph node involvement were not detected. Following thyroidectomy, the two patients are well and disease free with a follow up of 6, respectively 3 years. CONCLUSION: The incidence of incidentally thyroid microcarcinoma was not higher in patients with GD compared to other benign thyroid disorders described in the literature. Differentiated thyroid cancer found incidentally at surgery for hyperthyroidism seems to have a good prognosis. PMID- 18677915 TI - [Ultrasound guided percutaneous drainage in the treatment of local septic complications of chronic pancreatitis]. AB - AIM: To evaluate results of ultrasound guided percutaneous catheter drainage (PCD) for local septic complications of chronic pancreatitis. MATERIAL AND METHOD: Between 1999 and 2002, PCD was used in 19 patients with chronic pancreatitis for 6 infected acute pseudocysts, 3 post-necrotic abscesses, 3 abscesses following pancreatic resections, 6 abscesses with none of the precipitating factors present (no acute episode or recent surgical/endoscopic interventions) and a tuberculous peritoneal empyema. In 17 patients, collections were unilocular. RESULTS: Trocar technique was used in 14 patients and Seldinger technique in 5. Sixteen patients were successfully treated with PCD alone (mean drainage duration = 32 days) without abscess recurrence during the follow-up period (mean = 35.6 month). Surgical treatment was necessary in 3 patients. Pseudocyst recurrence occurred in another patient after 9 months. CONCLUSION: PCD should be considered as the initial therapy for chronic pancreatitis local septic complications. Surgery is reserved for patients in whom PCD fails. PMID- 18677916 TI - [PSA variations in persons with latent prostate cancer]. AB - The introduction and common use of serum PSA (Prostate Specific Antigen) has been demonstrated a useful index on latent prostate cancer diagnostic but in the same time has increased surgical intervention on histological forms with no eventual future evolution. Benign comportment of latent carcinomas being well known in advance, we correlated in vitro serum PSA from latent tumors, with the samples from a control group (prostates without signs of malignization). Levels of PSA were slightly elevated compared to age norms, mainly in cases with a large coexistent hypertrophy. Our reduced sample does not stand any statistic analysis, but this observation could eventually explain increased diagnostic and hyper treatment of non-important carcinomas from a clinical point of view. PMID- 18677917 TI - [Seven years of cochlear implant at the ENT Clinic of "Recuperare" Hospital Iasi]. AB - This paper presents our Cochlear Implant Departments results from the beginning until present time. Our implant list contains 161 candidates. The enlisted was made based on the criteria's for implant candidates' selection and we have implanted already 17 of them (11 children and 6 adults). Surgical interventions were done according to standard procedure - the approach being through mastoidectomy and posterior tympanotomy. We discuss the results obtained with cochlear implantation in each case. The outline idea is that the cochlear implant was a good choice in all our implanted cases and the patients had achieved at least the performances predicted before implantation for every particular situation. PMID- 18677918 TI - [Effectiveness of laparoscopic ovarian "drilling" in patients with polycystic ovarian syndrome and ovarian infertility]. AB - THE AIM of this study was to demonstrate the efficiency of laparoscopic ovarian drilling in the treatment of Polycystic Ovarian Syndrome (PCOS) associated with infertility. MATERIAL AND METHODS: We studied 54 patients with PCOS that also presented infertility and menorrhea. The PCOS was diagnosed by ultrasound in the 2nd Clinic of Obstetrics and Gynaecology in Iasi, using Bataglia and ASRM/ESHRE criteria. We selected 32 patients with PCOS and infertility, in which medical treatment with inductors of ovulation and Metformin failed and laparoscopy was indicated. In these patients we practiced bilateral laparoscopic ovarian drilling. RESULTS: In the next 3 months the menstrual cycle became regulated and 3 of the patients became pregnant. Other 10 patients became pregnant in the next 9 months after laparoscopic ovarian drilling. CONCLUSION: PCOS associated with infertility should be laparoscopy treated because the effect is faster and the fertility prognosis is better. PMID- 18677919 TI - [Caudal anesthesia in pediatric surgery]. AB - The study analyses the efficacy and the safety of caudal anesthesia for the intra and postoperative pain management in children. MATERIAL AND METHOD: A retrospective study was conducted over a five years period enrolling 150 children scheduled for various surgical procedures. We used the standard technique ("loss of resistance", "single shot", lateral decubitus). RESULTS: The most important age groups were 1 month - 1 year, 64 cases (42.66%) and 1-6 years - 71 cases (47.33%). We performed 144 caudal blocks associated with general anesthesia, 3 associated with sedation and 3 simple caudal blocks. In 129 cases, we used bupivacaine 0.25% as a single analgetic substance, in five cases bupivacaine 0.25% with lidocaine 1%, and in 16 cases bupivacaine 0.25% with fentanyl 1 mcg/kg. The quality of the analgesia was very good in 128 cases (85.33%), good in 12 (8%) and poor in 10 cases (6.66%). The morbidity rate was 13.33% (20 cases), the most frequent complication being urinary retention (8 cases - 5.33%). CONCLUSIONS: The caudal block is an efficient procedure for intra and postoperative analgesia in children. Associated with general anesthesia it is very useful for the management of the age groups 1 month - 1 year and 1-6 years. PMID- 18677920 TI - [Giant uterine fibroma. Case report]. AB - The presence of the giant abdominal-pelvic tumours is more and more rare because of the precocious methods of diagnosis. Sometimes the rare cases of giant uterine fibroma are met in medical practice and they need the use of atypical surgical techniques that are not entirely risk-free. In the present case, the giant uterine fibroma that was diagnosed (23/21/20 cm) was investigated clinically, and with imaging and pathologic techniques, then it underwent a surgical procedure in two stages, with major difficulties, because the entire anatomical structure of the pelvis was totally modified. PMID- 18677921 TI - Nutrigenomics in postgenomic era. AB - The last years modified major our conceptions about nutrition. These revolutionary changes were produced by implementation of new techniques of functional genomics. The nutrigenomics and nutrigenomics provide powerful approaches to unravel the complex relationships between bioactive molecules, genetic polymorphisms and biological system and can give rise to personalized nutrition and dietary recommendations. In monogenic diseases (phenylketonuria, galactosemia, lactose intolerance etc.) diet influence phenotypic expression and nutrigenomics will improve the prevention or treatment by the early identification of specific mutations or haplotype combinations that modulate dietary response in affected subjects. In the multifactor diseases, like cardiovascular diseases (CVD), obesity, type II diabetes mellitus or cancer, the nutrigenomics approach has begun to reveal that some of them are susceptible to dietary intervention and may modulate the onset and progression of disorders. PMID- 18677922 TI - [Structural changes of tumor microenvironment in liver metastases of colorectal carcinoma]. AB - The immunohistochemistry represents a very effective tool for describing the biological phenomena that characterize the primary or secondary neoplastic development. By using a complex immuno-morphologic approach, our study aims to develop a characterization of the metastatic liver microenvironment, based on the following features: similarities and differences in the proliferation activity, particular aspects of vascularity and immune response, the cellular dialogue generated at the level of the parenchyma - neoplastic tissue interface, the structure of the associated stroma. MATERIAL AND METHODS: Ten cases of liver metastases have been studied, including five treated exclusively by surgery (group 1) and five with recurrent metastases occurred after a previous treatment by thermonecrosis (group 2). The cases were investigated by routine histopathological exam and by immunohistochemistry. The choice of the antibodies was motivated by the follow-up of the parameter classes presented below: (i) proliferation/apoptosis; (ii) structure; (iii) angiogenesis and lymphangiogenesis; (iv) immunological reactivity; (v) cytokines. RESULTS: The elements defining the histopathologic and immunohistochemical patterns allowed a comparative evaluation of the metastasis models considered for the two groups of studied cases. The analysis of proliferation / apoptosis parameters proved a high aggressiveness in tumor proliferation for both types of metastases. Recurrent metastases showed an increased angiogenesis and a moderate lymphangiogenesis, as well as a massive matrix reshaping. The evaluation of the inflammatory infiltrate suggested that the development of the liver metastases is not accompanied by significant immunologic rejection, but it rather induces the tissue remodeling at the invasion border level. The recurrence corresponds to a typical interval of 6 months or longer, and is associated with intense fibrogenesis, angiogenesis, and inconsistent inflammatory infiltrate. CONCLUSIONS: Therefore, the efficiency of the thermonecrosis procedure with water vapors seems to drastically depend on the complete necrosis of the target tumor, plus the adjacent cells of the parenchyma in a neighborhood of some millimeters. Moreover, the therapy must include anti angiogenic resources as well as inflammation modulators, since the vascular supply and the leukocytic activity support the metastasis development. PMID- 18677923 TI - [Animal model of acute respiratory distress syndrome]. AB - Studying the animal models of acute respiratory distress syndrome (ARDS) represents a key factor in understanding the pathophysiology of this condition. OBJECTIVE: Creating a simple animal model of ARDS based on oleic acid injection, suitable for studying the alveolo-capillary membrane dysfunction. MATERIAL AND METHOD: 8 common rabbits weighting between 3 and 3.5 kg were subjected to peripheral vein injection of 0.5 ml of oleic acid. During the first 75 minutes chest radiographs were performed at each 15 minutes. At the end of this period an arterial blood-gas analysis was done, the animals were sacrificed and a complete autopsy and a microscopic examination of the lung were performed. RESULTS: Clinical, radiologic and histological features of ARDS were obtained in all cases after 75 minutes of oleic acid injection. CONCLUSION: Intravenous injection of oleic acid consistently induces ARDS in rabbits. The model is simple, reproducible and suitable for animal experimentation. PMID- 18677924 TI - [Ki-67 and p53 expressions versus differentiation degrees of colon adenocarcinoma]. AB - PURPOSE: The colon carcinoma prognostic is evaluated with respect to four categories of factors established by College of American Pathologists Consensus Statement 1999, among which a series of molecular markers is also included. Our study aimed to develop semiquantitative analysis of the immunohistochemistry pattern for proliferation and apoptosis markers, as well as of their connections with the histopathology subtypes of colon adenocarcinoma. MATERIAL AND METHODS: The group of study included 5 cases of well-differentiated adenocarcinoma, 5 cases of moderately differentiated adenocarcinoma and 5 cases of poor differentiated adenocarcinoma. The Ki-67 and p53 antibodies (DAKO) and the streptavidin-biotin method (LSAB Kit, DAKO) were used for the immunohistochemical staining. The semiquantitative analysis provided the Ki-67 and p53 indices, and subsequently their values were statistically correlated with the histologic differentiation degrees. RESULTS: All the cases presented positive immunohistochemistry reaction for Ki-67, the Ki-67 index varying on a large scale, between 5% and 95%. For p53, 10 out of the 15 cases (66%) presented positive immunohistochemistry reaction, with the index value between 22% and 95%, and 5 cases were negative. The statistical analysis revealed no significant difference between the Ki-67 indices of the three histopathologic subtypes (p = 0.674), whereas important differences appeared for the p53 indices (p = 0.007). CONCLUSION: Our findings support the correct understanding of the role played by the immunohistochemical markers, in the sense of the risk involved by considering a pre-definite hierarchy in the diagnosis strategy. PMID- 18677925 TI - [Gastric lymphoma. Morphological and immunohistochemical study]. AB - Most of extranodal lymphomas are localized in gastrointestinal tract, gastric lymphoma representing more than 50% of them. The role that Helicobacter pylori (H. pylori) plays in pathogenesis of gastric lymphoma has changed the therapeutic approach. AIMS: Description of morphological features and immunohistochemical pattern of gastric lymphomas from patients admitted in University Hospital "Sf. Spiridon" Iasi. MATERIAL AND METHODS: Thirty four gastric lymphomas were investigated using routine histopathological technics and immunohistochemical staining based on a large panel of antibodies: CD3, CD5, CD20, CD79a, CD23, CD30, cyclin-D1, BCL2, BCL6, ALK1, Ki67, CK-cocktail, anti-H. pylori. RESULTS: All gastric lymphomas were localized in the antrum, most of them being solitary and large-sized tumors. Ninety-seven percent were B-cell lymphomas, 41.17% were mucosa-associated lymphatic tissue lymphomas (MALT lymphomas), and the remaining were high grade lymphomas. Only one case was classified as peripheral T-cell lymphoma. Cytokeratin cocktail immunostaining improved the detection of typical lymphoepithelial lesions, which characterized exclusively the MALT lymphomas. The sensibility for H. pylori detection in gastric lymphoma cases was increased by 22% using anti-H. pylori antibodies. CONCLUSIONS: Immunohistochemistry is a diagnostic method for gastric lymphomas, being useful in identification of lymphoepithelial lesions, detection of H. pylori infection, and is mandatory for lymphomas classification according to WHO criteria. PMID- 18677926 TI - Flow cytometry-based quantification of cytokine levels in AML patients plasma and cell culture supernatants. AB - AIM: Bone marrow stromal cells (BMSCs) have been found to support leukemic cell survival; however, the mechanisms responsible are far from being elucidated yet. The main aim of the current study is to identify particular cytokine/chemokine patterns of acute myeloid leukemia (AML) cells, and, on a longer term, to correlate them with the patient outcome and response to therapy. Therefore, the influence of BMSCs on in vitro modulation of cytokine secretion (IL-1beta, TNF alpha, IL-10, IFN-gamma, IL-4, IL-5, and IL-2) by AML cells as well as the AML cells supportive capacity of BMSCs-derived soluble factors was investigated. MATERIAL AND METHODS: With this purpose, we used an in vitro experimental model consisting in the evaluation of the effect of BMSC confluent layers-conditioning medium (BMSC-CM) on M4/5 AML cell cultures. RESULTS: Our results show that BMSC CM from both AML patients and healthy subjects conferred a substantial beneficial effect on AML cells throughout the culture (p=0.0002 and 0.0020 respectively at 24 hours and p=0,0013 and 0,0030 respectively at 72 hours), with a temporary increase in AML cell viability conferred by BM plasma from AML patients. Significant differences were observed with respect to IL1 b secretion which was upregulated in AML cell cultures both after 24 and 72 hours following the addition of AML-BMSC-CM, in contrast to control-BMSC-CM. CONCLUSION: Our results suggest the contribution of BMSCs from AML patients to the generation of particular factors which may be key players involved in the in vivo maintenance of the malignant clone. PMID- 18677927 TI - [Ultrastructural aspects in the development of the human nephron--electron microscopy study]. AB - We have looked for electron microscopy aspects tissue fragments of human nephrons harvested from fetuses of 7,5-19 weeks to show ultrastructure aspects of the nephron during development. MATERIAL AND METHODS: We have used 9 fragments from human embryos; Four cases were aged of 7,5, 8, 12 and 17 weeks. The other 5 cases were between 18 and 19 weeks. Tissue fragments were fixed in glutaraldehyde at 4 degrees C and processed by the classic technique. RESULTS: The electron microscopy study shows the evolutive steps of the human nephron (stages I-IV), stages met for the 12-19 weeks embryo. For the cases under this age the meta nephrogenic blastema was dominant. In the IVth development stage, the cells in the proximal tubule show a higher development degree than the distal segments. CONCLUSION: These observations indicate that during early development the proximal segment is better developed and probably more functional than the distal segments. PMID- 18677928 TI - [The arcuate eminence of the petrous portion of the temporal bone. Descriptive features]. AB - The superior semicircular canal is an important landmark in the middle cranial fossa approach to the temporal bone and the internal acoustic meatus. This mark is frequently located by its topographic association to the arcuate eminence, a bony prominence located on the anterior surface of petrous portion of the temporal bone. In recent studies, otolaryngologists described that the arcuate eminence lies more posterior and laterally than the superior semicircular canal and stated that some eminences corresponded to sulci of the temporal lobe, of which most were traces of the occipitotemporal sulcus. Our work examines by dissection the relationship of the arcuate eminence to the superior semicircular canal, demonstrating a third origin of the arcuate eminence: well pneumatized temporal bones in the region of the epi-tympanic recess. PMID- 18677929 TI - [The effects of chronic exposure to cigarette smoke on vasomotor endothelial function of guinea pig pulmonary arteries]. AB - Chronic exposure to cigarette smoke on guinea-pigs induced a muscularization of pulmonary arterioles and endothelial dysfunction might be an early trigger of this vascular remodelling. Accordingly, the present study was aimed to evaluate the effect of a 12 weeks exposure to passive smoking on the pulmonary endothelial vasomotor function. MATERIAL AND METHOD: Vasodilator response of pulmonary arteries rings isolated in organ bath, precontracted with phenylephrine, was compared in the presence of cumulative doses (10(-9) to 10(-4) M) of endothelial dependent (acetylcholine, ACh and adenosine-diphosphate, ADP) and independent vasodilators (sodium nitroprusside, SNP), respectively. RESULTS: Our results suggest that chronic exposure of guinea-pigs to cigarette smoke induces the impairment only of NO-mediated endothelial response (vasodilation was 9.83 +/- 4.36 % for ACh 10(-5)M vs. 39.72 +/- 16.61 % in control, p = 0.005, respectively 36.64 +/- 7.21 % for ADP 10(-5)M vs. 55.53 +/- 13.51 %, p = 0.02). CONCLUSION: The in vitro study of pulmonary arteries vasomotor function in guinea pigs chronically exposed to cigarette smoke may represent a reliable and relevant experimental model for the assessment of pulmonary endothelial dysfunction in early stages of COPD. PMID- 18677930 TI - [Anatomo-clinical considerations regarding the celiac-mesenteric junction]. AB - AIM: This work points out the access ways - both the classical ones and the ones less known and used - to the origin and the first centimeters of the superior mesenteric artery (SMA), underlining the advantages and disadvantages of each of them. MATERIAL AND METHOD: Our work is based on dissection of 30 human bodies and 200 selective arteriographies. RESULTS: There are two classical ways to access the first part of SMA: inter-duodeno-pancreatic and the left thoraco-abdominal ways. CONCLUSIONS: An access way to the origin of the SMA is necessary for the revascularisation through different techniques of the visceral territory depending on the SMA, taking into account the fact that at this level we have the most frequent localization of the atheroma plaques, which leads to acute or chronic intestinal ischemia (manifest in the form of postprandial abdominal 'angor'). PMID- 18677931 TI - The accuracy of alveolar crest dimensions measurement for dental implants. In vitro study. AB - OBJECTIVES: The aim of this study was to evaluate the accuracy of radiological examination methods in quantitative preimplantar bone assessment. MATERIAL AND METHOD: The study was carried out on a dry cranium, totally edentate. There have been radiological appreciated the height of the alveolar process and its width, both to the maxilla and the mandible. Also, it has been established the distance between the free edge of the alveolar crest and the superior edge of the mandibular canal. The values radiological measured have then been compared with those obtained through direct measurement of these dimensions on the cranium. RESULTS: The results were performed in tables and diagrams. The greatest deviation from the real values was at the panoramic radiography, while the computer tomography was the most accurate examination among the radiological techniques used in this study. The panoramic radiography doesn't give information about the alveolar crest thickness, about it's declivity and the alveolar crest measurement generates significant errors. Linear tomography implies a difficult technique, but it has the advantage of a low irradiation,a wider accessibility and a lower cost than the CT. Computer tomography is the most accurate preimplantar evaluation technique but it has the disadvantage of a high patient radiation. CONCLUSIONS: The panoramic radiography represents an insufficient method for appreciating the preimplantar bone status. That's why it has to be supplemented with other radiological examinations, especially at the cases with alveolar crest atrophy, to avoid accidental penetration of adjacent structures. PMID- 18677932 TI - [New morpho-functional rehabilitation methods in cleft lip and palate]. AB - The interdisciplinary, complex therapeutic protocol of the cleft lip and palate patients, applied in the Clinic of Cranio-Maxillo-Facial Surgery of "I. Hatieganu" University of Medicine and Pharmacy Cluj-Napoca, involves the morphologic reconstruction as well as the functional rehabilitation. Functional rehabilitation is the aspect, which gives the esthetics, social and familial integration of the patient, offering good quality of life. METHOD: In the current study, a new method and concept of improving the phonetic function in the primary and secondary surgical steps, with the effect on muscle and bone, is presented. The new surgical techniques used comprise of the surgery of the levator veli palatini using the method designed by Sader, and bone distraction, during the same surgical procedure. The assessment of the phonetic results was performed using the NARSOM test. RESULTS: Following up the results of the techniques mentioned above, we consider that they improve extremely well the morphological status, while giving a functional and physiological support to the patient. CONCLUSION: Thus, they offer optimal conditions for the future progress of functional rehabilitation using specific speech therapy methods. PMID- 18677933 TI - [Study concerning the influence of the casting parameters of dental alloys on the cast prosthetic rehabilitation]. AB - The integration of a cast rehabilitation into the oral environment and its clinical longevity are mainly determined by the accuracy reproduction of the technological parameters. The aim of this study was to evaluate the structural defects induced by the modification of the technological conditions of casting the conventional and titanium alloys. MATERIAL AND METHOD: We used 35 cast prosthetic rehabilitation, divided into 7 groups. For each group we modified one parameter: the width of the wax pattern, the design and topography of the casting channels, the absence of the cleanliness of the wax pattern, the preparation of the investment material, the thermal system for the mold preparation and reduced quantity of metal used for casting. For the 7-th group, represented by titanium cast rehabilitation, we modified the design and dimensions of the casting channels. RESULTS: The changes in the technological conditions induced porosity, incomplete frameworks or plus on the prosthetic appliances. The fundamental theoretical knowledge of the clinician and dental technician and the strictly application of the technologies assure the premises of the satisfaction of the patient. PMID- 18677934 TI - [A combined surgical-orthodontic approach of a class II/1 malocclusion. A case report]. AB - This case report highlights the possibility of a non extraction orthodontic treatment, surgically assisted, in a Class II Division 1 malocclusions in order to reduce maxillary protrusion prior to the surgical advancement of the mandible for the correction of the skeletal deficiency. The combined orthodontic-surgical approach optimizes the profile while allowing the settling of a Angle Class I occlusion at the end of the entire treatment sequence. PMID- 18677935 TI - Synthesis and characterization of some new bishydrazones derived from isatin. AB - The scientific literature states that isatin and its derivatives exhibit antibacterial, antifungal, anticonvulsant, cytotoxic and anti-HIV activities. The aim of our study was to enlarge the number of isatin derivatives. MATERIAL AND METHOD: We synthesized some new hydrazones and then, the corresponding bishydrazones using salicylaldehyde. By condensing isatin and its derivatives (methyl-, nitro-, brom-) with hydrazine, corresponding hydrazones were obtained. The bishydrazones were synthesized by refluxing the obtained hydrazones with salicylaldehyde in methanolic solution. The structure of new compounds was confirmed by elemental and spectral analysis. The lipophilicity of the synthesized derivatives was expressed by clog P values. The antibacterial and antifungal activities of the compounds were determined using disk-diffusion method against Gram positive, Gram negative bacteria, and Candida albicans. RESULTS: New bishydrazines derived from isatin have been synthesized; their structures have been confirmed by elemental and spectral analysis. PMID- 18677936 TI - [Research concerning rutin semisynthetic derivatives. Synthesis, physico-chemical and microbiological characterisation of some new rutin-isoniazid derivatives]. AB - Isoniazid is one of the most used antituberculosis drugs, but its toxicity is high. In this study, four derivatives of rutin and isoniazid were prepared and their antibacterial and antimycotic activities were investigated. METHOD: Synthesis of these compounds started through reaction of rutin with fresh prepared sodium methoxide, subsequently treating with 1, 3-dichloro-2-propanol, 1 brom-3-chloropropane, 1, 2-dibrom-ethane or dibrommethane and finally, stirring with isoniazid, to afford crystalline, yellow, water soluble powders. RESULTS: The four compounds were marginally active against Staphylococcus aureus and Staphylococcus saprophyticus and were inactive against Escherichia coli. CONCLUSION: We synthesised four new derivatives of rutin and isoniazid; melting points, solubility, yields characterised these compounds, and their structure was confirmed by C, H, N elemental analysis and UV spectra. The compounds showed a modest antibacterial activity. We think that the toxicity of the new derivatives will be lower that of isoniazid toxicity. The next steps in our research will be the toxicological study and antituberculosis assay. PMID- 18677937 TI - Serum zinc and copper levels in some dermatological diseases. AB - Zinc (Zn) and copper (Cu) are essential metals for biochemical processes in the body. Many researchers have main interest in these two metals because it was establish that they are involved in immunological and inflammatory reactions. MATERIAL AND METHODS: The study comprised 49 patients of Dermatology Clinic from Iasi with different skin diseases (21 with psoriasis and 28 with varicose ulcer), during January - May 2006. The patients, both sexes, had ages between 19 and 81 years old. We determined serum Zn and Cu levels by atomic absorption spectrophotometry. The results were expressed in mg/L and we calculated Cu/Zn ratio, too; the values were compared with 50 healthy controls. RESULTS: In psoriasis, serum Zn and Cu mean levels (5.61 mg/L, respectively 1.27 mg/L) were higher as compared to controls (significance level p=0.04); the mean Cu/Zn ratio was 0.81 in psoriasis and 0.96 in control group. In varicose ulcer, mean levels were significantly increased (Zn-7.62 mg/L, Cu- 1.38 mg/L) compared to control group (Zn-0.81 mg/L, Cu-0.75 mg/L) and serum normal values; the mean Cu/Zn ratio was 1.00. 18 of all patients with psoriasis and varicose ulcer had higher serum Zn and Cu levels than normal values. PMID- 18677938 TI - [Production of chemoattractant-induced reactive oxygen species in human neutrophils--role of protein kinase C isoforms]. AB - The aim of our study was to characterize the priming effect of extracellular nucleotides on reactive oxygen species (ROS) production induced by formyl methionyl-leucyl-phenylalanine (fMLP), interleukin-8 (IL-8), leukotriene B4 (LTB4), and platelet activating factor (PAF). Also, we investigated the roles played by different protein kinase C (PKC) isoforms in nucleotide-induced priming. ROS production was determined by an isoluminol-based assay. MATERIAL AND METHOD: Nucleotide-induced priming was concentration- and time-dependent. The concentration of UTP able to cause maximal priming was 10 microM. When UTP (10 microM) was administered prior the agonist, the increase of the amplitude of the response reached the maximum at 1 minute of preincubation with the nucleotide. RESULTS: Calcium depletion of neutrophil caused significant inhibition of ROS production induced by all agonists tested, but did not affect the priming effect of the nucleotides. We tested the effect of several PKC inhibitors on the nucleotide-induced priming. GF 109203X (5 microM), an inhibitor of all neutrophil's PKC isoforms, or RO 31-8220 (5uM), an inhibitor of classical and novel PKC isoforms, abolished the responses induced by fMLP (10 nM) IL-8 (10 nM), LTB4 (100 nM) or PAF (100 nM). Go 6976 (100 nM), a selective inhibitor of classical PKC isoforms, had no effect on nucleotide-induced priming, suggesting that activation of these PKC isoforms does not play a role in the priming effect. Rottlerin (5 microM), a PKC delta inhibitor, almost abolished the effect of fMLP in the absence or in the presence of UTP, indicating that PKC delta is essential for the fMLP-induced effect; rottlerin also caused inhibition of ROS production induced by IL-8, LTB4 or PAF, however the priming effect of UTP was not affected for these chemoattractants. Our data suggest that classical PKC isoforms do not play a role in chemoattractant-induced ROS production. CONCLUSION: Although fMLP induced effect appears to be highly dependent on PKC delta activation, other chemoattractants are able to cause ROS production through PKC delta-independent mechanisms. PMID- 18677939 TI - [Monitoring system for cardiovascular parameters of drivers]. AB - In the world the road traffic accidents are in the 3rd place at WHO cause-of death statistics. One of the main causes of these accidents is driving fatigue as results of stress. The aim of this study is to monitoring the heart rate variability for identifying the grade of stress of the drivers. MATERIAL AND METHOD: For this stage of the study we design and build an device that recording the ECG signal and the heart rate. We analysed of line the heart rate variability for determining one possible link between the grade of fatigue and heart rate variability. The device use only two electrodes placed on the steering wheel. RESULTS: We use three categories of subjects: beginner drivers, amateur drivers with more than 3 year experience, and expert drivers. The monitoring is made for 30 minutes in intensive traffic. CONCLUSION: The preliminary results are satisfactory which determine us to continuing the research in this direction. PMID- 18677940 TI - [Superficial microwaves hyperthermia tumoral temperature distribution by using finite element analysis]. AB - The main aim of this study is to present a virtual method for tumor temperature distribution by using finite element analysis. MATERIAL AND METHODS: First step is geometrical synthesis that means tumor and neighbor structures three dimensional reconstruction using CT sections; the second represent temperature and material synthesis: temperature values application and tissues density, thermal conductivity, specific heat values application, then finite element analysis. RESULTS: Finite element analysis indicate tumor temperature distribution with maximum and minimum values as a result of microwave hyperthermia treatment. PMID- 18677941 TI - [International Congress of Dental Medicine. Celebration of Dental Faculty days, the 12-th edition, Iasi]. PMID- 18677943 TI - [Meaning of the Weber-Fechner law and the principle of displacement]. PMID- 18677942 TI - [Event-related potentials in different stages of the operation of visual working memory]. PMID- 18677944 TI - [Event-related potentials associated with a shift in the strategy of visual perception during recognition of a hierarchical stimulus]. PMID- 18677945 TI - [Visual-motor synchronization: analysis of the initiation and stable synchronization phases]. PMID- 18677946 TI - [The effect of craniotomy on the intracranial hemodynamics and cerebrospinal fluid dynamics in humans]. PMID- 18677947 TI - [The effect of motor activity on the formation and maintenance of the function of time perception in the course of ontogeny]. PMID- 18677948 TI - [Changes in the power of the low- and high-frequency bands of the heart rate variability spectrum in coronary heart disease patients with different severities of coronary atherosclerosis in the course of load tests]. PMID- 18677949 TI - [Functional reserves of the human body as assessed by Doppler ultrasonography]. PMID- 18677950 TI - [Cardiac and respiratory oscillations of the blood flow in microvessels of the human skin]. PMID- 18677951 TI - [Comparative analysis of the psychophysiological parameters of the system quanta of performance in physically trained and untrained subjects]. PMID- 18677952 TI - [Using molecular genetic methods for prognosis of aerobic and anaerobic performance in athletes]. PMID- 18677953 TI - [Mechanisms of changes in human hemodynamics under the conditions of microgravity and prognosis of postflight orthostatic stability]. PMID- 18677954 TI - [Effect of ecological factor on the functional state of adolescents]. PMID- 18677955 TI - [Integrated evaluation of the influence of a northern environment on human metabolism, physiological, and psychoemotional states]. PMID- 18677956 TI - [Prospects of studying the polymorphisms of key genes of the neurotransmitter systems: Communication II. The serotonergic system]. PMID- 18677957 TI - [Distribution of students according to profile of lateral organization of functions]. PMID- 18677958 TI - [The pumping function of the heart in athletic tourists during a PWC170 exercise]. PMID- 18677959 TI - [The neurohormonal mechanism of the effects of health promotion procedures on pregnant women]. PMID- 18677960 TI - [Changes in the beta-adrenergic and M-cholinergic modulating activities of the blood serum and urine in bronchial asthma]. PMID- 18677961 TI - [Direct development in the tailed amphibians (salamanders), its origin and evolution]. AB - Biphasic life circle including water larvae and terrestrial adults is the primitive character for recent amphibians. Respectively, the larval development type (sensu Zakhvatkin, 1975) is also primitive for them. However, many amphibians possess direct development, in which the most part of ontogenesis takes place in the egg, and a miniature copy of the adult adapted to the terrestrial mode of life comes into the world. Transition from the larval type to the direct one occurs several times independently in Apoda, Urodela, and Anura. Pathways and mechanisms of formation of the direct type, its evolutionary tendencies, and ontogenetic prerequisites of that transition are studied in the plethodontid urodelans herewith in details. It is observed that the entire process of initial formation and subsequent specialization of the direct type involves: 1) progressing displacement of ontogenesis into embryogenesis and loss of larval characters, 2) desynchronization of initially metamorphic transformations and processes with their progressive lost, 3) acceleration of the beginning of functional activity of the thyroid gland, and 4) subsiding of the role of thyroid hormones in the ontogenesis regulation. Transition to the direct development type involves similar kinds of ontogenetic transformations and regulations in both Anura and Urodela despite of their independent evolution. Respectively, mechanisms of that transition are universal for the amphibians. The ontogenetic prerequisites of that transition are shown to be either significant dissociability of the larval and adult stages of ontogenesis (in anurans and plethodontid urodelans), or absence of the extreme larval specializations and respective sharp differences between larvae and adults (in extant Apoda and extinct labyrinthodonts and seymouriamorphs). PMID- 18677962 TI - [Hyperbolic growth of marine and continental biodiversity through the phanerozoic and community evolution]. AB - Among diverse models that are used to describe and interpret the changes in global biodiversity through the Phanerozoic, the exponential and logistic models (traditionally used in population biology) are the most popular. As we have recently demonstrated (Markov, Korotayev, 2007), the growth of the Phanerozoic marine biodiversity at genus level correlates better with the hyperbolic model (widely used in demography and macrosociology). Here we show that the hyperbolic model is also applicable to the Phanerozoic continental biota at genus and family levels, and to the marine biota at species, genus, and family levels. There are many common features in the evolutionary dynamics of the marine and continental biotas that imply similarity and common nature of the factors and mechanisms underlying the hyperbolic growth. Both marine and continental biotas are characterized by continuous growth of the mean longevity of taxa, by decreasing extinction and origination rates, by similar pattern of replacement of dominant groups, by stepwise accumulation of evolutionary stable, adaptable and "physiologically buffered" taxa with effective mechanisms of parental care, protection of early developmental stages, etc. At the beginning of the development of continental biota, the observed taxonomic diversity was substantially lower than that predicted by the hyperbolic model. We suggest that this is due, firstly, to the fact that, during the earliest stages of the continental biota evolution, the groups that are not preserved in the fossil record (such as soil bacteria, unicellular algae, lichens, etc.) played a fundamental role, and secondly, to the fact that the continental biota initially formed as a marginal portion of the marine biota, rather than a separate system. The hyperbolic dynamics is most prominent when both marine and continental biotas are considered together. This fact can be interpreted as a proof of the integrated nature of the biosphere. In the macrosociological models, the hyperbolic pattern of the world population growth arises from a non-linear second order positive feedback between the demographic growth and technological development (more people - more potential inventors - faster technological growth - the carrying capacity of the Earth grows faster - faster population growth - more people - more potential inventors, and so on). Based on the analogy with macrosociological models and diverse paleontological data, we suggest that the hyperbolic character of biodiversity growth can be similarly accounted for by a non-linear second-order positive feedback between the diversity growth and community structure complexity. The feedback can work via two parallel mechanisms: 1) decreasing extinction rate (more taxa- higher alpha diversity, or mean number of taxa in a community - communities become more complex and stable - extinction rate decreases - more taxa, and so on) and 2) increasing origination rate (new taxa facilitate niche construction; newly formed niches can be occupied by the next "generation" of taxa). The latter possibility makes the mechanisms underlying the hyperbolic growth of biodiversity and human population even more similar, because the total ecospace of the biota is analogous to the "carrying capacity of the Earth" in demography. As far as new species can increase ecospace and facilitate opportunities for additional species entering the community, they are analogous to the "inventors" of the demographic models whose inventions increase the carrying capacity of the Earth. The hyperbolic growth of the Phanerozoic biodiverstiy suggests that "cooperative" interactions between taxa can play an important role in evolution, along with generally accepted competitive interactions. Due to this "cooperation", the evolution of biodiversity acquires some features of a self-accelerating process. Macroevolutionary "cooperation" reveals itself in: 1) increasing stability of communities that arises from alpha diversity growth; 2) ability of species to facilitate opportunities for additional species entering the community. PMID- 18677963 TI - [Long-term response of the plant species of four north-west caucasus alpine communities on their transfer into the new ecological conditions]. AB - Under consideration are plant species stability in respect to their occurrence in the foreign communities, and a role of abiotic factors in development of alpine comminities. The experiment was started in 1989 on Malaya Khatipara Mt (NW Caucasus) at the altitude of 2750 m. a. s.l. included reciprocal transplantation of tussock sites between four alpine communities. Significant changes in the tussock composition were observed due to better or worse acclimation of particular species, due to lost of some species, and because of invasion of aboriginal species from the surroundings into the transplanted sites. The most species involved appeared to be stable in respect to transplantations and some of them even better developed under new ecological conditions, the latter case indicating lack of coincidence of the aut- and sinecological optima. PMID- 18677964 TI - [Urbanization mechanisms in bird species: population systems transformations or adaptations at the individual level?]. AB - The present research deals with urbanization of wild bird and mammal species. Forms and mechanisms of population steadiness in the urban landscape have been examined. The urbanization process turned out to be a directed change of the population system forming de novo in the urbolandscape leading to a sustainable organization peculiar for the particular environment. The population organization of different types in urbolandscape is found to provide its stability under conditions of directed and fast changes accompanied with instability and heterogenous structure of habitats. It is shown that the same type of population organization meets the corresponding demands among different species settling in the urban environment. Its features are "openness" and "flowage" of the groups, far order of settlement levels and other units of population system, constant movements of the individuals between the groups as a respond to the signals of urboenvironment significant changes. The "urban" variant of the population system organization turns out to be opposite to that of the same species in the non urban habitats. After formation of the urban types by the species and successful developing of the town, the urban population becomes separated from the maternal local population and begins to exist independently in the urban landscape. The variety of adaptation aberrations in ecology, behavior, and mode of life of urban birds is the population system stability function in the urban landscape and is not a results of individual selection. It is shown that the urbanization process of the species goes firstly on the population level being the system structure transformation developed by the species towards the most stable state in the town (city) territory. Only after the appearance of stable urban population, the urban individuals show the rapid growth of different changes in ecology, behavior, mode of life that was traditionally described by naturalists as species adaptation to the city conditions. The key features of urban population stability/instability are described. Their application to closely related species allows us to distinguish potential urbanists from instable and vulnerable species that could be soon pushed out of the city. The application of corresponding criteria to the urban populations of such species constituting one guild allows us to predict if their developing in the given town would be successful or unsuccessful. The latter is very important since close species are, as a rule, ecologically indistinguishable in the urbanized landscapes. So one can not predict successful/unsuccessful urbanization taking into account the differences in the range of habitats, breeding success, and other external features. PMID- 18677965 TI - [Dicotyledons of cretaceous, palaeogene, and neogene. Adaptogenesis of the terminal phloem]. AB - A database containing information on the trait complex for terminal phloem of dicots leaf having been created during the last thirty years was analyzed. The typology of terminal phloem was verified. The monotypy of plant taxa up to the family level was confirmed. For orders and subclasses, the wide distribution of structural parallelisms was demonstrated. The lists of plant families with similar structures of the phloem terminals was elaborated. As a result of comparative analysis of these lists and paleobotanic data on the age of plant families, the specificity of the types of terminal phloem and the associated morphological and functional characteristics of the groups of families of the same evolutionary, age were revealed. Ancestral type prevails among the families belonging to Prodicots, which were emerged during Cretaceous. Symplastic type is characteristic to the families of Eudicots and biomes appeared in Paleogene, while the apoplastic type is characteristic to the families and biomes of Neogene. For each group to taxa, plant stem and root parasitism is characteristic for the crown taxa completing the structural sequences of trees and herbs. All these data allow discussion of the questions of climatic adaptogenesis of the character complex studied here, the impossibility of structural iterations in morphological evolution, and the reasons of similarities and differences in the evolutionary and contemporary zonal series of the terminal phloem in Dicots. PMID- 18677966 TI - Intrinsic and acquired drug resistance in malignant tumors. The main reason for therapeutic failure. AB - Drug resistance is the major reason for failure in cancer chemotherapy. Resistance may be either pre-existent (intrinsic resistance), or induced by drugs (acquired resistance), So far, no strategy has been found to overcome resistance, which is based on highly complex and individually variable biological mechanisms. In present clinical practice, drug resistance can only be recognized during treatment, after long lag times. Thus diagnostic tests are re quired, indicating resistance at an earlier stage, in order to avoid unnecessary medication, frequently associated with toxic side-effects. A number of new anti-cancer drugs are now available. In contrast to the unspecifically acting cytostatic chemotherapy, these compounds have targeted actions. However, as recent studies have shown, resistances and severe side-effects can also be found with targeted drugs. With the increasing number of new treatment regimens, the early diagnosis of resistance will optimize therapy, and indeed will be indispensable for individual cancer therapy. The resistance assays available for use in clinical practice should be integrated into cancer therapy. Research into this neglected area needs to be intensified. PMID- 18677967 TI - Validation of the APP23 transgenic mouse model of Alzheimer's disease through evaluation of risperidone treatment on aggressive behaviour. AB - Valid animal models are indispensable in the drug discovery pipeline for dementia. Transgenic APP23 mice model Alzheimer's disease patients' memory deficits and additionally present with various behavioural disturbances, such as aggressive behaviour. The present study investigated and confirmed significant sensitivity of the model to the aggression-lowering ability of the antipsychotic agent risperidone (CAS 106266-06-2). The sensitivity for such anti-aggressive action contributes to the therapeutic predictive validity of the APP23 model of Alzheimer's disease, which can be used as a pre-clinical screening tool for the identification of novel anti-aggressive agents. PMID- 18677968 TI - Effect of buspirone on the behavioral regulation of rats in low versus high anxiety conditions. AB - INTRODUCTION: Buspirone (CAS 33386-08-2) is reported to have anxiolytic effects in humans and is mostly described for mild anxiety. To further explore the effects of buspirone on different levels of anxiety, the effect of buspirone was evaluated in two different conditions of the open field which were distinguished as low and high anxiety (enclosed and exposed open field, respectively). MATERIALS AND METHODS: Twenty-eight albino Wistar rats (350-400 g) were tested in two different arena settings, an enclosed and an exposed open field. Fourteen animals were initially injected with 1 ml saline while the others (n = 14) received buspirone 3 mg/kg. RESULTS: The data showed clear differences in the two open-field settings, suggesting a higher anxiety level in the exposed open field. In addition, correlation analysis showed that the two anxiety tests measure different aspects of anxiety. Buspirone treatment reduced the behavioral activity in both the enclosed and exposed open-field, which is generally interpreted as an anxiogenic effect. However, buspirone increased the time in the center areas and decreased the frequencies in the outer regions. These behavioral changes are generally seen as an anxiolytic effect. Correlation analysis showed that buspirone treatment disrupted the relation between indices of anxiety. CONCLUSION: These results showed that in an open-field setting buspirone appears to have a dual effect. The reduced activity and increase in time spent in the center areas are indicative of both an anxiogenic and an anxiolytic effect, respectively. This was found in both open-field settings, suggesting that the effects of buspirone are independent of the anxiety level. PMID- 18677969 TI - Effect of ibudilast on non-specific symptoms in patients with chronic cerebral ischemia. Analysis of cerebral blood flow. AB - BACKGROUND AND PURPOSE: Many patients with chronic cerebrovascular disease complain that dizziness and depression negatively affect their daily lives. In these patients, ibudilast (CAS 50847-11-5) reportedly ameliorated dizziness. The efficacy of ibudilast was investigated and its effect on the cerebral blood flow (CBF) was recorded. METHODS: The study population consisted of 11 patients (male and female) with chronic cerebrovascular disease complaining of dizziness or depression. They received 30 mg of ibudilast orally per day. The grade of vertigo and depression at entry into this study and 2 and 6 months after the start of therapy was recorded. Their depressive state was scored with the Japan Stroke Scale-Depression Scale (JSS-D) and their cerebral blood flow (CBF) was measured before and approximately 3 months after the start of ibudilast therapy. RESULTS: At 6 months after the start of ibudilast therapy, all patients reported the resolution of dizziness; of the 6 patients with depression at entry, all experienced significant improvement. At 3 months, the CBF was significantly increased in the right frontal (p = 0.019) and occipital cortex (p = 0.004) with no significant changes in the cerebellar folia, subcortical gray matter (striatum and thalamus), and other cerebral cortices in the right cerebral hemisphere. There were no significant CBF changes in any areas of the left cerebral hemisphere. CONCLUSION: In patients treated with ibudilast, the amelioration of dizziness and depression was accompanied by a CBF increase in the right frontal and occipital cortices. These findings suggest that the right frontal and occipital cortices may be related to their dizziness and depression. PMID- 18677970 TI - Bioequivalence of two oral formulations of triflusal capsules in healthy volunteers. AB - Triflusal (CAS 322-79-2) is an antiplatelet agent related to salicylates used in several European and Latin American countries in the treatment of cardiovascular diseases. The aim of this paper was to evaluate the bioequivalence of triflusal derived from two preparations using both parent drug and metabolite pharmacokinetic data. The bioavailabolity was measured in 24 healthy male Caucasian volunteers following a single oral dose (600 mg) of the test or reference products in the fasting state. Blood samples were collected for 120 h. Plasma concentrations of triflusal and its metabolite 3-hydroxy-4 trifluoromethylbenzoic acid (HTB) were analyzed by high-performance liquid chromatography with UV and fluorescence detection, respectively. The non compartmental method was used for pharmacokinetic analysis. Log-transformed Cmax, AUC0-t and AUC0-infinity were tested for bioequivalence using ANOVA and Schuirmann's two-one sided t-test. Tmax was analyzed by nonparametric pharmacokinetic parameters of triflusal and HTB derived from the two formulations were nearly consistent with previous observations. Triflusal parameters derived from the test and reference drug were as follows: Cmax (16.85 +/- 11.41 vs 14.48 +/- 7.22 mg/l), AUC0-t (18.43 +/- 10.91 vs 16.22 +/- 7.58 mg/l per hour), Tmax (1 range 0.25-2h vs 0.875 range 0.25-1.5 h), and t(1/2) (0.49 +/- 00.27 vs 0.76 +/- 0.64). HTB parameters after test and reference formulation administration were as follows: Cmax (68.13 +/- 23.05 vs 65.51 +/- 19.44 mg/l), AUC0-t (2748.18 +/- 971.91 vs 2877.97 +/- 881.2 h x mg/l), AUC0-infinity (3350.15 +/- 1182.62 vs 3372.49 +/- 1110.35 h x mg/l), Tmax (2 range 1-10 h vs 2 range 0.75-12 h), and t(1/2) (42.19 +/- 7.82 vs 43.13 +/- 6.56 h). 90% of confidence intervals for the test/reference ratio of Cmax AUC0-t and AUC0-infinity derived from both triflusal and HTB were found within the range of 80%-125% acceptable for bioequivalence. No significant difference was found between the Tmax values for triflusal and HTB. It was concluded that the two preparations are bioequivalent and may be prescribed interchangeably. PMID- 18677971 TI - Validated high performance liquid chromatographic method for simultaneous determination of rosiglitazone, cilostazol, and 3,4-dehydro-cilostazol in rat plasma and its application to pharmacokinetics. AB - A high performance liquid chromatographic (HPLC) method for simultaneous determination of rosiglitazone, CAS 122320-73-4, RSG), cilostazol (CAS 73963-72 1, CLZ) and its active metabolite 3, 4-dehydro-cilostazol (DCLZ), using pioglitazone (PIO) as internal standard (IS), in rat plasma is described. The plasma was extracted with methyl t-butyl ether, the dry extract was reconstituted in mobile phase and the aliquot was injected. The eluent drugs were detected by UV at dual wavelength of 226 nm (RSG and DCLZ) and 257 nm (CLZ). The mobile phase consisting of acetonitrile:potassium di-hydrogen phosphate buffer (35:65 v/v) was used at the flow rate of 1.2 ml/min on a reverse phase C18 column. The absolute recovery was above 90% of all analytes over the concentration range of 25-2500 ng/ml for RSG and CLZ and 20-2000 ng/ ml for DCLZ. The relative standard deviation (RSD) of the inter-day and intra-day precision ranged from 2.8 to 8.4% and 0.9 to 5.9%, respectively. The method is simple, rapid, accurate and sensitive and was applied to pharmacokinetic studies. PMID- 18677972 TI - Effect of an orally active Th2 cytokine inhibitor, suplatast on "atopic cough" tosilate. AB - BACKGROUND: "Atopic cough" is a new clinical entity that presents with isolated chronic bronchodilator-resistant cough accepted in the Japanese Respiratory Society Guidelines for Management of Cough. The essential features are eosinophilic tracheobronchitis, increased cough reflex sensitivity and an atopic constitution. It has been suggested that activated helper T lymphocytes and the cytokines which are produced by these cells are involved in the pathogenesis, but the relationship between helper T cell-derived cytokines and the airway cough reflex sensitivity remains unknown. METHODS: The effect of an orally active Th2 cytokine inhibitor, suplatast tosilate (CAS 94055-76-2, IPD; 300 mg/day), on the cough response to inhaled capsaicin (CAS 404-86-4) was examined in ten patients with atopic cough. The capsaicin cough threshold, defined as the lowest concentration of capsaicin eliciting five or more coughs, was measured as an index of airway cough reflex sensitivity. The serum total immunoglobulin E (IgE) level and the peripheral blood eosinophil count were also determined after treatment with suplatast tosilate. RESULTS: The cough threshold measured after four weeks of treatment with suplatast tosilate was significantly increased compared to the value obtained with placebo, along with a decrease of the serum IgE level and peripheral eosinophil count. CONCLUSIONS: Th2 cytokines may increase the airway cough reflex sensitivity in patients with atopic cough. Oral administration of suplatast tosilate may be a novel therapy for atopic cough. PMID- 18677973 TI - Prevention of exercise-induced asthma by a fixed combination of disodium cromoglycate plus reproterol compared with montelukast in young patients. AB - BACKGROUND: The leukotriene inhibitor montelukast has been recommended against exercise-induced asthma (EIA), however, single-dose agents might be favourable in several aspects. OBJECTIVE: To compare the protective effects against EIA of a single inhalation of the combination disodium cromoglycate (DSCG, CAS 16110-51-3) and reproterol (REP, CAS 54063-54-6) with 3 days oral treatment of montelukast (MON, CAS 158966-92-8). METHODS: Open-label, cross-over, single-centre trial. Twenty-four 6 to 18-year-old children and adolescents, with reversible and stable airway obstruction, baseline FEV1 > or = 70%, predicted and proven EIA (i.e. a maximum decrease of FEV1 by > or = 20% compared with baseline) were treated with MON, orally for 3 days in the evening, or one single inhalation of DSCG/REP 20 min before the exercise challenge. The treatment sequence was randomised. The exercise test on a treadmill was performed under standardised conditions. RESULTS: 24 patients completed both periods. Both treatments clearly provided protection against EIA; however, protection of DSCG/REP was more pronounced than that of MON. This difference was statistically significant even if the data were adjusted for the increase in FEV1 between inhalation of DSCG/REP and challenge (DSCG/REP(adjusted). The nadir FEV1 level after exercise following prophylaxis with DSCG/REP was even higher than the pre-inhalation FEV1 value. From these data, protection indices of 66%, 81%, and 113% for MON, DSCG/REP(adjusted), and DSCG/REP(unadjusted), respectively, were estimated. CONCLUSIONS: Inhalation of DSCG/REP before exercise provides significantly better protection against EIA than three days treatment with MON. PMID- 18677974 TI - Pharmacokinetics of doxycycline and tissue concentrations of an experimental long acting parenteral formulation of doxycycline in Wistar rats. AB - Doxycycline hyclate (CAS 24390-14-5, doxycycline-h), an antibacterial with time dependent action, was formulated as a non-irritating long-acting parenteral formulation based on a beta-cyclodextrin: poloxamer-based matrix (doxycycline-h LA). Tissue and serum concentrations vs time profile were investigated after its subcutaneous injection to Wistar rats. Serum concentration profiles and key pharmacokinetic (PK) variables of doxycycline-h-LA were compared to the corresponding profiles and PK values obtained with an aqueous formulation of doxycycline-h administered either intramuscularly, orally or intravenously to Wistar rats. In all groups, the dose was 10 mg/kg. Doxycycline-h-LA showed outstanding bioavailability (951% or 477% if a correction formula is considered), as compared to the one obtained with an aqueous formulation (106-82%, respectively). Corresponding values for maximum serum concentration were 3.19 microg/ml and 3.00 microg/ml, respectively, and elimination half-lives were completely different: 42.49 h and 2.77 h for doxycycline-h-LA and the aqueous formulation, respectively. Considering minimal inhibitory concentrations of doxycycline for sensitive and resistant bacteria (from < or = 0.5 to > or =1.5 microg/ml), doxycycline-h-LA could be injected every 2 or 3 days, while aqueous doxycycline-h would require a dosing interval from 7.5 to 11 h. But if tissue concentrations are taken as braking points, the dosing interval will vary from 48 to 94 h. For doxycycline-h-LA, mean tissue:serum ratios were 2:1 for lungs, 9.8:1 for kidneys and 2.2:1 for intestine homogenates. These values are in close agreement with those found for the distribution of doxycycline in other species. PMID- 18677975 TI - An author's choice. PMID- 18677976 TI - PFOS phaseout pays off. PMID- 18677977 TI - Real-world pesticide mixtures harm salmon. PMID- 18677978 TI - EPA toxicity risk assessments in crisis. PMID- 18677979 TI - Refining predictions for water pathogens. PMID- 18677980 TI - Built to demolish, demolish to build. PMID- 18677981 TI - Mining for history in a Chinese lake. PMID- 18677982 TI - Arctic marine mammals: a tenuous future. PMID- 18677983 TI - Paradises invaded. PMID- 18677984 TI - A role for environmental engineering and science in preventing bioaerosol-related disease. PMID- 18677985 TI - Critical review: life-cycle inventory procedures for long-term release of metals. AB - Life-cycle assessment (LCA) is the method of inventorying, assessing, and interpreting environmental interventions caused by products and product systems through their life cycle. The ecotoxicity of metals has proven a challenge for LCA given metal characteristics such as reversibility of removal processes, speciation, and the effect on bioavailability and ecotoxic effect assessment. Our review focuses on the first part of the ecotoxic impact chain for metals, i.e., the release of metals from solid deposits. According to the principle of temporal justice, sustainability assessment tools such as LCA should accountfor emissions regardless of temporal location distribution. This is in LCA commonly interpreted as leaching until depletion of metals bound in solid wastes under the presumption that infinite time implies infinite weathering. This approach is risk conservative for metals and it hampers the use of LCA to assess remediation projects for soils and sediments contaminated by inorganic substances. We discuss metal significance and inventory issues in LCA, and review existing and proposed approachesto make LCA applicable to metal long-term emission. PMID- 18677986 TI - Policies for chemical hazard and risk priority setting: can persistence, bioaccumulation, toxicity, and quantity information be combined? AB - Existing methods used to screen chemical inventories for hazardous substances that may pose risks to humans and the environment are evaluated with a holistic mass balance modeling approach. The model integrates persistence (P), bioaccumulation (B), toxicity (T), and quantity (Q) information for a specific substance to assess chemical exposure, hazard, and risk. P and B are combined in an exposure assessment factor (EAF), P, B, and T in a hazard assessment factor (HAF), and P, B, T, and Q in a risk assessment factor (RAF) providing single values for transparent comparisons of exposure, hazard, and risk for priority setting. This holistic approach is illustrated using 200 Canadian Domestic Substances List(DSL) chemicals and 12 United Nations listed Persistent Organic Pollutants (POPs). Priority setting results are evaluated with those of multiple category-based screening methods employed by Environment Canada and applied elsewhere that use cutoff criteria in multiple categories (P, B, and T) to identify hazardous chemicals for more comprehensive evaluations. Existing methods have categorized the DSL chemicals as either higher priority (requiring further assessment; screened in) or lower priority (requiring no further action at this time; screened out). The priority setting results of the cutoff-based categorization are largely inconsistent with the proposed integrated method, and reasons for these discrepancies are discussed. Many chemicals screened out using existing methods have equivalent or greater risk potential than chemicals screened in. Decisions for screening assessments using binary classification on the basis of cutoff criteria can be flawed, and complementary holistic methods for priority setting evaluations such as the one proposed should be considered. PMID- 18677987 TI - Brown Superfund Basic research Program: a multistakeholder partnership addresses real-world problems in contaminated communities. AB - The NIEHS funds several basic and applied research programs, many of which also require research translation or outreach. This paper reports on a project by the Brown University Superfund Basic Research Program (SBRP), in which outreach and research translation teams collaborated with state regulatory agency personnel and community activists on a legislative initiative to mitigate the financial impacts of living in a contaminated community. The Environmentally Compromised Home Ownership (ECHO) program makes home equity loans of up to $25,000 available to qualified applicants. This collaboration provides a case study in community engagement and demonstrates how research translation and outreach activities that are clearly differentiated yet well-integrated can improve a suite of basic and applied research. Although engaging diverse constituencies can be difficult community-engaged translation and outreach have the potential to make research findings more useful to communities, address some of the social impacts of contamination, and empower stakeholders to pursue their individual and collectively held goals for remediation. The NIEHS has recently renewed its commitment to community-engaged research and advocacy, making this an optimal time to reflect on how basic research programs that engage stakeholders through research translation and outreach can add value to the overall research enterprise. PMID- 18677988 TI - Assessing the end-of-life impacts of buildings. AB - This paper builds on previous research on end of life of products by synthesizing some of the theories proposed in the literature and presenting a method for environmental decision-making related to buildings. This is achieved through different solutions, but most significantly through the use of hybrid life-cycle assessment and the definition of allocation boundaries in a way that decreases the uncertainty associated with technologicalforecasting. Results show that there is no significant difference between the results of two major end-of-life assessment approaches (attributional and consequential), and that the choice between the use of one or the other for buildings may not be a critical decision. Assessing the impacts of recycling polices requires accounting for product substitutions, market analysis, and the full supply chain impacts of the recycling chains. Increasing the recycling of concrete from deconstructed buildings from the current 27% rate to 50% could yield a 2-3% (2.7-5.6 million metric tons of CO2 equivalents) reduction in buildings' greenhouse gas emissions, or the equivalent of removing 408,000-847,000 typical cars from U.S. roads. PMID- 18677989 TI - Current and future linked responses of ozone and PM2.5 to emission controls. AB - Responses of ozone and PM2.5 to emission changes are coupled because of interactions between their precursors. Here we show the interdependencies of ozone and PM2.5 responses to emission changes in 2001 and 2050, with the future case accounting for both currently planned emission controls and climate change. Current responses of ozone and PM2.5 to emissions are quantified and linked on a daily basis for five cities in the continental United States: Atlanta, Chicago, Houston, Los Angeles, and NewYork. Reductions in anthropogenic NO(x) emissions decrease 24-h average PM2.5 levels but may either increase or decrease daily maximum 8-h average ozone levels. Regional ozone maxima for all the cities are more sensitive to NO(x) reductions than at the city center, particularly in New York and Chicago. Planned controls of anthropogenic NO(x) emissions lead to more positive responses to NO(x) reductions in the future. Sensitivities of ozone and PM2.5 to anthropogenic VOC emissions are predicted to decrease between 2001 and 2050. Ammonium nitrate formation is predicted to be less ammonia-sensitive in 2050 than 2001 while the opposite is true for ammonium sulfate. Sensitivity of PM2.5 to SO2 and NO(x) emissions changes little between 2001 and 2050. Both ammonium sulfate and ammonium nitrate are predicted to decrease in sensitivity to SO2 and NO(x) emissions between 2001 and 2050. The complexities, linkages, and daily changes in the pollutant responses to emission changes suggest that strategies developed to meet specific air quality standards should consider other air quality impacts as well. PMID- 18677990 TI - An assessment of fecal indicator bacteria-based water quality standards. AB - Fecal indicator bacteria (FIB) are commonly used to assess the threat of pathogen contamination in coastal and inland waters. Unlike most measures of pollutant levels however, FIB concentration metrics, such as most probable number (MPN) and colony-forming units (CFU), are not direct measures of the true in situ concentration distribution. Therefore, there is the potential for inconsistencies among model and sample-based water quality assessments, such as those used in the Total Maximum Daily Load (TMDL) program. To address this problem, we present an innovative approach to assessing pathogen contamination based on water quality standards that impose limits on parameters of the actual underlying FIB concentration distribution, rather than on MPN or CFU values. Such concentration based standards link more explicitly to human health considerations, are independent of the analytical procedures employed, and are consistent with the outcomes of most predictive water quality models. We demonstrate how compliance with concentration-based standards can be inferred from traditional MPN values using a Bayesian inference procedure. This methodology, applicable to a wide range of FIB-based water quality assessments, is illustrated here using fecal coliform data from shellfish harvesting waters in the Newport River Estuary, North Carolina. Results indicate that areas determined to be compliant according to the current methods-based standards may actually have an unacceptably high probability of being in violation of concentration-based standards. PMID- 18677991 TI - Nitrate contamination in groundwater on an urbanized dairy farm. AB - Urbanization of rural farmland is a pervasive trend around the globe, and maintaining and protecting adequate water supplies in suburban areas is a growing problem. Identification of the sources of groundwater contamination in urbanized areas is problematic, but will become important in areas of rapid population growth and development. The isotopic composition of NO3 (delta15N(NO3) and delta18O NO3), NH4 (delta15N(NH4)), groundwater (delta2H(wt) and delta18O(wt)) and chloride/bromide ratios were used to determine the source of nitrate contamination in drinking water wells in a housing development that was built on the site of a dairy farm in the North Carolina Piedmont, U.S. The delta15N(NO3) and delta18O NO3 compositions imply that elevated nitrate levels at this site in drinking well water are the result of waste contamination, and that denitrification has not significantly attenuated the groundwater nitrate concentrations. delta15N(NO3) and delta18O(NO3) compositions in groundwater could not differentiate between septic effluent and animal waste contamination. Chloride/ bromide ratios in the most contaminated drinking water wells were similar to ratios found in animal waste application fields, and were higher than Cl/Br ratios observed in septic drain fields in the area. delta18O(wt) was depleted near the site of a buried waste lagoon without an accompanying shift in delta2H(wt) suggesting water oxygen exchange with CO2. This water-CO2 exchange resulted from the reduction of buried lagoon organic matter, and oxidation of the released gases in aerobic soils. delta18O(wt) is not depleted in the contaminated drinking water wells, indicating that the buried dairy lagoon is not a source of waste contamination. The isotope and Cl/Br ratios indicate that nitrate contamination in these drinking wells are not from septic systems, but are the result of animal waste leached from pastures into groundwater during 35 years of dairy operations which did not violate any existing regulations. Statutes need to be enacted to protect the health of the homeowners that require well water to be tested prior to the sale of homes built on urbanized farmland. PMID- 18677992 TI - Trimethylamine as precursor to secondary organic aerosol formation via nitrate radical reaction in the atmosphere. AB - Amines in fine particulate matter have been detected and quantified during ambient studies of winter inversions in Logan, UT, using aerosol mass spectrometry. Amine-related compounds account for 0.5-6 microg m(-3) of fine particulate mass during some wintertime periods. The amine contributions sometimes show a clear diurnal pattern, reaching peak concentrations during the middle of the nightwhile decreasing during the morning and afternoon. Smog chamber reactions show that the reaction of tertiary amines with nitrate radical can account for this behavior in the atmosphere. The lower bound reaction rate of trimethylamine and nitrate radical is estimated at 4.4 x 10(-16) cm3/molecules/s with a conversion rate to the aerosol phase of approximately 65%. This suggests that amines could be a contributor to secondary organic aerosol formation in areas where nitrate radical is a significant player in oxidation chemistry. PMID- 18677993 TI - Carbonyl emissions from gasoline and diesel motor vehicles. AB - Carbonyls from gasoline-powered light-duty vehicles (LDVs) and heavy-duty diesel powered vehicles (HDDVs) operated on chassis dynamometers were measured by use of an annular denuder-quartz filter-polyurethane foam sampler with O-(2,3,4,5,6 pentafluorobenzyl)hydroxylamine derivatization and chromatography-mass spectrometry analyses. Two internal standards were utilized based on carbonyl recovery: 4-fluorobenzaldehyde for < C8 carbonyls and 6-fluoro-4-chromanone for > or = C8 compounds. Gas- and particle-phase emissions for 39 aliphatic and 20 aromatic carbonyls ranged from 0.1 to 2000 microg/L of fuel for LDVs and from 1.8 to 27 000 microg/L of fuel for HDDVs. Gas-phase species accounted for 81-95% of the total carbonyls from LDVs and 86-88% from HDDVs. Particulate carbonyls emitted from a HDDV under realistic driving conditions were similar to concentrations measured in a diesel particulate matter (PM) standard reference material. Carbonyls accounted for 19% of particulate organic carbon (POC) emissions from low-emission LDVs and 37% of POC emissions from three-way catalyst equipped LDVs. This identifies carbonyls as one of the largest classes of compounds in LDV PM emissions. The carbonyl fraction of HDDV POC was lower, 3.3 3.9% depending upon operational conditions. Partitioning analysis indicates the carbonyls had not achieved equilibrium between the gas and particle phases under the dilution factors of 126-584 used in the present study. PMID- 18677994 TI - Brominated flame retardants, polychlorinated biphenyls, and organochlorine pesticides in captive giant panda (ailuropoda melanoleuca) and red panda (Ailurus fulgens) from China. AB - Organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), and brominated flame retardants (BFRs) were investigated in captive giant and red panda tissues from China. The total concentrations of OCPs, PCBs, and polybrominated diphenyl ethers (PBDEs) in tissues ranged from 16.3 to 888 ng/g lipid weight (lw), 24.8 to 854 ng/g lw, and 16.4 to 2158 ng/g lw, respectively. p,p'-DDE and beta-HCH were major OCP contaminants. PCBs 99, 118, 153/132, 170, 180, and 209 were the major contributing congeners determined. Among PBDEs, congener BDE-209 was the most frequent and abundant, followed by BDE-206, BDE 208, BDE-207, BDE-203, BDE-47, and BDE-153. Decabromodiphenyl ethane (DeBDethane) was detected in 87 and 71% of the giant and red panda samples with concentrations up to 863 ng/g lw, respectively. The remarkable levels and dominance of BDE-209 and DeBDethane may relate to significant production, usage, or disposal of BFRs in China. The positive significant correlation between concentrations of PBDEs and PCBs in captive pandas may suggest that the exposure routes of PBDEs and PCBs to panda are similar. To our knowledge, this is the first report of the occurrence of DeBDethane in captive wildlife samples. Therefore, further studies are warranted to better understand DeBDethane production, transport, uptake, and toxicological effect. PMID- 18677995 TI - Mercury deposition in a polar desert ecosystem. AB - Trace metals have received considerable attention in the recent decades due to their potential toxic nature. Glacial snow and ice have been used extensively to elucidate historical changes in the atmospheric composition of trace metals and other compounds. Mercury concentrations in Antarctic ice have described changes in atmospheric mercury deposition during the transition from the Last Glacial Maximum to the Holocene, however the record of modern mercury deposition in Antarctica is limited. Here we present a record of net mercury deposition to Antarctic snow over the past two decades. Over decadal periods, mercury is conserved in the snowpack and is dependent on a regional oceanic source. Annual to subannual mercury concentrations in snow are to some extent preserved and show covariance with marine aerosols as evidenced by calcium concentrations. Aeolian inputs from exposed rock and soil also play a critical role in depositing mercury to Antarctic snow. Such identifications along with previous data illustrate that mercury transport directly from the glaciers may account for 25-65% of the total mercury concentration in proglacial streams and the surface waters of perennially ice-covered lakes. PMID- 18677996 TI - U, Pu, and Am nuclear signatures of the Thule hydrogen bomb debris. AB - This study concerns an arctic marine environment that was contaminated by actinide elements after a nuclear accident in 1968, the so-called Thule accident In this study we have analyzed five isolated hot particles as well as sediment samples containing particles from the weapon material for the determination of the nuclear fingerprint of the accident. We report that the fissile material in the hydrogen weapons involved in the Thule accident was a mixture of highly enriched uranium and weapon-grade plutonium and that the main fissile material was 235U (about 4 times more than the mass of 239Pu). In the five hot particles examined, the measured uranium atomic ratio was 235U/238U = 1.02 +/- 0.16 and the Pu-isotopic ratios were as follows: 24Pu/239Pu = 0.0551 +/- 0.0008 (atom ratio), 238Pu/239+240Pu = 0.0161 +/- 0.0005 (activity ratio), 241Pu/239+240Pu = 0.87 +/- 0.12 (activity ratio), and 241Am/ 239+240Pu = 0.169 +/- 0.005 (activity ratio) (reference date 2001-10-01). From the activity ratios of 241Pu/241Am, we estimated the time of production of this weapon material to be from the late 1950s to the early 1960s. The results from reanalyzed bulk sediment samples showed the presence of more than one Pu source involved in the accident, confirming earlier studies. The 238Pu/239+240PU activity ratio and the 240Pu/ 239Pu atomic ratio were divided into at least two Pu-isotopic ratio groups. For both Pu-isotopic ratios, one ratio group had identical ratios as the five hot particles described above and for the other groups the Pu isotopic ratios were lower (238Pu/ 239+240PU activity ratio approximately 0.01 and the 240Pu/P239Pu atomic ratio 0.03). On the studied particles we observed that the U/Pu ratio decreased as a function of the time these particles were present in the sediment. We hypothesis that the decrease in the ratio is due to a preferential leaching of U relative to Pu from the particle matrix. PMID- 18677997 TI - Isotope and ion selectivity in reverse osmosis desalination: geochemical tracers for man-made freshwater. AB - A systematic measurement of ions and 2H/1H, 7Li/6Li, 11B/10B, 18O/ 16O, and 87Sr/86Sr isotopes in feed-waters, permeates, and brines from commercial reverse osmosis (RO) desalination plants in Israel (Ashkelon, Eilat, and Nitzana) and Cyprus (Larnaca) reveals distinctive geochemical and isotopic fingerprints of fresh water generated from desalination of seawater (SWRO) and brackish water (BWRO). The degree of isotope fractionation during the passage of water and solutes through the RO membranes depends on the medium (solvent-water vs. solutes), chemical speciation of the solutes, their charge, and their mass difference. O, H, and Sr isotopes are not fractionated during the RO process. 7Li is preferentially rejected in low pH RO, and B isotope fractionation depends on the pH conditions. Under low pH conditions, B isotopes are not significantly fractionated, whereas at high pH, RO permeates are enriched by 20 per thousand in 11B due to selective rejection of borate ion and preferential permeation of 11B enriched boric acid through the membrane. The specific geochemical and isotopic fingerprints of SWRO provide a unique tool for tracing "man-made" fresh water as an emerging recharge component of natural water resources. PMID- 18677998 TI - Seven thousand years of records on the mining and utilization of metals from lake sediments in central China. AB - A 268 cm section of sediment core from Liangzhi Lake in Hubei province in central China was used to assess the use and accumulation of metals in the lake in the past 7,000 years. The concentrations of trace metals, including Cu, Pb, Ni, and Zn, and major elements, Ca, Fe, and Mg, in a 14C- dated segment of sediment core were analyzed. Historical trends on the input of metals to Liangzhi Lake from around 5000 BC to the present were recorded in the sediments, representing about 7,000 years of history on the mining and utilization of metals in central China. The concentrations of Cu, Ni, Pb, and Zn increased gradually from about 3000 +/- 328 BC, indicating the start of the Bronze Age in ancient China. During the period 467 +/- 257 to 215 +/- 221 AD, there was a rapid increase in the concentrations of these metals in the sediments, indicating enormous inputs of these metals at that time. This era corresponded to China's Warring States Period (475- 221 BC) and the early Han Dynasty (206 BC-220 AD), during which copper and lead were extensively used in making bronze articles such as vessels, tools, and weapons. From 1880 +/- 35 AD to the early 1900s, there was also a significant increase in the concentrations of metals such as Cu, Ni, and Pb, which probably reflected the metal emissions and utilization during the early period of industrial development and weapon manufacture during the wars in China. The Pb isotopic analysis showed that the surface and subsurface sediments had lower 206Pb/207Pb and 208Pb/ 207Pb ratios than the deeper layers, reflecting the additional input of Pb from mining activities that took place during the Bronze Age era and in modern times. This study provides direct evidence of the environmental impact of the mining and utilization of metals in the last 7,000 years in one of the important regions of Chinese civilization. PMID- 18677999 TI - Temporal trends of perfluoroalkyl compounds with isomer analysis in lake trout from Lake Ontario (1979-2004). AB - The temporal trends of perfluoroalkyl compounds (PFCs), including C7-C15 perfluorocarboxylates (PFCAs), perfluorosulfonates (PFSAs) and heptadecafluorooctane sulfonamide (PFOSA), were determined in lake trout collected between 1979 and 2004 from Lake Ontario. The average concentrations of total PFSAs (+/- standard error of the mean; range) increased from 20 ng g(-1) wet weight (+/- 4; 8-26) in 1979, peaked at 70 ng g(-1) (+/- 7; 58-91) in 1993, and were 46 ng g(-1) (+/- 10; 30-83) in 2004, with perfluorooctane sulfonate (PFOS) asthe most abundant PFC. The PFCAs exhibited similar temporal variation, with concentrations increasing from 1.4 ng g(-1) (+/- 0.1; 0.9-1.9) in 1979 to 9.4 ng g(1) (+/- 3.1; 3-17) in 1988, and were 6.8 ng g(-1) (+/- 1.0; 4.5-9.8) in 2004. Individual mean PFCA concentrations varied between 0.2 and 2 ng g(-1) (wet weight). Perfluorodecane sulfonate (PFDS) and PFOSA were the only compounds showing a declining trend in the past decade, after reaching a peak value in 1993. Branched C11 and C13 PFCA isomers were detected in the lake trout and confirmed in Niagara River suspended sediments, with trends in both matrices suggesting that declining emissions or use of products containing these isomers in part account for the observed PFCA trends in the mid-1990s. However, the most recent samples, comprised almost exclusively of linear isomers, indicate that current PFCA sources to Lake Ontario result from the telomerization process of linear telogens. PMID- 18678000 TI - Flame retardants in the atmosphere near the Great Lakes. AB - As part of the Integrated Atmospheric Deposition Network (IADN), air samples were collected at five sites around the Great Lakes (two urban, two rural, and one remote) every 12 days during 2005-2006, and the concentrations of polybrominated diphenylethers (PBDEs), 1,2-bis(2,4,6-tribromophenoxy)-ethane (TBE), Dechlorane Plus (DP), and decabromodiphenyl ethane (DBD PE) were measured. The highest mean concentrations of total PBDEs were found at the urban sites in Chicago and Cleveland (65 +/- 4 and 87 +/- 8 pg/m3, respectively), and the lowest at the remote site in Eagle Harbor (5.8 +/- 0.4 pg/m3). With the exception of Chicago, the atmospheric concentrations of BDE-47 and 99 (summed over the gas and particle phases) are decreasing rapidly with half-lives of approximately 2 years, but the concentrations of BDE-209 are not decreasing at any of the five sites. The atmospheric partial pressures of BDE-47 and 99 showed a strong Clausius-Clapeyron relationship with reciprocal atmospheric temperature. TBE, DBDPE and DP were detected at all sites, but with the exception of Chicago, there were insufficient data to determine temporal trends for these compounds. The influence of human population density and synoptic atmospheric transport patterns was explored to explain the relatively high concentrations of BDE-209 in Cleveland. PMID- 18678001 TI - Dechlorination of tetrachloroethylene in aqueous solutions using metal-modified zerovalent silicon. AB - The combination of zerovalent metal with a catalytic second metal ion (bimetallic materials) to enhance the dechlorination efficiency and rate of chlorinated compounds has received much attention. Bimetallic materials not only enhance the dechlorination process but also alter the reduction pathway and product distribution. In this study, the efficiency and rate of tetrachloroethylene (PCE) dechlorination by metal-modified zerovalent silicon was investigated as a potential reductant for chlorinated hydrocarbons under anoxic conditions. The X ray photoelectron spectroscopic (XPS) results showed that metal ions including Ni(II), Cu(II), and Fe(II) could be reduced to their zerovalent forms on the Si surface. The dechlorination of PCE obeyed the pseudo-first-order kinetics, and the pseudo-first-order rate constants (k(obs)) for PCE dechlorination followed the order Ni/Si > Fe/Si > Cu/Si. Addition of Cu(II) lowered the dechlorination efficiency and rate of PCE by Si, while the k(obs) values for PCE dechlorination in the presence of 0.1 mM Fe(II) and Ni(II) were 1.5-3.8 times higher than that by Si alone. In addition, the efficiency and rate of PCE dechlorination increased upon increasing the mass loading of Ni(II) ranging between 0.05 and 0.5 mM and then decreased when the Ni(II) loading was further increased to 1 mM. The scanning electron microscopic (SEM) images and electron probe microanalytical (EPMA) maps showed that the Ni nanoparticles deposited on the Si surface and aggregated to a large particle at 1 mM Ni(II), which clearly depicts that the Ni(II) loading of 0.5 mM is the optimal value to enhance the efficiency and rate of PCE dechlorination by Si. Also, the reaction pathways for PCE dechlorination changed from hydrogenolysis in the absence of Ni(II) to hydrodechlorination when Ni(II) concentrations were higher than 0.05 mM. Results obtained in this study reveal that the metal-deposited zerovalent silicon can serve as an environmentally friendly reductant for the enhanced degradation of chlorinated hydrocarbons for long-term performance. PMID- 18678002 TI - Octachlorodibenzodioxin formation on Fe(III)-montmorillonite clay. AB - Polychlorinated dibenzo-p-dioxins (PCDDs) are ubiquitous and highly toxic environmental contaminants found in surface and subsurface soils and in clay deposits. Interestingly, the congener profiles of such PCDDs are inexplicably dissimilar to those of known anthropogenic (e.g., pesticide manufacture, waste incineration) or natural (e.g., forest fire) sources. Characteristic features of soil or clay-associated PCDDs are the dominance of octachlorodibenzo-p-dioxin (OCDD) as the most abundant congener and very low levels of polychlorinated dibenzofurans (PCDFs). These propensities led to the hypothesis of in situ PCDD formation in soils and geologic clay deposits. In this study, we demonstrate the formation of OCDD on the naturally occurring and widely distributed clay mineral montmorillonite under environmentally relevant conditions. When pentachlorophenol (PCP)was mixed with Fe(III)-montmorillonite, significant amounts of OCDD were rapidly (minutes to days) formed (approximately 5 mg OCDD/kg clay) at ambient temperature in the presence of water. This reaction is initiated by single electron transfer from PCP to Fe(III)-montmorillonite thereby forming the PCP radical cation. Subsequent dimerization, dechlorination, and ring closure reactions result in formation of OCDD. This study provides the first direct evidence for clay-catalyzed formation of OCDD supporting the plausibility of its in situ formation in soils. PMID- 18678003 TI - Identifying competing aerobic nitrobenzene biodegradation pathways by compound specific isotope analysis. AB - Nitroaromatic compounds that contaminate soil and groundwater can be biodegraded by different, sometimes competing reaction pathways. We evaluated the combined use of compound-specific stable C and N isotope analysis to distinguish between enzymatic nitrobenzene oxidation by Comamonas sp. strain JS765 and partial reduction by Pseudomonas pseudoalcaligenes strain JS45 under aerobic conditions. Bulk 13C and 15N enrichment factors for nitrobenzene dioxygenation with JS765 were -3.9 per thousand +/- 0.09 per thousand (+/- 1sigma) and -0.75 per thousand +/- 0.09 per thousand, respectively. The corresponding primary apparent kinetic isotope effects (AKIE) of 1.0241 +/- 0.0005 for 13C and a secondary 15N AKIE of 1.0008 +/- 0.0001 are in very good agreement with the proposed enzymatic addition of dioxygen to the aromatic ring to form a cis-dihydrodiol in the rate-limiting step of nitrobenzene degradation. For the partial reduction pathway with JS45, epsilonC and epsilonN values were -0.57 per thousand +/- 0.06 per thousand and 26.6 per thousand +/- 0.7 per thousand. The 13C and 15N AKIEs amount to 1.0034 +/ 0.0003 and 1.0273 +/- 0.0008, respectively, and are consistent with the two electron reduction and dehydration of the aromatic NO2 group to nitrosobenzene. The combined evaluation of delta13C and delta15N changes in nitrobenzene, based on the isotope enrichment behavior found in this laboratory study, provide an excellent starting point for assessing of the extent of nitrobenzene biodegradation via competing pathways in contaminated environments. PMID- 18678004 TI - Effect of volatile-char interaction on the NO emission from coal combustion. AB - To clarify the effects of volatile-char interaction on the redistribution of fuel N to N2 during devolatilization and the reduction of NO through gas-solid reactions during combustion, two types of experiments were performed on a novel reactor. The separate combustion of volatile and char and the combustion of entrained pulverized coal, and the formation of NO was examined between 800 and 1100 degrees C by using four typical Chinese coals with different ranks. The effect of volatile-char interaction on fuel-N conversion to NO during combustion was elucidated through comparing the NO emissions from the two types of combustion experiments. The results show that the volatile-char interaction is more important in the redistribution of fuel-N to N2 during devolatilization than in the reduction of NO over 900 degrees C, and a contrary conclusion is obtained below 850 degrees C for all used coals. A specific parameter has been proposed to characterize the relative importance of the volatile-char interaction in the redistribution of fuel-N to N2 during devolatilization to the interaction in the reduction of NO to N2 during simulataneous combustion of volatile and char. The results are of significance for minimizing the NO formation in industrial combustion processes. PMID- 18678005 TI - Confounding impacts of iron reduction on arsenic retention. AB - A transition from oxidizing to reducing conditions has long been implicated to increase aqueous As concentrations, for which reductive dissolution of iron (hydr)oxides is commonly implicated as the primary culprit. Confounding our understanding of processes controlling As retention, however, is that reductive transformation of ferrihydrite has recently been shown to promote As retention rather than release. To resolve the role iron phases have in regulating arsenic concentrations, here we examine As desorption from ferrihydrite-coated sands presorbed with As(III); experiments were performed at circumneutral pH under Fe reducing conditions with the dissimilatory iron reducing bacterium Shewanella putrefaciens strain CN-32 over extended time periods. We reveal that with the initial phase of iron reduction, ferrihydrite undergoes transformation to secondary phases and increases As(III) retention (relative to abiotic controls). However, with increased reaction time, cessation of the phase transitions and ensuing reductive dissolution result in prolonged release of As(III) to the aqueous phase. Our results suggest that As(III) retention during iron reduction is temporally dependent on secondary precipitation of iron phases; during transformation to secondary phases, particularly magnetite, As(III) retention is enhanced even relative to oxidized systems. However, conditions that retard secondary transformation (more stable iron oxides or limited iron reducing bacterial activity), or prolonged anaerobiosis, will lead to both the dissolution of ferric (hydr)oxides and release of As(III) to the aqueous phase. PMID- 18678006 TI - Adaptation of aerobic, ethene-assimilating Mycobacterium strains to vinyl chloride as a growth substrate. AB - Contamination of drinking water source zones by vinyl chloride (VC), a known human carcinogen and common groundwater contaminant, poses a public health risk. Bioremediation applications involving aerobic, VC-assimilating bacteria could be useful in alleviating environmental VC cancer risk, but their evolution and activity in the environment are poorly understood. In this study, adaptation of ethene-assimilating Mycobacterium strains JS622, JS623, JS624, and JS625 to VC as a growth substrate was investigated to test the hypothesis that VC-assimilating bacteria arise from naturally occurring ethene-assimilating bacteria. VC consumption in the absence of microbial growth was initially observed in cultures grown in both ethene and 1/10-strength trypticase soy agar + 1% (w/v) glucose. After extended incubations (55-476 days), all strains commenced growth-coupled VC consumption patterns. VC-adapted cultures grown on 20 mM acetate subsequently retained their ability to assimilate VC. Three independent purity check methods (streak plates, 16S rRNA gene sequencing, and repetitive extragenic palindromic polymerase chain reaction) verified culture purity prior to and following VC adaptation. Overall, our results suggest that ethene-assimilating mycobacteria have a widespread ability to adapt to VC as a growth substrate. PMID- 18678007 TI - Isomer-specific biodegradation of methylphenanthrenes by soil bacteria. AB - It is assumed that bacteria generally degrade 2-methylphenanthrene (2MPhe) in preference to 1-methylphenanthrene (1MPhe), and that environmental biodegradation of methylated PAHs therefore can be described qualitatively by changes in relative concentrations of these isomers. Our objective was to investigate whether microbial phenanthrene degraders (Sphingomonas and Mycobacterium) show such isomer-specific PAH degradation. Eleven out of twenty-nine phenanthrene degraders could grow on methylphenanthrene. The mycobacteria grew only on 2MPhe, the sphingomonads grew mostly on 1MPhe, and one sphingomonad could utilize both substrates. Seven strains were tested in a two-phase system where 1MPhe and 2MPhe were supplied in heptamethylnonane. For these strains, a consistent description of biodegradation based on the 2MPhe/1MPhe diagnostic ratio would not be possible because three Mycobacterium and one Sphingomonas degraded 2MPhe faster than 1MPhe, another Sphingomonas degraded 1MPhe and 2MPhe at almost equal rates, and two Sphingomonas degraded 1MPhe faster than 2MPhe. Thus, environmental biodegradation of phenanthrenes may theoretically proceed with only minor changes in 2MPhe/1MPhe ratios if individual members of the degrader community have different isomer preferences. However, two soil microcosms polluted with bunker oil confirmed the general decline in 2MPhe/1MPhe ratio during oil biodegradation. PMID- 18678008 TI - Interactions of hydrophobic fractions of dissolved organic matter with Fe(3+) - and Cu(2+)-montmorillonite. AB - Interactions of dissolved organic matter (DOM) with clays can significantly affect a variety of soil processes. We studied adsorption and fractionation of hydrophobic acid (HoA) and hydrophobic neutral (HoN) fractions of DOM on Cu(2+)- and Fe(3+)-montmorillonite. Adsorption of both samples was higher on Fe(3+) montmorillonite than on Cu(2+)-montmorillonite. A pH increase of about one unit was recorded followed by HoA adsorption by Fe(3+)-montmorillonite. This suggested exchange of negatively charged DOM groups on surface hydroxyl groups of Fe(3+) montmorillonite surfaces. Adsorption of HoA on Cu(2+)-montmorillonite and HoN on Fe(3+)- and Cu(2+)-montmorillonites was governed mainly by van der Waals interactions. Spectroscopic analyses showed a distinct HoA fractionation by molecular size and aromaticity only by Fe(3+)-montmorillonite. On the basis of the pH measurements (increase in pH following adsorption of acid components) and enhanced DOM fractionation by molecular size and aromaticity we suggest that DOM reacted with Fe(3+)-montmorillonite similar to goethite. PMID- 18678009 TI - Modeling arsenic in the Patuxent Estuary. AB - A water quality model was developed to track the fate and transport of four arsenic species in the Patuxent Estuary: arsenate (As(V)), arsenite (As(III)), methylarsonate (MMA), and dimethylarsinate (DMA). Processes simulated include mass transport, solid-liquid partitioning with suspended solids, uptake and transformation of As(V) by phytoplankton, oxidation of As(III), demethylation of MMA and DMA, and settling/deposition/ resuspension of particulate arsenic in the water column. A sediment module was also developed and linked with the water column to generate fluxes of inorganic arsenic from the sediment bed. The arsenic model was calibrated using water quality data from the Patuxent Estuary over a period ranging from May 24, 1995 to October 29, 1997. Model results indicated that transformation of arsenic by phytoplankton is not a significant source of DMA to the lower Patuxent. Instead, results suggested that the primary source of methylated arsenic (DMA and MMA) to the lower estuary is beyond the downstream boundary (Chesapeake Bay). However, model results supported the hypothesis that flux of arsenic from the sediment is a significant source of inorganic arsenic to the lower estuary. PMID- 18678010 TI - Degradation of tertiary alkylamines during chlorination/chloramination: implications for formation of aldehydes, nitriles, halonitroalkanes, and nitrosamines. AB - Drinking water utilities are exploring the use of waters impacted by wastewater effluents and agricultural runoff to meet the demands of growing populations. Due to the elevated organic nitrogen concentrations in these waters, the pathways responsible for transformation of organic nitrogen into toxic nitrogenous disinfection byproducts during chlorine and chloramine disinfection are of current concern. Tertiary alkylamines are important functional groups in human waste products and various consumer products that can be released to drinking water supplies via wastewater effluents. We investigated degradation pathways for model tertiary alkylamines during chlorination and chloramination. Our results indicate that tertiary alkylamines degrade nearly instantaneously during chlorination to form aldehydes and secondary alkylamines quantitatively, with no significant regioselectivity. Similar results were observed during chloramination, but the observed degradation rates were much slower, with lower yields of aldehydes. As these major products were fairly stable, these results explain why tertiary amines are significant precursors of secondary nitrosamines during chloramination. Trichloronitromethane formed at very low yields during chlorination, but was not observed during chloramination; monochloronitromethane and dichloronitromethane were never detected. Despite the significant yields of aldehydes during chloramination, our results indicated low nitrile yields bythe reaction between chloramines and aldehydes. PMID- 18678011 TI - Nowcasting and forecasting concentrations of biological contaminants at beaches: a feasibility and case study. AB - Public concern over microbial contamination of recreational waters has increased in recent years. A common approach to evaluating beach water quality has been to use the persistence model which assumes that day-old monitoring results provide accurate estimates of current concentrations. This model is frequently incorrect Recent studies have shown that statistical regression models based on least squares fitting often are more accurate. To make such models more generally available, the Virtual Beach (VB) tool was developed. VB is public-domain software that prescribes site-specific predictive models. In this study we used VB as a tool to evaluate statistical modeling for predicting Escherichia coli (E. coli levels at Huntington Beach, on Lake Erie. The models were based on readily available weather and environmental data, plus U.S. Geological Service onsite data. Although models for Great Lakes beaches have frequently been fitted to multiyear data sets, this work demonstrates that useful statistical models can be based on limited data sets collected over much shorter time periods, leading to dynamic models that are periodically refitted as new data become available. Comparisons of the resulting nowcasts (predictions of current, but yet unknown, bacterial levels) with observations verified the effectiveness of VB and showed that dynamic models are about as accurate as long-term static models. Finally, fitting models to forecasted explanatory variables, bacteria forecasts were found to compare favorably to nowcasts, yielding adjusted coefficients of determination (adjusted R2) of about 0.40. PMID- 18678012 TI - Effect of pore-blocking background compounds on the kinetics of trace organic contaminant desorption from activated carbon. AB - This study examined the effect of pore-blocking (PB) background organic matter, which is known to hinder adsorption kinetics, on the rate of trace contaminant desorption. Adsorption, displaced desorption (DD) and nondisplaced desorption (NDD) kinetic tests were performed using powdered activated carbon (PAC) that was preloaded with natural organic matter (NOM). Since the NOM contained both strongly competing (SC) and PB components, the proposed model separated the contributions of the SC and PB NOM to the overall diffusion coefficient of the target contaminant. By factoring outthe SC NOM contribution, which increases the overall diffusion coefficient it was found that the relationship used to model the effect of PB NOM on adsorption kinetics could also describe desorption kinetics. The results highlighted the substantial influence of competitive SC NOM on the kinetics of adsorption and desorption. SC NOM competition aids contaminant removal by offsetting the undesirable effects of pore blocking on adsorption kinetics. However, for desorption events, PB NOM serves a practical benefit of reducing the rate of release of adsorbed micropollutants, while SC NOM counters that gain by both displacing contaminants and accelerating their diffusion. PMID- 18678013 TI - Modeling the importance of biota and black carbon as vectors of polybrominated diphenyl ethers (PBDEs) in the Baltic Sea ecosystem. AB - The POPCYCLING-Baltic model, a nonsteady state spatially resolved mass balance model of chemical transport in the Baltic Sea environment was modified to include black carbon (BC), dissolved organic carbon (DOC) and food-web bioaccumulation. The importance of these modifications to the transport of PBDE congeners BDE-47, 99, -153, and -209 was assessed by comparing time-series simulated with and without black carbon and biota between 1970 and 2005. Inclusion of black carbon improved the model fit to measurements from air, soil, and biota, and had a major effect on the mass balance. Modeled bulk concentrations of PBDEs in sediments and soils increased by a factor of 3 while concentrations in biota decreased by a factor of 2-5. Black carbon also doubled the recovery time of the system due to the limited availability of PBDEs for degradation. In comparison, the inclusion of biota had only a minor effect on the overall mass balance and recovery times. The modified model is constructed as a flexible matrix and can also be applied to persistent organic pollutants in other ecosystems besides the Baltic Sea. PMID- 18678014 TI - The effects of dual-domain mass transfer on the tritium-helium-3 dating method. AB - Diffusion of tritiated water (referred to as tritium) and helium-3 between mobile and immobile regions in aquifers (mass transfer) can affect tritium and helium-3 concentrations and hence tritium-helium-3 (3H/3He) ages that are used to estimate aquifer recharge and groundwater residence times. Tritium and helium-3 chromatographically separate during transport because their molecular diffusion coefficients differ. Simulations of tritium and helium-3 transport and diffusive mass transfer along stream tubes show that mass transfer can shift the 3H/3He age of the tritium and helium-3 concentration ([3H + 3He]) peak to dates much younger than the 1963 peak in atmospheric tritium. Furthermore, diffusive mass-transfer can cause the 3H/3He age to become younger downstream along a stream tube, even as the mean water-age must increase. Simulated patterns of [3H + 3He] versus 3H/3He age using a mass transfer model appear consistent with a variety of field data. These results suggest that diffusive mass transfer should be considered, especially when the [3H + 3He] peak is not well defined or appears younger than the atmospheric peak. 3H/3He data provide information about upstream mass transfer processes that could be used to constrain mass-transfer models; however, uncritical acceptance of 3H/3He dates from aquifers with immobile regions could be misleading. PMID- 18678015 TI - NMR determination of total carbonyls and carboxyls: a tool for tracing the evolution of atmospheric oxidized organic aerosols. AB - Nuclear magnetic resonance (NMR) spectroscopy is used to investigate the chemical composition of organic aerosol in terms of functional group distribution with a special focus on secondary organic aerosol (SOA) formation. The knowledge of the functional group composition is a benchmark for understanding how SOA components partition into the particulate phase and undergo chemical transformation. The paper presents a new chemical derivatization procedure coupled to proton NMR (1H NMR) analysis for the specific determination of total carbonylic groups in atmospheric aerosol samples, which couples with the procedure for determination of total carboxylic acid groups described in a previous work. A first deployment of the combined techniques for the analysis of PM10 samples collected in the Po Valley in the warm season shows that the concentration in the particulate phase of total carbonyls varies and covaries with respect to those of carboxylic acids and of less-oxidized functional groups. The proposed methodology provides the breakdown of the oxygenated fraction of the organic aerosol into major functional groups through well-established chemical methods and can be used to benchmark the more sensitive and widely used aerosol mass spectrometric techniques. PMID- 18678016 TI - A plume capture technique for the remote characterization of aircraft engine emissions. AB - A technique for capturing and analyzing plumes from unmodified aircraft or other combustion sources under real world conditions is described and applied to the task of characterizing plumes from commercial aircraft during the taxiing phase of the Landing/Take-Off (LTO) cycle. The method utilizes a Plume Capture and Analysis System (PCAS) mounted in a four-wheel drive vehicle which is positioned in the airfield 60 to 180 m downwind of aircraft operations. The approach offers low test turnaround times with the ability to complete careful measurements of particle and gaseous emission factors and sequentially scanned particle size distributions without distortion due to plume concentration fluctuations. These measurements can be performed for individual aircraft movements at five minute intervals. A Plume Capture Device (PCD) collected samples of the naturally diluted plume in a 200 L conductive membrane conforming to a defined shape. Samples from over 60 aircraft movements were collected and analyzed in situ for particulate and gaseous concentrations and for particle size distribution using a Scanning Particle Mobility Sizer (SMPS). Emission factors are derived for particle number, NO(x), and PM2.5 for a widely used commercial aircraft type, Boeing 737 airframes with predominantly CFM56 class engines, during taxiing. The practical advantages of the PCAS include the capacity to perform well targeted and controlled emission factor and size distribution measurements using instrumentation with varying response times within an airport facility, in close proximity to aircraft during their normal operations. PMID- 18678017 TI - Formation of N-nitrosamines from eleven disinfection treatments of seven different surface waters. AB - Formation of nine N-nitrosamines has been investigated when seven different source waters representing various qualities were each treated with eleven bench scale disinfection processes, without addition of nitrosamine precursors. These disinfection treatments included chlorine (OCl-), chloramine (NH2Cl), chlorine dioxide (ClO2), ozone (O3), ultraviolet (UV), advanced oxidation processes (AOP), and combinations. The total organic carbon (TOC) of the seven source waters ranged from 2 to 24 mg x L(-1). The disinfected water samples and the untreated source waters were analyzed for nine nitrosamines using a solid phase extraction and liquid chromatography-tandem mass spectrometry method. Prior to any treatment, N-nitrosodimethylamine (NDMA) was detected ranging from 0 to 53 ng x L(-1) in six of the seven source waters, and its concentrations increased in the disinfected water samples (0-118 ng x L(-1)). N-nitrosodiethylamine (NDEA), N nitrosomorpholine (NMor), and N-nitrosodiphenylamine (NDPhA) were also identified in some of the disinfected water samples. NDPhA (0.2-0.6 ng x L(-1)) was formed after disinfection with OCl-, NH2Cl, O3, and MPUV/OCl-. NMEA was produced with OCl- and MPUV/OCl-, and NMor formation was associated with O3. In addition, UVtreatment alone degraded NDMA; however, UV/ OCl- and AOP/OCl- treatments produced higher amounts of NDMA compared to UV and AOP alone, respectively. These results suggest that UV degradation or AOP oxidation treatment may provide a source of NDMA precursors. This study demonstrates that environmental concentrations and mixtures of unknown nitrosamine precursors in source waters can form NDMA and other nitrosamines. PMID- 18678018 TI - Comparison of selection methods to deduce natural background levels for groundwater units. AB - Establishment of natural background levels (NBL) for groundwater is commonly performed to serve as reference when assessing the contamination status of groundwater units. We compare various selection methods to establish NBLs using groundwater quality data for four hydrogeologically different areas in the highly populated and developed subcatchment Western River Rhine, The Netherlands: selection of old groundwater (before 1945), of tritium-free groundwater (i.e., infiltrated before 1950), and of groundwater having no agricultural contamination by NO3 and SO4. Differences as well as similarities in percentile values for Cl, NH4, and SO4 concentrations are observed among the selection methods as well as the spatial units, pointing out that selection of the data setis a crucial step in deducing NBLs. The following general points of attention are deduced: (1) reference to composition of recharge water (rain or river infiltrate) is necessary to confirm the statistical outcomes, (2) old analyses are affected by conservation errors after sampling for redox-sensitive solutes and may be obtained by selective sampling, (3) old analyses are the only direct reference for NBLs for groundwater units having only anthropogenically influenced, young groundwater at present, and (4) establishment of a priori percentile values as maximum NBL is not right and confirmation by additional process-based insight in the controls on water composition is necessary. PMID- 18678019 TI - Novel chamber to measure equilibrium soil-air partitioning coefficients of low volatility organic chemicals under conditions of varying temperature and soil moisture. AB - The need to determine soil-air partitioning coefficients (K(SA)) of low volatility organic chemicals as a measure of their distribution in the soil surface after release into the environment resulted in the development of a novel chamber system, which has been filed for patent. A major advantage of this pseudo static system is that sufficient time can be factored into the experiment to ensure that the system has achieved equilibrium. In a highly precise method, the air is collected in adsorption tubes and subsequently liberated in a thermodesorption system for the quantitation of the adsorbed compound. The precision of the method is great enough that even the effects of temperature and soil moisture on the soil-air partitioning of very low-volatility compounds can be quantified. Because of analytical detection limits, quantitation of these influences has not been possible to date. Functionality of the setup was illustrated by measurements on the fungicide fenpropimorph. K(SA) values of fenpropimorph displayed a negative relationship with temperature and soil moisture. The type of application (spraying or incorporation) and the use of formulated compounds was shown to have a major impact on the measured K(SA) values. Comparison with calculations using an estimation method revealed that the use of experimentally determined K(SA) values will facilitate a more adequate consideration of volatilization in recent model approaches. PMID- 18678020 TI - Underpotential deposition-anodic stripping voltammetric detection of copper at gold nanoparticle-modified ultramicroelectrode arrays. AB - The sensitive detection of copper (II) at gold nanoparticle-modified ultramicroelectrode arrays (UMEAs) is reported. Gold nanoparticles were electrodeposited onto the UMEAs surface by applying a constant positive potential of 1.6 V for 20 min in a 20-nm gold nanoparticle solution. This process significantly increases the electrode area without losing the UMEAs analytical features. Underpotential deposition-anodic stripping voltammetry of copper (II) with such modified UMEAs was performed and showed a high increase in sensitivity (25.9 +/- 1.3 nC x micro-1) and a broader linear range of response (0-10 microM) compared with those values obtained using bare UMEAs (7.5 +/- 0.6 nC x microM(-1) and 0-2 microM, respectively). The copper content of acid extracts of contaminated soils was successfully determined with the modified UMEAs and results are in good agreement with those obtained using the ICP-AES standard method. Overall, this work shows an alternative easy-to-use novel miniaturized device for the rapid and reliable determination of copper in soil samples whose application could be readily extended to other heavy metals of environmental interest. PMID- 18678021 TI - Surfactants as bubble surface modifiers in the flotation of algae: dissolved air flotation that utilizes a chemically modified bubble surface. AB - In this paper we present an investigation into the use of bubbles modified with surfactants in dissolved air flotation (DAF). Bubble modification was investigated by dosing surfactants of varying character into the saturator of a DAF unit in turn. The cell removal efficiency only improved when using a cationic surfactant where optimum removal of Microcystis aeruginosa cells was obtained when using 0.0022-0.004 mequiv L(-1) surfactant. However, the magnitude of the removal differed according to the hydrophobicity of the surfactant. Typically, the more efficiently the surfactant adsorbed at the bubble interface, the better the removal efficiency. When the dose to saturator ratio was kept constant and the recycle ratio varied, the removal efficiency improved with increasing recycle ratio, reaching a maximum removal efficiency of 87% for M. aeruginosa. This value was comparable with that predicted by a theoretical model. The bubble collection efficiency of a maximum of two cells per bubble was constant irrespective of the influent cell number or recycle ratio. Treatment of additional species in this way revealed a relationship between increasing size and both increasing removal efficiency and decreasing surfactant dose, which is supported by theoretical relationships. PMID- 18678022 TI - Ozonation of ciprofloxacin in water: HRMS identification of reaction products and pathways. AB - Degradation products formed during ozonation of an aqueous solution of the antibiotic ciprofloxacin in a bubble column are analyzed by HPLC-UV high resolution mass spectrometry. Based on the identification of the reaction products, reaction pathways are proposed starting with (1) degradation at the piperazinyl substituent, (2) degradation at the quinolone moiety with formation of isatin analogues, and (3) degradation at the quinolone moiety with formation of anthranilic acid analogues. Unlike H2O2 addition (10 microM), pH (3,7, and 10) strongly affects degradation product formation during ozonation. Degradation at the quinolone core is favored at pH 7. Addition of t-butanol, a hydroxyl radical scavenger, ruled out formation of isatin and anthranilic acid analogues. Because the carboxylic group and the keto group at the quinolone moiety are essential for antibacterial activity, degradation at pH 7 seems to be promising for reduction of bacterial resistance against quinolones in contaminated water. PMID- 18678023 TI - Reduced toxicity of olive mill waste waters by oxidative coupling with biomimetic catalysis. AB - Large quantities of environmentally toxic olive mill waste waters (OMWW) result from olive oil production worldwide. A synthetic water-soluble meso-tetra(2,6 dichloro-3-sulfonatophenyl)porphyrinate of iron(III) chloride (FePha) was used as biomimetic catalystto oxidatively couple toxic phenols in OMWW fractions obtained by micro-, ultra-, and nanofiltration, and reverse osmosis. The occurrence of oxidative coupling in different OMWW size-fractions was assessed by high performance size exclusion chromatography (HPSEC), before and after conformational disruption with acetic acid, and measurements of proton spin lattice relaxation time in the rotating frame (T1(rho)H) through 13C-CPMAS-NMR spectroscopy. The concurrent reduction in toxicity of OMWW size-fractions brought about by the FePha treatment was monitored by an algal bioassay. HPSEC chromatograms of OMWW samples subjected to catalyzed coupling showed apparent weight-average molecular weight (Mwa) values varying from 18 to 185% larger than for control. Moreover, when such FePha-treated fractions were added to acetic acid prior to HPSEC, the Mwa values still ranged from 14 to 162% larger than for control fractions similarly treated with acetic acid. This evidence of polymerization among toxic phenols was confirmed by T1(rho)(H) values which were significantly enhanced by the FePha treatment, thereby indicating an increased conformational rigidity of OMWW materials. These molecular changes were reflected in a significantly reduced toxicity exerted on microalgae by the OMWW size fractions subjected to catalyzed oxidative couplings. Our results suggest that OMWW can be effectively treated with a biomimetic catalyst to induce oxidative phenol polymerization and reduce their toxicity before amendments to soils or other disposal means. PMID- 18678024 TI - Hydrothermal synthesis and photocatalytic activity of zinc oxide hollow spheres. AB - ZnO hollow spheres with porous crystalline shells were one-pot fabricated by hydrothermal treatment of glucose/ZnCl2 mixtures at 180 degrees C for 24 h, and then calcined at different temperatures for 4 h. The as-prepared samples were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy and nitrogen adsorption-desorption isotherms. The photocatalytic activity of the as-prepared samples was evaluated by photocatalytic decolorization of Rhodamine B aqueous solution at ambient temperature. The results indicated that the average crystallite size, shell thickness, specific surface areas, pore structures, and photocatalytic activity of ZnO hollow spheres could be controlled by varying the molar ratio of glucose to zinc ions (R). With increasing R, the photocatalytic activity increases and reaches a maximum value at R = 15, which can be attributed to the combined effects of several factors such as specific surface area, the porous structure and the crystallite size. Further results show that hollow spheres can be more readily separated from the slurry system by filtration or sedimentation after photocatalytic reaction and reused than conventional powder photocatalyst. After many recycles for the photodegradation of RhB, the catalyst does not exhibit any great loss in activity, confirming ZnO hollow spheres is stability and not photocorroded. The prepared ZnO hollow spheres are also of great interest in solar cell, catalysis, separation technology, biomedical engineering, and nanotechnology. PMID- 18678025 TI - Sensitizers on inorganic carriers for decomposition of the chemical warfare agent yperite. AB - Sulfur-containing compounds, such as mercaptans, alkali sulfides, alkali sulfites, and alkali thiosulfates, are byproducts of industrial processes and pollutants of waste and natural waters. Other sulfur-containing compounds such as yperite are dangerous chemical weapons. Efficient photocatalytic decomposition of these molecules is a process that can find applications in emergency situations or for the controlled destruction of chemical warfare stockpiles. A series of heterogeneous photocatalysts consisting of a metal phthalocyanine or 2,4,6 triphenylpyrylium as photoactive components encapsulated inside the cavities of zeolite Y or the mesoporous channels of MCM-41 or supported on silica or titania silica was tested for the photocatalytic decomposition of yperite. Two types of photoreactors, either an open reactor naturally aerated or a closed quartz tube with a constant airflow using UV or visible ambient light were used. These tests demonstrated that iron and manganese phthalocyanine and 2,4,6-triphenylpyrylium embedded in NaY or titania-silica can be suitable solid photocatalysts for the degradation of yperite using UV and visible irradiation. PMID- 18678026 TI - Essential explanation of the strong mineralization performance of boron-doped diamond electrodes. AB - Electrochemical oxidation of p-nitrophenol was examined using differentanodic materials, including T/boron-doped diamond (BDD), Ti/SnO2-Sb/PbO2, and Ti/SnO2-Sb anodes. The results demonstrated that Ti/BDD anodes had a much stronger mineralization performance than the other two anodes. Furthermore, it was found that hydroxyl radicals could mainly exist as free hydroxyl radicals at BDD anodes, which could react with organic compounds effectively. This implied that the dominant mechanism for a much higher mineralization capacity of BDD anodes would be attributed to the existence of free hydroxyl radicals in the BDD anode cell rather than adsorbed hydroxyl radicals on the BDD anode. To further corroborate this hypothesis, electrochemical oxidation of p-substituted phenols (p-nitrophenol, p-hydroxybenzaldehyde, phenol, p-cresol, and p-methoxyphenol) was examined at the Ti/BDD, Ti/SnO2-Sb/ PbO2, and Ti/SnO2-Sb anodes, respectively. The study revealed that for Ti/BDD electrodes, the degradation rate of p substituted phenols (k) increased with the increase of Hammett's constant (sigma), which confirmed the dominance of free hydroxyl radicals at BDD anodes and its effective reaction with organics therein. For Ti/SnO2-Sb/PbO2 electrodes, the degradation rate of p-substituted phenols (k) increased with the increase of initial surface concentration gamma (representing the adsorption capacity of phenols to electrode surface), which indicated that organic compounds mainly reacted with adsorbed hydroxyl radicals at PbO2 anodes. For Ti/SnO2-Sb electrodes, however, k increased with the increase of the integrated parameter S (representing the effects of both sigma and gamma), which implied that organic compounds reacted with both adsorbed hydroxyl radicals and free hydroxyl radicals at SnO2 anodes. PMID- 18678027 TI - Polyoxometalate-enhanced oxidation of organic compounds by nanoparticulate zero valent iron and ferrous ion in the presence of oxygen. AB - In the presence of oxygen, organic compounds can be oxidized by zerovalent iron or dissolved Fe(II). However, this process is not a very effective means of degrading contaminants because the yields of oxidants are usually low (i.e., typically less than 5% of the iron added is converted into oxidants capable of transforming organic compounds). The addition of polyoxometalate (POM) greatly increases the yield of oxidants in both systems. The mechanism of POM enhancement depends on the solution pH. Under acidic conditions, POM mediates the electron transfer from nanoparticulate zerovalent iron (nZVI) or Fe(II) to oxygen, increasing the production of hydrogen peroxide, which is subsequently converted to hydroxyl radical through the Fenton reaction. At neutral pH values, iron forms a complex with POM, preventing iron precipitation on the nZVI surface and in bulk solution. At pH 7, the yield of oxidant approaches the theoretical maximum in the nZVI/O2 and the Fe(II)/O2 systems when POM is present, suggesting that coordination of iron by POM alters the mechanism of the Fenton reaction by converting the active oxidant from ferryl ion to hydroxyl radical. Comparable enhancements in oxidant yields are also observed when nZVI or Fe(II) is exposed to oxygen in the presence of silica-immobilized POM. PMID- 18678028 TI - Bactericidal effect of zero-valent iron nanoparticles on Escherichia coli. AB - Zero-valent iron nanoparticles (nano-Fe0) in aqueous solution rapidly inactivated Escherichia coli. A strong bactericidal effect of nano-Fe0 was found under deaerated conditions, with a linear correlation between log inactivation and nano Fe0 dose (0.82 log inactivation/mg/L nano-Fe0 x h). The inactivation of E. coli under air saturation required much higher nano-Fe0 doses due to the corrosion and surface oxidation of nano-Fe0 by dissolved oxygen. Significant physical disruption of the cell membranes was observed in E. coli exposed to nano-Fe0, which may have caused the inactivation or enhanced the biocidal effects of dissolved iron. The reaction of Fe(II) with intracellular oxygen or hydrogen peroxide also may have induced oxidative stress by producing reactive oxygen species. The bactericidal effect of nano-Fe0 was a unique property of nano-Fe0, which was not observed in other types of iron-based compounds. PMID- 18678029 TI - Design of BDD-TiO2 hybrid electrode with P-N function for photoelectroatalytic degradation of organic contaminants. AB - P-N hybrid electrode of boron-doped diamond (BDD) and TiO2 were designed and fabricated via selective deposition of TiO2 onto BDD electrode. This hybrid electrode exhibit high photoelectrocatalytic activities toward degradation of acid orange II (AOII) and 2, 4-dichloropheonl (2,4-DCP) due to the P-N effect and high electrocatalytic and photocatalytic activities of BDD electrode and TiO2 particles. The structures of TiO2 and BDD were confirmed by Raman spectra analysis. Atom force microscopy and scanning electron microscopy showed that the TiO2 deposits consist of adherent nanomicro-sized particles, scattered on the BDD substrate. AOII and 2,4-DCP in a solution can be efficiently degraded at the hybrid electrode in the photoelectrocatalysis (PEC) process. Effect of applied bias potentials and solution pH values on AOII and 2,4-DCP degradation were investigated. In the electro-oxidation process, some intermediates such as phenols were detected and they accumulated with the reaction evolution based on the analysis of UV-vis and GC-MS variation. By contrast, phenols intermediates will be degraded with the reaction evolution in the photoelectrocatalysis process. And, organic aromatic and aliphatic carboxylic acids were detected. Furthermore, different degradation mechanism of AOII and 2,4-DCP in the electro oxidation, photocatalysis, and photoelectrocatalysis is proposed. PMID- 18678030 TI - Pretreatment of Hanford medium-curie wastes by fractional crystallization. AB - Acceleration of the schedule for decontamination of the Hanford site using bulk vitrification requires implementation of a pretreatment operation. Medium-curie waste must be separated into two fractions: one is to go to a waste treatment and immobilization plant and a second, which is low-activity waste, is to be processed by bulk vitrification. The work described here reports research on using fractional crystallization for that pretreatment. Sodium salts are crystallized by evaporation of water from solutions simulating those removed from single-shell tanks, while leaving cesium in solution. The crystalline products are then recovered and qualified as low-activity waste, which is suitable upon redissolution for processing by bulk vitrification. The experimental program used semibatch operation in which a feed solution was continuously added to maintain a constant level in the crystallizer while evaporating water. The slurry recovered at the end of a run was filtered to recover product crystals, which were then analyzed to determine their composition. The results demonstrated that targets on cesium separation from the solids, fractional recovery of sodium salts, and sulfate content of the recovered salts can be achieved by the process tested. PMID- 18678031 TI - Degradation of lead-contaminated lignocellulosic waste by Phanerochaete chrysosporium and the reduction of lead toxicity. AB - Lead, as one of the most hazardous heavy metals to the environment interferes with lignocellulosic biomass bioconversion and carbon cycles in nature. The degradation of lead-polluted lignocellulosic waste and the restrain of lead hazards by solid-state fermentation with Phanerochaete chrysosporium were studied. Phanerochaete chrysosporium effectively degraded lignocellulose, formed humus and reduced active lead ions, even at the concentration of 400 mg/kg dry mass of lead. The highest lignocellulose degradation (56.8%) and organic matter loss (64.0%) were found at the concentration of 30 mg/kg of lead, and at low concentration of lead the capability of selective lignin biodegradation was enhanced. Microbial growth was delayed in polluted substrate at the initial stage of fermentation, and organic matter loss is correlated positively with microbial biomass after 12 day fermentation. It might be because Phanerochaete chrysosporium developed active defense mechanism to alleviate the lead toxicity. Scanning electron micrographs with energy spectra showed that lead was immobilized via two possible routes: adsorption and cation exchange on hypha, and the chelation by fungal metabolite. The present findings will improve the understandings about the degradation process and the lead immobilization pathway, which could be used as references for developing a fungi-based treatment technology for metal-contaminated lignocellulosic waste. PMID- 18678033 TI - Model-centered approach to early planning and design of an eco-industrial park around an oil refinery. AB - Industrial symbiosis promises environmental and economic gains through a utilization of the waste of some processes as a resource for other processes. Because of the costs and difficulties of transporting some wastes, the largest theoretical potential for industrial symbiosis is given when facilities are colocated in an eco-industrial park (EIP). This study proposes a model-centered approach with an eight-step procedure for the early planning and design of an eco industrial park considering technical and environmental factors. Chemical process simulation software was used to model the energy and material flows among the prospective members and to quantify the benefits of integration among different firms in terms of energy and resources saved as compared to a reference situation. Process simulation was based on a combination of physical models of industrial processes and empirical models. The modeling allows for the development and evaluation of different collaboration opportunities and configurations. It also enables testing chosen configurations under hypothetical situations or external conditions. We present a case study around an existing oil and gas refinery in Mongstad, Norway. We used the approach to propose the colocation of a number of industrial facilities around the refinery, focused on integrating energy use among the facilities. An EIP with six main members was designed and simulated, matching new hypothetical members in size to the existing operations, modeling material and energy flows in the EIP, and assessing these in terms of carbon and hydrogen flows. PMID- 18678032 TI - Photoreactive TiO2/carbon nanotube composites: synthesis and reactivity. AB - Electron-hole recombination limits the efficiency of TiO2 photocatalysis. We have investigated the efficacy with which anatase/carbon nanotube (CNT) composite materials reduce charge recombination and enhance reactivity. We synthesized nanostructured assemblies composed of different proportions of anatase (5 or 100 nm) and either single-or multi-walled CNTs. The composites were prepared using a simple low temperature process in which CNTs and anatase nanoparticles were dispersed in water, dehydrated at 80 degrees C, and dried at 104 degrees C. The structures of the various TiO2/CNT composites were characterized by scanning electron microscopy (SEM) and their function was tested by phenol oxidation. Charge recombination was compared by measuring the photoluminescence spectra of select composites. We found that a nanostructured composite assembled from the 100 nm anatase and single-walled CNTs (SWCNTs) exhibited enhanced and selective photocatalytic oxidation of phenol in comparison to both pure anatase and Degussa P25. A mechanism for the enhanced reactivity is proposed in which electrons are shuttled from TiO2 particles to the SWCNTs as a result of an optimal TiO2/ CNT arrangement that stabilizes charge separation and reduces charge recombination. In addition, the SWCNT assembly provides better catalyst-support (dispersal and connection) than multi-walled CNTs. PMID- 18678034 TI - Efficiency dilution: long-term exergy conversion trends in Japan. AB - This analysis characterizes century-scale trends in exergy efficiency in Japan. Exergy efficiency captures the degree to which energy inputs (such as coal) are converted into useful work (such as electricity or power to move a vehicle). This approach enables the estimation of net efficiencies which aggregate different technologies. Sectors specifically analyzed are electricity generation, transport, steel production, and residential space heating. One result is that the aggregate exergy efficiency of the Japanese economy declined slightly over the last half of the 20th century, reaching a high of around 38% in the late 1970s and falling to around 33% by 1998. The explanation for this is that while individual technologies improved dramatically over the century, less exergy efficient ones were progressively adopted, yielding a net stabilization or decline. In the electricity sector, for instance, adoption of hydropower was followed by fossil-fired plants and then by nuclear power, each technology being successively less efficient from an exergy perspective. The underlying dynamic of this trend is analogous to declining ore grades in the mining sector. Increasing demand for exergy services requires expended utilization of resources from which it is more difficult to extract utility (e.g., falling water versus coal). We term this phenomenon efficiency dilution. PMID- 18678035 TI - Activated carbon cloth as anode for sulfate removal in a microbial fuel cell. AB - By employing the sulfate-reducing bacterium Desulfovibrio desulfuricans we demonstrate the possibility of electricity generation in a microbialfuel cell (MFC) with concomitant sulfate removal. This approach is based on an in situ anodic oxidative depletion of sulfide produced by D. desulfuricans. Three different electrode materials, graphite foil (GF), carbon fiber veil (CFV), and high surface area activated carbon cloth (ACC), were evaluated for sulfide electrochemical oxidation. In comparison to CFV and GF electrodes, ACC was a superior materialfor sulfide adsorption and oxidation and showed significant potential for harvesting energy from sulfate-rich solutions in the form of electricity. Sulfate (3.03 g dm(-3)) was removed from a bacterial suspension, which represented 99% removal. A maximum power density of 0.51 mW cm(-2) (normalized to geometric electrode area) was obtained with a one-chamber, air breathing cathode and continuous flow MFC operated in batch mode at 22 degrees C. PMID- 18678036 TI - Exergy analysis of integrated waste management in the recovery and recycling of used cooking oils. AB - Used cooking oil (UCO) is a domestic waste generated daily by food industries, restaurants, and households. It is estimated that in Europe 5 kg of UCO are generated per inhabitant, totalling 2.5 million metric tons per year. Recovering UCO for the production of biodiesel offers a way of minimizing and avoiding this waste and related pollution. An exergy analysis of the integrated waste management (IWM) scheme for UCO is used to evaluate such a possibility by accounting for inputs and outputs in each stage, calculating the exergy loss and the resource input and quantifying the possible improvements. The IWM includes the collection, pretreatment, and delivery of UCO and the production of biodiesel. The results show that the greatest exergy loss occurs during the transport stages (57%). Such exergy loss can be minimized to 20% by exploiting the full capacity of collecting vans and using biodiesel in the transport stages. Further, the cumulative exergy consumption helps study how the exergy consumption of biodiesel can be further reduced by using methanol obtained from biogas in the transesterification stage. Finally, the paper discusses how increasing the collection of UCO helps minimize uncontrolled used oil disposal and consequently provides a sustainable process for biodiesel production. PMID- 18678037 TI - Copper oxide-based model of persistent free radical formation on combustion derived particulate matter. AB - We have found that environmentally persistent free radicals (PFRs) are formed by adsorption of substituted aromatic molecular precursors on the surface of cupric oxide-containing particles at temperatures between 100 and 400 degrees C. This temperature range corresponds to the conditions in the postflame, cool zone of combustion, and thermal processes. Depending upon the nature of the precursor and the adsorption temperature, both substituted phenoxyl and semiquinone radicals are formed. The PFRs are formed through a mechanism of initial physisorption, followed by chemisorption via elimination of water or hydrogen chloride, and electron transfer resulting in the simultaneous reduction of Cu(II) to Cu(I) and formation of the PFR. The PFRs are still observable by electron paramagnetic resonance (EPR) after exposure to air for more than a day. Their lifetimes under vacuum appear to be infinite. Other redox-active transition metals such as iron are expected to also mediate or catalyze the formation of PFRs. The properties of the observed radicals are consistent with radicals previously observed on airborne and combustion-generated particulate matter. We propose a catalytic biochemical cycle for both the particle-associated semiquinone and phenoxyl PFRs that result in the formation of hydroxyl radical and other reactive oxygen species (ROS). This suggests that combustion-generated, particle-associated PFRs may be responsible for the oxidative stress resulting in cardiopulmonary disease and probably cancer that has been attributed to exposure to airborne fine particles. PMID- 18678038 TI - Decline in perfluorooctanesulfonate and other polyfluoroalkyl chemicals in American Red Cross adult blood donors, 2000-2006. AB - In 2000, 3M Company, the primary global manufacturer, announced a phase-out of perfluorooctanesulfonyl fluoride (POSF, C8F17SO2F)-based materials after perfluorooctanesulfonate (PFOS, C8F17SO3-) was reported in human populations and wildlife. The purpose of this study was to determine whether PFOS and other polyfluoroalkyl concentrations in plasma samples, collected in 2006 from six American Red Cross adult blood donor centers, have declined compared to nonpaired serum samples from the same locations in 2000-2001. For each location, 100 samples were obtained evenly distributed by age (20-69 years) and sex. Analytes measured, using tandem mass spectrometry, were PFOS, perfluorooctanoate (PFOA), perfluorohexanesulfonate (PFHxS), perfluorobutanesulfonate (PFBS), N-methyl perfluorooctanesulfonamidoacetate (Me-PFOSA-AcOH), and N-ethyl perfluorooctanesulfonamidoacetate (Et-PFOSA-AcOH). The geometric mean plasma concentrations were for PFOS 14.5 ng/mL (95% CI 13.9-15.2), PFOA 3.4 ng/ mL (95% CI 3.3-3.6), and PFHxS 1.5 ng/mL (95% CI 1.4-1.6). The majority of PFBS, Me-PFOSA AcOH, and Et-PFOSA-AcOH concentrations were less than the lower limit of quantitation. Age- and sex-adjusted geometric means were lower in 2006 (approximately 60% for PFOS, 25% for PFOA, and 30% for PFHxS) than those in 2000 2001. The declines for PFOS and PFHxS are consistent with their serum elimination half-lives and the time since the phase-out of POSF-based materials. The shorter serum elimination half-life for PFOA and its smaller percentage decline than PFOS suggests PFOA concentrations measured in the general population are unlikely to be solely attributed to POSF-based materials. Direct and indirect exposure sources of PFOA could include historic and ongoing electrochemical cell fluorination (ECF) of PFOA, telomer production of PFOA, fluorotelomer-based precursors, and other fluoropoly-mer production. PMID- 18678039 TI - Salmon olfaction is impaired by an environmentally realistic pesticide mixture. AB - Many of the salmon-producing waterways of the world contain pesticides known to harm olfactory sensory neurons (OSNs) that are critically important throughout the salmon lifecycle. The ability of OSNs to retain functionality after exposure to complex pesticide mixtures remains unknown. Here we show that a 96-h exposure to an environmentally realistic concentration of a mixture made from the ten most frequently occurring pesticides in British Columbia's Nicomekl River reduced the OSN responses of rainbow troutto a behaviorally relevant odorant. Odor-evoked responses were not altered by exposure to one-fifth of the realistic concentration, and this may have been due an upregulation in detoxification enzymes, since glutathione-S-transferase activity reached a maximum (> 32% above control) at this concentration. Mixture exposure did not help to prevent OSN impairment from a second, brief (5 min) exposure to a higher (20 x) concentration of the mixture, suggesting longer-term, low-concentration exposures may not prevent damage from brief, high-concentration pulse exposures. This study demonstrates that environmentally observed pesticide mixtures can injure salmon olfactory tissue, and by extension, contribute to the threatened and endangered status of many salmon stocks. PMID- 18678040 TI - Alveolar air and urine analyses as biomarkers of exposure to trihalomethanes in an indoor swimming pool. AB - The exposure of workers and swimmers at an indoor swimming pool to trihalomethanes (THMs) as a consequence of water chlorination was evaluated by analyzing alveolar air and urine samples. Environmental monitoring of THMs in water and ambient air was also performed in order to assess the possible correlation between environmental and biological samples. The sampling was done concurrently, taking the urine and alveolar air samples before and after the work shift for 15 workers and the swimming activity for 12 swimmers. A high THM uptake was observed in alveolar air and urine of subjects exposed, with chloroform being the most abundant THM. Mean chloroform levels in alveolar air and urine before exposure were 4 microg/ m3 and 475 ng/L, respectively. After 2 h of exposure, concentration increases of ca. 8 times in alveolar air and 2 times in urine were observed in workers. After 1 h swimming, the increases found in swimmers were ca. 20 and 3 times in alveolar air and urine, respectively. High increases have also been observed in bromodichloromethane levels. We have obtained excellent correlations between the chloroform concentrations found in the swimming pool ambient air/alveolar air, and between the urine/ alveolar air of the participants after exposure (r > 0.9). In conclusion, alveolar air provides better response sensitivity and shorter reaction time to external exposure than urine, being therefore the most sensitive biomarker. PMID- 18678041 TI - High percentage inorganic arsenic content of mining impacted and nonimpacted Chinese rice. AB - Two approaches were undertaken to characterize the arsenic (As) content of Chinese rice. First, a national market basket survey (n = 240) was conducted in provincial capitals, sourcing grain from China's premier rice production areas. Second, to reflect rural diets, paddy rice (n = 195) directly from farmers fields were collected from three regions in Hunan, a key rice producing province located in southern China. Two of the sites were within mining and smeltery districts, and the third was devoid of large-scale metal processing industries. Arsenic levels were determined in all the samples while a subset (n = 33) were characterized for As species, using a new simple and rapid extraction method suitable for use with Hamilton PRP-X100 anion exchange columns and HPLC-ICP-MS. The vast majority (85%) of the market rice grains possessed total As levels < 150 ng g(-1). The rice collected from mine-impacted regions, however, were found to be highly enriched in As, reaching concentrations of up to 624 ng g(-1). Inorganic As (As(i)) was the predominant species detected in all of the speciated grain, with As(i) levels in some samples exceeding 300 ng g(-1). The As(i) concentration in polished and unpolished Chinese rice was successfully predicted from total As levels. The mean baseline concentrations for As(i) in Chinese market rice based on this survey were estimated to be 96 ng g(-1) while levels in mine-impacted areas were higher with ca. 50% of the rice in one region predicted to fail the national standard. PMID- 18678042 TI - Fenton-like reaction catalyzed by the rare earth inner transition metal cerium. AB - Cerium (Ce) is a rare earth metal that is not known to have any biological role. Cerium oxide materials of several sizes and shapes have been developed in recent years as a scaffold for catalysts. Indeed even cerium oxide nanoparticles themselves have displayed catalytic activities and antioxidant properties in tissue culture and animal models. Because of ceria's ability to cycle between the +3 and +4 states at oxygen vacancy sites, we investigated whether cerium metal would catalyze a Fenton-like reaction with hydrogen peroxide. Indeed, cerium chloride did exhibit radical production in the presence of hydrogen peroxide, as assessed by relaxation of supercoiled plasmid DNA. Radical production in this reaction was also followed by production of radical cation of 2,2'-azinobis-(3 ethylbenzthiazoline-6-sulfonic acid) (ABTS). Radical scavengers and spin traps were capable of competing with ABTS for radicals produced in this cerium dependent Fenton-like reaction. Electron paramagnetic resonance experiments reveal both hydroxyl radical and superoxide anion in a reaction containing cerium and hydrogen peroxide. Based on these results we propose that cerium is capable of redox-cycling with peroxide to generate damaging oxygen radicals. PMID- 18678043 TI - An environmental estrogen alters reproductive hierarchies, disrupting sexual selection in group-spawning fish. AB - There is global concern regarding the potential impacts of endocrine-disrupting chemicals (EDCs) on the health of wildlife and humans. Exposure to some estrogens, at concentrations found in the environment impairs reproductive function and behavior. However, nearly all work on endocrine disruption has investigated the effects of exposure on individuals and there is an urgent need to understand impacts on populations. Many fish have mating systems with complex social structures and it is not known whether EDCs will exaggerate or buffer the reproductive skews caused by the dominance hierarchies that normally occur for these species. This study investigated the impact of exposure to the pharmaceutical estrogen ethinylestradiol (EE2) on reproductive hierarchies and sexual selection in group-spawning fish. Breeding zebrafish were exposed to environmentally relevant concentrations of EE2, and effects were determined on reproductive output, plasma androgen concentrations (in males), and reproductive success through microsatellite analyses of the offspring. Reproductive hierarchies in breeding colonies of zebrafish were disrupted by exposure to EE2 at a concentration that did not affect the number of eggs produced. The effect was a reduction in the skew in male paternity and increased skew in female maternity. This disruption in the reproductive hierarchy in group spawning fish, if it occurs in the wild, has potentially major implications for population genetic diversity. Reproductive success in male zebrafish was associated with elevated plasma concentrations of the male sex hormone 11-ketotestosterone and this hormone was suppressed in EE2-exposed males. PMID- 18678044 TI - Ingested microscopic plastic translocates to the circulatory system of the mussel, Mytilus edulis (L). AB - Plastics debris is accumulating in the environment and is fragmenting into smaller pieces; as it does, the potential for ingestion by animals increases. The consequences of macroplastic debris for wildlife are well documented, however the impacts of microplastic (< 1 mm) are poorly understood. The mussel, Mytilus edulis, was used to investigate ingestion, translocation, and accumulation of this debris. Initial experiments showed that upon ingestion, microplastic accumulated in the gut. Mussels were subsequently exposed to treatments containing seawater and microplastic (3.0 or 9.6 microm). After transfer to clean conditions, microplastic was tracked in the hemolymph. Particles translocated from the gut to the circulatory system within 3 days and persisted for over 48 days. Abundance of microplastic was greatest after 12 days and declined thereafter. Smaller particles were more abundant than larger particles and our data indicate as plastic fragments into smaller particles, the potential for accumulation in the tissues of an organism increases. The short-term pulse exposure used here did not result in significant biological effects. However, plastics are exceedingly durable and so further work using a wider range of organisms, polymers, and periods of exposure will be required to establish the biological consequences of this debris. PMID- 18678045 TI - [Pain management in the family doctor's office]. PMID- 18678046 TI - [Patients with epilepsy in the family doctor's office]. PMID- 18678047 TI - [Warning symptom TIA--role of the general practitioner]. PMID- 18678048 TI - [Transient global amnesia: an often misconceived disorder]. PMID- 18678049 TI - [Challenging pharmacotherapy in dementia]. PMID- 18678050 TI - [Bipolar disorders]. PMID- 18678051 TI - [Behavioral therapy in primary care]. PMID- 18678052 TI - [Acute headache: what you should not at all fail to do]. PMID- 18678053 TI - [Could it be depression? Early symptoms and forewarnings]. PMID- 18678054 TI - [Depression: treatment in the family doctor's office]. PMID- 18678055 TI - [Therapy of Parkinson's disease 2008]. PMID- 18678056 TI - [Diagnosis and treatment of anxiety disorders]. PMID- 18678057 TI - [Myasthenia gravis--diagnosis, therapy and special problems]. PMID- 18678058 TI - [Successful cardiac intervention--anxiety continues]. PMID- 18678059 TI - [Treatment of the restless legs syndrome]. PMID- 18678060 TI - [Effect of expectancies on negative mood regulation]. AB - The role of expectancies regarding negative mood regulation in improving the mood states of undergraduate students when they experience both negative and positive life events was investigated. In Study 1, a Japanese language scale for measuring expectancies for negative mood regulation (NMR scale) was developed. In Study 2, undergraduates (n = 95) completed the NMR scale twice within an interval of five weeks. They also completed a self-rating depression scale and a scale of life events in interpersonal and achievement domains. The results indicated that participants with high NMR scores did not experience depression when they faced negative events, if they concurrently experienced positive life events. These results suggest that expectancies for negative mood regulation alleviate the negative mood states by using a mood regulation strategy moderated by positive life events, and predict the level of depression. PMID- 18678061 TI - [Adult mother-daughter relationships and psychological well-being: attachment to mothers, depressive symptoms, and self-esteem]. AB - This study examined how daughter's reported quality of their mother-daughter relationships during childhood and adulthood is related to their psychological well-being (depressive symptoms and self-esteem). A cross-sectional sample of 363 women, age 26 to 35 years, completed questionnaires. The association between the quality of daughters' relationships with their mothers and their psychological well-being depended on the daughters' marital and parental status. Regression estimates suggested that among single daughters and married daughters with children, childhood attachment dimensions (avoidance and anxiety) significantly contributed to psychological well-being, even after controlling for the effects of current closeness and excessive dependence. Current closeness, and excessive care seeking and care giving to their mother contributed to the psychological well-being of single daughters and married daughters without children, even after controlling for the effects of childhood attachment. PMID- 18678062 TI - [Visual illusion of a vertical gradient: A pitched visual environment and a pitched floor]. AB - We investgated visual illusions of a vertical gradient by using minitures of a road in three experiments. The independent variables were the pitched visual pattern around the road and the pitched floor on which the observer stood. The dependent variable was the angle of the road that the observer adjusted to appear to be level in depth. In Experiment 1, the experimental room was pitched independently of the pitched floor on which the observer stood. Pitching the room was found to be effective for the illusion, but pitching the floor was not. In Experiment 2, the horizontally striped pattern of the sidewalls was.pitched in the range of +/- 30 degrees and the height of the sidewalls was also varied. The illusion was found to be maximal for patterns pitched at +/- 10 degrees, but the sidewall height did not influence the illusion. In Experiment 3, six visual patterns that were produced by combining horizontal and vertical stripes were pitched at +/- 10 degrees. The results showed that the lattice pattern was the most effective, and the horizontal stripe was more effective than the vertical, in contributing to the illusion. PMID- 18678063 TI - [Independent resource of each hemisphere modulates selective attention]. AB - Based on the load theory and the assumption that each hemisphere has independent resources, we examined the effects of perceptual load in each hemisphere on the compatibility effect. In Experiments 1, and 2ab, two letter-strings were presented to the left and right visual-fields with a distracter, which was presented on the center of the screen. Two conditions were prepared by pairing a letter-string which contained a target with one which did not. Right-handed participants were asked to identify the target in the letter-strings while ignoring the distracter. The results showed that the compatibility effect was larger when the perceptual load of the letter-string which did not contain a target was low. This suggests that the residual resources of the hemisphere where the target was not projected facilitated the processing of the distracter. In Experiment 3, two letter-strings were presented to both hemispheres. The results showed that the compatibility effect was constant, irrespective of the perceptual load of the letter-string. Our findings suggested that selective attention is modulated by the resources of each hemisphere. PMID- 18678064 TI - [Semantic priming within and between the visual fields: an event-related brain potential study]. AB - To investigate interhemispheric transfer of language information, event-related brain potentials (ERPs) were recorded during performance of a semantic matching task in which prime and target word pairs were sequentially presented to either the left (LVF) or the right (RVF) visual field. A posterior slow negative shift prior to the target presentation developed over both hemispheres for LVF primes, whereas for RVF primes the negative shift declined over the right hemisphere. An N400 attenuation was observed for targets semantically related to the primes and was predominant over the left parietal site. The N400 priming effect was significantly reduced for LVF targets preceded by RVF primes, compared to other patterns of presentation. These findings indicate that semantic priming is attenuated in the right hemisphere when information is transferred from the left hemisphere. PMID- 18678065 TI - [Infants' responses to the still-face situation at five and nine months: focusing on sex differences]. AB - This study investigated developmental changes and sex differences in infants' responses to a female stranger during a face-to-face interaction, still-face, and reunion phase. Twenty-two infants (11 boys and 11 girls) were observed at five and nine months of age. At five months, the infants smiled less in the still-face phase, while at nine months the infants smiled less at each subsequent phase. Girls, but not boys, smiled more at nine months than five months. Girls' gazing toward the social partner's face decreased during the still-face and reunion phases. Girls gazed toward the social partner's face more than the boys, independent of the phases and age. The findings support the view the effect of a still-face on infants is robust at both five and nine months and for both sexes. But the effect remained in the reunion phase only for the nine-month-olds. These findings suggest that girls have more interest in their social partner than boys. PMID- 18678066 TI - [Influence of odor cueing on the retrieval processes for autobiographical memories]. AB - This study investigated the influence of odor cueing on the retrieval process for autobiographical memories. In Experiment 1, forty two participants were asked to retrieve autobiographical memories in response to a label cue only, or a label cue with a congruent odor cue, or label cue with an incongruent odor cue. The results showed that in the label cue only condition and the incongruent odor cue condition, the latency with a concurrent task was slower than with no concurrent task. However, this was not observed in the congruent odor cue condition. In order to verify the results of Experiment 1, in Experiment 2, twenty participants were investigated with regard to autobiographical memories cued by odors having a high or low rate of identification. The results showed that memories cued by odors with a high rate of identification were retrieved faster. These results suggest that dual processing by odor and label promotes retrieval processes for autobiographical memories. PMID- 18678067 TI - [Influence of evaluations and coping behavior in relationships with supervisors on the psychological stress responses of subordinates: A comparison between full time workers and students working part-time jobs]. AB - This research examined workers' and supervisors' evaluations of their relationships and coping behaviors as related to the workers' psychological stress responses. The participants were full-time workers and students working part-time jobs. This study focused on both informal and formal relationships. The results showed that full-time workers who evaluated their informal relationships with their supervisors as being more negative and perceived their supervisors' evaluations as being more positive had greater psychological stress responses. The psychological stress responses of students working part-time jobs were not significantly associated with their evaluations or their supervisors' evaluations. The results for full-time workers indicate that supervisors' positive evaluations of their informal relationships increased the subordinates' psychological stress responses. The proportion of variance attributed to the coping behavior of full-time workers was lower than for students working part time jobs. PMID- 18678068 TI - [Subjective probability of reward receipt and the magnitude effect in probability discounting]. AB - Previous research suggested that larger probabilistic rewards were discounted more steeply than smaller probabilistic rewards (the magnitude effect). This research tests the hypothesis that the magnitude effect reflects the extent to which individuals distrust the stated probability of receiving different amounts of rewards. The participants were 105 college students. Probability discounting of two different amounts of rewards (5 000 yen and 100 000 yen) and the subjective probability of reward receipt of the different amounts (5 000 yen, 100 000 yen and 1 000 000 yen) were measured. The probabilistic 100 000 yen was discounted more steeply than the probabilistic 5 000 yen. The subjective probability of reward receipt was higher in the 5 000 yen than in the 100 000 yen condition. The proportion of subjective probability of receiving 5 000 yen to that of receiving 100 000 yen was significantly correlated with the proportion of degree of probability discounting for 5 000 yen to that for 100 000 yen. These results were consistent with the hypothesis stated above. PMID- 18678069 TI - [Nursing response to health care in France, concerning a discipline]. PMID- 18678070 TI - [Nursing clinic, a reality that needs to be developed]. PMID- 18678071 TI - [Intimacy of nursing]. PMID- 18678072 TI - [Production of nursing writing]. PMID- 18678073 TI - [Nursing competence close by a public in great danger]. PMID- 18678074 TI - [Protocols, references to nurses, quality of collective autonomy and personal dependence]. PMID- 18678075 TI - [Groups which analyse clinical practice, an opportunity for framing nursing to reinforce the management of nursing]. PMID- 18678076 TI - [Radioscopy:of the nursing profession in the twenty first century]. PMID- 18678077 TI - [Nursing movements, professional representation and social conflicts]. PMID- 18678078 TI - [Current problems in nursing]. PMID- 18678079 TI - [Thought and thinking: splendor and misery of their proper roles]. PMID- 18678080 TI - [Nurses: practice and redefinition; an opportunity to mark dimensions in nursing?]. PMID- 18678081 TI - [Conditions for the emergence of a scientific and professional discipline]. PMID- 18678082 TI - [Science of nursing: epistemologic reflections on a project of discipline ]. PMID- 18678083 TI - [Reflections on the history of nurses in Quebec]. PMID- 18678084 TI - [Practice, role and position of nurses in primary care in six European countries, in Ontario and in Quebec]. PMID- 18678085 TI - [New roles for nurses in Ireland]. PMID- 18678086 TI - [Lessons learned from a French experience: nursing science at the UFR in Bobigny]. PMID- 18678087 TI - [Improbable interdisciplinary science and techniques of physical and sports activities]. PMID- 18678088 TI - [Universality of development of nurses: promises and shadows]. PMID- 18678089 TI - [Development of universities and research in nursing: a goal for the profession, health and society?]. PMID- 18678090 TI - [Universities for the formation of nurses in France: an opportunity for professionals, usage and development of public health?]. PMID- 18678091 TI - [Multiple challenges for the nursing profession and research on nursing care in France: registering the process in the course of mutations in research and public health]. PMID- 18678092 TI - [Elements of synthesis in the works of Congress: from the point of view of knowledge, university training of nurses and research]. PMID- 18678093 TI - [Allergen challenge used to diagnose food allergy--oral allergy syndrome]. AB - Food allergy is an immune mediated unwanted side effect to food. All types of hypersensitivity can be involved; the most prevalent is IgE mediated one. The diagnosis is based on the history, skin tests with allergens and detection of specific IgE. These procedures are not highly predictive for clinical reactivity to food allergens, true clinical reactivity can be confirmed by allergen challenge. Oral allergy syndrome is represented by mostly subjective symptoms in oral cavity, usually triggered by fruit or vegetable allergens, cross-reacting with pollen allergens. This cross-reactivity is based on protein homology and immunologic similarity. To confirm the diagnosis of food allergy, the ,,golden standard" is the double blind placebo controlled food challenge. The basic features of this test are discussed, particularly in the diagnosis of the oral allergy syndrome. Controlled food challenge is a useful test for patient's diagnosis confirmation, evaluation of other diagnostic tests or evaluation of food allergenicity. PMID- 18678094 TI - [Gene expression changes during insulin resistance and "diabesity" in insulin sensitive tissues and possibilities of their regulation]. AB - Type 2 diabetes mellitus (T2DM) gains considerable and pandemic proportions and becomes noticeable problem in a social and economic sphere. In spite of all effort, exact mechanism of T2DM origin has not been elucidated yet. Studying of transcriptom is one possibility how to explain pathophysiological processes in insulin-sensitive tissues. Obtained data can serve as a base for predicting of new therapeutic targets of this disease. This overall review introduces crucial genes whose level of products changes during T2DM. The article gives notice to a diet composition, which is an important environmental factor, which is able to influence a disease outbreak. Not only the role of fats, but also influence of some plant compounds, which would be able to serve as an alternative to present prophylaxis or treatment of T2DM, have been discussed. PMID- 18678095 TI - [Biological and non-biological elimination therapy of acute liver failure. Experimental study on large laboratory animal]. AB - BACKGROUND: Development of biological and non-biological artificial liver devices in the previous 20 years enabled effective treatment of acute liver failure (ALF) of patients waiting for liver transplantation or for spontaneous liver parenchyma regeneration. Aim of the study was the evaluation of the effectiveness of biological (BAL - bioartificial liver) and non-biological (FPSA - Fractionated plasma separation and adsorption) methods in the treatment of experimental ALF on large laboratory animal. METHODS AND RESULTS: Surgical model of ALF with liver devascularization in pigs (weight 25-40 kg) was provided following monitoring of ALF markers (AST, ALT, bilirubin, ammoniac, glycaemia, INR) including intracranial pressure (ICP). Control group included animals without treatment of ALF. Results of both experimental groups were compared and statistically worked out with that of controls by T-test and Mann-Whitney non-parametric test by EXCEL and QUATRO. BAL group: 10 pigs (weight 30 +/- 5 kg) with ALF were treated by BAL with isolated hepatocytes. When plasma bilirubin was compared, significant differences (p < 0.05) in 6 and 9 hours interval were found favouring BAL group (18.1 vs. 13.1, 22.9 vs. 13.2 mmol/l). The value of ICP in both groups was no significant. Prometheus group: 14 pigs weight 35 kg (35 +/- 5 kg) with the identical ALF were treated by Prometheus (FPSA). Level of serum bilirubin in experimental group when compared to control group was significantly lower (p < 0.01) at 6 hour interval 12.81 +/- 6.54 vs. 29.84 +/- 9.99 at 9 hour 11.94 +/- 4.14 vs. 29.95 +/- 12.36 and at 12 hour 13.88 +/- 6.31 vs. 26.10 +/- 12.23 mmol/l. No significant difference in serum ammonia level was found. ICP was significantly different from 9 hour to 12 hour interval in favour of FPSA group (p < 0.01): 9 hour 19.1 +/- 4.09 vs. 24.1 +/- 2.85, 10 hour 21.9 +/- 3.63 vs. 25.1 +/- 2.19, 11 hour 22.5 +/- 3.98 vs. 26.3 +/- 3.50 and 12 hour 24.0 +/- 4.66 vs. 29.8 +/- 5.88 mm Hg. CONCLUSIONS: Significant improvement of bilirubin and ICP levels resulting from the treatment with fractionated plasma separation and adsorption (Prometheus) were observed in the case of experimental ALE Except the bilirubin levels, bioartificial liver provided by O. liver Performer with isolated hepatocytes did not bring any significant improvement of laboratory markers, including ICP. PMID- 18678096 TI - [Relation between insulin resistance and amount of visceral adipose tissue]. AB - BACKGROUND: Association of obesity with metabolic and cardiovascular complications depends on the adipose tissue distribution. The role of intraabdominal, i.e. visceral, adipose tissue in pathogenesis of insulin resistance is still not elucidated. The aim of this study was to investigate the relation between insulin resistance and contribution of visceral and subcutaneous fat in a group of women with a wide range of body weight. METHODS AND RESULTS: 62 women (age 21-66 years) among which 32 were non-obese and 30 obese (BMI > 30 kg/m2) were examined. The amount of visceral and subcutaneous fat was evaluated using computerized tomography, total body fat evaluated using bioimpedance, and the degree of insulin resistance was evaluated using glucose disposal (M) during euglycemic hyperinsulinemic clamp. Obese women had lower insulin sensitivity than non-obese (5.88 +/- 2.17 vs 3.32 +/- 1.44 mg/min/kg, p <0.001) and higher absolute amount of visceral fat. However, the relative amount of visceral fat (related to total body fat or subcutaneous fat) was not different between the two groups. In the entire study group, the magnitude of insulin sensitivity did correlate with absolute amount of total and visceral fat, but no correlation with relative amount of visceral fat was found. CONCLUSIONS: The results suggest that the absolute amount of fat, either total or visceral, is a stronger predictor of the degree of insulin resistance than the relative contribution of visceral fat. PMID- 18678097 TI - [Gene expression profiling in prediction of tumor response to neoadjuvant concomitant chemoradiotherapy in patients with locally advanced rectal carcinoma: pilot study]. AB - BACKGROUND: Neoadjuvant concomitant chemoradiotherapy has become a standard treatment of locally advanced rectal adenocarcinomas (LARA). It can reduce tumor volume, thus increases a feasibility of sphincter-sparing surgery, shows less acute toxicity, improves local control rate. It is based on fluoropyrimidines (5 fluorouracil, capecitabine) with concurrent radiotherapy. The aim of the study was to evaluate the capability of gene expression method to identify nonresponders (NR) pretherapeutically. METHODS AND RESULTS: 17 patients with LARA, clinical stage II, III according to IUCC were enrolled into our pilot study. Response to therapy was determined clinically by transrectal ultrasonography and CT/MRI before and after therapy and histopathologically by TRG (tumor regression grade) according to Mandard. Patients with TRG 1-2 were included to responders group (R) and patients with TRG 4-5 composed NR group. Gene expression levels of 440 genes were obtained by low-density oligonucleotide microarrays. Gene expression data analysis based on SAM (Significance Analysis of Microarrays) and t-test methods identified 8 genes (RB1, RBBP4, HYOUI, JUNB, MDM4, CANX, MMP2, TCF7L2) significantly upregulated in NR. CONCLUSIONS: Validation of identified changes on the mRNA level (Real-Time PCR) and on protein level (immunohistochemistry) is ongoing. We suggest that low-density oligonucleotide microarray technology could contribute to individualize the therapy of patients with LARA. PMID- 18678098 TI - [Conduct disorders in seven-year-old children--results of ELSPAC study. 3. Postnatal development]. AB - BACKGROUND: In the European Longitudinal Study of Pregnancy and Childhood (ELSPAC), the pediatric observation of the cohort of 6100 7-year-old children described 4.4% of them as having conduct disorders. METHODS AND RESULTS: The predominant markers such as strong negativism, aggressivity, attention deficit, non-adequate reactions, hyperactivity, disorders of psychomotoric development, school maturation delay, poorer school results and social adaptation described children behavioral problems. The first signs of conduct disorders were often discovered already at the 18th month and the 3rd year of children's age. The acceptance of standardized criteria of possible conduct disorders in the routine pediatric practice, and the special psychologic and/or psychiatric care for children with the confirmed diagnosis should be recommended as soon as in their toddler age. CONCLUSIONS: Possible causal associations between prenatal exposure to nicotine and conduct disorders in childhood are another strong reason for the primary prevention and smoking cessation in antenatal clinics. PMID- 18678099 TI - [Is the assessment of serum creatinine reliable?]. AB - Chemical and enzymatic methods are used to measure creatinine in serum and urine. Chemical methods are mostly based on the reaction of creatinine with alkaline picrate (Jaffe reaction). The Jaffe reaction is not specific for creatinine, the same reaction resulting in Jaffe-like chromogens show many interfering substances (ascorbic acid, bilirubin, proteins, ketones, cephalosporins and other drugs). Chemical and enzymatic methods show similar accuracy and day-to-day precision. Chemical methods are cheaper than enzymatic methods. Enzymatic methods require low sample volume and are not affected by the interfering substances as the chemical methods. Presented case report shows an unusual occurrence of drug interference in the enzymatic creatine deaminase procedure. Biological factors (circadian rhythm, pregnancy, hemodialysis, transplantation, stress, exercise), analytical and preanalytical factors (pH, glucose, pyruvate, bilirubin, fatty acids, sample storage and sample collection - gel tubes) and biological variability of creatinine play significant role in the creatinine examination. PMID- 18678100 TI - [What should we strive for and what should we omit?]. PMID- 18678101 TI - [Professor R. Cihak-anatomist and embryologist--on his 80th birthday]. PMID- 18678102 TI - [Nobel Prize laureates]. PMID- 18678103 TI - Communicating health messages. PMID- 18678104 TI - As health bodies begin to engage with the press in earnest. PMID- 18678106 TI - Get out and run! Get caught! Just don't get caught. PMID- 18678105 TI - Marketing health, influencing behaviour. PMID- 18678107 TI - Mental pictures: citizen or consumer? PMID- 18678108 TI - The educational preparation of student nurses as communicators of health and wellbeing. PMID- 18678109 TI - The disorder of health information. PMID- 18678110 TI - 'Those terrible issues'. PMID- 18678111 TI - Sun safety: what are the health messages? AB - Solar ultraviolet radiation (UVR) is a human carcinogen. The UK and the worldwide incidence of cutaneous (skin) melanoma are rising. It is now the second most common cancer in 15-34 year-olds in the UK, increasing by 49% in 1991-2000. The UK government prioritized skin cancer in 1992 in their public health strategy The Health of the Nation, and set the target for reducing the yearly rise in skin cancer incidence by 2005. In 1998 in the Saving Lives: Our Healthier Nation strategy, targets were set to reduce the death rate from cancer, including skin cancer, by 20% by 2010. As a consequence of these targets, since the mid-1990s public health campaigns to reduce sun exposure and skin cancer incidence in the UK have used specific interventions for at-risk groups and the mass media to reach the general population. Evidence indicates a shift in knowledge, with further work required to increase change in behaviour and attitudes. An associate health problem is vitamin D deficiency, which is prevalent across certain UK population groups. The main childhood health problem associated with vitamin D deficiency is the development of rickets, with evidence indicating those at high risk are exclusively breastfed dark-skinned infants, the elderly, those with dark skin pigmentation, and certain cultural groups that cover their bodies in clothing. Adolescents are the group with the highest incidence of low vitamin D status in the UK, with children of Asian descent at higher risk than Caucasian children. Increasing the outdoor activity levels of children in winter and during the cooler hours in summer is one way of increasing safe sun exposure. It is imperative to educate the general public on the risk of vitamin D deficiency, and offer advice on safe sun exposure, dietary sources of vitamin D, and dietary supplements. More research is required on recommendations for daily vitamin D supplements to determine optimal vitamin D levels in different population groups. There also needs to be reliable and accessible evidence on the links between vitamin D deficiency and lack of sun exposure, with sun safe messages adjusted accordingly. PMID- 18678112 TI - Understanding mental distress in film and media: a new agenda? AB - This paper surveys some recent developments in media criticism and recent developments in film and media representations of mental distress. Focusing on a representations drawn from various forms of media, the paper argues that media and film images of mental distress are in many cases 'positive' and sympathetic, although they can also contain sexist, racist and other problematic elements that are not commonly identified by anti-stigma campaigners. It also suggests that while still valid in many ways, existing anti-stigma criticism tends to focus on a rather undifferentiated notion of 'violence to others' as the sole criterion against which media images are judged. Finally, the paper proposes that critics and campaigners pay closer attention to how the particular form or genre of any media text influences its treatment of psychological distress. PMID- 18678113 TI - Health information: a case of saturation or 57 channels and nothing on? AB - The claim is made by many that we have reached a point where we are exposed to too much information. This potential phenomenon is particularly prominent in the health field where there is a suggestion that the volume of available information has increased significantly and more importantly that such volume has had detrimental effects on both the overall quality of such information and the ability of people to process and use it. This paper explores the nature and validity of these claims. Within the context of the notion of an 'information society', it outlines a range of concerns that have been expressed in relation to this increase, including the simple problem of overload, the potential for less robust information to enter the system and thus the overall quality of available information to decline. This excess of information is seen to act against the benefits that are sought--information can be invalid and people may not have time to reflect and act on excessive information loads. This can result in irrationality or disinformation. The suggestion is made, however, that these concerns are largely unsupported by empirical evidence and are potentially the basis of a panic over the entry of alternative perspectives on health. The pessimistic perspectives are thus balanced by more constructive and optimistic views on this growth and opening up of information production and potential consumption. Seeing information creation as organic and pluralistic, it is suggested that increased information volume can actually be a constructive phenomenon. The paper concludes with the contention that it is unrealistic to expect a return to former circumstances of controlled and limited information flows. Rather, a series of more pragmatic suggestions is offered within existing circumstances, including differentiating between information rich and poor health areas, addressing structural issues like information access and health literacy, and working towards organizing health information so that it is of a high quality, is physically accessible, relevant to the needs and literacy of groups, and in a usable form. PMID- 18678114 TI - Public health and Web 2.0. AB - This article examines the nature and role of Web 2.0 resources and their impact on health information made available though the Internet. The transition of the Web from version one to Web 2.0 is described and the main features of the new Web examined. Two characteristic Web 2.0 resources are explored and the implications for the public and practitioners examined. First, what are known as 'user reviews' or 'user testimonials', which allow people to comment on the health services delivered to them, are described. Second, new mapping applications that take advantage of the interactive potential of Web 2.0 and provide tools to visualize complex data are examined. Following a discussion of the potential of Web 2.0, it is concluded that it offers considerable opportunities for disseminating health information and creating new sources of data, as well as generating new questions and dilemmas. PMID- 18678115 TI - Implementation of dietary and general lifestyle advice among women with polycystic ovarian syndrome. AB - AIMS: Central obesity and insulin resistance are strongly implicated in the etiology of polycystic ovarian syndrome (PCOS) and their reduction is therefore a central treatment focus. Weight loss has been consistently successful in reducing insulin resistance and restoring ovulation and fertility in women with PCOS. The purpose of the current study is to investigate the general attitudes towards dieting and exercise in women with PCOS, together with the extent of implementation of the dietary and lifestyle advice that these women are given as part of their treatment. General knowledge of patients about their condition (PCOS) has also been assessed. METHODS: Fifty three pre-menopausal women, over the age of 18 years, with a confirmed diagnosis of PCOS, were recruited from the Endocrinology Department, Middlesex Hospital, London. Subjects were interviewed individually using an interview-guided questionnaire. RESULTS: Sixty four per cent of the subjects were overweight (BMI > 25 kg/m2) and 55% of those were obese (BMI > 30 kg/m2). All subjects identified the importance of weight control in the management of their condition. However, only nine out of the 35 overweight women had actually been referred to a dietician and only 22 reported taking moderate exercise at least once a week in order to improve their health. The main sources of information on weight management were the internet and their consultant endocrinologist, who they normally see only twice a year. CONCLUSION: In spite of the fact that weight loss and weight maintenance are absolutely vital in the treatment of PCOS, our study has shown that the support given to PCOS patients to help reduce and control their weight is inadequate and needs to be improved. PMID- 18678116 TI - Risk-based regulatory system and its effective use in health and social care. AB - The regulatory landscape of the health and adult social care system in England is constantly changing. The establishment of a new regulator, the Care Quality Commission (CQC), will help to integrate the regulation of the health and adult social care sector together and strengthen the safety and quality assurance for patients and service users. It will have the challenge of regulating a much bigger sector and wider variety of organizations than its predecessor bodies- i.e. Healthcare Commission, Commission for Social Care Inspection and Mental Health Act Commission. An effective and efficient regulatory approach is needed by the new regulator to achieve the objectives, without increasing the regulatory burden on the services. A risk-based regulatory approach may help to deal with those challenges by proactively identifying and assessing any key risks to the quality and safety of these services. There are a number of perceived benefits of adapting such a risk-based approach by the regulator, both in terms of effectiveness and efficiency, as experienced by many other regulators in the UK and abroad. PMID- 18678117 TI - Understanding neonatal jaundice: UK practice and international profile. AB - Over the last 25 years there have been considerable advances in the treatment and technologies used in the care of newborn infants. Most of these advances are related to the care of the premature infants and there have been few changes in the management of conditions commonly seen in term infants. Neonatal jaundice is one of the commonest neonatal disorders and has been recognized since early history. Early neonatal jaundice is usually caused by the physiological destruction of red blood cells in the infant and its importance lies in the ability of the bilirubin pigment so produced to cross the blood brain barrier resulting in neurotoxicity. Prolonged neonatal jaundice (after 14 days of age) may be an indication of an underlying liver disorder. The approach to neonatal jaundice has remained largely unchanged over the last two to three decades. We continue to rely on visual inspection to assess the severity of early neonatal jaundice. We have technology that is effective in reducing the level of bilirubinaemia but in the UK there is no clear consensus as to the level at which jaundice should be treated. We do not have a standardized approach to the management of prolonged jaundice and there is potential for infants with significant liver problems to be diagnosed at a relatively late stage. Some countries (for example the US) have professional bodies who have introduced guidelines to ensure a standardized approach to the jaundice infant. We have little information about neonatal jaundice treatment in other parts of the developing world. PMID- 18678118 TI - Efficacy of activated charcoal in yellow oleander poisoning. PMID- 18678119 TI - Is fresh frozen plasma effective for thrombocytopenia in adults with dengue fever? A prospective randomised double blind controlled study. AB - RATIONALE: Thrombocytopenia is a common problem which causes concern and complications in dengue fever. If proven effective, intravenous fresh frozen plasma is a simple and widely available therapeutic option to manage thrombocytopenia. OBJECTIVE: To test the efficacy of fresh frozen plasma (FFP) on thrombocytopenia in patients with dengue fever. DESIGN: 109 serologically confirmed dengue patients with platelet counts <40 000/mm3 were randomised into two groups. Group A (treatment) comprised 53 patients and group B (control) 56 patients. Group A received an intravenous infusion of 3 units (600 ml) of FFP over 90 minutes. Group B received an intravenous infusion of an equal volume of isotonic saline over the same period. The primary outcome measure was the difference between pre- and post-interventional platelet counts at 12, 24 and 48 hours. RESULTS: Following Intervention, the mean platelet count was significantly higher in Group Athan in Group B at 12 hours (p=0.04; t-test). The mean platelet counts continued to be higher in Group A than in Group B at 24 and 48 hours post intervention, but the differences were not statistically significant. CONCLUSIONS: In dengue patients with thrombocytopenia, infusion of 600 ml FFP may contribute to a significant increase in platelet count in the first 12 hours, but not thereafter. PMID- 18678120 TI - Sinhala translation of child behaviour checklist: validity and reliability. AB - OBJECTIVE: To translate the child behaviour checklist (CBCL) into Sinhala and validate it for assessment of mental health status of children aged 5-10 years. DESIGN AND SETTING: Translation/back-translation method was used to translate the English CBCL into Sinhala. Each item in the Sinhala CBCL (CBCL-S) was rated by mental health professionals to determine semantics, content, and conceptual validity types. To ascertain criterion validity, total scores obtained for CBCL-S by administering it to parents or parent surrogates of 49 girls and 80 boys aged 5-10 years attending the specialist psychiatry clinics and 69 boys and 69 girls in the same age group from the community were compared with clinical diagnoses by a child psychiatrist. Receiver operator characteristic curves were drawn to obtain the cut-off points in CBCL-S for boys and girls separately. RESULTS: Semantics, content, and conceptual and criterion validity of CBCL-S were satisfactory. At the cut-off level of 39, CBCL-S had a sensitivity of 90% and a specificity of 88% for boys and a sensitivity of 89% and a specificity of 92% for girls. Internal consistency, test-retest reliability, and inter-interviewer reliability of CBCL-S were satisfactory. INTERPRETATION: CBCL-S is a valid and reliable instrument to measure mental health status of Sinhalese children aged 5 10 years in Sri Lanka. PMID- 18678121 TI - The operational cost of a urology unit. AB - OBJECTIVES: To assess the operational cost of a urology unit, individual cost for certain index operations in urology, and to develop a framework to audit finances of a unit. DESIGN: A financial audit. SETTING: Urology unit in a teaching hospital. METHODS: Data of cost in providing urology services during one month were collected. It included three main areas: ward, operating theatre and outpatient clinic. Direct costs included staff wages, drugs, consumables, investigations and food. Indirect expenses such as administration, water, electricity and cleaning services were also calculated. For each type of operation a relative value was assigned depending on the nature of the operation. When direct expenses were not available, the hospital was divided into different cost centres and apportioning of the cost was done accordingly. RESULTS: The monthly operational cost of running a 19 bed urology unit with three operating sessions a week was Rs. 1 294 259. Staff wages constituted 61.2% of the cost. The cost of performing a pyelolithotomy was Rs. 18 669. Transurethral resection of the prostate (TURP) was done at a cost of Rs. 21 271. CONCLUSION: When the basic principles and the framework are understood, clinicians can perform financial audit and cost analysis of their units. PMID- 18678122 TI - Validity and reliability of Tamil version of Strengths and Difficulties Questionnaire self-report. AB - BACKGROUND: Child and adolescent mental health services are scarce in Sri Lanka. Moreover there are no validated instruments to measure the mental health problems of Tamil speaking adolescents in Sri Lanka. Hence, strengths and difficulties questionnaire (SDQ) self-report was translated into Tamil and validated in the District of Jaffna, Sri Lanka. SDQ self-report was translated from English to Tamil using standard translation and back translation method. The Tamil version of the SDQ self-report was validated against a gold standard (ie. diagnosis made by a consultant psychiatrist). Both clinic and community samples were included in the validation study. Thirty-five adolescents (11-16 years) attending a psychiatric clinic and 91 adolescents from the community were included. The consultant psychiatrist made the diagnosis independent of the SDQ responses. Receiver operating characteristics (ROC) curve was used to measure the validity of SDQ. Reliability was measured using internal consistency. RESULTS: The ROC curve indicated high validity for the identification of adolescents with mental health problems: area under the curve 0.87 (95% - CI = 0.79-0.93). International cut-off point of 15 gave a sensitivity of 69.4% and specificity of 92.2%. Cronbach's alpha values (0.67-0.78) were good in all subscales and total difficulties scale. CONCLUSION: The SDQ self-report Tamil version can be used effectively for screening of adolescents to identify mental health problems as well as for research purposes. PMID- 18678123 TI - A case of imported malaria: the first report of a Plasmodium malariae infection in Sri Lanka after 37 years. AB - We report a case of Plasmodium falciparum and P. malariae mixed infection in a patient who had been living in Malawi. This is the first case of P. malariae reported in Sri Lanka in 4 decades. The presence of both parasites was confirmed by microscopy and polymerase chain reaction (PCR). The history strongly indicated that the infection had been acquired from Malawi. The patient had liver dysfunction and a transient glomerulonephritis, both of which subsided with antimalarial treatment. PMID- 18678124 TI - An unusual variant of malignant struma ovarii. AB - Struma ovarii is a highly specialised form of mature ovarian teratoma. When thyroid tissue constitutes 50% or more of the ovarian neoplasm, the tumour is termed struma ovarii. Malignant struma ovarii is rare and makes up 0.1-0.3% of all ovarian teratomas [2]. We describe a case of follicular variant of papillary carcinoma arising from struma ovarii with elevated CA-125 and pseudo-Meig syndrome. PMID- 18678125 TI - Diagnosis and management of depression. AB - Depression is a disabling illness which decreases quality of life and productivity. The point prevalence of major depression ranges from 5% to 9% among adults in primary care settings. Both pharmacological and psychological treatment are effective in treating depression. PMID- 18678126 TI - Depression in the physically ill. AB - The risk of depression is increased in many medical and surgical conditions. Specific serotonin reuptake inhibitors are generally recommended as first line treatment in the physically ill. Care must be taken in prescribing antidepressants in the physically ill, because of side-effects, hepatic and renal impairment, and potential for drug interactions. PMID- 18678127 TI - Depression in children and adolescents. AB - Among children and adolescents, the prevalence of depressive symptoms is about 15%. Clinically significant depression occurs in 5%. Depression in children overlaps with other childhood emotional and behavioural disorders. Depression in children is multifactorial and includes biological, psychological and social factors. Tricyclic antidepressants such as imipramine are ineffective in children and have limited effect in adolescents. SSRIs are also of limited value. For total health benefit in the treatment of depression, cognitive behaviour therapy (CBT) is found to be more beneficial than SSRIs. PMID- 18678128 TI - Serum carotenoids in eight Sri Lankan children. PMID- 18678129 TI - Well men clinic service in the primary health care setting. PMID- 18678130 TI - Multiple autoimmune phenomena preceding a lymphoproliferative disorder. PMID- 18678131 TI - Clinical laboratory practitioners speak out on Capitol Hill. PMID- 18678132 TI - Re: Clin Lab Sci 21(1) F O Delwiche. April 8, 2008. Articles about medline and PUBMED. PMID- 18678133 TI - Re: Clin Lab Sci 21(2) D D Beck. May 20, 2008. Progress of the DCLS. PMID- 18678134 TI - The doctorate in clinical laboratory science: an executive summary. AB - This article describes, in informational bullets, the concept of the doctorate in clinical laboratory science. The intent of the article is to support the marketing of these new practitioners and to provide the conceptual frame and links to data for proposals required to implement educational programs for them. PMID- 18678135 TI - The effects of over-anticoagulated blood on hematocrit values by the microcentrifuge method. AB - OBJECTIVE: To determine equivalency of hematocrit results by three methods. DESIGN: A total of 101 whole blood samples in EDTA tubes were analyzed in this repeated measures study. SETTING: East Carolina University's clinical laboratory science program, Greenville NC. PARTICIPANTS: The blood specimens were from adult patients at Nash General Hospital in Rocky Mount NC who had a CBC performed. MAIN OUTCOME MEASURE: Hematocrit values from a whole blood sample with EDTA anticoagulant performed by a Sysmex XE-2100 and by microcentrifuge with two different types of capillary tubes (i.e., heparinized and non-heparinized) filled from the EDTA tubes. RESULTS: The hematocrit means of the total sample for the three methods were 36.2%, 35.4%, and 35.6% for the Sysmex XE-2100, non heparinized capillary tubes, and heparinized capillary tubes, respectively. Pearson correlation coefficient (pairwise) analyses produced significant r-values at an alpha of .01 for all three method comparisons. CONCLUSIONS: Based on statistically significant Pearson (pairwise) correlation coefficients, the hematocrit values by all three methods can be considered relatively equivalent. The differences between methods are quite small and would be clinically insignificant, thus likely not altering clinical decisions. Though this study was conducted under somewhat ideal conditions relative to the blood specimens selected, the results indicate that the additional dilution produced in a heparinized capillary tube when being filled from an EDTA-anticoagulated tube is not sufficient to produce clinically different microhematocrit results as compared to using the recommended non-heparinized capillary tube. PMID- 18678137 TI - Validation study: clarity multistrip urocheck. AB - OBJECTIVE: To test the interchangeability of a previously untested urine reagent strip, Clarity (RAC Medical), with the gold standard, Multistix (Bayer). DESIGN: Seventy-six urine samples were tested with both the comparator and the gold standard urine reagent strips. Pairs of reagent strips were analyzed in the Clinitek Analyzer, recording the following: leukocytes, nitrite, urobilinogen, protein, pH, blood, specific gravity, ketone, bilirubin, glucose, and color. Data was assessed using statistical comparison of ordinal data (chi-square, Fisher's Exact, kappa, and weighted kappa). This study was approved by the Indiana University-Purdue University at Indianapolis Institutional Review Board. SETTING: The study took place at Wishard Health Services, Indianapolis IN. PATIENTS OR OTHER PARTICIPANTS: All urine tested was obtained from patients of the primary care clinic at Wishard Health Services. INTERVENTIONS: n/a. PRIMARY OUTCOME MEASURE: The ability for both reagent strips to generate (statistically significant) identical readings across all 11 measurements for each sample. RESULT: Kappa values were deemed the best indicator to consistently examine the reproducibility of all 11 measurements of the Clarity versus the Multistix. Ten of eleven measurements were concluded to be non-reproducible by the Clarity strips; nitrite readings achieved a kappa value above 0.85, whereas all other readings achieved kappa values well below the acceptable limits of this investigation (ranging from 0.00 to 0.65). CONCLUSION: There was a lack of statistically significant agreement between the results of both products and therefore it was concluded that both products cannot be used interchangeably. PMID- 18678136 TI - Growth inhibition of human colon cancer cells by plant compounds. AB - OBJECTIVE: Evidence is accumulating that compounds of plant origin (phytochemicals) exert anti-cancer effects. The purpose of this study was to determine if resveratrol, cinnamaldehyde, and piperine (from red grapes, cinnamon, black pepper respectively) have anti-proliferative effects on colon cancer. DESIGN: Quantitative effects of each phytochemical on concentration responses and time courses of proliferation of cultured human colon cancer cells (DLD-1) were assessed. SETTING: Research was performed at Saint Louis University. MAIN OUTCOMES MEASURES: Responses were measured by spectrophotometry of surviving cells stained by a dye method. RESULTS: Phytochemicals displayed anti proliferative effects on DLD-1 cells in concentration- and kinetic-dependent manners. Cinnamaldehyde offered statistically significant effects at 24 hours [200 microM], 48 hours [100 - 200 microM], and 72 hours [200 microM]. Piperine displayed a trend towards anti-proliferation at 24 hours and statistically significant inhibition at 48 and 72 hours [100 - 200 microM]. Resveratrol displayed significant anti-proliferative effects at 24 hours [50-200 microM], 48 hours [10-200 microM], and 72 hours [10-200 microM]. CONCLUSION: Cinnamaldehyde, piperine, and resveratrol offer significant in vitro anti-proliferative effects on cultured human colon cancer cells. While each phytochemical exhibited significant anti-proliferative effects, resveratrol results were most impressive in that lower concentrations administered at regular intervals were significantly effective. These results taken together with everyday dietary availability of concentrations used in this study strongly suggest that regular intake of low doses of these phytochemicals offer preventive effects against colon cancer. PMID- 18678138 TI - Consumer satisfaction to laboratory test interpretation by the ASCLS response team. AB - OBJECTIVE: To assess consumer satisfaction to responses to laboratory test interpretations as provided by the American Society for Clinical Laboratory Science (ASCLS) Consumer Response Team. Additional information studied included demographics, whether a response to the question was received, and the respective discipline related to the question. DESIGN: A computerized questionnaire was sent to 339 participants who had sent questions concerning laboratory test results to the ASCLS consumer website (www.ascls.org) in May 2007. A total of99 completed questionnaires (29.3%) provided usable data for analysis. SETTING: Participants answered the questionnaire via electronic mail and results were summarized in Zoomerang. PARTICIPANTS: Participants were national and international consumers who had sent a question regarding their laboratory results to the ASCLS website. Individuals were 18 years of age or older. Participation was voluntary and anonymous. MAIN OUTCOME MEASURES: Consumer satisfaction, measured by eleven satisfaction statements, with laboratory interpretations by the ASCLS Response Team averaged 4.0 on the five-point Likert scale: 1 = Strongly disagree to 5 = Strongly agree. Overall satisfaction of the website itself was 4.2 on the five point Likert scale 1 = Poor to 5 = Excellent. RESULTS: The majority of respondents were female (71.1%) and ranged in age from 36-64 years (71.7%). Seventy-six percent of respondents reported they had received an answer to their laboratory test question. The most frequent disciplines for questions received were in chemistry, immunology, and hematology, respectively. CONCLUSIONS: This study indicates consumers of the ASCLS website were very satisfied with the clinical laboratory scientist volunteers' responses. The ASCLS Consumer Response Team model is contributing to the advancement of healthcare by providing this important service to the public. PMID- 18678139 TI - Factors that impact clinical laboratory scientists' commitment to their work organizations. AB - OBJECTIVE: To assess the predictive ability of various aspects of the work environment for organizational commitment. METHODS: A questionnaire measuring three dimensions of organizational commitment along with five aspects of work environment and 10 demographic and work setting characteristics was sent to a national, convenience sample of clinical laboratory professionals. SETTING AND PARTICIPANTS: All persons obtaining the CLS certification by NCA from January 1, 1997 to December 31, 2006. Only respondents who worked full-time in a clinical laboratory setting were included in the database. MAIN OUTCOME MEASURES: Levels of affective, normative, and continuance organizational commitment, organizational support, role clarity, role conflict, transformational leadership behavior of supervisor, and organizational type, total years work experience in clinical laboratories, and educational level of respondents. Questionnaire items used either a 7-point or 5-point Likert response scale. RESULTS: Based on multiple regression analysis for the 427 respondents, organizational support and transformational leadership behavior were found to be significant positive predictors of affective and normative organizational commitment. Work setting (non-hospital laboratory) and total years of work experience in clinical laboratories were found to be significant positive predictors of continuance organizational commitment. Overall the organizational commitment levels for all three dimensions were at the neutral rating or below in the slightly disagree range. CONCLUSIONS: The results indicate a less than optimal level of organizational commitment to employers, which were predominantly hospitals, by CLS practitioners. This may result in continuing retention problems for hospital laboratories. The results offer strategies for improving organizational commitment via the significant predictors. PMID- 18678140 TI - Lower limit of assay sensitivity: an under-recognised and significant problem in von Willebrand disease identification and classification. AB - von Willebrand disease (VWD) is the most common inherited bleeding ailment, and is characterised by low levels of, or abnormal function in, the plasma protein von Willebrand factor (VWF). However, the laboratory testing process is problematic because of both the heterogeneity of VWD and the limitations in the tests used to identify reduced or abnormal VWF. OBJECTIVE: This study reports on the lower levels of sensitivity for the different assays used in the diagnostic process for VWD and their significance in the diagnostic identification and classification ofVWD. METHODS: The RCPA Haematology QAP is an international external quality assurance (EQA) program that includes VWF/VWD testing within one of its special haemostasis modules. Over the past 10 years, over 50 samples have been distributed to participants, including five samples devoid of VWF and derived from either true Type 3 VWD patients or else from commerciallypurchased VWF deficient plasma. Samples were tested blind by study participants, who report back both numerical values (for VWF and Factor VIII:C) and an interpretation regarding whether or not VWD is suggested by laboratory findings, and if so, the probable VWD subtype. RESULTS: Returned data indicates that the lower level of sensitivity (LLS) tends to be around 5-10U/dL for Factor VIII:C, VWF antigen(VWF:Ag), VWFcollagen binding (VWF:CB), and VWF 'activity' (VWF:Act), but canreach 20U/dL or more for VWF ristocetin cofactor (VWF:RCo). There does not appear to be any improvement over the past decade despite ongoing automation of methodology, and indeed, automation does not seem to provide better LLS performance. CONCLUSIONS: Limitations in the LLS of VWD testing have significant implications in terms of the identification and classification of an individual's VWD, given that these laboratory assays are used to identify VWD andhelp characterise functional VWF discordance, and that the majority of severe VWD subtypes have levels of VWF below 20U/dL. Thus, laboratories will sometimes be unable to distinguish whether VWF deficient samples derive from Type 3 VWD or severe Type 1 VWD or even Type 2 VWD. PMID- 18678141 TI - Telomerase and other novel approaches to bladder cancer detection. AB - The current protocols in place for bladder cancer screening are cystoscopy and urine cytology. Cytology does not have an adequate sensitivity in low-grade malignancy and has limited utility in the screening and management of bladder cancer patients. Urine tumor markers aimed at detection of cancer via voided urine are an attractive alternative to cytology. Currently, FDA-approved tumor marker assays lack the characteristics of an ideal test and have yet to revolutionize bladder cancer detection. Novel tumor markers, not yet FDA approved, have the potential to change disease management algorithms that currently include voided urine cytology. Telomerase, an enzyme present in greater than 80% of all cancer cells, has the potential to be a successful bladder tumor marker for cancer surveillance and monitoring. PMID- 18678142 TI - [New responsibilities for nursing]. PMID- 18678143 TI - [Nursing responsibilities of the school health nurse: healthy in school]. PMID- 18678144 TI - [Ethics of geriatric nursing: between autonomy and nursing care responsibility]. PMID- 18678145 TI - [Alternative ambulatory care concepts:--1: Bielefeld model. Young and old together in one house]. PMID- 18678146 TI - [The "breast care nurse" of the Heidelberg University Gynecologic Clinic: nurse specialist supports women with breast cancer]. PMID- 18678147 TI - [Case management in the context of the expert organization hospital: responsibility for process control]. PMID- 18678148 TI - [Combined case and individual management in neonatology: promoting the competence of parents]. PMID- 18678149 TI - [Health promotion and salutary nursing: patient centered counseling in nursing]. PMID- 18678150 TI - [Step by step toward nursing care competence--15: Back saving nursing care]. PMID- 18678151 TI - [Intercommunication and information flow. An explorative study about ward rounds and patients' documentation]. AB - In modern patient interprofessional communication is an impor tant factor of good outcome. The aim of this study was to analyse the intercommunication during ward rounds and information passed by patients' documentation on an internal and geriatric medicine ward. Beside frequent interruptions the ward rounds showed a restricted flow of information that is based and targeted on the chief physician Nursing staff felt excluded from the informational flow. Regarding patients' documentation staff complained about lack of information and illegible notes. Availability of written information was found to be problematic. A team orientated approach could help to improve interprofessional communication in the future. Besides the importance of carefully performed documentation as a reliable form of communication, communicative contribution of the nursing staff has to be upvalued. PMID- 18678152 TI - [Incomplete nursing performance due to lack of time: unusual dismissal actually not justified]. PMID- 18678153 TI - [From case to case: continual improvement]. PMID- 18678154 TI - [Recognizing and understanding illnesses: sweating as the leading symptom]. PMID- 18678155 TI - [Study on electroacupuncture at shuigou (GV 26) for treatment of mild and moderate shock: a multicentral and randomized controlled trial]. AB - OBJECTIVE: To probe into the effect of electroacupuncture (EA) at Shuigou (GV 26) on mild and moderate shock. METHODS: With 3-center randomized control study method, 276 cases were assigned to an EA plus medicine group and a medication group, 138 cases in each group. They were treated respectively with western medicine plus EA at Shuigou (GV 26), and simple western medicine. Their curative effects were observed after treatment for 6 hours. RESULTS: The blood pressure was immediately increased in the EA plus medicine group and the increase of blood pressure was earlier than that in the medication group (P<0.001). The markedly effective rate of 52.9% in the EA plus medicine group was significantly higher than 18.1% in the medication group (P<0.001). CONCLUSION: The therapeutic effect of EA at Shuigou (GV 26) plus western medicine on mild and moderate shock is better than that of simple western medicine. PMID- 18678156 TI - [A comparative study of effects of electroacupuncture with different stimulation parameters on medicine-induced abortion]. AB - OBJECTIVE: To observe the effect of electroacupuncture (EA) with different stimulation parameters on medicine-induced abortion. METHODS: One hundred and nine cases of early pregnancy who asked medicine-induced abortion were allocated to an EA group A (n = 37), an EA group B (n = 38) and a medication group (n = 34). Within 30-60 min after oral administration of Misoprostol, in the EA group A, EA was given at bilateral Hegu (LI 4) and Sanyinjiao (SP 6) with cluster waves of 100 Hz and in the EA group B, EA was given at Hegu (LI 4) for 20 min and then at Sanyinjiao (SP 6) for 5 min with continuous waves of 50 Hz. EA was not given to the medication group. The complete abortion rate, duration of eliminating embryonic sac, colporrhagia lasting time and abdominal pain condition were recorded. RESULTS: The complete abortion rate was 91.9% in the EA group A and 86.8% in the EA group B, which were higher than 82.4% in the medication group, with no significant differences between the 3 groups (P>0.05); the duration of eliminating embryonic sac and the colporrhagia lasting time in the two EA groups were significantly shorter than those in the medication group (P<0.05, P<0.01); alleviation of abdominal pain in the EA group B was better than the medication group (P<0.01) and the EA group A (P<0.05). CONCLUSION: Different stimulation parameters of EA have different effects on abortion. PMID- 18678157 TI - [Effects of acupuncture under guidance of qi street theory on endocrine function in the patient of epilepsy]. AB - OBJECTIVE: To observe effects of acupuncture under guidance of qi street theory on endocrine function in the patient of epilepsy. METHODS: Sixty cases of epilepsy were randomly divided into an acupuncture-medicine group and a medication group, 30 cases in each group. The medication group were treated with Valpromide, and the acupuncture-medicine group were treated with Valpromide and acupuncture at main points Renying (ST 9), Fengfu (GV 16), Tianzhu (BL 10). Epilepsy scores and plasma total cortisol (CORT), estradiol (E2), pregnendione (P), and prolactin (PRL) levels before and after treatment were observed. Results The total effective rates were 96.7% in the two groups with no significant difference between the two groups (P>0.05); the epilepsy score before treatment was (13.58+/-4.01) points and after treatment was (8.30+/-4.52) points in the acupuncture-medicine group, and (12.97+/-3.91) points and (7.86+/-4.90) points in the medication group, respectively, with significant improvement after treatment in the score in the two groups (P<0.05). After treatment, plasma CORT, E2, P and PRL levels in the acupuncture-medicine group and plasma CORT, E2, P levels in the medication group had significant changes (P<0.05 or P<0.01). There was a significant difference between the two groups in CORT level after treatment (P<0.05). CONCLUSION: Acupuncture under guidance of qi street theory is an effective method for improving endocrine function in the patient of epilepsy. PMID- 18678158 TI - [Treatment of acute injury of soft tissue around shoulder joint by exercise needling and electroacupuncture as main]. AB - OBJECTIVE: To search for an effective method for treatment of acute injury of soft tissue around shoulder joint. METHODS: One hundred and twenty cases were randomly divided into an observation group, a control group 1 and a control group 2. The observation group were treated by exercise needling, electroacupuncture (EA) at Jianyu (LI 15), Jianliao (TE 14), Jianzhen (SI 9), etc. and TDP; the control group 1 were treated with EA at the same points as those in the observation group; the control group 2 were treated with oral administration of Fenbid slow-released capsules. The therapeutic effects were assessed after the first session of treatment, after one therapeutic course and one month after all the treatment by visual analog scale (VAS). RESULTS; The total effective rate of 100.0% and the cured-markedly effective rate of 95.0% in the observation group were better than 90.0%, 75.0% in the control group 1 and 70.0% and 45.0% in the control group 2 (P<0.01 or P<0.05); the VAS scores after the first treatment, after one therapeutic course and one month after all the treatment in the observation group were significantly lower than those in the two control groups (P<0.05). CONCLUSION: The combined treatment of exercise needling, EA and TDP is a good rehabilitation method for acute injury of soft tissue around shoulder joint. PMID- 18678159 TI - [Effect of acupuncture at three acupoints of eye on Bell palsy]. AB - OBJECTIVE: To probe the effect of acupuncture at three acupoints of eye on Bell palsy. METHODS: Seventy-six cases were randomly divided into a routine acupuncture group and a Yan three needling group, 38 cases in each group. The routine acupuncture group were treated with electroacupuncture (EA) at routinely selected acupoints including Yifeng (TE 17), Dicang (ST 4), etc. and the Yan three needling group were treated by EA at the routinely selected acupoints combined with acupuncture at three acupoints of eye including Jingming (BL 1), Shangming, Chengqi (ST 1). The intensity on 0.05 ms in the intensity/time (I/t) curve for frontal ventral fronto-occipital muscle and orbicular muscle of mouth at the affected side was used for assessment criteria of course of disease, and frontal ventral fronto-occipital muscle restoring the raising eyebrow action and orbicular muscle of mouth restoring to House-Brackmann grade I and II were regarded as the therapeutic time limit. RESULTS: Routine EA treatment combined with acupuncture at the 3 acupoints of eye could significantly increase clinical therapeutic effect on Bell palsy with a cured rate of 89.5%, which was better than 65.8% in the routine acupuncture group (P<0.05), and the therapeutic cycle was shorted. CONCLUSION: Acupuncture at the 3 acupoints of eye can significantly improve Bell palsy and promote recovery of functions of facial nerves. PMID- 18678160 TI - [Observation on therapeutic effect of blood-letting puncture combined with red hot needle therapy on knee osteoarthritis]. AB - OBJECTIVE: To observe the therapeutic effect of blood-letting puncture combined with red-hot needle therapy on knee osteoarthritis. METHODS: One hundred and twenty-nine cases were randomly divided into a bloodletting puncture plus red-hot needle therapy group (n=73) and a routine acupuncture group (n=56). The blood letting puncture plus red-hot needle therapy group were treated by blood-letting puncture at Weizhong (BL 40), and red-hot needle pricking Heding (EX-LE 2), Dubi (ST 35), Xiyan (EX-LE 5), Yinlingquan (SP 9), Yanglingquan (GB 34), Xuanzhong (GB 39) and Ashi points, twice each week, 4 times constituting one course. The routine acupuncture group were treated by routine acupuncture at the same points as those for red-hot needle pricking, once each day, two weeks constituting one course. RESULTS: After treatment for 2 courses, the joint pain score and the illness serious index were 2.68+/-0.88 and 4.25+/-1.02, and 4.68+/-1.89 and 7.46+/-2. 13 in the two groups, respectively, with very significant differences before and after treatment in the two groups (P<0.01), the former being better than the later (P<0.05). The clinical cured rate and the total effective rate were 37.0% and 94.5% in the blood-letting puncture plus red-hot needle therapy group and 19.6% and 89.3% in the routine acupuncture group, with a very significant difference between the two groups(P<0. 01). CONCLUSION: Blood-letting puncture combined with red-hot needle therapy has obvious therapeutic effect on knee osteoarthritis. PMID- 18678161 TI - [Observation on therapeutic effect of shu-needling therapy as main on cervical spondylosis of nerve root type]. AB - OBJECTIVE: To explore a better method for treatment of cervical spondylosis of nerve root type. METHODS: One hundred and two cases were randomly divided into an observation group and a control group, 51 cases in each group. The observation group were treated with Shu-needling, i. e. the needles were deeply inserted into the cervical vertebrae of corresponding Jiaji (EX-B 2), and the control group were treated with routine needling method. Electroacupuncture was used in the two groups. RESULTS: The cured rate and the total effective rate were 68.6% and 98.0% in the observation group and 47.1% and 84.3% in the control group, with significant differences between the two groups (both P<0.05). CONCLUSION: Shu needling therapy as main has a better therapeutic effect on cervical spondylosis of nerve root type. PMID- 18678162 TI - [Study on surface infrared radiation spectrums of danzhong (CV 17) in the patient of hyperplasia of mammary glands]. AB - OBJECTIVE: To compare differences of surface infrared radiation spectrums between Danzhong (CV 17) A high-sensitive PHE and non-acupoint control point in the patient of hyperplasia of mammary glands. METHODS: 201 surface infrared spectrograph was used to detect infrared radiation spectrums of Danzhong (CV 17) and non-acupoint control point at 1.5-16.0 microm wave band in the patient of hyperplasia of mammary glands. RESULTS: The shape of the infrared spectrums of Danzhong (CV 17) was similar to that of the non-acupoint control point, but with lower radiation intensity. Of the 59 wavelength spots detected, 13 (6.75-8.25 microm, 9.00 microm, 9.25 microm, 9.75 microm, 13.25-13.75 microm) had significant differences in infrared radiation intensity between Danzhong (CV 17) and non-acupoint control point (P<0.05). CONCLUSION: The intensity of infrared radition of Danzhong (CV 17) is lower and Qimen (LR 14) is higher than that of the non-acupoint control point in the patient with hyperplasia of mammary glands, showing that different channels are at different states of deficiency and excess under pathological condition of hyperplasia of mammary glands. PMID- 18678163 TI - [Comparative study on effects of different acupuncture manipulation methods at neiguan (PC 6) on hand spasm in the patient of stroke]. AB - OBJECTIVE: To observe therapeutic effects of different needling methods at Neiguan (PC 6) on hand spasm in the patient of stroke. METHODS: One hundred and seventy-two cases were randomly divided into a simple acupuncture group (n=36), a slowly twirling needle group (n = 34), a rapidly twirling needle group (n = 30), a simple lifting-thrusting group (n = 35) and a lifting-thrusting-twirling needle group (n = 37). Acupuncture was given at Neiguan (PC 6), twice each day, 5 min each time, and in combination with rehabilitation exercise. Modified Ashworth scale and Fugl-Meyer (FMA) hand function score were used for assessment of therapeutic effects. RESULTS: The transient effective rate and the effective rate after treatment for 8 weeks for hand spasm were 66.7% and 55.6% in the simple acupuncture group, 82.4% and 85.3% in the slowly twirling needle group, 96.7% and 90.0% in the rapidly twirling needle group, 80.0% and 82.9% in the simple lifting thrusting group and 94.6% and 91.9% in the lifting-thrusting-twirling needle group, the rapidly twirling needle group and the lifting-thrusting-twirling needle group being better than other 3 groups (P<0.01). After treatment, the grades for Ashworth scale and FMA scores in the lifting-thrusting-twirling needle group and the rapidly twirling needle group were better than those in other 3 groups (P<0.05, P<0.01), with no significant difference between the two groups (P>0.05). CONCLUSION: Acupuncture at Neiguan (PC 6) has a very definite transient effect, and the lifting-thrusting-twirling needling manipulation and the rapidly twirling needling manipulation have the best inhibitive effect on spasm. PMID- 18678164 TI - [Effects of catgut embedding at "Zusanli" (ST 36) and "Shenshu" (BL 23) on morphine analgesic tolerance and locomotor sensitization in the rat]. AB - OBJECTIVE: To compare effects of catgut embedding at "Zusanli" (ST 36) and "Shenshu" (BL 23) on Morphine analgesic tolerance and locomotor sensitization induced by chronic Morphine administration and the mechanism. METHODS: The rats were randomly divided into a model group, a non-acupoint group, a Shenshu group and a Zusanli group. The rats, except those in the model group, were pretreated with acupoint catgut-embedding 10 days before the first Morphine injection. The Morphine-tolerance model was established and the pain threshold was detected by hot-plate test every day. Locomotor activities were recorded after the first Morphine injection and Morphine-challenging 1 week after withdrawal of Morphine. The positive neurons of nitric oxide synthetase (NOS) were showed by NADPH-d histochemical method. RESULTS: Compared with the non-acupoint group, catgut embedding at "Zusanli" (ST 36) could attenuate the Morphine analgesic tolerance and the increase of locomotor activities in rats. Meanwhile, the expression of NOS positive neurons in nucleus accumbens septi and dorsal striatum decreased in the Zusanli group. There were no significant differences between the Shenshu group and the non-acupoint group in the analgesic threshold and locomotor sensitization, but the expression of NOS positive neurons in the striatum region significantly decreased. CONCLUSION: Catgut embedding at "Zusanli" (ST 36) can attenuate Morphine analgesic tolerance and reverse formation of locomotion sensitization induced by chronic Morphine administration, which are possibly related with inhibition of the expression of NOS positive neurons in nucleus accumbens septi and dorsal striatum. PMID- 18678165 TI - [Effect of acupuncture on target action of ligustrazine in treatment of toxic deafness induced by gentamicin in guinea pigs]. AB - OBJECTIVE: To explore an effective method for treating Gentamicin-induced deafness and the mechanism. METHODS: Guinea pigs were randomly divided into 5 groups: normal control group (group A), model group (group B), Ligustrazine group (group C), acupuncture group (group D) and Ligustrazine plus acupuncture group (group E). The group C, D and E were treated respectively by simple Ligustrazine, simple acupuncture at "Tinggong" (SI 19), "Yifeng" (TE 17), and "Waiguan" (TE 5), and Ligustrazine plus acupuncture. Ten days later, the auditory brainstem response (ABR) thresholds for the wave III , apoptosis and expressions of Bcl-2 and Bax proteins in the organ of Corti of the guinea pig were detected. RESULTS: In the group E, the ABR threshold was significantly lower than that in the group C (P<0.05), and apoptotic cells, the expression of Bax protein and the ratio of Bax/Bcl-2 were lower than those in the group C and D in the organ of Corti, and Bcl-2 protein expression was increased. CONCLUSION: Acupuncture at "Tinggong" (SI 19), "Yifeng" (TE 17), and "Waiguan" (TE 5) has a certain target-synergistic action on Ligustrazine and can increase therapeutic effect of Ligustrazine on Gentamicin-induced deafness, which are possible related with the inhibition of apoptosis, down-regulation of Bax expression and up-regulation of Bcl-2 expression. PMID- 18678166 TI - [Survey of factors of the patient with lumbago seeking for acupuncture treatment and observation on the therapeutic effect of acupuncture at early days]. AB - OBJECTIVE: To explore factors of the patient with lumbago seeking for acupuncture treatment and to observe the therapeutic effect of acupuncture at early days. METHODS: The factors of the patient with lumbago seeking for acupuncture treatment were investigated by questionnaire in 45 cases of lumbago. The patients enrolled were treated by acupuncture and moxibustion for 1-2 sessions with Shenshu (BL 23), Dachangshu (BL 25), Mingmen (GV 4), Yaoyangguan (GV 3) selected. Changes of pain, restlessness score and ODI before and after treatment were used for assessment of clinical therapeutic effect at early days. RESULTS: Recommendation (12 cases, 26.7%), believing Chinese medicine (10 cases, 22.2%), reputation of hospital (6 cases, 13.3%), public praise of doctors (5 cases, 11.1%) were main causes for the patient with lumbago seeking for acupuncture treatment. ODI, pain score and restlessness score were 11.0+/-7.3, 5.0+/-1.9, 4.8+/-2.1 after treatment and 12.6+/-7.7, 6.5+/-1.8, 6.7+/-2.1 before treatment, respectively, with significant differences before and after treatment (all P < 0.05). The assess ment help score before and after treatment were 7.1+/-1.5 and 7.2+/-2.3, respectively, with no significant difference before and after treatment (P>0.05). CONCLUSION: Good reputation of hospital and doctors, and good public praise of acupuncture and moxibustion are main external factors for the patient with lumbago seeking for acupuncture treatment, and the therapeutic effect of acupuncture and moxibustion on lumbago at early days is identical to the patient's expectation. PMID- 18678167 TI - [Study on relative problems of acupuncture and moxibustion for treatment of simple obesity]. AB - There are many methods of acupuncture and moxibustion for treatment of simple obesity with definite therapeutic effects, so, they are followed with interest. However, their therapeutic effects are very different for different individuals, and there are many factors influencing therapeutic effects. The author consults 31 papers in publication about Chinese medicine, acupuncture and moxibustion at home in recent years, and in combination with own clinical practice and experience of many years on acupuncture and moxibustion for slimming, and ponders over the relative problems of syndrome differentiation treatment, therapeutic course, rebound, diet control, sports, psycho-therapy, etc. in clinical treatment, and raises personal some knowledge, so as to probe to increasing clinical effects of acupuncture and moxibustion on simple obesity and provide reference material for clinical treatment in future. PMID- 18678168 TI - [Samples of professor Xie Xi-liang's direct moxibustion]. AB - Prof. XIE Xi-liang in his early years was a disciple of Mr. CHENG Dan-an. He has been engaged in clinical work and teaching for over 60 years with rich experience, and he pays attention to application of acupuncture, moxibustion and medicine for different persons, especially pays attention to moxibustion methods, such as direct moxibustion, with less points selected, and good results, convenient manipulation, moxibustion of one session only for 3-5 min. He teaches the patients and their family for self-moxibustion at home, saving time. This method has better short-term and long-term therapeutic effects for hypo-immune patients, pain diseases, and it is suitable to treatment of diseases and health care. PMID- 18678169 TI - [A brief talk on characteristics of Japanese collateral puncture]. AB - The characteristics of Japanese collateral puncture are expounded from history evolution and technical feature. The position of pricking blood and blood-letting volume for the three Japanese collateral puncture, Japanese collateral puncture at Jing-well points, at fine collaterals and at the skin are emphatically discussed and briefly compared with Chinese pricking blood therapy, so as to better develop the collateral puncture therapy. PMID- 18678170 TI - [Contributions of professor Xue Chong-cheng in acupuncture and neuropsychiatry]. AB - Professor Xue Chong-cheng devotes himself in clinical and research of integrative works of TCM, acupuncture, neuropsychiatry, neurosurgery and medicopsychology for more than 70 years. He firstly confirmed acupoints are identical with nerve and motor points, found propagating meridian sensation on phantom limbs of acquired and congenital amputates. The sensation can pass over fresh incision wound and it exits and disappears with the cortical sensation. He proposed a central theory that a meridians model is present in the brain. He suggested examination of general afferent system, cortical sensation and deep pain of syringomyelia with acupuncture as they are absent in routine methods but still present during needling. He firstly reported meridian type of sensory epilepsy and it was recognized by the Epilepsy Center of the USA. He treated psychosis with electroacupuncture convulsive therapy, the dosage of current used was less than 4% of the conventional method. The Journal of Psychiatry of USA recognized it is a dramatic progress for more than 40 years. He compiled first integrative medical questionnaire for assessment of pain. According to TCM theories and modern method of standardization he established inventories and national norms of personality and constitution for the corresponding examinations. They fulfill the gap of China. He firstly proposed the model of TCM is temporo-spatio-socio-psycho biological. Now he is nearly ninety years old but is still struggling on the first line. PMID- 18678171 TI - [Study on the origin of Dou Han-qing's academic thought of acupuncture and moxibustion]. AB - The origin of Dou Han-qing's academic thought of acupuncture and moxibustion is studied by analysis on relative contents in the books Weisheng Baojian, Jisheng Bacui and Zhenjiu Sishu Zhenjing Zhinan and the two papers "On reinforcing reducing manipulation" and "Questions and answers of qi and blood", holding that DOU Han-qing's academic thought of acupuncture and moxibustion is influenced more by Suwen (Plain Questions) and Nanjing (Classic on Medical Problems), not by Lingshu (Miraculous Pivot). PMID- 18678172 TI - [Acupuncture and moxibustion treatment of anxiety neurosis and study on characteristics of acupoint selection]. AB - Review and summarize the published literature since 1994 on treatment of anxiety neurosis with acupuncture and moxibustion, so as to provide evidence for standard acupuncture and moxibustion treatment program. It is indicated that Baihui (GV 20), Neiguan (PC 6), Shenmen (HT 7), Sanyinjiao (SP 6), Yintang (EX-HN 3), etc. were the most commonly selected for clinical treatment of anxiety neurosis. Acupuncture and moxibustion treatment of anxiety neurosis stresses the special function of acupoints in acupoint selection, reflecting the therapeutic characteristics of "treatment based on the brain" and "treatment based on the heart" for mental disorders, and emphasizing the important position of TCM Zang- and Fu-organs syndrome differentiation in treatment of anxiety neurosis. PMID- 18678173 TI - Catholics using contraceptives: religion, family planning, and interpretive agency in rural Mexico. AB - Research on how religion shapes contraceptive practices and fertility has paid insufficient attention to how people interpret religious teachings. This study draws on ethnographic fieldwork in Degollado, Mexico, to describe generational and social-contextual differences in how women interpret and use religious doctrine to achieve their fertility desires without jeopardizing their standing as devout Catholics. Contrasting the family planning beliefs and practices of young Mexican women with those of older women (many of whom are the younger women's parents and in-laws), in a rural town in which the religious regulation of everyday life is pervasive, reveals how a common set of religious teachings and principles can be used to guide two different generational strategies for fertility regulation. The ethnographic data presented here highlight the creativity with which people use religious frameworks to justify their behavior. Research exploring how religion--and culture more broadly--influences fertility and contraceptive use should give greater attention to the dynamic interplay between cultural beliefs and institutions, social context, and interpretive agency. (STUDIES PMID- 18678174 TI - Fertility transitions in developing countries: progress or stagnation? AB - Over the past quarter-century,fertility has declined rapidly in many developing countries. Projections typically assume that this trend will continue until replacement level is reached. Recent evidence suggests, however, that ongoing fertility declines may have slowed or stalled in a number of countries in transition. This study examines the pace of fertility change in developing countries that have multiple Demographic and Health Surveys to determine whether ongoing transitions are decelerating or stalling. The main findings are that in sub-Saharan African countries, the average pace of decline in fertility was lower around 2000 than in the mid-1990s and that more than half the countries in transition in this region have stalled. PMID- 18678175 TI - Rates of induced abortion in Iran: the roles of contraceptive use and religiosity. AB - Iran has experienced a dramatic decline in fertility in recent decades, but limited access to legal abortion continues to lead many women whose pregnancies are unwanted or mistimed to undergo clandestine, unsafe abortions. No official data on the abortion rate in Iran have been collected, however. This study uses the 2000 Iran Demographic and Health Survey to estimate the abortion rate for the country as a whole and for specific regions, and to explore the role of contraceptive use and religiosity in explaining regional variations in abortion rates. We estimate the total abortion rate for the country to be 0.26 abortions per married woman, and the annual general abortion rate to be 7.5 abortions per 1,000 married women aged 15-49. We find that the negative effect of modern contraceptive use on the abortion rate is 51 percent greater than the negative effect of religiosity, and we highlight the implications of these findings for policies on reproductive health and family PMID- 18678176 TI - Intimate partner violence and interference with women's efforts to avoid pregnancy in Jordan. AB - This study examines the association between intimate partner violence (IPV) and women's experience of interference with their attempts to avoid pregnancy among 353 women surveyed at reproductive health clinics throughout Jordan. Approximately 20 percent of respondents indicated that their husbands or someone else had interfered. Among those others than husbands who were identified, mothers-in-law were the most frequently mentioned, followed by the respondents' mothers and sisters-in-law. Multivariate logistic regression was used to control for potential confounders in determining whether each of the three measures of intimate partner violence (physical violence, sexual violence, and controlling behaviors) was significantly associated with having an increased risk of experiencing interference, as were several sociodemographic variables: nonconsanguineous marriage, residence with in-laws, and rural residence. Physicians, nurses, and family planning counselors must be made aware of the challenges that women may face from their families when they attempt to regulate their fertility. PMID- 18678177 TI - Clinic-based surveillance of adverse pregnancy outcomes to identify induced abortions in Accra, Ghana. AB - Reliable measures of induced abortion remain elusive, especially when the public perception is that the procedure is immoral or improper. This study draws on interviews using a modified preceding birth technique (PBT) with women attending antenatal and maternity clinics in Accra to compare rates of adverse pregnancy outcomes (stillbirths, miscarriages, and induced abortions) with rates from a household maternity history and the Ghana Demographic and Health Survey. The reports from the antenatal clinics produced some of the highest rates for adverse outcomes of pregnancy. In light of the generally high coverage of antenatal services found even in developing countries, the method based on the PBT holds promise for the improvement of reports of miscarriage and abortion worldwide. PMID- 18678178 TI - Wrong-site surgery: we're not doing all that we can. AB - *Adverse events are rarely due to any single error, but rather to a series of errors. *Patient involvement is a critical component in efforts to minimize wrong site, wrong side surgeries. *Do not assume you have been given correct information; double check it every step of the way. PMID- 18678179 TI - Christiana slashes sepsis mortality rate. AB - *Special packet for staff members includes treatment algorithm, care management guidelines, and ED treatment order set. *Strategies stress proper antibiotic administration, rapid fluid resuscitation, and early central venous catheter placement. *Rapid response teams become vectors to administer treatment outside the ED. PMID- 18678180 TI - How do you get your board on board? AB - *Show your board members the evidence - that means, the bad news as well as the good. *Select the appropriate 'storytellers' to go before the board; real-life examples will make a stronger impression. *Make sure the board understands the 'big picture' in terms of organizational quality and safety performance. PMID- 18678181 TI - SHEA estimates preventable HAIs. AB - *Perhaps as many as 82% of deaths from bloodstream infections could be prevented, according to SHEA study. *Compliance with timing of antibiotic administration to prevent surgical site infection is cited as needing improvement. *Removal of the catheter to prevent urinary tract infections is 'sometimes easier said than done,' says expert. PMID- 18678182 TI - New care model transforms facility. AB - *Nursing role is redefined; RNs now act more as case managers, with LPNs handling task-oriented activities. *Central nurses' station removed; alcoves created for atmosphere conducive to communication and collaboration. *Lean tool used to establish improvement targets and realize a reduction in errors. PMID- 18678183 TI - Medical and veterinary doctors, social scientists and agricultural researchers meet to carry forward the fight against cysticercosis, a neglected and fatal disease of the poor. PMID- 18678184 TI - Veterinary education in South Africa: the classes of 1934 & 1935. AB - The Class of 1934 included 2 graduates who created milestones for the veterinary profession in South Africa. Jack Boswell was the first Onderstepoort graduate to start his own private practice without ever joining the government service. George van der Wath has the distinction of being the only South African veterinarian to become Chairman of the prestigious South African Wool Board. Ashton Tarr was President of the South African Veterinary Medical Association from 1966-1969. Concise descriptions are given of the varied life histories of the 14 members of the Classes of 1934 and 1935. All except Boswell initially joined government service, one serving mainly in the Colonial Service before eventually returning to South Africa. Three spent their entire careers in the South African Veterinary (Field) Services, finally occupying very senior positions in that division. One ended his career lecturing at a university. Lambrechts was the first veterinarian to occupy the 'resurrected' post of Director of Veterinary Services reserved for field veterinarians. Only one of the graduates opted for research, but went farming after obtaining a DVSc degree. Three spent the greater part of their careers in private practice, Thiel from as early as 1937. Two went into municipal (public health) service, one becoming director of an abattoir. Only one saw military service in World War II. Two died before they were 50 years old. Unfortunately, virtually nothing is known about Erasmus' career. At 97 Thiel holds the distinction of being the oldest Onderstepoort graduate. PMID- 18678185 TI - Epidemiology, disease and control of infections in ruminants by herpesviruses--an overview. AB - There are at least 16 recognised herpesviruses that naturally infect cattle, sheep, goats and various species of deer and antelopes. Six of the viruses are recognised as distinct alphaherpesviruses and 9 as gammaherpesviruses. Buffalo herpesvirus (BflHV) and ovine herpesvirus-1 (OvHV-1) remain officially unclassified. The prevalence of ruminant herpesviruses varies from worldwide to geographically restricted in distribution. Viruses in both subfamilies Alphaherpesvirinae and Gammaherpesvirinae cause mild to moderate and severe disease in respective natural or secondary ruminant hosts. Accordingly, the economic and ecological impact of the viruses is also variable. The molecular characteristics of some members have been investigated in detail. This has led to the identification of virulence-associated genes and construction of deletion mutants and recombinant viruses. Some of the latter have been developed as commercial vaccines. This paper aims to give an overview of the epidemiology and pathogenesis of infection by these viruses, immuno-prophylaxis and mechanisms of recovery from infection. Since there are 128 ruminant species in the family Bovidae, it is likely that some herpesviruses remain undiscovered. We conclude that currently known ruminant alphaherpesviruses occur only in their natural hosts and do not cross stably into other ruminant species. By contrast, gammaherpesviruses have a much broader host range as evidenced by the fact that antibodies reactive to alcelaphine herpesvirus type 1 have been detected in 4 subfamilies in the family Bovidae, namely Alcelaphinae, Hippotraginae, Ovibovinae and Caprinae. New gammaherpesviruses within these subfamilies are likely to be discovered in the future. PMID- 18678186 TI - A serological prevalence survey of Brucella abortus in cattle of rural communities in the province of KwaZulu-natal, South Africa. AB - A serological survey of Brucella abortus in cattle originating from communal grazing areas of Kwa Zulu Natal was carried out between March 2001 and December 2003. The survey was designed as a 2-stage survey, considering the diptank as the primary sampling unit. In total 46 025 animals from 446 diptanks of 33 magisterial districts were sampled and tested using the Rose Bengal test and Complement Fixation Test. The apparent prevalence at district level was adjusted for clustering, diagnostic test sensitivity and specificity, and mapped using ArcView version 3.3. The prevalence of brucellosis in communal grazing areas of Kwa-Zulu Natal was found to be 1.45 % (0.84-2.21%) and varied from 0 to 15.6 % between magisterial districts. In 19 of the 33 magisterial districts no serological reactors were observed. A large variation in prevalence was found within diptank areas. Brucellosis was found to be most prevalent in the northeastern area of the province. The findings of the survey are discussed. PMID- 18678187 TI - The occurrence of dermatosparaxis in a commercial Drakensberger cattle herd in South Africa. AB - Dermatosparaxis is a heritable collagen dysplasia causing skin extensibility and fragility. In Belgian Blue cattle this mutation has been described as a 3 base pair (bp) change followed by a 17bp deletion in the gene coding for procollagen 1 N-Proteinase (pNPI). An outbreak in a commercial Drakensberger herd in South Africa followed the introduction in late 2000 of a 3-year-old bull that developed skin lesions in 2001 and was culled in 2002. Some of his offspring were similarly affected, 1 of which was kept as a breeding bull after his sire's death. Two affected calves were referred to the Onderstepoort Veterinary Academic Hospital in October 2005. Detailed examination revealed only skin abnormalities limited to the lateral extremities of the thorax, abdomen and pelvis, viz. either acute lacerations of varying sizes, slow healing defects or thin scars in chronic cases. During a subsequent farm visit, 13 animals with similar wounds were seen in the herd of 146 animals. Electron microscopic examination of skin biopsies revealed haphazard arrangement and loose packing of dermal collagen fibrils within collagen fibres. The fibrils showed size variation and slightly irregular outlines on cross-section, consistent with mild dermatosparaxis. DNA samples of affected calves were analysed using primers designed to amplify the region of the pNPI gene that contained the mutation described in Belgian Blue cattle, but this mutation could not be demonstrated in any of the animals tested. It is concluded that a form of dermatosparaxis with a different gene mutation from that described in Belgian Blue cattle exists in Drakensberger cattle in South Africa. This possibly also explains the milder and more delayed clinical signs and the milder dermal collagen ultrastructural abnormalities. PMID- 18678188 TI - Estimating exposed pulp lengths of tusks in the African elephant (Loxodonta africana africana). AB - Captive and wild African elephants frequently suffer tusk fractures. Several institutions shorten the tusks of captive elephants to reduce fractures and injury as a result of behaviour within enclosures. Fracturing or coronal amputations that expose pulp lead to pain for the elephant. Estimating coronal pulp lengths may thus help to minimise the risk of pulp exposure during amputations. We aimed to determine the length of the pulp beyond the lip margin from an external tusk characteristic. Tusks collected from elephants in Namibia and the Kruger National Park had similar morphological relationships. This statistical property allowed us to correct for missing data in our data sets. Pulp volume and pulp length correlated with tusk circumference at the lip. Even so, the circumference at the lip could not predict the length of the pulp in the crown external to the lip. Our findings suggest that tusks, irrespective of sex or age, amputated further than 300 mm from the lip should not expose pulp. PMID- 18678189 TI - Occurrence of Theileria parva infection in cattle on a farm in the Ladysmith district, KwaZulu-Natal, South Africa. AB - Theileria parva causes widespread morbidity and mortality in cattle in endemic regions. An outbreak of theileriosis occurred on a farm near Ladysmith in KwaZulu Natal, South Africa, which is not a declared Corridor disease-infected area. A survey of Red Brangus cattle from all age groups and areas of the farm was performed. Transmission of the parasite from infected animals on the farm to susceptible animals by tick transmission and tick-stabilate injection, was attempted. The survey indicated high numbers of animals with antibody titres to T. parva but only 6 infected animals, based on real-time PCR and RLB analysis. The transmission experiments failed to transmit the parasite. The study shows the difficulty in elucidating a source of infection and determining the dynamics of new infections in a herd where multiple possible sources are present and treatment with tetracyclines has taken place. PMID- 18678190 TI - The use of electrochemically activated saline as a uterine instillation in pony mares. AB - Twelve pony mares were randomly assigned to either a control or a treatment group and inseminated with fresh, raw semen from a single stallion of known fertility in a cross-over trial design. Pregnancy was diagnosed by transrectal ultrasound 12-14 days post-ovulation and then terminated by administration of a luteolytic dose of cloprostenol. Treatment mares received a uterine instillation of 100 ml of electrochemically activated (ECA) saline 4-12 hours post-insemination. Control mares received no treatment post-insemination. Per cycle pregnancy rate was 58.3 % in the control group and 50 % in the treatment group. There was no statistical difference (P = 1.000) in pregnancy rate between the 2 groups. The principles of ECA and applications of ECA saline are discussed. PMID- 18678191 TI - A comparison of the oral application and injection routes using the onderstepoort biological products fowl typhoid vaccine, its safety, efficacy and duration of protection in commercial laying hens. AB - This study was undertaken to establish whether the Onderstepoort Biological Products Fowl Typhoid (OBPft) vaccine registered as an injectable vaccine was effective and safe when administered orally to commercial layers. Its efficacy and duration of protection were compared with application by intramuscular injection. Commercial brown layer hens were used as they were found to be highly susceptible to Salmonella gallinarum infections. In the vaccine safety trial birds were euthanased at timed intervals spanning 4 weeks postvaccination. Necropsies were performed and samples were taken and tested. No clinical signs or mortalities could be attributed to the OBPft vaccine nor could active shedding of the vaccine strain be detected. Slight pathological changes were noted with both routes of vaccination; however, these changes were transient, returning to normal within the observation period. The injected groups showed a better serological response with the rapid serum plate agglutination (RSPA) test than the orally vaccinated groups. In the duration of protection trial, birds were challenged at 3-8-week intervals post-vaccination. All unvaccinated birds died. Protection 8 and 16 weeks after vaccination was above 60 %,by 24 weeks after challenge, the vaccine protection was below 30 %. It was found that there was no significant difference (P < 0.05) in the protection offered by either the oral or injected route of vaccination with the OBPft vaccine. PMID- 18678192 TI - A serological survey of African horse sickness in Botswana. AB - A retrospective serological survey of African horse sickness (AHS) in Botswana covering a 10-year period (1995-2004) is reported. The survey involved horses showing clinical symptoms of the disease; the horses had not been vaccinated against AHS. Over the period surveyed, serological evidence suggestive of infection with AHS virus (AHSV) was found in 99 clinical cases out of which 41.4% (41/99) cases were found during the 1st half (1995-1999) and 58.6 % (58/99) cases were found in the 2nd half of the survey period (2000-2004). These serological findings are discussed in relation to AHSV serotypes isolated from diseased horses in Botswana before and during the period of this serological survey. PMID- 18678193 TI - Acute normovolaemic haemodilution--2 case studies. AB - Acute normovolaemic haemodilution (ANH) is a technique used to preserve a patient's owns red blood cells and reduce the incidence of heterogeneous blood transfusion. This paper describes the use of the technique in a dog and a kitten. A significant benefit of ANH can be shown in the canine case presented. The dog lost 1800 ml of blood during surgery but the haematocrit was only reduced to 33% 6 hours after the end of surgery. The kitten, however, did not benefit from ANH. It lost a small volume of blood during surgery and developed complications. This paper also describes some of the potential complications that may occur. To the best of my knowledge, this is the 1st clinical description of ANH in a dog and a cat. PMID- 18678194 TI - Acute lead intoxication in a pregnant mare. AB - Lead (Pb) intoxication in horses is usually a chronic phenomenon with clinical signs associated with central nervous dysfunction. This report gives details of a case of acute Pb intoxication in a 9-year-old American Saddlebred mare with severe, progressive and ultimately fatal neurological deterioration. During the 4 days of hospitalisation, clinical signs progressed from intermittent headshaking and depression to severe, continuous, uncontrollable manic behaviour. At autopsy, three grey-coloured, hard metal particles were present in the gastrointestinal tract and subsequently found to contain 2614 ppm Pb. Lead concentrations in the brain, liver, stomach and kidney were 29, 4, 6 and 2 ppm wet weight, respectively. PMID- 18678195 TI - Efficacy of 3 anthelmintics in communally grazed sheep as reflected by faecal egg count reduction tests in a semi-arid area of South Africa. AB - A survey was conducted on the occurrence of anthelmintic resistance of nematodes in communally grazed sheep in a semi-arid area near Mafikeng, South Africa, from January to March 2006. In the herds belonging to twelve smallholder sheep farmers, the efficacies of albendazole, levamisole and closantel were tested by faecal egg count reduction tests where 80 % efficacy was considered the cut off for anthelmintic resistance. The results of the faecal egg count reduction tests showed more than 80% efficacy with all the drugs used in most cases, but there were notable exceptions. In 1 case, closantel had an efficacy of 72%, albendazole had an efficacy of 68% and levamisole showed efficacies of 58%, 60% and 75% respectively on 4 farms. The occurrence of anthelmintic resistance in this farming sector is of concern and steps should be taken to prevent its further spread and development to avoid a situation developing as on numerous commercial sheep farms in South Africa where resistance is common. PMID- 18678196 TI - Perioperative nursing in the spectrum of nursing practice. PMID- 18678197 TI - The remains of the body: human tissue, competence and consent in an age of profit. AB - Over the past few decades human tissues and fluids have increasingly become of interest to health-oriented research due to their potential use in the development of new diagnostic tools, drugs and treatment modalities. They have also become valuable commodities that figure prominently in the recovery of hormones for cosmetic purposes, the production of proteins and in a whole range of uses in the biopharmacological industry. Unfortunately, current understanding of the ethical and legal status of human tissue and fluids, and of the conditions under which they may be recovered and used, is somewhat uneven. The aim of this presentation is to outline the ethical and legal considerations that must be met if a recovery and use protocol is to meet appropriate standards. PMID- 18678198 TI - Clinical implementation of a scrubless chlorhexidine/ethanol pre-operative surgical hand rub. AB - The objective of surgical scrubbing is to reduce the bioburden on the hands of the surgical team in hope that if gloves are punctured or torn, the number of bacteria released at the operation site will be minimal and therefore reduce the risk of site infection. Long procedures with scrubbing and soaping can, however, be counterproductive because with repetition they tend to cause skin abrasions, damages and injuries without further reducing the risk of bacterial release. Within a general review of OR processes, it was decided to substitute to the standard surgical scrub a "new" scrubless pre-op surgical hand rub procedure. This article summarizes the results. PMID- 18678199 TI - Oh, by the way, the patient is pregnant! AB - Approximately one to three per cent of pregnant women undergo surgery that is unrelated to their pregnancy. In Canada this represents about 5,000 patients each year that present unique challenges to the perioperative nurse and the entire surgical team. Approximately five to ten per cent of these patients are involved in trauma, which causes 46.3% of maternal deaths. A small percentage of elective procedures are carried out in the first trimester, before the patient herself is aware of the procedure. The majority of procedures are required for urgent and emergent conditions that require surgery despite the risks to the mother and fetus. This article will discuss perioperative care of the non-obstetric pregnant patient and to introduce a nursing care guideline that can be used as a quick reference tool. The care discussed in the appended Guideline focuses on the pregnant condition and is to be used in conjunction with routine perioperative care practices. Semi-elective and urgent surgery is not contraindicated by pregnancy, although anesthetic and surgical approaches must be modified to promote the safety of mother and her fetus. If possible, the surgery should be postponed to the second trimester. By this time major systems of the fetus are formed and the uterus does not yet infringe on abdominal structures and manipulation may be kept to a minimum. In the first trimester, spontaneous abortion is the greatest risk at 12%. This decreases to less than five per cent in the second and third trimesters. Pre-term labor presents the greatest risk in the second and third trimesters. The most common need for surgery in pregnancy is associated with appendicitis, biliary tract disease, intestinal obstruction, urinary calculi and trauma. PMID- 18678201 TI - Caveat emptor. PMID- 18678200 TI - Every inch... PMID- 18678202 TI - Does Aggrenox (aspirin/dipyridamole) reduce the risk of recurrent stroke to a greater degree than aspirin alone? PMID- 18678203 TI - Are thiazide diuretics an effective treatment for hypertension in patients with chronic kidney disease? PMID- 18678204 TI - June 2008 is Oklahoma DVT Awareness Month. AB - A concurrent resolution recognizing the importance of raising public awareness of deep-vein thrombosis (DVT) was passed by the state legislature and signed on February 12, 2008, declaring June 4, 2008, Oklahoma DVT Awareness Day and June 2008, Oklohoma DVTAwareness Month. As part of this effort, the Oklahoma State Medical Association is actively involved in the education of health professionals about DVT. This resolution was co-authored by State Senator Johnnie Crutchfield, and State Representatives Greg Piatt and Dr. Doug Cox. PMID- 18678205 TI - Refocusing emergency department utilization. PMID- 18678206 TI - Hawai'i air quality monitoring assessment: some effects of Hawai'i's smoke-free work and public places law. AB - In November 2006, the Smoke-Free Work and Public Places Law passed to protect people from secondhand smoke in Hawai'i. An air-quality monitoring assessment to determine the difference this law made in air quality was conducted at 15 bars/restaurants. Levels of particulate matter (PM2.5) at enclosed (indoor) venues fell 90% after implementation of the law while partially enclosed restaurants/bars were all below the EPA 24 hour average limit both before and after the law. PMID- 18678207 TI - Substantial systolic improvement and reverse cardiac remodeling in elderly-onset idiopathic dilated cardiomyopathy > or = 65 years of age. AB - Significant systolic improvement and reverse remodeling in dilated cardiomyopathy (DCM) are well known, however they have been rarely described among elderly subjects. The authors retrospectively reviewed 86 patients with a diagnosis of DCM seen at a clinic during April-November 2005. The authors found 18 patients with elderly-onset idiopathic DCM (age of onset > or = 65 years, mean age 71.8 +/ 6.2 years), who had substantial improvement in left ventricular ejection fraction (LVEF) > or = 20 units (%). During a mean follow-up of 8.6 +/- 5.5 years, mean LVEF and left ventricular end-diastolic diameter improved from 30.6 +/- 7.9% to 58.3 +/- 6.5% (p < 0.0001) and 57.5 +/- 7.0 mm to 44.6 +/- 5.5 mm (p < 0.0001), respectively Fifteen of the 18 patients (83%) had a history of hypertension. Systolic blood pressure at the initial referral clinic correlated with improved contractility (p = 0.0275, r = 0.52). The eighteen patients found in our small patient population suggest that substantial systolic improvement and reverse remodeling is seen in elderly patients with idiopathic DCM. PMID- 18678208 TI - Medical licensure in the United States: past, present, and future. PMID- 18678209 TI - The hematologist's approach to myelodysplastic syndrome. PMID- 18678210 TI - Issues in medical malpractice XXIV. PMID- 18678211 TI - [The ACCLAIM trial]. PMID- 18678212 TI - [Changes in patient survival and quality of life after heart transplantation]. AB - Heart transplantation was performed firstly in 1967, but it became a valuable option in the 1980s, due to the availability of cyclosporine and of the technique for rejection monitoring by means of serial endomyocardial biopsies. Post transplant survival improved over the years, mainly due to a reduction in early mortality for infection or acute rejection. Expected 1-year and 5-year survivals are around 85% and 70%, respectively. During the past 20-30 years, better therapies for heart failure have been developed, leading to restriction of heart transplant candidacy to truly refractory heart failure. On the contrary, the criteria for donor acceptance have been liberalized, due to the discrepancy between heart transplant candidates and available organs. It must be kept in mind that renal and/or hepatic insufficiency that may be a consequence of heart failure, pulmonary hypertension, and donor age, all remain risk factors for mortality after transplantation. In order to maintain and possibly improve the results of heart transplantation, effective strategies to increase safely the donor pool are of utmost importance. Moreover, long-term post-transplant recipients present new challenges to research and clinical practice. Mechanical circulatory support devices represent a surgical bridge or an alternative to transplantation; their expansion is limited by costs, organizational burden, and by patient difficulties in accepting this therapy. PMID- 18678213 TI - [Obstructive sleep apnea syndrome and cardiovascular diseases]. AB - Obstructive sleep apnea (OSA) syndrome is one of the most common respiratory disorders in humans. There is emerging evidence linking OSA to vascular disease, particularly hypertension. The underlying pathophysiological mechanisms that link OSA to cardiovascular diseases such as hypertension, congestive heart failure and atrial fibrillation are not entirely understood, although they certainly include mechanical events, increased sympathetic activity and oxidative stress. This review will examine the evidence and mechanisms linking OSA syndrome to cardiovascular disease. PMID- 18678214 TI - [The digitalis: should we forget it?]. AB - For more than 200 years digitalis has been considered of paramount importance in the treatment of heart failure and atrial fibrillation. The IN-CHF Italian registry shows that prescriptions were reduced from 63.3% in the period 1995-1999 to 40% in the period 2000-2005, a very different trend compared to prescriptions of angiotensin-converting enzyme inhibitors, angiotensin receptor antagonists, and beta-blockers. The commercial value of digitalis is much lower than other drugs and it does not seem to be of interest for the pharmaceutical companies. Unfortunately, this is a logical trend in the business world. For many years the major indications of digitalis have been heart failure and atrial fibrillation. The most important study on efficacy of digitalis in the treatment of heart failure was the DIG trial (1997), which showed no difference in mortality when compared to placebo but significant beneficial effects in reducing hospital admission rates. Many post-hoc researches evaluated blood levels of digitalis and proved beneficial effects also on mortality when digoxinemia values were 0.5-0.9 ng/ml. In conclusion, digitalis should still be considered effective for the treatment of heart failure; therefore prescription modality as well as the range of normal values of digoxinemia should be updated. PMID- 18678215 TI - [The homecare management of chronic heart failure followed up by hospital cardiologists: a pilot study]. AB - BACKGROUND: The management of patients with chronic heart failure turns to new diagnostic and therapeutic approaches, focusing on an integrated network among general practitioners and cardiologists. METHODS: In order to improve heart failure patient care, we developed a pilot study to assess the feasibility of a homecare program including visits performed by hospital cardiologists on request of general practitioners. RESULTS: This pilot study confirmed the feasibility of the collaborative project among general practitioners and cardiologists and the suitability of home visits performed on request. In 80/96 visits (85%) the cardiologist confirmed that home visit could be shared. Preliminary epidemiological data showed an elderly population (mean age 82 years) with prevalence of females (60%); hypertension in 46%, diabetes in 27%, and infrequent access to outpatient visits. CONCLUSIONS: This integrated management showed good results in terms of an improvement of cooperation among general practitioners and cardiologists, and allowed to verify a very high level of diagnostic accuracy of general practitioners. PMID- 18678218 TI - [Insulin therapy may hasten congestive heart failure in cardiac patients: case series and review of the literature]. AB - We describe the appearance, within a few weeks from the beginning of insulin therapy, of signs and symptoms of congestive heart failure in three diabetic patients with known cardiac disease (one with aortic stenosis, one with hypertensive heart disease and one with ischemic heart disease), who however had never previously shown signs of heart failure. Reduction of the dose of insulin, along with a moderate diuretic therapy, led to resolution of clinical condition. Sodium retention and increased vascular permeability by insulin could be the causes of this phenomenon. In clinical practice it is necessary to remember that the beginning of insulin treatment could exacerbate left ventricular dysfunction to an overt heart failure. Future studies should evaluate whether insulin therapy plays a negative role in the long-term prognosis of patients with asymptomatic left ventricular dysfunction, as it has already been hypothesized in those with overt heart failure. PMID- 18678217 TI - [Effects of pretreatment with amiodarone infusion in patients with persistent atrial fibrillation submitted to external electrical cardioversion: a single center experience]. AB - BACKGROUND: Several studies demonstrated the efficacy of amiodarone pretreatment in achieving bet-ter outcomes after electrical cardioversion of atrial fibrillation. In the majority of cases, oral amiodarone for at least 1 month was administered, with the result of lengthening of pre-cardioversion time. Only one study in the literature reported high-dose amiodarone infusion, showing an increase in the incidence of slow arrhythmias. The aim of this study was to test the efficacy of pretreatment infusion of a single dose of amiodarone few hours before electrical cardioversion in restoring sinus rhythm and reducing the incidence of early arrhythmic recurrences. METHODS: The study was retrospective. We analyzed a population of 155 patients with persistent atrial fibrillation, from May 2003 to November 2005. The first group of 86 patients was treated with amiodarone at the dose of 4 mg/kg in 30 min, few hours before electrical cardioversion; the second group of 69 patients was treated with electrical cardioversion without pharmacological pretreatment. The two groups were homogeneous for age, sex, coronary artery disease, duration of arrhythmia, atrial dimensions, left ventricular ejection fraction, and paddle position for electrical cardioversion. RESULTS: There were no significant differences between the two groups in terms of efficacy of cardioversion (95.3 vs 91.3%, p = NS). Pretreatment with bolus of amiodarone significantly reduced the incidence of immediate recurrence (3.5 vs 17.4%, p < 0.05) and the mix of immediate and early recurrence (19.7 vs 33.3%, p < 0.05). There were no significant differences in the incidence of late recurrences (17.4 vs 13%, p = NS). There were no significant bradyarrhythmias in the two groups. Amiodarone pretreatment did not reduce energy delivery to obtain cardioversion. CONCLUSIONS: Amiodarone pretreatment with intravenous bolus few hours before electrical cardioversion reduces short-term recurrences of atrial fibrillation. It does not reduce energy delivery of electrical cardioversion and does not increase the incidence of slow arrhythmias. Randomized prospective studies are warranted to confirm these findings. PMID- 18678219 TI - [Partial anomalous pulmonary venous drainage: early diagnosis and complications after surgical repair of a rare pathology difficult to identify]. AB - Partial anomalous pulmonary venous connection is a rare congenital heart defect and it is usually difficult to identify by transthoracic echocardiography alone. Here we report a case in a newborn, identified by echocardiographic imaging techniques with subcostal views, to detect the anomalous venous return. Our case is an uncommon one, as regards both its anatomy and early diagnosis. Surgical repair can be safely managed by means of multiple techniques with low morbidity. This correction may be associated with complications such as superior vena cava occlusion and sinus node dysfunction. PMID- 18678220 TI - [Multislice computed tomography and conventional coronary angiography in diffuse coronary artery ectasia]. PMID- 18678221 TI - [The SMASH-VT trial]. PMID- 18678216 TI - [The Italian SurveY on carDiac rEhabilitation 2008 (ISYDE 2008): study presentation]. AB - In this paper, the Italian Society of Cardiac Rehabilitation and Prevention (GICR) presents the third survey on the status of cardiac rehabilitation (CR) in Italy. The Italian SurveY on carDiac rEhabilitation 2008 (ISYDE 2008) is a multicenter, observational study aimed at identifying the number and characteristics of Italian CR facilities, both in terms of health operators and interventions. Clinical records of all patients consecutively discharged within the whole network--composed of up to 200 CR units--from January 28 to February 10, 2008 will also be reviewed for diagnosis of admission, comorbidities, rehabilitation programs, and drug therapy, in order to obtain a snapshot of current implementation strategies in daily clinical practice. The survey will adopt a web-based methodology for data provision and transmission. Preliminary results of the survey are expected in the late summer 2008. PMID- 18678222 TI - [Cardiomyopathies 2008. Rethinking the issue in light of the recent American Heart Association and European Society of Cardiology statements]. PMID- 18678223 TI - [Diagnosis and treatment of non-ST-elevation acute coronary syndromes. Common clinical lessons and a few differences between the ESC and the ACC/AHA guidelines published in 2007]. PMID- 18678224 TI - [Beyond dyssynchrony: what are the factors determining the response to cardiac resynchronization therapy?]. AB - Although cardiac resynchronization therapy is currently used for treatment of refractory heart failure in patients with low ejection fraction and cardiac dyssynchrony, there is a substantial number of non-responders. This indicates that, in addition to cardiac dyssynchrony, there are other factors affecting response to cardiac resynchronization therapy. Pre-implant identification of these factors appears of crucial importance in order to finalize the resynchronization treatment to those patients who have the highest probability of a positive response. In this review the main non-dyssynchrony determinants of response to cardiac resynchronization therapy are presented and discussed. PMID- 18678225 TI - [Prophylactic use of cardiac implantable defibrillators in patients with severe left ventricular dysfunction: how to deal with decision making among guidelines, clinical practice, ethical problems, and limited economic resources]. AB - Sudden cardiac death (SCD) is usually due to ventricular tachycardia/fibrillation and represents one of the most important medical and socio-economical problems in western countries. It accounts for approximately 1 life/1000 subjects/year. New and effective treatments are necessary to reduce such dramatic event. During the last decade implantable cardioverter-defibrillators (ICDs) showed to be an effective tool to reduce both total and SCD mortality either when used for secondary or primary SCD prevention. At present, ICD implantation guidelines suggest to implant an ICD in all the patients on the basis of a left ventricular ejection fraction < or =30-35% only. This scarcely sensitive and specific criterion implies the necessity to implant very costly devices in a wide number of patients to save only few lives. A more accurate patient selection is desirable either from a clinical or ethical or economic point of view. Fortunately, this appears to be possible using well known and proven epidemiological, clinical and risk stratification data. On the basis of such data, more selective ICD implantation criteria might be used in older patients or in patients with significant comorbidity or in those patients identified at very low risk of SCD. PMID- 18678226 TI - [Percutaneous rheolytic thrombectomy with AngioJet for pulmonary embolism: methods and results in the experience of a high-volume center]. AB - BACKGROUND: Pulmonary embolism is a common disease with significant mortality. Anticoagulant and thrombolytic therapies are a mainstay in the management of acute pulmonary embolism, especially with hemodynamic compromise. However, systemic drugs cannot achieve timely and effective treatment of acute pulmonary embolism in all patients. Recently, rheolytic thrombectomy has been proven to be effective in the treatment of massive thrombosis in vein grafts. The purpose of this study was to describe the rheolytic thrombectomy procedure and to evaluate the efficacy of thrombus removal using the AngioJet catheter in patients with acute massive and submassive pulmonary embolism. METHODS: Thirty consecutive patients (16 men, 14 women, mean age 67.9 +/- 13.5 years) with massive or submassive pulmonary embolism were treated with rheolytic thrombectomy. The Miller index and the systolic pulmonary pressure were evaluated pre- and post rheolytic thrombectomy. In-hospital results were analyzed according to the learning curve of operators. RESULTS: Technical success was achieved in 93.3% of patients. Eleven patients received adjunctive loco-regional thrombolytic agent (alteplase) or abciximab. A significant improvement in the obstruction, perfusion and Miller indexes and in the systolic pulmonary pressure was observed after rheolytic thrombectomy (p <0.0001) in all patients. The achievement of a larger experience by the operators was associated with a less procedural complication rate (i.e. post-procedural increase in creatinine, cardiac arrest during the procedure, post-procedural hemoptysis, and in-hospital mortality). Total in hospital mortality occurred in 5 patients (16.7%), 60% of them presented with cardiogenic shock. All patients were alive at the 12-month follow-up, except one who died after 3 months for myocardial infarction. CONCLUSIONS: Percutaneous rheolytic thrombectomy using the AngioJet catheter may be a valid treatment option for patients with massive or submassive pulmonary embolism with rapid and significant hemodynamic improvement and encouraging results at early and long term follow-up. Greater laboratory and operator experience yield better clinical results. PMID- 18678227 TI - [Percutaneous rheolytic thrombectomy for the treatment of acute pulmonary embolism]. PMID- 18678228 TI - [An unusual type of dyspnea]. PMID- 18678229 TI - [Bivalirudin and cardiac surgery: two case reports]. AB - Immune reactions to heparin and in particular heparin-induced thrombocytopenia are not rare and potentially fatal complications of heparin treatment. These conditions are frequently underdiagnosed in cardiac surgery. Moreover, few data are available in the literature about the use of alternative anticoagulants to heparin in this setting. We describe the successful use of bivalirudin in 2 patients with hypersensitivity to heparin who underwent cardiac surgery. PMID- 18678230 TI - [Saddle embolus through foramen ovale patency]. PMID- 18678231 TI - Different effects of spinally applied prostaglandin D2 on responses of dorsal horn neurons with knee input in normal rats and in rats with acute knee inflammation. AB - Prostaglandin D2(PGD2) is the most produced prostanoid in the CNS of mammals, and in behavioral experiments it has been implicated in the modulation of spinal nociception. In the present study we addressed the effects of spinal PGD2 on the discharge properties of nociceptive spinal cord neurons with input from the knee joint using extracellular recordings in vivo, both in normal rats and in rats with acute inflammation in the knee joint. Topical application of PGD2 to the spinal cord of normal rats did not influence responses to mechanical stimulation of the knee and ankle joint except at a high dose. Specific agonists at either the prostaglandin D2 receptor 1 (DP1) or the prostaglandin D2 receptor 2 (DP2) receptor had no effect on responses to mechanical stimulation of the normal knee. By contrast, in rats with inflamed knee joints either PGD2 or a DP1 receptor agonist decreased responses to mechanical stimulation of the inflamed knee and the non-inflamed ankle thus reducing established inflammation-evoked spinal hyperexcitability. Vice versa, spinal application of an antagonist at DP1 receptors increased responses to mechanical stimulation of the inflamed knee joint and the non-inflamed ankle joint suggesting that endogenous PGD2 attenuated central sensitization under inflammatory conditions, through activation of DP1 receptors. Spinal application of a DP2 receptor antagonist had no effect. The conclusion that spinal PGD2 attenuates spinal hyperexcitability under inflammatory conditions is further supported by the finding that spinal coapplication of PGD2 with prostaglandin E2 (PGE2) attenuated the PGE2-induced facilitation of responses to mechanical stimulation of the normal joint. PMID- 18678232 TI - Ethnobotanical study of medicinal plants used by Paliyar tribals in Theni district of Tamil Nadu, India. AB - An ethnobotanical survey was carried out among the Paliyar tribals in various tribal villages of Theni district, Tamil Nadu, India during December 2004 to January 2006. A total of 101 species of ethnomedicinal plants belonging to 90 genera and 48 families were reported with the help of standardized questionnaires among 15 tribal informants between the ages of 26 to 82. The study shows a high degree of ethnobotanical novelty and the use of plants among the Paliyars reflects the revival of interest in traditional folk medicine. PMID- 18678233 TI - Comparative genomics approach to the expression of figalpha, one of the earliest marker genes of oocyte differentiation in medaka (Oryzias latipes). AB - We analyzed molecular cascades of sex differentiation in medaka gonads by examining the transcriptional regulation of the oocyte-expressed gene, figalpha. We first confirmed that figalpha is one of the earliest marker genes of oocyte differentiation by quantitative RT-PCR and in situ hybridization. Expression of putative figalpha target genes, zpc4 and zpb, followed that of figalpha. A meiosis-specific gene, scp3, showed expression temporally and spatially similar to figalpha. To characterize the cis-regulatory sequences of figalpha, we compared genomic organizations of vertebrate figalpha genes. Both number and sequence homology of the C-terminal exons showed divergence, suggesting their less important roles. In the frog, Xenopus tropicalis, and in many teleosts, figalpha is located between hexokinase 2 and beta-adducin. We compared this genomic region for potential cis-regulatory elements and found no DNA stretches with high homology. In spite of this lack of sequence similarities, fluorescent protein transgenes surrounded with figalpha flanking sequences from the compact genomes of fugu or Tetraodon faithfully reproduced the endogenous expression of figalpha in the medaka oocytes, indicating conserved regulatory mechanisms. PMID- 18678234 TI - Structural features and gene-expression profiles of actin homologs in Porphyra yezoensis (Rhodophyta). AB - The marine red alga Porphyra yezoensis contains an actin gene family consisting of at least four isoforms (PyACT1, 2, 3 and 4). The amino acid identity between isoforms exceeds 83%, and each contains a putative nuclear export signal (NES). We scanned the sequences for amino acids in regions homologous to the intermonomeric interface of actin filaments. Few residues expected to engage in cross-linking were conserved between the four isoforms. The results of the sequence analyses suggest that PyACT2 probably functions in the nucleus as a monomer (G-actin) or in other unconventional forms. In addition, the distribution and position of the introns were different from those in florideophycean actin genes. The expression level of PyACT3 in matured gametophytes was significantly higher than in those in a vegetative state, although the mRNA was detected at similar levels in both apical and basal parts of thalli. The expression levels of PyACT2 and 4, on the other hand, did not change significantly between the matured and vegetative gametophytes. The PyACT3 may serve as a molecular marker for monitoring thallus maturation in this species. PMID- 18678235 TI - Time-dependent acetylsalicylic acid effects on liver CYP1A and antioxidant enzymes in a rat model of 7,12-dimethylbenzanthracene (DMBA)-induced mammary carcinogenesis. AB - 7,12-Dimethylbenzanthracene (DMBA) is an abundant environmental contaminant, which undergoes bioactivation, primarily by the CYP1 family, both in liver and extra-hepatic tissues. Dietary acetylsalicylic acid (ASA) has been recently reported to inhibit DMBA-mediated mammary tumour formation in rats. Chemopreventive substances may reduce the risk of developing cancer by decreasing metabolic enzymes responsible for generating reactive species (phase I enzymes) and/or increasing phase II enzymes that can deactivate radicals and electrophiles. To test these hypotheses, Sprague-Dawley female rats were orally administered ASA as lysine acetylsalicylate (50 mg per capita/day for 21 days in water), DMBA (10 mg per capita in olive oil on day 7, 14, and 21), ASA and DMBA in combination, and vehicles only, respectively. Six rats for each group were sacrificed on day 8, 15, and 22. The DMBA-mediated increase in hepatic CYP1A expression and related activities was not significantly affected by ASA, which, conversely, enhanced in a time-dependent manner the liver reduced glutathione content (up to 52%) and the activity of NAD(P)H-quinone oxidoreductase (up to 34%) in DMBA-treated rats. It is proposed that the positive modulation of the hepatic antioxidant systems by ASA may play a role in the chemoprevention of mammary tumourigenesis induced by DMBA in the female rat. PMID- 18678236 TI - Pharmacokinetic evaluation of indomethacin ethyl ester-loaded nanoencapsules. AB - Goals were to evaluate indomethacin ethyl ester-nanoencapsules (IndOEt-NC) pharmacokinetics in rats and the in vivo ester conversion to indomethacin (IndOH). After i.v. and oral administration exclusively IndOH was detected in plasma. The AUC(IndOEt-NC)/AUC(IndOH) ratio after i.v. dosing was 0.68, accounting for dose and molecular weight differences, probably due to increased IndOH clearance after IndOEt-NC administration (alpha=0.05). The results confirm that antiedematogenic activity reported for IndOEt-NC is due to IndOH. Encapsulation did not protect the ester which in vivo is rapidly released and converted to IndOH, acting as a pro-drug. PMID- 18678237 TI - Phytochemistry and in vitro pharmacological activities of South African Vitex (Verbenaceae) species. AB - AIM OF THE STUDY: The in vitro phytochemical and pharmacological investigation of the non-volatile extracts of five South African Vitex species (Verbenaceae); V. obovata ssp. obovata, V. obovata ssp. wilmsii, V. pooara, V. rehmannii and V. zeyheri were investigated in order to validate their traditional use to treat a wide range of ailments such as malaria, wounds, skin diseases and body pains. MATERIAL AND METHODS: The antimicrobial activity was assessed using the minimum inhibitory concentration assay. Through bioactivity-guided fractionation, the fraction responsible for the antimicrobial activity was determined. The toxicity profile, anti-oxidant and anti-inflammatory activity was evaluated using the tetrazolium cellular viability, 2,2-diphenyl-1-picrylhydrazyl and 5-lipoxygenase assays respectively. The antimalarial activity of the extracts and isolated compound from V. rehmannii was also investigated on the chloroquine-resistant Gambian FCR-3 strain of Plasmodium falciparum using the tritiated hypoxanthine incorporation assay. RESULTS: Mostly good antimicrobial inhibition was evident against Gram-positive bacteria (0.02-8.00 mg/ml) and lower activity against the Gram-negative bacteria and the yeast (0.50-8.00 mg/ml). The fraction responsible for antimicrobial activity of V. rehmannii was purified to give a labdane diterpene as an inseparable epimeric mixture of 12S,16S/R-dihydroxy-ent-labda 7,13-dien-15,16-olide. Cirsimaritin was also isolated and identified from V. rehmannii. All the species, apart from V. zeyheri, exhibited scavenging activity (IC50: 22.14+/-1.74 to 33.06+/-1.68 microg/ml) in the anti-oxidant assay. None of the species displayed any anti-inflammatory activity at 100 microg/ml. All the extracts and the labdane diterpene exhibited good antimalarial activity, with the labdane diterpene being the most active (IC50: 2.39+/-0.64 microg/ml). The test extracts were shown to be highly toxic, displaying safety index values ranging from 0.53 to 2.59. CONCLUSION: Of all the pharmacological investigations, the antimalarial and antimicrobial activity exhibited greatest activity and may provide a scientific basis for the ethnomedical use of Vitex species. PMID- 18678238 TI - Anti-inflammatory activity of the extracts and fractions from Erigeron multiradiatus through bioassay-guided procedures. AB - ETHNOPHARMACOLOGICAL RELEVANCE: Erigeron multiradiatus (Lindl.) Benth., an herb that grows in the alpine and subalpine meadow of Qinghai-Tibet plateau, has been widely used as a folk remedy by the native people for treatment of various inflammatory ailments. AIM OF THE STUDY: In order to isolate and identify the active components of Erigeron multiradiatus for anti-inflammatory activity, a preliminary phytochemical study and a bioassay-guided fractionation and purification process was performed. MATERIAL AND METHODS: The dry whole plant Erigeron multiradiatus was extracted with 50% ethanol and then separated into CHCl(3), n-BuOH, and aqueous fractions. The anti-inflammatory activities of each fraction were investigated using two in vivo inflammation models. RESULTS: These results exhibited varying degrees of anti-inflammatory activities and the n-BuOH fraction showed the strongest anti-inflammatory activities. The n-BuOH fraction was then subjected to separation and purification using macroporous resins column chromatography and Sephadex LH-20 leading to two flavonoids glucuronides identified as scutellarein-7-O-beta-glucuronide and apigenin-7-O-beta glucuronide. Furthermore, LC-MS/MS identification and quantification of isolated compounds were also performed. CONCLUSION: Scutellarein-7-O-beta-glucuronide and apigenin-7-O-beta-glucuronide were considered as major components and principally responsible for the anti-inflammatory activity of Erigeron multiradiatus. Thus the results of our study provide a scientific basis for the utilization of Erigeron multiradiatus in traditional Tibetan medicine. PMID- 18678239 TI - In vitro anticancer screening of South African plants. AB - AIM OF THE STUDY: The purpose of the present study is to evaluate South African plants for their anticancer activity. MATERIALS AND METHODS: Plant species were collected throughout South Africa and voucher specimens were deposited and identified at the South African National Biodiversity Institute. Plant extracts were prepared and screened for in vitro anticancer activity against a panel of three human cell lines (breast MCF7, renal TK10 and melanoma UACC62) at the CSIR. Plant extracts that exhibited anticancer activity against these three human cell lines were screened by the NCI against sixty human cancer cell lines organized into sub-panels representing leukaemia, melanoma, cancer of the lung, colon, kidney, ovary, central nervous system, breast and prostate. RESULTS: A total of 7500 plant extracts were screened for in vitro anticancer activity against breast MCF7, renal TK10 and melanoma UACC62 human cell lines between the period 1999 and 2006. Hits were classified into four categories based on their total growth inhibition of the cell lines. A hit rate of 5.9% was obtained for extracts which showed moderate activity and these were screened by the NCI against a panel of sixty human cancer cell lines. The extracts of plant species with limited published information for their anticancer properties were subjected to bioassay guided fractionation and the active constituents isolated and identified. The largest number of plant specimens in this study was from the family Asteraceae, which is rich in sesquiterpene lactones. CONCLUSIONS: Although the extracts of the plants were randomly selected, 68% of these plant species which were hits in the screening programme are reported to be used medicinally. Based on our data, it appears that unrelated medicinal use of the source plants may serve as an initial guide to selection of plants for anticancer screening. PMID- 18678240 TI - Retardation in somatosensory cortex development induced by postnatal BrdU treatment in mice. AB - Cerebral dysgeneses are in the background of several neurological and mental disturbances. The aim of the present study was to investigate structural and activity changes following disturbed postnatal neuronal development in mice. Newborn C57Bl6 mice were exposed to 5-bromo-2'-deoxyuridine (BrdU: daily 50 microg/g body weight) during a period between postnatal days P0-P5 or P0-P11, respectively, and neuronal malformation and malfunctioning of somatosensory (barrel field) cortex was analyzed in adolescent animals. Alterations in histological architecture of interneuronal and glial elements were studied and correlated with electrophysiological modifications. Between P30 and P35 days litters underwent ex vivo electrophysiological experiments to examine the changes in basic excitability and in synaptic efficacy. Parallel immunohistochemistry was performed to detect BrdU, GABA and GFAP. There were no BrdU immunopositive cell nuclei in control animals, but marked staining was observed in both BrdU treated groups. Lessening in the number of GABAergic neurons was observed in the treated groups. GFAP immunohistochemical analysis has shown an increased number of activated astroglial cells in treated animals. Reduction of the number of GABAergic neurons was observed in the treated groups. Electrophysiological recordings on cortical slices showed increased excitability in the treated groups. PMID- 18678241 TI - Prenatal exposure to a non-steroidal anti-inflammatory drug or saline solution impairs sciatic nerve morphology: a stereological and histological study. AB - The toxic effect of non-steroidal anti-inflammatory drugs (NSAIDs) during development has been widely investigated. While it has been shown that these drugs impair central nervous development and compromise the neural activity, the effects of these substances on the development of peripheral nerves are still not clarified. In the present study, sciatic nerves withdrawn from three experimental groups of 4-week-old rats, prenatally exposed to either saline solution, or diclofenac sodium, and controls not exposed to any substance, were evaluated in terms of axon number, cross-sectional area of axon and myelin sheet thickness as well as of the ultrastructure of nerve fibers. Comparisons of stereological estimations among these three groups showed that axon number and mean axon cross sectional area, but not average myelin sheet thickness, were significantly decreased in rats that were exposed to both diclofenac sodium and also to the saline solution, in comparison of the control group. Electron microscope analysis revealed, in both treated groups, deterioration of myelin sheaths that was more pronounced in rats that were exposed to diclofenac sodium. Altogether, these findings show that the prenatal administration of both diclofenac sodium and saline solution impairs peripheral nervous system development, thus suggesting that this potential teratogenic effect should be also taken into consideration in the clinical use of these substances in pregnant patients. PMID- 18678242 TI - Mindfulness meditation training effects on CD4+ T lymphocytes in HIV-1 infected adults: a small randomized controlled trial. AB - Mindfulness meditation training has stress reduction benefits in various patient populations, but its effects on biological markers of HIV-1 progression are unknown. The present study tested the efficacy of an 8-week Mindfulness-based stress reduction (MBSR) meditation program compared to a 1-day control seminar on CD4+ T lymphocyte counts in stressed HIV infected adults. A single-blind randomized controlled trial was conducted with enrollment and follow-up occurring between November 2005 and December 2007. A diverse community sample of 48 HIV-1 infected adults was randomized and entered treatment in either an 8-week MBSR or a 1-day control stress reduction education seminar. The primary outcome was circulating counts of CD4+ T lymphocytes. Participants in the 1-day control seminar showed declines in CD4+ T lymphocyte counts whereas counts among participants in the 8-week MBSR program were unchanged from baseline to post intervention (time x treatment condition interaction, p=.02). This effect was independent of antiretroviral (ARV) medication use. Additional analyses indicated that treatment adherence to the mindfulness meditation program, as measured by class attendance, mediated the effects of mindfulness meditation training on buffering CD4+ T lymphocyte declines. These findings provide an initial indication that mindfulness meditation training can buffer CD4+ T lymphocyte declines in HIV-1 infected adults. CLINICAL TRIALS REGISTRATION: clinicaltrials.gov, Identifier: NCT00600561. PMID- 18678243 TI - Peripheral cytokines profile in Parkinson's disease. AB - Higher levels of proinflammatory cytokines are found in Parkinson's disease (PD) patient's brains and inflammation is thought to be a major contributor to the neurodegeneration. During the inflammatory process, microglial release of proinflammatory cytokines act on the endothelium of blood-brain barrier (BBB) cells to stimulate upregulation of adhesion molecules. Consequently, this upregulation leads to the recruitment of passing T cells and monocytes, which express the counter receptors, that then go on to release more cytokines [Whitton, P.S., 2007. Inflammation as a causative factor in the aetiology of Parkinson's disease, Br. J. Pharmacol. 50, 963-976; Kortekaas, R., Leenders, K.L., Van Oostrom, J.C., Vaalburg, W., Bart, J., Willemsen, A.T., Hendrikse, N.H., 2005. Blood-brain barrier dysfunction in parkinsonian midbrain in vivo, Ann. Neurol. 57, 176-179]. In addition, a systemic inflammatory response results in the production of cytokines which circulate in the blood and communicate with neurons within the brain. Thus, a central inflammatory reaction interacts with peripheral blood mononuclear cells (PBMCs) modulating immune activity. The present study investigates levels of production and expression of cyto/chemokines by PBMCs in PD patients. Basal and LPS-induced levels of MCP-1, RANTES, MIP 1alpha, IL-8, IFNgamma, IL-1beta and TNFalpha were significantly higher in PD patients than in HC subjects (p<0.001), as determined by RT-PCR and Elisa methods. Cyto/chemokine levels were significantly correlated with UPDRS III and H/Y stage (p<0.001). The Pearson's correlation coefficient (R) was also used to assess the strength of the relationship between NF-kappaBp65 levels and all studied cyto/chemokines and between NF-kappaBp65, UPDRS III and H/Y score in PD patients. The overall results strengthen and extend the knowledge of the peripheral dysregulation in the cytokine network associated with PD. PMID- 18678244 TI - Specific real-time PCR assays for the detection and quantification of Mycoplasma mycoides subsp. mycoides SC and Mycoplasma capricolum subsp. capripneumoniae. AB - Contagious bovine pleuropneumonia and contagious caprine pleuropneumonia are two severe respiratory infections of ruminants due to infection by Mycoplasma mycoides subsp. mycoides SC (MmmSC) and Mycoplasma capricolum subsp. capripneumoniae (Mccp), respectively. They are included in the OIE list of notifiable diseases. Here we describe the development of rapid, sensitive, and specific real-time PCR assays for the detection and quantification of MmmSC and Mccp DNA. MmmSC PCR primers were designed after whole genome comparisons between the published sequence of MmmSC strain type PG1(T) and the sequence of an M. mycoides subsp. mycoides large colony strain. For Mccp, previously published conventional PCR primers were applied. SYBR green was used as a detection agent for both assays. The assays specifically detected the targeted species in both cultures and clinical samples, and no cross-amplifications were obtained from either heterologous mycoplasma strain cultures or European field samples. The sensitivity of these new assays was estimated at 3-80 colony forming units per reaction and 4-80fg of DNA, representing a 2-3log increase in sensitivity compared to established conventional PCR tests. These new real-time PCR assays will be invaluable for application in various fields such as direct detection in diagnostic laboratories. PMID- 18678245 TI - Comparison of two twin-screw extruders for continuous granulation. AB - A comparison was made between two twin-screw extruders (APV Baker and Leistritz Micro) used for continuous wet granulation. Both extruders had similar screw configurations, based on the length-to-diameter ratio of the screws, existing out of a conveying zone of 20 D, i.e. 20 times the screw diameter and a granulation zone of 4D. The kneading blocks in the granulation zone were 2.2 and 2.5D for the Leistritz and APV extruders, respectively. An experimental design was used to investigate the influence of process parameters (total input rate and screw speed) and extruder type on granule and tablet quality. Dicalcium phosphate and alpha-lactose monohydrate were used as water-insoluble and water-soluble excipients, respectively. For dicalcium phosphate, the amount of fines (<125 microm), median granule size and granule friability were significantly influenced by extruder type and total input rate. For lactose, the amount of oversized agglomerates and median granule size were significantly affected by extruder type and total input rate. The granule formulations were properly agglomerated on both the extruders, although the extruder type had an important influence on the granule properties, which was more pronounced for dicalcium phosphate. This study shows that a given formulation cannot simply be interchanged between the two extruders without further work on the geometrics of the extruders. PMID- 18678246 TI - Expression of matrix metalloproteinase-2 and -9 and of tissue inhibitor of matrix metalloproteinase-1 in liver regeneration from oval cells in rat. AB - Oval cells participate in liver regeneration when hepatocyte replication is impaired. These precursor cells proliferate in periportal regions and organize in ductules. They are surrounded by a basement membrane, the degradation of which by matrix metalloproteinases (MMP) might trigger their terminal differentiation into hepatocytes. We studied the expression of MMP-2 and MMP-9 and that of one of their tissue inhibitors (TIMP-1) in a model of hepatic regeneration from precursor cells. Regeneration was induced by treating rats with 2 acetylaminofluorene followed by partial hepatectomy. MMP-2 and MMP-9 hepatic expression paralleled oval cell number with a peak at day 9-14 after hepatectomy. They were mainly detected in oval cells. TIMP-1 mRNA and oncostatin M receptor mRNA, a major regulator of TIMP-1 synthesis, markedly increased from day 1 after surgery until day 9 and then declined; they were mainly detected in interlobular bile duct cells and oval cells until day 14. In agreement with the in vivo data, the WB-F344 liver precursor cell line expressed MMP-2 and MMP-9, as well as TIMP 1 and oncostatin M receptor. These data suggest that (a) early increased TIMP-1 synthesis by biliary and oval cells favors basement membrane deposition around proliferating ductular structures through MMP inhibition, (b) delayed increased MMP expression, concomitant to decreased TIMP-1 synthesis, leads to basement membrane degradation, preceding oval cell differentiation, (c) the oncostatin M pathway might play a major role in increased TIMP-1 synthesis. PMID- 18678247 TI - T-cells behaving badly: structural insights into alloreactivity and autoimmunity. AB - T-cells play a critical role in protective immunity, with their broad receptor repertoire capable of engaging diverse foreign pMHC landscapes. While the versatility and specificity of this MHC-restricted response is the hallmark of adaptive immunity, unwanted TCR interactions can profoundly effect the health of the host leading for instance to allograft rejection or autoimmunity. In allogeneic transplantation, such adverse reactions can occur by an indirect pathway when the TCR interacts with self-MHC molecules presenting allogeneic MHC derived peptides. Direct T-cell alloreactivity involves recognition of the allogeneic molecule itself either through molecular mimicry or by novel pMHC binding modes. By contrast, auto-reactive TCRs are considered to interact in a manner distinct from cognate pMHC interactions. Here we review recent advances in the field, focusing on structural data pertaining to alloreactivity and auto reactivity and discuss implications for T-cell mediated transplant rejection and autoimmune disorders. PMID- 18678248 TI - The Aspergillus nidulans stress response transcription factor StzA is ascomycete specific and shows species-specific polymorphisms in the C-terminal region. AB - Orthologues of the Aspergillus nidulans gene stzA were identified and characterised in an additional 19 fungi. These orthologues were restricted to, and found within all the Pezizomycotina subphyla of the Ascomycota, for which data are available, but not the Saccharomycotina or Taphrinomycotina subphyla. Intron analysis indicated that both intron loss and gain have occurred in this gene. The orthologous proteins demonstrate considerable size variation (between 663 and 897 amino acids); with almost all this variability accounted for by a hyper-variable region that is carboxy terminal to the zinc finger region. The Hypocrea jecorina orthologue (ACE1) has the binding site 5'AGGCA. There is evidence of competition, or interaction, between the ACE1/StzA and AreA binding sites in promoters of stzA and its orthologues, as well as genes involved in the metabolism of amino acids. The A. nidulans and A. fumigatus cpcA promoters have seven potential ACE1/StzA binding sites, six of which are highly conserved in position. Two very closely positioned sites are conserved across 14 of the 19 fungi analysed. Potential CpcA binding sites (5'TGAC/GTCA) have been identified between -50 and -170bp of the ATG start in the promoters of 16 of the stzA orthologues. PMID- 18678249 TI - Protein technologies. PMID- 18678251 TI - Axiomatic methods, dopamine and reward prediction error. AB - The phasic firing rate of midbrain dopamine neurons has been shown to respond both to the receipt of rewarding stimuli, and the degree to which such stimuli are anticipated by the recipient. This has led to the hypothesis that these neurons encode reward prediction error (RPE)-the difference between how rewarding an event is, and how rewarding it was expected to be. However, the RPE model is one of a number of competing explanations for dopamine activity that have proved hard to disentangle, mainly because they are couched in terms of latent, or unobservable, variables. This article describes techniques for dealing with latent variables common in economics and decision theory, and reviews work that uses these techniques to provide simple, non-parametric tests of the RPE hypothesis, allowing clear differentiation between competing explanations. PMID- 18678250 TI - Message in a nucleus: signaling to the transcriptional machinery. AB - Tissue differentiation and signal transduction involve dramatic changes in gene expression. These changes can be brought about by the expression or activation of sequence-specific transcription factors. In order to regulate their target genes, such factors must navigate the intricate chromatin environment and engage the complex basal transcriptional machinery. We discuss three mechanisms through which signaling pathways can interact with complexes that alter chromatin structure or recruit RNA polymerase II. Signals that promote differentiation may alter the properties of such transcriptional regulatory complexes by incorporating tissue-specific subunits. Alternatively, adaptor subunits specialized to interact with specific transcription factors may allow a single complex to respond to multiple signals. Finally, individual regulatory proteins may integrate a variety of signals, allowing crosstalk between pathways. PMID- 18678252 TI - The role of fMRI in cognitive neuroscience: where do we stand? AB - Functional magnetic resonance imaging (fMRI) has quickly become the most prominent tool in cognitive neuroscience. In this article, I outline some of the limits on the kinds of inferences that can be supported by fMRI, focusing particularly on reverse inference, in which the engagement of specific mental processes is inferred from patterns of brain activation. Although this form of inference is weak, newly developed methods from the field of machine learning offer the potential to formalize and strengthen reverse inferences. I conclude by discussing the increasing presence of fMRI results in the popular media and the ethical implications of the increasing predictive power of fMRI. PMID- 18678253 TI - Spiking networks for Bayesian inference and choice. AB - Systems neuroscience traditionally conceptualizes a population of spiking neurons as merely encoding the value of a stimulus. Yet, psychophysics has revealed that people take into account stimulus uncertainty when performing sensory or motor computations and do so in a nearly Bayes-optimal way. This suggests that neural populations do not encode just a single value but an entire probability distribution over the stimulus. Several such probabilistic codes have been proposed, including one that utilizes the structure of neural variability to enable simple neural implementations of probabilistic computations such as optimal cue integration. This approach provides a quantitative link between Bayes optimal behaviors and specific neural operations. It allows for novel ways to evaluate probabilistic codes and for predictions for physiological population recordings. PMID- 18678254 TI - Signalling mechanisms underlying development and maintenance of dopamine neurons. AB - Majority of the brain dopamine (DA) neurons reside in a distinct area in the midbrain and project axons into the striatum and frontal cortex to control central nervous system (CNS) functions such as movement, motivation and mood. Age associated specific loss of DA neurons particularly in the midbrain region substantia nigra pars compacta (SNpc) causes Parkinson disease (PD), an incurable condition characterized by rigidity, involuntary and slowed movement affecting about 1% of people over the age of 60 years. Dopamine neurons appear to be one of the most sensitive types of neurons to both intrinsic and extrinsic stressors in the brain. Here we summarize how transcription factors, growth factors and in particular neurotrophic factors are used to make and maintain DA neurons. We also discuss mechanisms that underlie their specific vulnerability and highlight current state of art in drug development. PMID- 18678255 TI - From biophysics to cognition: reward-dependent adaptive choice behavior. AB - In neurobiological studies of various cognitive abilities, neuroscientists use mathematical models to fit behavioral data from well-controlled experiments and look for neural activities that are correlated with parameters in those models. The pinpointed neural correlates are often taken as evidence that a given task is performed according to the prescription of the applied model, and the relevant brain areas encode parameters of such a model. However, to go beyond correlations toward causal understanding, it is necessary to elucidate at multiple levels the neural circuit mechanisms of cognitive processes. This review focuses on recent studies of reward-based decision-making that have begun to tackle this challenge. PMID- 18678256 TI - Differentially expressed genes in resistant and susceptible Bombyx mori strains infected with a densonucleosis virus. AB - We investigated variations in the gene expression of Bombyx mori following infection with a densonucleosis virus (BmDNV-Z). Two B. mori near-isogenic lines, Jingsong and Jingsong.nsd-Z.NIL, which are highly susceptible and completely resistant to BmDNV-Z, respectively, were used in this study. The infection profiles of BmDNV-Z in the midguts of the B. mori Jingsong and Jingsong.nsd-Z.NIL larvae revealed that the virus invaded the midguts of both of these strains. However, its proliferation was notably inhibited in the midgut of the resistant strain. By using the suppression subtractive hybridization method, three cDNA libraries were constructed to compare BmDNV-Z responsive gene expression between the two silkworm lines. In total, 151 differentially expressed genes were obtained. Real-time qPCR analysis confirmed that 11 genes were significantly up regulated in the midgut of the Jingsong.nsd-Z.NIL strain following BmDNV-Z infection. Our results imply that these up-regulated genes might be involved in B. mori immune responses against BmDNV infection. PMID- 18678257 TI - Hippocampal atrophy and abnormal brain development following a prolonged hyperthermic seizure in the immature rat with a focal neocortical lesion. AB - In rats subjected to a focal cortical lesion soon after birth, hyperthermia at P10 induces a prolonged epileptic seizure, often followed by temporal lobe epilepsy in the adult. To determine whether brain damage and notably hippocampal atrophy occur early on in this model, whole brain as well as hemispheric, cortical, subcortical and hippocampal volumes was measured in non-lesioned and lesioned rat pups, 2 days (P12) and 12 days (P22) after the hyperthermic seizure. All pups with a cortical lesion showed reductions in whole brain and in ipsilateral hemispheric, cortical and hippocampal volumes at P12, which persisted at P22 in pups having also sustained a prolonged hyperthermic seizure at P10. Limiting the duration of the seizure with Diazepam prevented the hippocampal atrophy. Thus, a prolonged hyperthermic seizure in immature brain with a subtle neocortical lesion impairs normal brain development, and the duration of the seizure appears to be a key factor in generating hippocampal atrophy. PMID- 18678258 TI - Heterologous high-level E. coli expression, purification and biophysical characterization of the spine-associated RapGAP (SPAR) PDZ domain. AB - Spine-associated RapGAP (SPAR) is a 1783 residue, multidomain scaffolding protein which is a component of the NMDA receptor/PSD-95 complex in the post-synaptic density (PSD) of dendritic spines. Using a parallel expression screening approach, we identified a strategy to solubly express the SPAR PDZ domain in Escherichia coli. We show that maltose binding protein is required for the production of solubly expressed protein. We also show that small changes in construct length (2-5 residues) result in differential susceptibilities of the expressed proteins to proteolytic digestion, required for the expression tag removal. This has allowed us to identify a large-scale E. coli expression and purification protocol that results in the production of mg quantities of the SPAR PDZ domain. This is the first time that any of the multiple SPAR functional domains have been expressed in E. coli in quantities suitable for biophysical and biochemical studies, allowing us to investigate the role of the PDZ domain in SPAR function within the PSD. PMID- 18678259 TI - Functional expression in Bacillus subtilis of mammalian NADPH-cytochrome P450 oxidoreductase and its spore-display. AB - The technology for over-expressing NADPH-cytochrome P450 reductase (CPR), a diflavin-containing enzyme, offers the opportunity to develop enzymatic systems for environmental detoxication and bioconversions of drugs, pesticides and fine chemicals. In this study, Bacillus subtilis was chosen to express rat CPR (rCPR) because of its capacities for high protein production and spore formation. rCPR was expressed in B. subtilis DB104 under the transcriptional control of an IPTG inducible fusion promoter of P(groE) and P(tac). The expressed rCPR was released into the culture medium after sporulation by autolysis of the host cell. It was associated with and displayed on the spore surfaces; this was confirmed by measuring rCPR activity in purified spores and analyzing its accessibility to anti-rCPR antibodies using flow cytometry. The spore-displayed rCPR was able to reduce cytochrome c and ferricyanide, and also assisted in the O-deethylation of 7-ethoxyresorufin and 7-ethoxy-4-trifluoromethylcoumarin (EFC) by human cytochrome P450 1A2, indicating that it was functionally active. Spore surface display of rCPR in B. subtilis appears to be useful for preparing cytochrome P450 related enzymes, and spore biocatalysts of rCPR are likely to have wide biotechnological applications. PMID- 18678260 TI - Investigation of the influence of carbon dioxide concentrations on cerebral physiology by susceptibility-weighted magnetic resonance imaging (SWI). AB - Breathing carbogen (5% CO2 / 95% O2) dramatically increases cerebral blood flow (CBF), which induces a blood oxygenation level dependent (BOLD) related vascular signal change due to the concomitantly increased oxyhemoglobin concentration in the veins. However, carbogen often causes discomfort due to its forced strong and deep breathing which also may lead to severe motion artifacts in magnetic resonance imaging. In this study, susceptibility-weighted imaging (SWI) was performed with CO2 levels of 0, 1.67%, 3.33% and 5% to measure the induced BOLD signal changes in venous vessels and brain tissue. Susceptibility-weighted imaging data from 15 healthy subjects and one patient with a brain tumor were acquired. The signal magnitude of cortical veins increased relative to pure oxygen by 3.5+/-3.8%, 10.3+/-4.5%, and 22.7+/-8.8% for CO2 concentrations of 1.67%, 3.33%, and 5%, respectively. Significant signal changes were detected in segmented white matter for 5% CO2, and gray matter for both 3.3% and 5% CO2. The influence of motion artifacts was clearly traceable by the broadening of the signal distribution in segmented tissue. Heterogeneous signal changes were observed in the patient for the same tumor regions at both 3.33% and 5% CO2. Signal phase values of white and gray matter changed only very slightly with increasing CO2. Based on our findings we recommend the reduction of CO2 concentration to about 3% when using a mixture of O2 and CO2. All subjects also reported highly improved breathing comfort at 3.3% CO2 as compared to 5%. The marginal phase change of white and gray matter supports the assumption that deoxygenated blood alone does not explain the commonly observed phase difference between the two tissues. PMID- 18678261 TI - On the construction of an inter-subject diffusion tensor magnetic resonance atlas of the healthy human brain. AB - Voxel based morphometry (VBM) has been increasingly applied to detect diffusion tensor (DT) image abnormalities in patients for different pathologies. An important requisite for a robust VBM analysis is the availability of a high dimensional non-rigid coregistration technique that is able to align both the spatial and the orientational DT information. Consequently, there is a need for an inter-subject DTI atlas as a group specific reference frame that also contains this orientational DT information. In this work, a population based DTI atlas has been developed that incorporates such orientational DT information with high accuracy and precision. The proposed methodology for constructing such an atlas is compared with a subject based DTI atlas, in which a single subject is selected as the reference image. Our results demonstrate that the population based atlas framework is more accurate with respect to the underlying diffusion information. PMID- 18678262 TI - Look who's talking: the deployment of visuo-spatial attention during multisensory speech processing under noisy environmental conditions. AB - In a crowded scene we can effectively focus our attention on a specific speaker while largely ignoring sensory inputs from other speakers. How attended speech inputs are extracted from similar competing information has been primarily studied in the auditory domain. Here we examined the deployment of visuo-spatial attention in multiple speaker scenarios. Steady-state visual evoked potentials (SSVEP) were monitored as a real-time index of visual attention towards three competing speakers. Participants were instructed to detect a target syllable by the center speaker and ignore syllables from two flanking speakers. The study incorporated interference trials (syllables from three speakers), no-interference trials (syllable from center speaker only), and periods without speech stimulation in which static faces were presented. An enhancement of flanking speaker induced SSVEP was found 70-220 ms after sound onset over left temporal scalp during interference trials. This enhancement was negatively correlated with the behavioral performance of participants -- those who showed largest enhancements had the worst speech recognition performance. Additionally, poorly performing participants exhibited enhanced flanking speaker induced SSVEP over visual scalp during periods without speech stimulation. The present study provides neurophysiologic evidence that the deployment of visuo-spatial attention to flanking speakers interferes with the recognition of multisensory speech signals under noisy environmental conditions. PMID- 18678263 TI - Pleistocene separation of mitochondrial lineages of Mytilus spp. mussels from Northern and Southern Hemispheres and strong genetic differentiation among southern populations. AB - Smooth-shelled mussels, Mytilus spp., have an antitropical distribution. In the Northern Hemisphere, the M. edulis complex of species is composed of three genetically well delineated taxa: M. edulis, M. galloprovincialis and M. trossulus. In the Southern Hemisphere, morphological characters, allozymes and intron length polymorphisms suggest that Mytilus spp. populations from South America and Kerguelen Islands are related to M. edulis and those from Australasia to M. galloprovincialis. On the other hand, a phylogeny of the 16S rDNA mitochondrial locus demonstrates a clear distinctiveness of southern mussels and suggests that they are related to Mediterranean M. galloprovincialis. Here, we analysed the faster-evolving cytochrome oxidase subunit I locus. The divergence between haplotypes of populations from the two hemispheres was confirmed and was found to predate the divergence between haplotypes of northern M. edulis and M. galloprovincialis. In addition, strong genetic structure was detected among the southern samples, revealing three genetic entities that correspond to (1) South America and Kerguelen Island, (2) Tasmania, (3) New Zealand. Using the trans Arctic interchange as a molecular clock calibration, we estimated the time since divergence of populations from the two hemispheres to be between 0.5 million years (MY) and 1.3 MY (average 0.84 MY). The contrasting patterns observed for the nuclear and the organelle genomes suggested two alternative, complex scenarios: two trans-equatorial migrations and the existence of differential barriers to mitochondrial and nuclear gene flow, or a single trans-equatorial migration and a view of the composition of the nuclear genome biased by taxonomic preconception. PMID- 18678264 TI - Olfaction as a model system for the neurobiology of mammalian short-term habituation. AB - Olfaction represents an ideal model system for the study of mammalian habituation given that it is an anatomically relatively simple system with strong reciprocal connections to the limbic system, driving both reflexive and non-reflexive (motivated) behaviors that are easily quantifiable. Data are reviewed here demonstrating short-term habituation of the odor-evoked heart-rate orienting reflex described according to the criteria for habituation outlined by Thompson and Spencer [Thompson, R. F., & Spencer, W. A. (1966). Habituation: A model phenomenon for the study of neuronal substrates of behavior. Psychological Reviews, 73(1), 16-43]. A necessary and sufficient mechanism of short-term habituation is then described, which involves a metabotropic glutamate receptor mediated depression of afferent input to the piriform (primary olfactory) cortex. Finally, evidence for, and a mechanisms of, dishabituation of the orienting reflex and cortical adaptation are described. PMID- 18678265 TI - Auditory trace fear conditioning requires perirhinal cortex. AB - The hippocampus is well-known to be critical for trace fear conditioning, but nothing is known about the importance of perirhinal cortex (PR), which has reciprocal connections with hippocampus. PR damage severely impairs delay fear conditioning to ultrasonic vocalizations (USVs) and discontinuous tones (pips), but has no effect on delay conditioning to continuous tones. Here we demonstrate that trace auditory fear conditioning also critically depends on PR function. The trace interval between the CS offset and the US onset was 16s. Pre-training neurotoxic lesions were produced through multiple injections of N-methyl-D aspartate along the full length of PR, which was directly visualized during the injections. Control animals received injections with phosphate-buffered saline. Three-dimensional reconstructions of the lesion volumes demonstrated that the neurotoxic damage was well-localized to PR and included most of its anterior posterior extent. Automated video analysis quantified freezing behavior, which served as the conditional response. PR-damaged rats were profoundly impaired in trace conditioning to either of three different CSs (a USV, tone pips, and a continuous tone) as well as conditioning to the training context. Within both the lesion and control groups, the type of cue had no effect on the mean CR. The overall PR lesion effect size was 2.7 for cue conditioning and 3.9 for context conditioning. We suggest that the role of PR in trace fear conditioning may be distinct from some of its more perceptual functions. The results further define the essential circuitry underlying trace fear conditioning to auditory cues. PMID- 18678266 TI - Effect of estrogen and its antagonist on the expression of arginine vasotocin (AVT) and its oxytocic-like receptor VT3 in the shell gland of Japanese quail, Coturnix coturnix japonica. AB - Avian neurohypophysial hormone arginine vasotocin (AVT) is known to regulate shell gland contractility during oviposition. While studying the role of estrogen in the expression and regulation of AVT and its oxytocic-like receptor VT3, using in situ hybridization and immunohistochemistry, it was observed that the expression of AVT and its receptor was not detected in the shell gland of sexually immature Japanese quail. However, administration of estrogen to these birds not only stimulates the growth and activity (as assessed by increased mucosal fold length, total protein content and alkaline phosphatase level) of the shell gland but also upregulates the expression of AVT and VT3. Further, administration of estrogen antagonist tamoxifen to sexually mature bird shows opposite results. On the other hand, localization of ir-AVT, observed in the ovary of sexually mature bird, was not detected in the estrogen treated sexually immature quail. It is concluded that estrogen not only affects the growth and differentiation of avian oviduct, but also regulates the expression of shell gland AVT and its receptor VT3. Present findings suggest that the locally synthesized AVT acts in a paracrine way to upregulate VT3 receptor and thus facilitates the endocrine function of neurohypophysial AVT during oviposition. PMID- 18678267 TI - Daily rhythms of body temperature and heat production of sibling Mastomys species from different ecosystems--the response to photoperiod manipulations. AB - We compared body temperature (T(b)) and metabolic rates, measured as oxygen consumption (VO(2)), daily rhythms of two sibling species of the genus Mastomys. We also studied their responses to long day (16L: 8D, LD) and short day (8L: 16D, SD) photoperiod manipulations at a constant ambient temperature of 26+1 degrees C. We noted significant differences in T(b) and VO(2) daily rhythm patterns, under SD and LD-acclimation between the sibling species. These differences explain adaptation to the climatic conditions that prevail in the different ecosystems where these species live. To the best of our knowledge, this is the first time that physiological differences between the two siblings are measured by using chronobiological methods. PMID- 18678268 TI - Hornet silk proteins in the cocoons produced by different Vespa species inhabiting Japan. AB - We compared the components of hornet silk - a fibrous protein occurring in the cocoons produced by hornet larvae - among 6 species of the genus Vespa inhabiting Japan: V. simillima, V. dybowskii, V. crabro, V. mandarinia, V. ducalis, and V. analis. From the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE), N-terminal amino acid sequence analysis, and 5'-RACE, it was found that the major component proteins composing hornet silk could be divided into 6 groups. Among these 6 proteins, 5 proteins were common to the hornet silks of all 6 Vespa species. The SDS-PAGE major band corresponding to the remaining protein was observed only in the hornet silks of V. mandarinia and V. ducalis. This correspondence between V. mandarinia and V. ducalis can probably be explained in terms of the phylogenetic relationships of the Vespa species. PMID- 18678269 TI - Molecular cloning and characterization of proliferating cell nuclear antigen (PCNA) from Chinese shrimp Fenneropenaeus chinensis. AB - The proliferating cell nuclear antigen gene was cloned from Fenneropenaeus chinensis (FcPCNA). The full-length cDNA sequence of FcPCNA encodes 260 amino acids showing high identity with PCNAs reported in other species. FcPCNA expressed especially high in proliferating tissues of shrimp such as haematopoietic tissue (HPT) and ovary. In order to understand the response of HPT to bacteria and virus challenge, mRNA level of FcPCNA in HPT was analyzed after shrimp were challenged by Vibrio anguillarum and white spot syndrome virus (WSSV). FcPCNA expression in HPT of shrimp was responsive to WSSV and Vibrio challenge, but different expression profiles were obtained after challenge by these two pathogens. The data provide additional information to understand the defense mechanisms of shrimp against virus and bacteria. PMID- 18678270 TI - Pretreatment with the Gram-positive bacterial cell wall molecule peptidoglycan improves bacterial clearance and decreases inflammation and mortality in mice challenged with Pseudomonas aeruginosa. AB - The objective of this study was to determine if inflammatory tolerance and enhancement of innate immune function could be induced by the Gram-positive cell wall component peptidoglycan (PGN). Male mice (C57BL6/J or C3H/HeJ, 8-12 weeks of age) were given intraperitoneal injections of 1mg PGN on 2 consecutive days. The mice were then challenged with lipopolysaccharide (LPS) or live Pseudomonas aeruginosa (1 x 10(8) colony-forming units) 2 days after the second pretreatment. Mice pretreated with PGN had diminished plasma concentrations of TNFalpha and IFNgamma and elevated concentrations of IL-10 in response to a subsequent LPS or Pseudomonas challenge when compared to untreated controls. Bacterial clearance was improved in mice pretreated with PGN, and mortality in response to a subsequent Pseudomonas challenge was significantly attenuated. PGN pretreatment of LPS-unresponsive mice (C3H/HeJ) verified that the effect of PGN pretreatment was not due to any LPS contamination. We have previously demonstrated that PGN pretreatment induced resistance to a Gram-positive bacterial challenge. The present study extends those results by showing that exposure to the Gram-positive bacterial cell wall component peptidoglycan also induces cross-tolerance to LPS and non-specifically enhances innate immune function in that PGN-pretreated mice had increased resistance to Gram-negative bacterial challenge. PMID- 18678271 TI - Quantification of bacterial internalization by host cells using a beta-lactamase reporter strain: Neisseria gonorrhoeae invasion into cervical epithelial cells requires bacterial viability. AB - Neisseria gonorrhoeae can invade into cervical epithelial cells to overcome this host defense barrier. We developed a beta-lactamase reporter system that allowed us to quantify at the single cell level if a host cell internalized a viable or nonviable microorganism. We autodisplayed beta-lactamase on the surface of FA1090 [FA1090Phi(bla-iga')] and demonstrated by confocal fluorescence microscopy and flow cytometry that FA1090Phi(bla-iga') cleaved the beta-lactamase substrate CCF2 AM loaded into host cells only when gonococci were internalized by these host cells. While FA1090Phi(bla-iga') adhered to almost all ME180 cells, viable N. gonorrhoeae were internalized by only a subset of cells during infection. Nonviable gonococci adhered to, but were not internalized by ME180 cells, and failed to recruit F-actin to sites of adherent bacteria. Overall, we show that epithelial cell invasion is a dynamic process that requires viable N. gonorrhoeae. We demonstrate the advantages of the beta-lactamase reporter system over the gentamicin protection assay in quantifying bacterial invasion. The reporter system that we have developed can be adapted to studying the internalization of any bacterial species into any host cell. PMID- 18678272 TI - Retinoic acid prevents Chlamydia pneumoniae-induced foam cell development in a mouse model of atherosclerosis. AB - Chlamydia pneumoniae, a common respiratory pathogen, has been associated with cardiovascular disease. C. pneumoniae infection accelerates atherosclerotic lesion development in hyperlipidemic animals. Retinoic acid, an anti-oxidant, inhibits infection of endothelial cells by C. pneumoniae. The present study demonstrated that retinoic acid suppresses the acceleration of foam cell lesion development induced by C. pneumoniae in hyperlipidemic C57BL/6J mice. Retinoic acid treatment had no effect on foam cell lesion development in uninfected animals. Lung infection and duration was decreased in treated mice, suggesting one mechanism by which retinoic acid reduces C. pneumoniae-accelerated foam cell lesion formation in hyperlipidemic mice. PMID- 18678273 TI - c-Jun N-terminal kinase pathways in diabetes. AB - Type 2 diabetes develops from insulin resistance and has become a worldwide epidemic. The c-Jun N-terminal kinases have been considered as signaling molecules linking inflammation and insulin resistance. Genetic disruption of c Jun N-terminal kinase-1 gene prevents the development of insulin resistance in obese and diabetic mice. Inhibition of c-Jun N-terminal kinases by a small cell permeable peptide improves insulin sensitivity in mice. Hepatic inhibition of c Jun N-terminal kinases using a dominant-negative protein or knockdown of c-Jun N terminal kinase-1 gene by RNA interference reduces blood glucose and insulin levels and enhances hepatic insulin signaling in mice. Recent evidence demonstrates that the hepatic c-Jun N-terminal kinase pathway plays an important role in lipid and lipoprotein homeostasis in mice. This review discusses recent advances in our understanding of the role of c-Jun N-terminal kinase pathway in metabolic control and its potential as a target for the treatment of type 2 diabetes. PMID- 18678274 TI - On the stage division mechanism in pharmaceuticals development processes. AB - This paper contributes to the understanding of a recent trend taking place in the pharmaceutical industry, whereby large companies develop a meaningful number of alliances with smaller firms, to share stages of drug development processes. Small firms tend to operate in earlier stages and intuitively, this may suggest that this is because they are more efficient, or more willing, than larger firms to take such early risk. In exchange, larger firms would afford the higher development costs of later phases. In this article we argue that the intuition appears correct if large companies have enough resources to develop in-house all potentially interesting projects, but not necessarily so when they are resource constrained. PMID- 18678275 TI - Drug discovery in the era of Facebook--new tools for scientific networking. AB - Social networking is beginning to make an impact on the drug discovery process. While bioinformatics and chemoinformatics underpin research at a scientific level, rapid communication between individual researchers across continents now allows the global exchange of ideas, tools and technologies. Networking at this level of speed and reach is quite a recent phenomenon. It facilitates the development of common interests, accelerates technology transfer and increases cooperative and competitive behaviour. In this review, we critically evaluate different web based networking approaches as effective resources for the drug discovery scientist. We also ask whether social networking sites will evolve into serious and credible resources for the drug discovery community. PMID- 18678276 TI - Targeted therapy of cancer using diphtheria toxin-derived immunotoxins. AB - The mortality rate in cancer patients demands novel therapy. One of the novel approaches developed in recent decades includes immunotoxins. Cancer cells frequently have specific growth factor receptors/antigens overexpressed on their surface; this is the principle of selective targeting of immunotoxins. Ligands recognizing these receptors and antigens can be conjugated to modified toxins. Continuous efforts are being made (i) to investigate molecules exclusively expressed on cancer cells, (ii) to improve the specificity and efficacy of these immunotoxins, (iii) to eliminate side effects (iv) to decrease immunogenicity and (v) to improve pharmacokinetics and ensure better drug delivery. PMID- 18678277 TI - Integrating the discovery pipeline for novel compounds targeting ion channels. AB - Many ion channels are attractive therapeutic targets for the treatment of neurological or cardiovascular diseases; there is a continuous need for selective channel antagonists and/or agonists. Recently, several technologies have been developed that make exploration of the enormous diversity of venom-derived peptidic toxins more feasible. Integration of exogenomics with synthetic methods such as diselenide or fluorous bridges, backbone spacers, and N-to-C cyclization provides an emerging technology that promises to accelerate discovery and development of natural products based compounds. These drug-discovery efforts are complemented by novel approaches to modulate the activities of ion channels and receptors. Taken together, these technologies will advance our knowledge and understanding of ion channels and will accelerate their expansion as targets for first-in-class therapeutics. PMID- 18678279 TI - Cytotoxic and genotoxic effects of methotrexate in germ cells of male Swiss mice. AB - Methotrexate (MTX) is an anti-metabolite drug widely used in the treatment of neoplastic disorders, rheumatoid arthritis and psoriasis. Developed as an analogue of folic acid, it inhibits purine and pyrimidine synthesis that accounts for its therapeutic efficacy as well as for its toxicities. MTX has narrow therapeutic index and its toxicity has been reported in various organ systems including gastrointestinal, haematologic and central nervous system. The objective of the present study is to investigate the germ cell toxicity induced by MTX in male Swiss mice. MTX was administered intraperitoneally (ip) at the doses of 5, 10, 20 and 40 mg/kg to mice (20-25 g) weekly once (wk) for 5 and 10 weeks. The animals were sacrificed 1 week after receiving the last treatment of MTX. The germ cell toxicity was evaluated using testes weight (wt), sperm count, sperm head morphology, sperm comet assay, histology, TUNEL and halo assay in testis. MTX treatment significantly reduced the sperm count and increased the occurrence of sperm head abnormalities in a dose dependent manner. It induced the testicular toxicity as evident from the histology of testis. Sperm comet, TUNEL and halo assay in testis also revealed significant DNA damage after MTX treatment. On the basis of the present study, it can be concluded that MTX induced germ cell toxicity in mice. PMID- 18678278 TI - The impact of enzyme engineering upon natural product glycodiversification. AB - Glycodiversification of natural products is an effective strategy for small molecule drug development. Recently, improved methods for chemo-enzymatic synthesis of glycosyl donors has spurred the characterization of natural product glycosyltransferases (GTs), revealing that the substrate specificity of many naturally occurring GTs as too stringent for use in glycodiversification. Protein engineering of natural product GTs has emerged as an attractive approach to overcome this limitation. This review highlights recent progress in the engineering/evolution of enzymes relevant to natural product glycodiversification with a particular focus upon GTs. PMID- 18678280 TI - The ultrastructure of the digestive cells of Argulus japonicus, Thiele 1900 (Crustacea: Branchiura). AB - The ultrastructure of the cells of the digestive system of Argulus japonicus is described with the use of transmission electron microscopy. Specimens of Argulus japonicus were collected from the Vaal Dam in South Africa and fixed in Todd's fixative. The samples were post fixed in osmium tetroxide and embedded in resin. The anterior midgut is composed mostly of R cells while the enteral diverticula are composed mainly of R cells in the proximal diverticules and of F cells in the distal diverticula. The posterior midgut is composed of very large papilliform B cells and of R cells. The R cells in the anterior midgut probably absorb nutrients including lipids. The F cells are filled mostly with rough endoplasmic reticulum, suggesting enzyme synthesis, while the B cells portrayed endocytotic vesicles, indicating intracellular digestion of predigested food. The R cells of the posterior midgut are less active than cells present in the anterior midgut. E cells and peritrophic membrane were not observed. PMID- 18678281 TI - Fascicle switching generates a chiasmal neuroarchitecture in the embryonic central body of the grasshopper Schistocerca gregaria. AB - The central body is a prominent neuropilar structure in the midbrain of the grasshopper and is characterized by a fan-shaped array of fiber columns, which are part of a chiasmal system linking anterior and posterior commissures. These columns are established during embryogenesis and comprise axons from cell clusters in the pars intercerebralis, which project to the central body via the so-called w, x, y, z tracts. Up to mid-embryogenesis the primary axon scaffold in both the brain and ventral nerve cord comprises a simple orthogonal arrangement of commissural and longitudinal fiber pathways. No chiasmata are present and this pattern is maintained during subsequent development of the ventral nerve cord. In the midbrain, individual axons entering the commissural system from each of the w, x, y, z tracts after mid-embryogenesis (55%) are seen to systematically de fasciculate from an anterior commissure and re-fasciculate with another more posterior commissure en route across the midline, a feature we call "fascicle switching". Since the w, x, y, z tracts are bilaterally symmetrical, fascicle switching generates chiasmata at stereotypic locations across the midbrain. Choice points for leaving and entering fascicles mark the anterior and posterior positions of each future column. As the midbrain neuropil expands, the anterior and posterior groups of commissures condense, so that the chiasmata spanning the widening gap between them become progressively more orthogonally oriented. A columnar neuroarchitecture resembling that of the adult central body is already apparent at 70% of embryogenesis. PMID- 18678282 TI - Targeted therapeutics for cancer treatment: major progress towards personalised molecular medicine. PMID- 18678283 TI - A single early-life seizure impairs short-term memory but does not alter spatial learning, recognition memory, or anxiety. AB - The impact of a single seizure on cognition remains controversial. We hypothesized that a single early-life seizure (sELS) on rat Postnatal Day (P) 7 would alter only hippocampus-dependent learning and memory in mature (P60) rats. The Morris water maze, the novel object and novel place recognition tasks, and contextual fear conditioning were used to assess learning and memory associated with hippocampus/prefrontal cortex, perirhinal/hippocampal cortex, and amygdala function, respectively. The elevated plus maze and open-field test were used to assess anxiety associated with the septum. We report that sELS impaired hippocampus-dependent short-term memory, but not spatial learning or recall. sELS did not disrupt performance in the novel object and novel place recognition tasks. Contextual fear conditioning performance suggested intact amydgala function. sELS did not change anxiety levels as measured by the elevated plus maze or open-field test. Our data suggest that the long-term cognitive impact of sELS is limited largely to the hippocampus/prefrontal cortex. PMID- 18678284 TI - Seven large variations in the extent of RNA editing in plant mitochondria between three ecotypes of Arabidopsis thaliana. AB - Most RNA editing sites in flowering plant mitochondria are located in coding regions of mRNAs and are usually essential for correct gene expression. Although accordingly little variation should be tolerated, editing sites appear and disappear even between closely related flowering plant species. To investigate whether such editing site variations also occur within species, we analyzed 379 RNA editing sites in the three ecotypes Columbia, Landsberg erecta and C24 of Arabidopsis thaliana. While all editing sites as such are conserved, we identify seven RNA editing sites with 40-60% differences in effective editing between individual ecotypes. These quantitative variations show that the extent of RNA editing in plant mitochondria is very flexible and can change even more rapidly than the evolution of species. The ecotype-specific variations of the RNA editing extent are Mendelian-inherited and can now be used to follow and identify the nuclear loci responsible for these RNA editing phenotypes. PMID- 18678285 TI - Expression of the TrkA or TrkB receptor tyrosine kinase alters the double-strand break (DSB) repair capacity of SY5Y neuroblastoma cells. AB - In the childhood tumor neuroblastoma, high expression of the TrkA neurotrophin receptor is associated with a favorable prognosis and a lack of structural chromosomal changes, whereas TrkB is expressed in aggressive neuroblastomas demonstrating high genomic instability. The ability to repair DNA double-strand breaks (DSBs) is considered a central determinant of chromosomal stability with nonhomologous end joining (NHEJ) being the major pathway of DSB repair in vertebrates. Here, we used the SH-SY5Y human neuroblastoma cell line ectopically expressing either TrkA or TrkB as a model system to analyze the impact of Trk receptor expression on NHEJ-mediated DSB repair. In a cell-free NHEJ assay, SY5Y TrkA cells displayed a significantly higher efficiency for NHEJ compared to SY5Y TrkB cells. To detect possible underlying mechanisms, gene expression data (Affymetrix U95A microarray chips) obtained from the same SY5Y-TrkA/TrkB model system were reanalyzed focussing on genes involved in DNA repair. Expression of XRCC4, a central component of NHEJ, was significantly upregulated in SY5Y-TrkA compared to SY5Y-TrkB cells. Expression data were confirmed using real-time PCR and western blotting. Additionally, XRCC4 expression was enhanced in most primary neuroblastomas with high TrkA expression. The TrkA-induced increase in NHEJ activity could be reverted by XRCC4 knock-down, confirming the induction of XRCC4 by TrkA to be essential for the observed phenotype. Our data provide the first evidence for a functional relationship between tyrosine kinase receptor signaling and NHEJ-mediated DSB repair in cancer cells, potentially contributing to their genomic stability. PMID- 18678286 TI - PI3K-Akt signaling regulates basal, but MAP-kinase signaling regulates radiation induced XRCC1 expression in human tumor cells in vitro. AB - As demonstrated recently, ionizing radiation (IR) can mediate phosphorylation of DNA-PKcs in human tumor cells through stimulation of the PI3K/Akt pathway. It is also known that DNA-PKcs directly interacts the X-ray repair cross-complementing group 1 protein (XRCC1) involved in base excision repair (BER). Therefore, in the present study we investigated the role of PI3K/Akt activity and DNA-PKcs on XRCC1 expression/stabilization. In contrast to the DNA-PKcs-deficient glioblastoma cell line MO59J, the DNA-PKcs-proficient counterpart MO59K as well as human lung adenocarcinoma A549 cells presented a high basal level of XRCC1 expression. Radiation doses of 3-12Gy did not stimulate a further enhanced expression of XRCC1 in DNA-PKcs-proficient cells (MO59K and A549) within 180min post irradiation. However, a marked induction of XRCC1 expression was apparent in DNA PKcs-deficient MO59J cells. Targeting of DNA-PKcs as well as PI3K/Akt pathway by specific kinase inhibitors and/or siRNA reduced basal XRCC1 expression in un irradiated DNA-PKcs-proficient cells to the level observed in DNA-PKcs-deficient cells. Reduction of basal expression of XRCC1 by XRCC1-siRNA, AKT-siRNA as well as DNA-PKcs inhibitor facilitated IR-induced XRCC1 expression. XRCC1 expression induced by irradiation, however, was independent of PI3K/Akt signaling, but dependent of MAPK-ERK1/2. By immuno-precipitation experiments and confocal microscopy a complex formation of XRCC1 and DNA-PKcs was shown. Applying gamma H2AX foci analysis it was shown that basal expression of XRCC1 is important for the repair of IR-induced DNA-double strand breaks (DNA-DSBs). These data indicate that IR-induced XRCC1 expression is dependent on the expression level of DNA-PKcs and basal activity status of PI3K/Akt signaling. Likewise, potential of IR induced XRCC1 expression depends on its basal expression level. PMID- 18678287 TI - Clinical and molecular characterization of 40 patients with Noonan syndrome. AB - Noonan syndrome (NS, OMIM 163950) is an autosomal dominant disorder, with a prevalence at birth of 1:1000-1:2500 live births, characterized by short stature, facial and skeletal dysmorphisms, cardiovascular defects and haematological anomalies. Missense mutations of PTPN11 gene account for approximately 50% of NS cases, while molecular lesions of other genes of the RAS/MAPK pathway -KRAS, SOS1 and RAF1 - play a minor role in the molecular pathogenesis of the disease. Forty patients were enrolled in the study with a PTPN11 mutation detection rate of 31.5%, including a novel missense mutation, Phe285Ile, in a familial case with high intrafamilial phenotypic variability. All patients negative for PTPN11 mutations were further screened for mutations of the KRAS, SOS1, and RAF1 genes, revealing a Thr266Lys substitution in SOS1 in a single patient, a newborn with a subtle phenotype, characterized by facial dysmorphisms and a mild pulmonic stenosis. PMID- 18678288 TI - Caititu: a tool to graphically represent peptide sequence coverage and domain distribution. AB - Here we present Caititu, an easy-to-use proteomics software to graphically represent peptide sequence coverage and domain distribution for different correlated samples (e.g. originated from 2D gel spots) relatively to the full sequence of the known protein they are related to. Although Caititu has a broad applicability, we exemplify its usefulness in Toxinology using snake venom as a model. For example, proteolytic processing may lead to inactivation or loss of domains. Therefore, our proposed graphic representation for peptides identified by two dimensional electrophoresis followed by mass spectrometric identification of excised spots can aid in inferring what kind of processing happened to the toxins, if any. Caititu is freely available to download at: http://pcarvalho.com/things/caititu. PMID- 18678289 TI - Polymorphisms in the estrogen receptor genes are associated with hip fractures in Chinese. AB - INTRODUCTION: Hip fractures (HF) are a major cause of public health burden with strong genetic determination. However, the true causal genes remain largely unknown. MATERIALS AND METHODS: Based on the important biological role of estrogens in bone homeostasis, this study aimed to investigate whether the estrogen receptor genes, ESR1 and ESR2, affect the onset of HF in 700 elderly Chinese subjects (350 with osteoporotic HF and 350 healthy controls). We genotyped 32 SNPs in total and examined their associations both by the single-SNP and haplotype tests. RESULTS: We identified two novel SNPs of ESR1, rs3020314 and rs1884051, were significantly associated with HF (rs3020314: P=0.0004, OR=1.66, 95%CI: 1.25-2.18; rs1884051: P=0.0004, OR=1.46, 95%CI: 1.19-1.81). We firstly detected significant association of ESR2 with HF (rs960070: P=0.0070, OR=1.43, 95%CI: 1.10-1.86). Haplotype analyses corroborated our single-SNP results. CONCLUSION: Our findings have important implications for understanding the pathology of osteoporotic fractures. Independent replication studies are needed to validate our results and explore the most possible functional variants for molecular studies. PMID- 18678290 TI - Usefulness of self-reported leisure-time physical activity to predict long-term survival in patients with coronary heart disease. AB - Self-reported leisure-time physical activity level correlates well with both cardiovascular (CV) and non-CV mortality in subjects without coronary heart disease (CHD). The impact of leisure-time physical activity on long-term outcomes has not been well studied in patients with preexisting CHD, who are often physically limited because of symptoms, medications, and co-morbid conditions. The aim was to determine the long-term prognostic value of self-reported leisure time physical activity in a large CHD cohort. Leisure-time physical activity was evaluated using a self-administered questionnaire and categorized using a 4-level scale (sedentary, mild, moderate, and strenuous) in 14,021 of 24,958 subjects from the Coronary Artery Surgery Study Registry with suspected or proven CHD who underwent cardiac catheterization from 1974 to 1979. Median long-term follow-up was 14.7 years (interquartile range 9.8 to 16.2). Clinical outcomes were evaluated according to physical activity level and adjusted for potential confounders. Long-term all-cause and CV mortality progressively increased from most to least active subjects, with sedentary patients showing a 1.6-fold increase in mortality for both these outcomes (hazard ratio [HR] 1.63, 95% confidence interval [CI] 1.34 to 1.97, p <0.0001 for all-cause mortality). Similar trends were noted for men and women and in adjusted models, although HRs were attenuated after adjusting for age, gender, smoking, hypertension, diabetes mellitus, total cholesterol, body mass index, and ejection fraction (adjusted HR 1.23, 95% CI 1.01 to 1.49, p = 0.03 for all-cause mortality; adjusted HR 1.25, 95% CI 0.99 to 1.57, p = 0.05 for CV mortality). In conclusion, leisure-time physical activity independently predicted long-term survival in men and women with chronic stable CHD. PMID- 18678291 TI - Relation of epicardial and pericoronary fat to coronary atherosclerosis and coronary artery calcium in patients undergoing coronary angiography. AB - Fat surrounding coronary arteries might aggravate coronary artery disease (CAD). We investigated the relation between epicardial adipose tissue (EAT) and pericoronary fat and coronary atherosclerosis and coronary artery calcium (CAC) in patients with suspected CAD and whether this relation is modified by total body weight. This was a cross-sectional study of 128 patients with angina pectoris (61 +/- 6 years of age) undergoing coronary angiography. EAT volume and pericoronary fat thickness were measured with cardiac computed tomography. Severity of coronary atherosclerosis was assessed by the number of stenotic (> or =50%) coronary vessels; extent of CAC was determined by the Agatston score. Patients were stratified for median total body weight (body mass index [BMI] 27 kg/m(2)). Overall, EAT and pericoronary fat were not associated with severity of coronary atherosclerosis and extent of CAC. In patients with low BMI, those with multivessel disease had increased EAT volume (100 vs 67 cm(3), p = 0.04) and pericoronary fat thickness (9.8 vs 8.4 mm, p = 0.06) compared with those without CAD. Also, patients with severe CAC had increased EAT volume (108.0 vs 69 cm(3), p = 0.02) and pericoronary fat thickness (10.0 vs 8.2 mm, p value = 0.01) compared with those with minimal/absent CAC. In conclusion, EAT and pericoronary fat were not associated with severity of coronary atherosclerosis and CAC in patients with suspected CAD. However, in those with low BMI, increased EAT and pericoronary fat were related to more severe coronary atherosclerosis and CAC. Fat surrounding coronary arteries may be involved in the process of coronary atherosclerosis, although this is different for patients with low and high BMIs. PMID- 18678292 TI - Safety of diagnostic coronary angiography during uninterrupted therapeutic warfarin treatment. AB - Long-term warfarin therapy is assumed to increase bleeding and access site complications after coronary angiography and it is often recommended to postpone invasive procedures to reach international normalized ratio (INR) levels <1.8. To assess the safety and feasibility of diagnostic coronary angiography during uninterrupted warfarin therapy, we retrospectively analyzed all consecutive patients (n = 258) on warfarin therapy referred for diagnostic coronary angiography in 2 centers with long experience in uninterrupted warfarin therapy during coronary angiography and in 1 center with a policy of preprocedural warfarin pause. An age- and gender-matched control group (n = 258) with similar disease presentation (unstable or stable symptoms) was collected from each center. Radial access was used in 56% of patients in the warfarin group and in 60% of controls (p = 0.21). There was no difference in access site and bleeding complications (1.9% vs 1.6%) or major adverse cardiovascular and cerebrovascular events (0.4% vs 0.8%) between the warfarin group and their controls. Warfarin was interrupted in 80 patients (31%), and bridging therapy was used in 24 of these patients (30%). INR levels were higher in the uninterrupted warfarin group (2.3 vs 1.9, p <0.001), but the incidence of access site complications was not higher (1.7%) than in patients (n = 80) with a warfarin pause (2.5%) or in patients with pause and bridging therapy (8.3%). Need for blood transfusions (n = 2) occurred only in patients with bridging therapy. Access site complications were more common in the 22 patients with supratherapeutic anticoagulation (INR >3) than in patients with therapeutic periprocedural INR (9.1% vs 1.5%, p <0.05). In conclusion, a simple strategy of performing coronary angiography during uninterrupted therapeutic warfarin anticoagulation is a tempting alternative to bridging therapy and is likely to lead to considerable cost savings. PMID- 18678293 TI - Assessment of culprit and remote coronary narrowings using optical coherence tomography with long-term outcomes. AB - Much currently known information about vulnerable plaque stems from postmortem studies that identified several characteristics making them prone to rupture, including the presence of a thin fibrous cap and a large lipid core. This study used optical coherence tomography (OCT) to assess culprit and remote coronary narrowings and investigate whether intracoronary OCT in living patients was able to visualize morphologic features associated with vulnerable plaque in postmortem studies. Twenty-three patients successfully underwent OCT before percutaneous coronary intervention. The culprit lesion and mild to moderate coronary narrowings remote from the target stenosis were investigated. Using OCT, the culprit lesion was found to be fibrous in 39.1%, fibrocalcific in 34.4%, and lipid rich in 26.1% of cases. Two patients met criteria for thin-cap fibroatheroma (TCFA; defined as the presence of a signal-rich fibrous cap covering a signal-poor lipid/necrotic core with cap thickness <0.2 mm). Most plaques at remote segments were proximal to the culprit lesion (73.9%) and predominantly fibrous and lipid rich. OCT identified 7 TCFA lesions in 6 patients with a mean cap thickness of 0.19 +/- 0.05 mm, extending for 103 degrees +/- 49 degrees of the total vessel circumference. At 24 months of clinical follow-up, the only event occurred in a patient with in-stent restenosis who underwent repeated percutaneous revascularization. There were no clinically apparent plaque rupture-related events in the 6 patients found to have remote TCFA. This study showed that OCT can be safely applied to image beyond the culprit lesion and can detect in vivo morphologic features associated with plaque vulnerability using retrospective pathologic examination. In conclusion, detection of TCFA, particularly in stable patients, is desirable and may principally allow for early intervention and prevention of adverse events. PMID- 18678294 TI - Comparison of increased aspirin dose versus combined aspirin plus clopidogrel therapy in patients with diabetes mellitus and coronary heart disease and impaired antiplatelet response to low-dose aspirin. AB - The effects of therapy with aspirin 300 mg/day and with combined aspirin 100 mg/day plus clopidogrel 75 mg/day on platelet function were compared in patients with diabetes mellitus and coronary artery disease and impaired antiplatelet responses to aspirin 100 mg/day. The study population consisted of 151 outpatients with type II diabetes mellitus and coronary artery disease who were taking aspirin 100 mg/day. Of the 151 patients, a subgroup of subjects with impaired aspirin response were selected on the basis of the results of platelet aggregometry. Nonresponsiveness to aspirin was defined as mean aggregation > or =69% with 3 micromol/L adenosine diphosphate and mean aggregation > or =70% with 2 micromol/L collagen. Aspirin semiresponders were defined as meeting 1 but not both of these criteria. Nonresponders and semiresponders were randomized equally to aspirin 300 mg/day and aspirin 100 mg/day plus clopidogrel 75 mg/day, and aggregation tests were repeated after 2 weeks. Sixty of the 151 patients with diabetes (40%) were found to respond to aspirin inadequately. Platelet aggregation induced by adenosine diphosphate and collagen decreased significantly after aspirin 300 mg/day or combined therapy. Combined treatment was found to have a stronger inhibitory effect on platelet aggregation induced by adenosine diphosphate than aspirin 300 mg/day (p = 0.002). Impaired aspirin response was resolved by increasing the aspirin dose or adding clopidogrel to aspirin (p <0.0001 for each). However, desired platelet inhibition was achieved in significantly more patients by combined treatment than by aspirin 300 mg/day (p <0.05). In conclusion, aspirin 100 mg/day does not inhibit platelet function adequately in a significant number of patients with diabetes mellitus and coronary artery disease. Increasing the aspirin dose to 300 mg/day or adding clopidogrel to aspirin can provide adequate platelet inhibition in a significant number of those patients with impaired responses to low-dose aspirin. PMID- 18678295 TI - Comparison of higher clopidogrel loading and maintenance dose to standard dose on platelet function and outcomes after percutaneous coronary intervention using drug-eluting stents. AB - Adequate antiplatelet therapy is paramount for good clinical outcomes in patients undergoing percutaneous coronary intervention (PCI). The purpose of this study was to determine whether a high-dose regimen of clopidogrel in patients undergoing PCI is superior to standard dosing. A total of 119 patients undergoing PCI were blindly randomized in 2:1 fashion to receive clopidogrel loading 600 mg on the table immediately before PCI and 75 mg 2 times/day for 1 month (high-dose group) versus standard dosing (300 mg loading and 75 mg/day; low-dose group). Platelet aggregation was measured using light transmission aggregometry at baseline, 4 hours, and 30 days. The composite of cardiovascular death, myocardial infarction, and target vessel revascularization was studied at 30 days in addition to major and minor bleeding. Baseline characteristics and baseline platelet aggregation were similar in the 2 groups. Percent inhibitions of platelet activity were 41% and 27% in the high-dose group versus 19% and 10% in the low-dose group at 4 hours and 30 days (p = 0.046 and 0.047, respectively). Composite clinical end points were 10.3% in the high-dose group and 23.8% in the low-dose group (p = 0.04). No difference was noted in major or minor bleeding. In conclusion, a higher loading and maintenance dose of clopidogrel in patients undergoing PCI results in superior platelet inhibition and decreased cardiovascular events without increasing bleeding complications. PMID- 18678296 TI - Long-term outcomes after percutaneous coronary intervention of bifurcation narrowings. AB - The optimal approach to percutaneous coronary intervention (PCI) of bifurcation lesions remains unclear, reflecting lack of long-term follow-up and heterogeneity of lesions encountered. We evaluated the long-term outcome of patients undergoing bifurcation PCI followed in the prospective bifurcation registry at the University Health Network, Toronto, Ontario, Canada. Of 526 patients undergoing bifurcation PCI between November 2003 and March 2005, most (n = 406) were treated by main vessel stenting only (n = 266) or crush/culotte stenting (n = 140). After median follow-up of 26.5 months, major adverse cardiac events (MACEs) and Canadian Cardiovascular Society class > or =2 angina occurred in 28.5% and 22.3% of patients in these groups, respectively (p = 0.190), whereas MACE rates were 20.8% for main vessel stenting and 18.7% for crush/culotte stenting (p = 0.670). A low bifurcation angle was associated with better outcomes in the crush/culotte group but had no effect on outcome of patients treated with main vessel stenting only. Use of crush/culotte techniques independently predicted freedom from MACEs or Canadian Cardiovascular Society class > or =2 angina compared with main vessel stenting only (odds ratio 0.55, 95% confidence interval 0.32 to 0.94, p = 0.029). In conclusion, the use of crush/culotte stenting is safe, with efficacy and MACE rates being similar to main vessel stenting alone. Our observations regarding the effect of lesion characteristics such as bifurcation angle and extent of side branch disease on outcome underscore the need for randomized trials that are inclusive of patients with complex side branch disease. PMID- 18678297 TI - Impact of early and delayed stroke on in-hospital and long-term mortality after isolated coronary artery bypass grafting. AB - Stroke after coronary artery bypass grafting (CABG) is an infrequent, yet devastating complication with increased morbidity and mortality. We sought to determine risk factors for early (intraoperatively to 24 hours) and delayed (>24 hours to discharge) stroke and to identify their impact on long-term mortality after CABG. We studied 4,140 consecutive patients who underwent isolated CABG from 1992 to 2003. Long-term survival data (mean follow-up 7.4 years) were obtained from the National Death Index. Independent predictors for stroke and in hospital mortality were determined by multivariate logistic regression analysis including all available preoperative, intraoperative, and postoperative risk factors. Independent predictors for long-term mortality were determined by multivariate Cox regression analysis. One hundred two patients (2.5%) developed early stroke and 36 patients (0.9%) delayed stroke. Independent predictors for early stroke were age, recent myocardial infarction, smoking, femoral vascular disease, body mass index, reoperation for bleeding, postoperative sepsis and/or endocarditis, and respiratory failure, whereas those for delayed stroke were female gender, white race, preoperative renal failure, respiratory failure, and postoperative renal failure. Early stroke was an independent predictor for in hospital (odds ratio 3.49, 95% confidence interval [CI] 1.56 to 7.80, p = 0.002) and long-term (hazard ratio 1.70, 95% CI 1.30 to 2.21, p <0.001) mortalities. Delayed stroke was not an independent predictor for in-hospital (odds ratio 0.90, 95% CI 0.23 to 3.51, p = 0.878) or long-term (hazard ratio 0.66, 95% CI 0.38 to 1.17, p = 0.156) mortality. In conclusion, risk factors for early in-hospital stroke differ from those of delayed in-hospital stroke after CABG. Early stroke is an independent predictor for in-hospital and long-term mortalities, suggesting the need for a more frequent follow-up and appropriate pharmacologic therapy after discharge. PMID- 18678298 TI - The Editor's Roundtable: Endothelial dysfunction in cardiovascular disease. PMID- 18678299 TI - Comparison of serum lipid values in subjects with and without the metabolic syndrome. AB - Insulin resistance is supposed to be the basis of metabolic syndrome (MS), although it is difficult to measure. The ratio of triglyceride (TG) to high density lipoprotein (HDL) has been proposed as a surrogate marker of insulin resistance in overweight subjects. The aim of the present study was to assess the accuracy of the TG/HDL ratio for the diagnosis of MS. Data of 18,778 active workers (77.6% men) enrolled in 3 insurance companies in Spain were collected from their annual health examinations. Mean age was 42.2 +/- 10.7 years. MS was assessed according to modified Adult Treatment Panel III criteria. Prevalences of MS were 18.8% in men and 6.1% in women. Mean value of the TG/HDL ratio was 2.50 +/- 2.2 and increased in parallel to the number of MS components present. Subjects with MS had a ratio that was 2 times higher compared with those without (5.10 vs 2.03, p <0.001). Receiver operating characteristic curves were performed to assess the capability of the TG/HDL ratio to contribute to a diagnosis of MS and 80% sensitivity and 78% specificity were obtained for values >2.75 in men and >1.65 in women. In conclusion, the TG/HDL ratio is a feasible and accurate measurement for assessment of MS in healthy subjects. We propose cut-off values of 2.75 for men and 1.65 for women for a diagnosis of MS. PMID- 18678300 TI - Effects of adding prescription omega-3 acid ethyl esters to simvastatin (20 mg/day) on lipids and lipoprotein particles in men and women with mixed dyslipidemia. AB - Prescription omega-3 acid ethyl esters (P-OM3) are commonly used for treatment of very high triglyceride levels, often in combination with a statin, to lower persistent hypertriglyceridemia. This randomized, crossover trial evaluated 6 weeks of combination therapy with simvastatin 20 mg/day plus P-OM3 4 g/day or placebo in 39 men and women (average age 58 years) with a triglyceride concentration 200 to 600 mg/dl and non-high-density lipoprotein (non-HDL) cholesterol greater than their National Cholesterol Education Program treatment goals after a 5-week diet lead-in. Non-HDL cholesterol decreased from baseline (209 mg/dl) by 40% for P-OM3 + simvastatin compared with 34% for placebo + simvastatin (p <0.001). Favorable changes for P-OM3 + simvastatin versus placebo + simvastatin were also observed for very low-density lipoprotein (VLDL) cholesterol (-42% vs -22%), triglyceride (-44% vs -29%), total cholesterol (-31% vs -26%), HDL cholesterol (+16% vs +11%), apolipoprotein B (-32% vs -28%), total cholesterol:HDL cholesterol ratio (-39% vs -33%), triglyceride:HDL cholesterol ratio (-51% vs -37%), and systolic (-5.0 vs 0.3 mm Hg) and diastolic (-3.3 vs 1.8 mm Hg) blood pressures (p <0.05 for all). VLDL particle concentration and size decreased and LDL particle size increased significantly more with P-OM3 + simvastatin than with placebo + simvastatin (all p <0.05). Changes in LDL cholesterol, LDL particle concentration, HDL particle size and concentration, and apolipoprotein A-I did not differ significantly between treatments. In conclusion, P-OM3 + simvastatin appears to be a useful therapeutic option for the management of mixed dyslipidemia. PMID- 18678301 TI - Efficacy and safety of a potent and selective peroxisome proliferator activated receptor alpha agonist in subjects with dyslipidemia and type 2 diabetes mellitus. AB - The weak peroxisome proliferator activated receptor-alpha (PPAR-alpha) agonists gemfibrozil and fenofibrate achieve only small increases in high-density lipoprotein (HDL) cholesterol. CP-778,875 is a more potent PPAR-alpha agonist developed to produce greater HDL cholesterol increases. This randomized, multicenter, double-blinded, placebo-controlled study evaluated the efficacy and safety of CP-778,875 in subjects with mixed dyslipidemia and type 2 diabetes. Eight-six subjects with low HDL cholesterol (< or =45 mg/dl for men and < or =55 mg/dl for women) and increased triglycerides (150 to 500 mg/dl) who had coexisting type 2 diabetes were randomized. Subjects received CP-778,875 doses of 0.5, 2, or 6 mg/day or placebo for 6 weeks. Any other lipid-altering therapy was stopped at screening. The primary end point was percent change in HDL cholesterol from baseline. The 2-mg/day dose of CP-778,875 significantly increased HDL cholesterol by 14%. The 2-mg dose also increased concentrations of apolipoprotein (apo) A-I, HDL(2) cholesterol, and HDL(3) cholesterol by 13%, 12%, and 19%, respectively. An unusual dose-response pattern was observed in that at 6 mg/day CP-778,875 only increased HDL cholesterol by 3% and decreased HDL(2) cholesterol by 24%. Fasting triglyceride levels were significantly decreased to a similar extent (26%) by all 3 doses of CP-778,875. CP-778,875 significantly increased homocysteine levels. There was no significant relation between change in homocysteine and change in apoA-I or HDL cholesterol. No subjects developed myopathy. In conclusion, CP-778,875 2 mg/day significantly increased HDL cholesterol, significantly lowered fasting triglycerides, and increased apoA-I and HDL subfractions. The clinical relevance of the increase in homocysteine levels is unknown. PMID- 18678302 TI - Indexes of kidney function and coronary artery and abdominal aortic calcium (from the Framingham Offspring Study). AB - It is uncertain whether moderate chronic kidney disease (CKD) or measures of kidney function are associated with subclinical atherosclerosis as represented by coronary artery calcium (CAC) or abdominal aortic calcium (AAC). We used logistic and linear regression analyses to relate CKD (glomerular filtration rate <60 ml/min/1.73 m(2)), cystatin C (cysC), and microalbuminuria (MA) with CAC and AAC obtained using multidetector computed tomography in Framingham Heart Study Offspring participants (mean age 59 years, 55.3% women). Increased CAC and AAC were defined as > or =90th percentile age- and gender-specific cutpoints based on a healthy referent sample. Major cardiovascular disease risk factors were accounted for in multivariable models. Of 1,179 participants, 1,174 had AAC measurements and 1,147 had CAC measurements, 6.3% had CKD, and 8.3% had MA. CKD was not associated with CAC (multivariable-adjusted odds ratio [OR] for CKD 1.18, 95% confidence interval 0.59 to 2.36, p = 0.63) or AAC (multivariable-adjusted OR for CKD 1.11, 95% confidence interval 0.61 to 2.04, p = 0.73). CysC was associated with CAC in age- and gender-adjusted but not in multivariable models (age- and gender-adjusted OR for log cysC per SD increment and CAC 1.19, 95% confidence interval 1.01 to 1.41, p = 0.04; multivariable-adjusted OR 1.14, 95% confidence interval 0.95 to 1.38, p = 0.15). MA was not associated with CAC (OR 0.81, 95% confidence interval 0.41 to 1.61, p = 0.54). Neither cysC nor MA was significantly associated with AAC in age- and gender- or multivariable-adjusted models. In conclusion, CKD, cysC, and MA are not associated with CAC or AAC when accounting for cardiovascular disease risk factors. PMID- 18678303 TI - Comparison of benefits and mortality in cardiac resynchronization therapy in patients with atrial fibrillation versus patients in sinus rhythm (Results of the Spanish Atrial Fibrillation and Resynchronization [SPARE] Study). AB - The efficacy of cardiac resynchronization therapy (CRT) in patients with atrial fibrillation (AF) and the need for atrioventricular junction ablation in these patients is controversial. The aim of the study was to analyze CRT results in patients with permanent AF. A total of 470 consecutive patients who underwent CRT in 6 centers were included in this study. Of these patients, 126 (27%) had permanent AF. Patients were evaluated at baseline and 12 months. No difference was found in the magnitude of improvement experienced by patients with AF compared with those in sinus rhythm (SR) with respect to quality of life, distance in 6-minute walking test, and left ventricular reverse remodeling. Despite the beneficial effects of CRT, death from refractory heart failure at 12 months was higher in patients with AF (17 of 126; 13.5%) than those in SR (14/344; 4.1%; p <0,001). Furthermore, permanent AF was an independent predictive factor for mortality from refractory heart failure (hazard ratio 5.4, 95% confidence interval 1.9 to 15.1). In conclusion, patients with AF treated with CRT who survived at the 12-month follow-up had the same functional improvement and remodeling as those in SR. However, AF was an independent risk factor for mortality from heart failure after CRT implantation. PMID- 18678304 TI - Frequency, duration, and predictors of newly-diagnosed atrial fibrillation following dual-chamber pacemaker implantation in patients without a previous history of atrial fibrillation. AB - The Silent Atrial Fibrillation Detection With Stored EGMs (SAFE) registry is the first prospective study to characterize the incidence, duration, and predictors of atrial high-rate episodes (AHREs) in patients without previous clinically diagnosed atrial fibrillation after dual-chamber pacemaker implantation. Patients underwent in-clinic device interrogation at 2 weeks and then 3, 6, and 12 months after implantation. An AHRE was defined as an atrial tachyarrhythmia with an atrial rate > or =180 beats/min lasting > or =5 minutes. During the first month after pacemaker implantation, AHREs were identified in 68 patients (4.6%). Thirty four of these patients (50%) did not have AHREs beyond the first month after implantation; these patients were older than those who continued to have AHREs (79 +/- 9 vs 74 +/- 12 years, p = 0.05). Within the first 6 months after pacemaker implantation, 150 patients (10%) experienced a total of 455 AHREs, of which only 28 (6%) prompted clinical visits. The 6-month freedom from an AHRE was 82%. Overall, 75% of patients with AHREs had multiple episodes; most AHREs lasted 5 to 60 minutes. In conclusion, 6 months after dual-chamber pacemaker implantation, 10% of patients experienced > or =1 AHRE, usually lasting 5 to 60 minutes. Some patients had AHREs only within the first 30 days after implantation, suggesting the possibility of transient atrial proarrhythmia related to lead insertion. PMID- 18678305 TI - Gender differences in patients admitted with advanced decompensated heart failure. AB - Broad population studies of patients with stable ambulatory heart failure have associated female gender with better age-adjusted survival. This study investigated whether there are gender-specific differences in clinical presentation, response to intensive medical therapy, and outcomes in patients admitted with advanced (cardiac index <2.4 L/min/m(2)) decompensated heart failure (ADHF). We reviewed 278 consecutive patients (age 54 +/- 12 years, cardiac index 1.7 +/- 0.4 L/kg/m(2), pulmonary capillary wedge pressure 26 +/- 9 mm Hg, serum creatinine 1.4 +/- 0.8 mg/dl) with ADHF treated with intensive medical therapy guided by pulmonary artery catheter in a dedicated heart failure intensive care unit from 2000 to 2006. Compared with men (n = 226), women (n = 52) had similar baseline characteristics with the exception of a higher prevalence of nonischemic cause. No differences in medical therapy on admission, during intensive medical therapy, or at discharge were observed. Intensive medical therapy was associated with significant hemodynamic improvement independent of gender. All-cause mortality and heart failure rehospitalization rates were similar between genders. However, adjusted for cause, women with ischemic cardiomyopathy had higher all-cause mortality rates (50% vs 37%, hazard ratio 1.95, 95% confidence interval 0.98 to 3.90, p = 0.05) and those with nonischemic cardiomyopathy had lower all-cause mortality rates (19% vs 40%, hazard ratio 0.40, 95% confidence interval 0.17 to 0.96, p = 0.01) than men. In conclusion, women presenting with ADHF had baseline characteristics and response to therapy similar to men. Overall outcomes were similar between men and women, although subgroup analysis suggested better survival for women with a nonischemic cause. PMID- 18678306 TI - Patterns of structural and functional remodeling of the left ventricle in chronic heart failure. AB - Patients with heart failure show a wide variety of alterations in left ventricular (LV) volume, mass, and function. The purpose of this study was to define the common patterns of LV structural and functional remodeling and consider their clinical implications in patients with chronic heart failure. Two dimensional echocardiograms obtained during the screening phase of a study involving patients (n = 315) with chronic heart failure were used to calculate LV volume, mass, geometry, and ejection fraction (EF). Inclusion required the diagnosis of heart failure in symptomatic patients on medical therapy. Measures of LV size or function were not used as inclusion or exclusion criteria. Plots of EF against LV end-diastolic volume (EDV) showing an inverse curvilinear relation allowed a description of 4 remodeling patterns. Pattern A (n = 66) was defined as normal EDV (<91 ml/m(2)) and normal EF (> or =50%); 65% of these patients showed LV hypertrophy or concentric remodeling. Pattern B (n = 65) was defined as normal EDV and depressed EF; hypertrophy or concentric remodeling was present in 63%. Pattern C (n = 175) was defined as increased EDV and depressed EF; eccentric hypertrophy was present in 94%. Pattern D (n = 9) was defined as increased EDV and normal EF; eccentric hypertrophy was present in 88%. In conclusion, these patterns of remodeling encompass a wide spectrum of geometric changes with different clinical and pathophysiologic features and possibly different management strategies. PMID- 18678307 TI - Prognostic value of N-terminal pro-type-B natriuretic peptide and Doppler left ventricular diastolic variables in patients with chronic systolic heart failure stabilized by therapy. AB - Prognostication of patients with chronic heart failure (HF) stabilized by therapy may be difficult. Therefore, the aim was to evaluate whether combined assessment of plasma N-terminal pro-B natriuretic peptide (NT-pro-BNP) and Doppler left ventricular (LV) diastolic variables was relevant to the prognosis of patients with stable HF. Outpatients with LV systolic HF (ejection fraction < or =45%), classified using clinical criteria as decompensated (n = 94) and stable HF (n = 219), underwent a complete Doppler echocardiographic study. NT-pro-BNP was measured together with mitral wave velocities, E wave deceleration time, and tissue Doppler early septal annular velocity. Median follow-up was 22 months. Freedom from all-cause mortality or HF hospitalization at 24 months was worst (44%) in patients with decompensated HF, intermediate (58%) in patients with stable HF with NT-pro-BNP higher than the median (>1,129 pg/ml), and best (92%) in patients with lower NT-pro-BNP (log-rank p <0.0001). In patients with stable HF, NT-pro-BNP >1,129 pg/ml (hazard ratio [HR] 2.84, p = 0.003), E wave deceleration time <150 ms (HR 2.31, p = 0.004), and tissue Doppler early septal annular velocity <8 cm/s (HR 2.18, p = 0.01) were predictors of the end point at multivariate analysis. The addition of Doppler LV diastolic variables and NT-pro BNP significantly improved the chi-square test for outcome prediction (from 14.4 to 46.4). In conclusion, NT-pro-BNP and spectral and tissue Doppler variables of LV diastolic dysfunction added independent and incremental contributions to prognostic stratification of patients with stable HF. PMID- 18678308 TI - Comparison of the prognostic usefulness of N-terminal pro-brain natriuretic Peptide in patients with heart failure with versus without chronic kidney disease. AB - In patients with chronic heart failure (CHF), N-terminal pro-brain natriuretic peptide (NT-pro-BNP) predicted poor outcome. Clinical predictors of NT-pro-BNP and its usefulness in the presence of chronic kidney disease (CKD) are largely unknown. A total of 341 patients with stable CHF were enrolled, of whom 183 (54%) had CKD. During a follow-up of 620 +/- 353 days, 57 patients (17%) experienced a cardiac event (cardiac death, need for extracorporeal assist device, or urgent cardiac transplantation), and 64 patients (20%) were rehospitalized because of worsening CHF. NT-pro-BNP was related to New York Heart Association functional class (R = 0.44, p <0.001) and inversely related to ejection fraction (R = -0.52, p <0.001) and glomerular filtration rate (R = -0.32, p <0.001). A cardiac event was independently predicted by NT-pro-BNP (hazard ratio [HR] 1.56, p <0.001), ejection fraction (HR 0.95, p = 0.018), and serum sodium (HR 0.89, p = 0.004). Using receiver-operator characteristic analysis, NT-pro-BNP > or =1,474 pg/ml best separated patients with or without cardiac events. In patients without CKD, outcome was significantly worse in patients with NT-pro-BNP >1,474 pg/ml in comparison to patients with NT-pro-BNP <1,474 pg/ml (event-free survival rate 0% vs 75%; p <0.001). In patients with CKD, outcome was also significantly worse in subjects with NT-pro-BNP >1,474 pg/ml in comparison to those with NT-pro-BNP <1,474 pg/ml (event-free survival rate 48% vs 93%; p <0.001). NT-pro-BNP independently predicted rehospitalization caused by worsening CHF (HR 1.26, p = 0.023), and a cut-off value of 1,474 pg/ml also separated patients with poor and intermediate prognosis in the CKD and non-CKD groups. In conclusion, NT-pro-BNP independently predicted morbidity and mortality in patients with CHF with and without CKD. PMID- 18678309 TI - Accuracy of the flow convergence method for quantification of aortic regurgitation in patients with central versus eccentric jets. AB - Proximal isovelocity surface area (PISA) has been proposed as a quantitative method to assess the severity of aortic regurgitation (AR). Yet the accuracy of this method in patients with eccentric AR jets is unknown. The aims of this study were to compare the accuracy of the PISA method for the quantification of AR severity in patients with central versus eccentric AR jets and to verify whether imaging from the left parasternal instead of the apical window improves the accuracy of the PISA method in patients with eccentric jets. Fifty patients with AR (21 with central jets and 29 with eccentric jets) underwent PISA and phase contrast cardiac magnetic resonance (CMR) measurements of AR volume. In patients with eccentric AR jets, PISA measurements obtained from the left parasternal and apical windows were compared. In patients with central AR jets, CMR- and PISA derived AR volumes were similar (28 +/- 19 vs 30 +/- 20 ml, p = 0.34), were strongly correlated (r = 0.92, p <0.0001), and differed minimally from each other (by 2 +/- 8 ml). In patients with eccentric AR jets, PISA-derived AR volumes underestimated those measured by CMR (38 +/- 22 vs 51 +/- 27 ml, bias -13 +/- 20 ml) and were correlated only fairly (r = 0.69, p <0.001). Imaging from the left parasternal window eliminated the differences between CMR- and PISA-derived AR volumes (51 +/- 27 vs 53 +/- 26 ml, p = 0.24) and improved the correlation between the 2 measures (r = 0.95). In conclusion, in patients with eccentric AR jets imaged from the apical window, the PISA method significantly underestimated AR severity. This was no longer the case when imaging was performed from the left parasternal instead of the apical window. PMID- 18678310 TI - Exercise-induced changes of functional mitral regurgitation in asymptomatic or mildly symptomatic patients with idiopathic dilated cardiomyopathy. AB - It has remained unclear why functional mitral regurgitation (MR), even if it is of a mild degree, has prognostic importance in patients with idiopathic dilated cardiomyopathy (IDC). Exercise-induced changes in functional MR, which might be a clue to this question, have not been fully clarified. Thus, in this study, semisupine exercise echocardiography was performed on 32 asymptomatic or mildly symptomatic patients with IDC (29 men, mean age 45 +/- 14 years). The mean ejection fraction was 28 +/- 10% (range 13% to 45%). The effective regurgitant orifice (ERO) area of MR was measured, as well as echocardiographic parameters including mitral valve geometry. ERO at rest was associated best with systolic mitral tenting area (r(S) = 0.85, p <0.001). Functional MR did not newly appear during exercise in 9 subjects without MR at rest. In the remaining 23 subjects with functional MR at rest, all showed exacerbations of MR, with a median ERO of 10.5 mm(2) (interquartile range 6.3 to 16.5) to 18.7 mm(2) (interquartile range 9.5 to 29.3) (p <0.001). An increase in ERO was correlated best with the enlargement of tenting area (r(S) = 0.90, p <0.001) and was the strongest independent determinant of exercise duration (beta = -0.55, p = 0.002, multiple R(2) = 0.46). In conclusion, functional MR complicated with IDC was significantly exacerbated during exercise, with mitral valve deformation, which was strongly related to exercise intolerance; thus, the clinical impact of functional MR in patients with IDC could be more serious than can be expected by its degree at rest. PMID- 18678311 TI - Comparison and results of transcatheter closure of patent ductus arteriosus using the swivel-disk device versus plug occluder in children. AB - The transcatheter closure of certain types of patent ductus arteriosus (PDA) remains a challenge. The investigators report initial clinical experience with 25 patients who underwent PDA occlusion with the use of a swivel-disk device (SDD) or a plug occluder (PO). The patients were divided into 2 groups: in the SDD group, 12 patients aged 1 to 2 years with type A PDA underwent attempted closure using the SDD, a modified Amplatzer duct occluder with a very low profile retention disk that can adapt itself at different PDA insertion angles. The mean PDA diameter was 4.8 +/- 1.5 mm (range 3.8 to 8). In the PO group, 13 patients aged 0.5 to 3 years with type C, D, or E PDA underwent attempted occlusion with the PO, a tubular occluder made of Nitinol wire mesh. The mean PDA diameter was 4.2 +/- 3.5 mm (range 1.2 to 9). The 2 occluders are filled with Dacron patches. The mean device diameters were 6.5 +/- 1.2 and 6.8 +/- 2.2 mm (range 4 to 11) in the SDD and PO groups, respectively. Complete angiographic closure was seen in 24 of 25 patients. The deployment of 4 5-PDA5 coils abolished a residual shunt associated with hemolysis in a 5-month-old patient with a large type C PDA after the implantation of an 11-mm PO. No other complications were observed. In conclusion, the SDD and the PO are promising additions to the armamentarium for PDA closure. PMID- 18678313 TI - Endothelial dysfunction in impaired fasting glycemia, impaired glucose tolerance, and type 2 diabetes mellitus. AB - The aim of this study was to evaluate whether abnormal endothelial function is present in early stages of diabetes, such as impaired glucose tolerance (IGT) and impaired fasting glucose (IFG). Endothelial function was assessed by measuring flow-mediated dilatation and nitrate-induced dilatation of the brachial artery using high-resolution ultrasound. Fasting serum lipid levels were determined, and glucose and insulin values in response to a 75-g oral glucose load were also measured. The results showed the following new findings: (1) compared with subjects with normal glucose tolerance, those with IFG and IGT had impaired flow mediated dilatation, more remarkable in subjects with type 2 diabetes mellitus than those with IFG and IGT, and (2) flow-mediated dilatation was inversely and strongly related to the extent of hyperglycemia. In conclusion, endothelial dysfunction is present in subjects with IGT and IFG, indicating endothelial damage in these stages. PMID- 18678312 TI - Relation of aortic wall thickness and distensibility to cardiovascular risk factors (from the Multi-Ethnic Study of Atherosclerosis [MESA]). AB - To determine the relation between aortic wall thickness (WT) and aortic distensibility (AD) with traditional cardiovascular risk factors in the Multi Ethnic Study of Atherosclerosis (MESA) cohort, 1,053 participants in MESA who underwent cardiac magnetic resonance imaging were consecutively selected for the measurement of aortic WT and AD. Double inversion-recovery fast spin-echo images of the thoracic aorta were obtained to measure average and maximum WT. AD was measured at the same level using a gradient-echo cine sequence. Average and maximum WT were positively correlated with increasing age, and AD was inversely related to age (p <0.01). Compared with normotensive participants, those with hypertension had significantly greater mean average WT (2.45 vs 2.23 mm, p <0.01) and maximum WT (3.61 vs 3.41 mm, p <0.01) and lower AD (0.15 vs 0.2 mm Hg(-1), p <0.01). In multiple regression analysis, older age and hypertension were significantly associated with higher mean average WT, while older age, male gender, and higher blood pressure were associated with higher mean maximum WT. AD was inversely related to older age, hypertension, current smoking, African American ethnicity, and lower high-density lipoprotein cholesterol level. In conclusion, in the MESA cohort, older age and higher blood pressure were associated with higher aortic WT and lower AD. Decreased AD was further associated with current smoking, African American ethnicity, and higher high density lipoprotein cholesterol level. PMID- 18678314 TI - Feasibility of a new generation three-dimensional echocardiography for right ventricular volumetric and functional measurements. AB - Right ventricular (RV) dimensions and function are of diagnostic and prognostic importance in cardiac disease. Because of the peculiar morphology of the right ventricle, 2-dimensional echocardiography has several limitations in RV evaluation. Recently, new 3-dimensional transthoracic echocardiographic software adapted for RV morphology was introduced. The aims of this study were to evaluate the feasibility of 3-dimensional RV analysis in a large population and to compare and correlate 3-dimensional RV data with classic 2-dimensional and Doppler parameters, including tricuspid annular plane systolic excursion and peak systolic velocity on Doppler tissue imaging, RV fractional shortening area, RV stroke volume (by the Doppler method), and pulmonary arterial systolic pressure. Two hundred subjects were studied: 48 normal controls and 152 patients with valvular heart disease (104 patients), idiopathic dilated cardiomyopathy (20 patients), or pulmonary hypertension (28 patients). The mean times for 3 dimensional acquisition and 3-dimensional reconstruction were 3 +/- 1 and 4 +/- 2 minutes, respectively. Imaging quality was good in most cases (85%). The mean RV diastolic and systolic volumes were 103 +/- 38 and 46 +/- 28 ml, respectively. The RV ejection fraction (RVEF) was correlated negatively with pulmonary arterial systolic pressure and positively with tricuspid annular plane systolic excursion, peak systolic velocity, and fractional shortening area. The pathologic group was characterized by larger RV volumes and lower RVEFs. Three-dimensional echocardiography clearly showed that in the pathologic group, patients with pulmonary hypertension had the largest RV volumes and the lowest RVEFs and that those with idiopathic dilated cardiomyopathy were characterized by RVEFs lower than those of patients with valvular disease. In conclusion, this new quantitative 3-dimensional method to assess RV volumes and function is feasible, relatively simple, and not time consuming. Data obtained with 3-dimensional analysis are well correlated with those obtained by 2-dimensional and Doppler methods and can differentiate normal and pathologic subjects. PMID- 18678315 TI - Heart rate reduction is probably not the main beneficial mechanism by which beta blockade improves outcome in patients with systolic chronic heart failure. PMID- 18678316 TI - Dofetilide for the prevention of postoperative atrial fibrillation after coronary surgery: is it a useful routine prophylaxis? PMID- 18678318 TI - Regarding bundle branch block in patients with heart failure. PMID- 18678319 TI - Relation of adult-onset asthma to coronary heart disease and stroke. PMID- 18678320 TI - Maternally inherited Birk Barel mental retardation dysmorphism syndrome caused by a mutation in the genomically imprinted potassium channel KCNK9. AB - We describe a maternally transmitted genomic-imprinting syndrome of mental retardation, hypotonia, and unique dysmorphism with elongated face. We mapped the disease-associated locus to approximately 7.27 Mb on chromosome 8q24 and demonstrated that the disease is caused by a missense mutation in the maternal copy of KCNK9 within this locus. KCNK9 is maternally transmitted (imprinted with paternal silencing) and encodes K(2P)9.1, a member of the two pore-domain potassium channel (K(2P)) subfamily. The mutation fully abolishes the channel's currents--both when functioning as a homodimer or as a heterodimer with K(2P)3.1. PMID- 18678339 TI - Exercise and other indirect challenges to demonstrate asthma or exercise-induced bronchoconstriction in athletes. AB - The prevalence of exercise-induced bronchoconstriction is reported to be high among recreational and elite athletes, yet diagnosis is often symptom-based. Indirect challenges such as the laboratory exercise challenge provide objective criteria for proper diagnosis and treatment. However, a standardized protocol using appropriate exercise intensity, duration, and dry air inhalation is often not implemented, and thus a false-negative test may result. This article reviews and describes the symptom-based diagnosis, the exercise challenge, and other indirect challenges such as eucapnic voluntary hyperpnea, hypertonic saline inhalation, and inhaled powdered mannitol as methods to diagnose and evaluate exercise-induced bronchoconstriction. Advantages and disadvantages of each diagnostic procedure are presented. PMID- 18678321 TI - Germline mutations and variants in the succinate dehydrogenase genes in Cowden and Cowden-like syndromes. AB - Individuals with PTEN mutations have Cowden syndrome (CS), associated with breast, thyroid, and endometrial neoplasias. Many more patients with features of CS, not meeting diagnostic criteria (termed CS-like), are evaluated by clinicians for CS-related cancer risk. Germline mutations in succinate dehydrogenase subunits SDHB-D cause pheochromocytoma-paraganglioma syndrome. One to five percent of SDHB/SDHD mutation carriers have renal cell or papillary thyroid carcinomas, which are also CS-related features. SDHB-D may be candidate susceptibility genes for some PTEN mutation-negative individuals with CS-like cancers. To address this hypothesis, germline SDHB-D mutation analysis in 375 PTEN mutation-negative CS/CS-like individuals was performed, followed by functional analysis of identified SDH mutations/variants. Of 375 PTEN mutation negative CS/CS-like individuals, 74 (20%) had increased manganese superoxide dismutase (MnSOD) expression, a manifestation of mitochondrial dysfunction. Among these, 10 (13.5%) had germline mutations/variants in SDHB (n = 3) or SDHD (7), not found in 700 controls (p < 0.001). Compared to PTEN mutation-positive CS/CS like individuals, those with SDH mutations/variants were enriched for carcinomas of the female breast (6/9 SDH versus 30/107 PTEN, p < 0.001), thyroid (5/10 versus 15/106, p < 0.001), and kidney (2/10 versus 4/230, p = 0.026). In the absence of PTEN alteration, CS/CS-like-related SDH mutations/variants show increased phosphorylation of AKT and/or MAPK, downstream manifestations of PTEN dysfunction. Germline SDH mutations/variants occur in a subset of PTEN mutation negative CS/CS-like individuals and are associated with increased frequencies of breast, thyroid, and renal cancers beyond those conferred by germline PTEN mutations. SDH testing should be considered for germline PTEN mutation-negative CS/CS-like individuals, especially in the setting of breast, thyroid, and/or renal cancers. PMID- 18678340 TI - Asthma and the elite athlete: summary of the International Olympic Committee's consensus conference, Lausanne, Switzerland, January 22-24, 2008. AB - Respiratory symptoms cannot be relied on to make a diagnosis of asthma and/or airways hyperresponsiveness (AHR) in elite athletes. For this reason, the diagnosis should be confirmed with bronchial provocation tests. Asthma management in elite athletes should follow established treatment guidelines (eg, Global Initiative for Asthma) and should include education, an individually tailored treatment plan, minimization of aggravating environmental factors, and appropriate drug therapy that must meet the requirements of the World Anti-Doping Agency. Asthma control can usually be achieved with inhaled corticosteroids and inhaled beta(2)-agonists to minimize exercise-induced bronchoconstriction and to treat intermittent symptoms. The rapid development of tachyphylaxis to beta(2) agonists after regular daily use poses a dilemma for athletes. Long-term intense endurance training, particularly in unfavorable environmental conditions, appears to be associated with an increased risk of developing asthma and AHR in elite athletes. Globally, the prevalence of asthma, exercise-induced bronchoconstriction, and AHR in Olympic athletes reflects the known prevalence of asthma symptoms in each country. The policy of requiring Olympic athletes to demonstrate the presence of asthma, exercise-induced bronchoconstriction, or AHR to be approved to inhale beta(2)-agonists will continue. PMID- 18678342 TI - Mind the widening gap: have improvements in asthma care increased asthma disparities? PMID- 18678344 TI - Correlation of IL-31 serum levels with severity of atopic dermatitis. PMID- 18678343 TI - Prenatal exposure to a farm environment modifies atopic sensitization at birth. AB - BACKGROUND: Previous cross-sectional surveys have suggested that maternal exposure to animal sheds during pregnancy exerted a protective effect on atopic sensitization in children lasting until school age. OBJECTIVE: We sought to evaluate the effects of maternal exposure to animal sheds and other farm-related exposures during pregnancy on cord blood IgE levels in a prospective birth cohort. METHODS: Pregnant women living in rural areas in Austria, Finland, France, Germany, and Switzerland were recruited in the third trimester of pregnancy. Information on maternal farm-related exposures, nutrition, and health during pregnancy was obtained by means of interviews. Specific IgE levels for food and common inhalant allergens were assessed in cord blood of 922 children and peripheral blood samples of their mothers. RESULTS: Different sensitization patterns in cord blood of farm and nonfarm children were observed. In multivariable analysis consumption of boiled, but not unboiled, farm milk during pregnancy was positively associated with specific IgE to cow's milk independently from maternal IgE. In contrast, there was an inverse relationship between maternal exposure to animal sheds and cord blood IgE levels against seasonal allergens (adjusted odds ratio, 0.38; 95% CI, 0.21-0.70). This association was not confounded by maternal IgE levels. Maternal contact with hay enhanced the protective effect of exposure to animal sheds on IgE levels to grass pollen in cord blood. CONCLUSIONS: Maternal exposure during pregnancy influences atopic sensitization patterns in cord blood. The (microbial) context of allergen contact possibly modifies the risk of atopic sensitization. PMID- 18678346 TI - Scrutinizing ethics as applied to clinical research. PMID- 18678345 TI - Anti-TNF-alpha (infliximab) therapy for severe adult eosinophilic esophagitis. PMID- 18678347 TI - Asthma and athletics. PMID- 18678349 TI - The level of insulin growth factor-1 receptor expression is directly correlated with the tumor uptake of (111)In-IGF-1(E3R) in vivo and the clonogenic survival of breast cancer cells exposed in vitro to trastuzumab (Herceptin). AB - INTRODUCTION: Our objective was to define the relationships between tumor uptake of [(111)In]-IGF-1 and [(111)In]-IGF-1(E3R), an analogue which does not bind insulin growth factor-1 (IGF-1) binding proteins (i.e., IGFBP-3), and the level of IGF-1 receptor (IGF-1R) expression on human breast cancer (BC) xenografts in athymic mice, as well as the feasibility for tumor imaging. A second objective was to correlate IGF-1R (and HER2 density) with the cytotoxicity of trastuzumab in the absence/presence of IGFBP-3 or the IGF-1R tyrosine kinase inhibitor, AG1024. METHODS: The tumor and normal tissue uptake of [(111)In]-IGF-1 and [(111)In]-IGF-1(E3R) were determined at 4 h postinjection in mice implanted subcutaneously with MDA-MB-231, H2N, HR2 or MCF-7/HER2-18 human BC xenografts (8.5x10(4), 1.4x10(4), 4.0x10(4) and 1.0x10(5) IGF-1R/cell, respectively). The effect of co-injection of IGF-1 (50 microg) or IGFBP-3 (2 or 25 microg) was studied. The relationship between tumor uptake of [(111)In]-IGF-1(E3R) and IGF-1R density was examined. MicroSPECT/CT imaging was performed on mice with MCF-7/HER2 18 tumors injected with [(111)In]-IGF-1(E3R). The surviving fraction of BC cells exposed to trastuzumab (67.5 mug/ml) in the absence/presence of IGFBP-3 (1 microg/ml) or the IGF-1R kinase inhibitor, AG1024 (1 or 5 microg/ml), was determined. RESULTS: [(111)In]-IGF-1 was specifically taken up by MCF-7/HER2-18 xenografts; tumor uptake was decreased twofold when co-injected with IGF-1 (1.9+/ 0.1 vs. 1.0+/-0.1 %ID/g). Co-injection of IGBP-3 decreased kidney uptake of [(111)In]-IGF-1 up to twofold and increased circulating radioactivity threefold. There was a strong linear correlation (r(2)=0.99) between the tumor uptake of (111)In-IGF-1(E3R) and IGF-1R density. Tumor uptake ranged from 0.4+/-0.05 %ID/g for H2N to 2.5+/-0.5 %ID/g for MCF-7/HER2-18 xenografts. MCF-7/HER2-18 tumors were visualized by microSPECT/CT. Resistance of BC cells to trastuzumab was directly associated with IGF-1R expression, despite co-expression of HER2. The resistance of HR2 cells could be partially reversed by IGFBP-3 or AG1024. CONCLUSION: Imaging of IGF-1R expression using [(111)In]-IGF-1(E3R) may be useful for identifying HER2-positive tumors in BC patients that are resistant to trastuzumab through this mechanism. PMID- 18678350 TI - Evaluation of a bromine-76-labeled progestin 16alpha,17alpha-dioxolane for breast tumor imaging and radiotherapy: in vivo biodistribution and metabolic stability studies. AB - INTRODUCTION: Progesterone receptors (PRs) are present in many breast tumors, and their levels are increased by certain endocrine therapies. They can be used as targets for diagnostic imaging and radiotherapy. METHOD: 16alpha,17alpha-[(R)-1' alpha-(5-[(76)Br]Bromofurylmethylidene)dioxyl]-21-hydroxy-19-norpregn-4-ene-3,20 dione ([(76)Br]16alpha,17alpha-[(R)-1'-alpha-(5-bromofurylmethylidene)dioxyl]-21 hydroxy-19-norpregn-4-ene-3,20-dione (3)), a PR ligand with relative binding affinity (RBA)=65 and log P(o/w)=5.09+/-0.84, was synthesized via a two-step reaction, and its tissue biodistribution and metabolic stability were evaluated in estrogen-primed immature female Sprague-Dawley rats. RESULTS: [(76)Br]16alpha,17alpha-[(R)-1'-alpha-(5-bromofurylmethylidene)dioxyl]-21-hydroxy 19-norpregn-4-ene-3,20-dione 3 was synthesized in 5% overall yield with specific activity being 200-1250 Ci/mmol. [(76)Br]16alpha,17alpha-[(R)-1'-alpha-(5 bromofurylmethylidene)dioxyl]-21-hydroxy-19-norpregn-4-ene-3,20-dione 3 demonstrated high PR-mediated uptake in the target tissue uterus (8.72+/-1.84 %ID/g at 1 h) that was reduced by a blocking dose of unlabeled progestin R5020, but the nonspecific uptake in blood and muscle (2.11+/-0.14 and 0.89+/-0.16 %ID/g at 1 h, respectively) was relatively high. [(76)Br]16alpha,17alpha-[(R)-1'-alpha (5-bromofurylmethylidene)dioxyl]-21-hydroxy-19-norpregn-4-ene-3,20-dione 3 was stable in whole rat blood in vitro, but it was not stable in vivo due to the fast metabolism that occurred in the liver, resulting in the formation of a more polar radioactive metabolite and free [(76)Br]bromide. The level of free [(76)Br]bromide in blood remained high during the experiment (2.11+/-0.14 %ID/g at 1 h and 1.52+/-0.24 %ID/g at 24 h). The tissue distribution of [(76)Br]16alpha,17alpha-[(R)-1'-alpha-(5-bromofurylmethylidene)dioxyl]-21-hydroxy 19-norpregn-4-ene-3,20-dione 3 at 1 and 3 h was compared with that of the (18)F analogs, [(18)F]FFNP fluoro furanyl norprogesterone (FFNP) 1 and ketal 2. CONCLUSION: [(76)Br]16alpha,17alpha-[(R)-1'-alpha-(5 bromofurylmethylidene)dioxyl]-21-hydroxy-19-norpregn-4-ene-3,20-dione 3 may have potential for imaging PR-positive breast tumors at early time points, but it is not suitable for imaging at later times or for radiotherapy. PMID- 18678351 TI - The role of technetium-99m-labeled octreotide acetate scintigraphy in suspected breast cancer and correlates with expression of SSTR. AB - PURPOSE: To evaluate the value of (99m)Tc-octreotide acetate (hereafter, (99m)Tc octreotide) somatostatin receptor (SSTR) scintigraphy in the detection of primary breast cancer and to correlate with expression of SSTRs. MATERIALS AND METHODS: Fifty-four female and 1 male patients (range, 17-77 years; mean age, 48 years) with palpable breast lesion were included in this study. (99m)Tc-octreotide and (99m)Tc-MIBI scintigraphy were undertaken in all patients, and the region of interest was drawn around each lesion. Tumor uptake was measured and expressed as the ratio of tumor to normal tissue activity (T/NT). Final clinical diagnosis was confirmed by histopathological analysis. Expression of SSTR1-5 mRNA was measured with RT-PCR in 15 patients with malignant neoplasm, and protein level of SSTR-2 and SSTR-5 was measured using immunohistochemical staining in 15 patients with malignant neoplasm and 18 patients with benign lesion. RESULTS: Thirty-five patients were confirmed to have infiltrative ductal breast carcinoma, 1 patient with cellular cancer, 1 patient with adenocarcinoma and 18 patients had benign lesions. The sensitivity, specificity, positive predictive value and negative predictive value of (99m)Tc-octreotide in detection of primary lesion were 91.8%, 22.2%, 71.8% and 57.1%, respectively. No significant correlation was observed between T/NT of (99m)Tc-octreotide and (99m)Tc-MIBI. SSTR mRNA subtypes were variably expressed. SSTR3 was the most highly expressed, followed by SSTR1, SSTR2 and SSTR5; SSTR4 was the least expressed in the level of mRNA of SSTR. Significant correlation was shown between T/NT and the expression of SSTR2 mRNA (r=0.73, P<.01). CONCLUSION: (99m)Tc-octreotide acetate scintigraphy was sensitive for the detection of primary lesion of breast cancer; however, nonspecific breast tissue uptake hampered the specificity and clinical value in the detection of lymph node metastasis. Five subtypes of SSTR mRNA and protein SSTR2 and SSTR5 were expressed variably in breast cancer due to tumor heterogeneity. (99m)Tc-octreotide imaging may hold promise in the evaluation of the level of SSTR2 in vivo. PMID- 18678352 TI - Enhancement of somatostatin-receptor-targeted (177)Lu-[DOTA(0)-Tyr(3)]-octreotide therapy by gemcitabine pretreatment-mediated receptor uptake, up-regulation and cell cycle modulation. AB - INTRODUCTION: Clinical studies of patients treated with somatostatin-receptor (sstr)-targeted [DOTA(0)-Tyr(3)]-octreotide (DOTATOC) labeled with (177)Lu and (90)Y have shown overall response rates in the range of 9-33%. This study evaluates the potential for combination therapy with gemcitabine in an effort to improve clinical outcomes. METHODS: Human pancreatic adenocarcinoma Capan-2, rat pancreatic cancer AR42J and human small cell lung cancer NCI-H69 cells were each treated with 1 microg/ml gemcitabine for 4 days followed by replacement of the medium alone for four additional days. Cell cycle and direct receptor-uptake studies were performed with (177)Lu-DOTATOC after the total 8-day treatment as described. Cell viability and apoptosis experiments were performed to study the effects of gemcitabine pretreatment and (177)Lu-DOTATOC radionuclide therapy. Parallel control studies were performed with receptor-non-targeted (177)Lu-DOTA and DOTATOC. RESULTS: Cells treated with gemcitabine for 4 days showed a down regulation of sstr expression as determined by (177)Lu-DOTATOC uptake. However, after 4 days of additional growth in absence of gemcitabine, the uptake of (177)Lu-DOTATOC was 1.5-3 times greater than that of the untreated control cells. In gemcitabine-pretreated Capan-2 cells, 84% of the cell population was in the G(2)M phase of the cell cycle. Due to sstr up-regulation and cell cycle modulations, synergistic effects of gemcitabine pretreatment were observed in cell viability and apoptosis assays. (177)Lu-DOTATOC resulted in two to three times greater apoptosis in gemcitabine-pretreated Capan-2 cells compared to the untreated cells. CONCLUSION: Gemcitabine pretreatment up-regulates sstr expression and acts as a radiosensitizer through cell cycle modulation. The rational combination of gemcitabine and sstr-targeted radiopharmaceuticals represents a promising chemoradiation therapeutic tool with great potential to improve clinical outcomes and, thus, merits further study. PMID- 18678353 TI - Radiolabeling of annexin A5 with (99m)Tc: comparison of HYNIC-Tc vs. iminothiolane-Tc-tricarbonyl conjugates. AB - In the perspective of expanding the use of annexin A5 (anx A5) as radioactive tracer of cell death in vivo, we recently described its radiolabeling with (99m)Tc-tricarbonyl [(99m)Tc(H(2)O)(3)(CO)(3)](+) via the mercaptobutyrimidyl group (anx A5-SH). The aim of the present article was to compare this new method with the HYNIC strategy (anx A5-HYNIC), recognized at present as the reference for the radiolabeling of proteins with (99m)Tc. Similar radiolabeling yields and better chemical stability were obtained with the [anx A5-SH-(99m)Tc-tricarbonyl] complex. Since the [anx A5-HYNIC-(99m)Tc(tricine)(2)] conjugate shows isomeric forms which can affect the biological properties whereas [anx A5-SH-(99m)Tc tricarbonyl] is less or not prone to such drawback, the latter seems superior to the former. Furthermore, (anx A5-SH) is readily obtained via commercial sources of Traut's reagent whereas (anx A5-HYNIC) is not. The results provide encouraging evidence in the development of anx A5-labeled reagent for apoptose imaging. PMID- 18678354 TI - [(11)C]Choline positron emission tomography/computed tomography for staging and restaging of patients with advanced prostate cancer. AB - INTRODUCTION: To evaluate [(11)C]Choline positron emission tomography (PET)/computed tomography (CT) for staging and restaging of patients with advanced prostate cancer and to compare the diagnostic performance of PET, CT and PET/CT. METHODS: Forty-five consecutive patients with advanced prostate cancer underwent [(11)C]Choline-PET/CT between 5/2004 and 2/2006. RESULTS: Overall, 295 lesions were detected: PET alone, 178 lesions; diagnostic CT, 221 lesions; PET/CT (low-dose CT), 272 lesions; PET/CT (diagnostic CT), 295 lesions. Two thirds of the lesions were located in the bone; one third in the prostate, lymph nodes, periprostatic tissue and soft tissue (lung, liver). The use of diagnostic CT did not result in a statistically significant difference with respect to lesion localization certainty and lesion characterization (P=.063, P=.063). PET-negative but PET/CT-positive lesions were mostly localized in the bone (78%, 91/117) as were PET-positive and CT-negative lesions (72%, 53/74). Of the latter, 91% (48/53) represented bone marrow and 9% (5/53) cortical involvement. CONCLUSIONS: Staging and restaging with [(11)C]Choline PET/CT in patients with advanced prostate cancer improve the assessment of local and regional recurrent as well as metastatic disease including skeletal manifestations. [(11)C]Choline PET/CT (with a low-dose CT) results in improved localization and lesion characterization. [(11)C]Choline PET/CT provides an added value for skeletal manifestations. [(11)C]Choline PET/CT changed disease management in 11 (24%) of 45 patients with advanced prostate cancer. PMID- 18678355 TI - N(3)-Substituted thymidine analogues V: synthesis and preliminary PET imaging of N(3)-[(18)F]fluoroethyl thymidine and N(3)-[(18)F]fluoropropyl thymidine. AB - INTRODUCTION: [(18)F]-Labeled analogues of thymidine have demonstrated efficacy for PET imaging of cellular proliferation. We have synthesized two [(18)F] labeled N(3)-substituted thymidine analogues, N(3)-[(18)F]fluoroethyl thymidine (N(3)-[(18)F]-FET) and N(3)-[(18)F]fluoropropyl thymidine (N(3)-[(18)F]-FPrT), and performed preliminary PET imaging studies in tumor-bearing mice. METHODS: Thymidine was converted to its 3',5'-O-bis-tetrahydropyranyl ether, which was then converted to the N(3)-ethyl and propyl-substituted mesylate precursors. Reactions of these mesylate precursors with n-Bu(4)N[(18)F] or K[(18)F]/kryptofix followed by acid hydrolysis and HPLC purification yielded N(3)-[(18)F]-FET and N(3)-[(18)F]-FPrT, respectively. Subcutaneous (sc) xenografts of H441 human non small cell lung cancer were established in two groups of mice (each n=6). Micro PET images of the tumor-bearing animals were acquired after intravenous injection of N(3)-[(18)F]-FET or N(3)-[(18)F]-FPrT (3700 KBq/animal). RESULTS: The radiochemical yields were 2-12% (d.c.) for N(3)-[(18)F]-FET and 30-38% (d.c.) for N(3)-[(18)F]-FPrT. Radiochemical purity was >99% and calculated specific activity was >74 GBq/mumol at the end of synthesis. The accumulation of N(3)-[(18)F]-FET and N(3)-[(18)F]-FPrT in the tumor tissue at 2 h postinjection was 1.81+/-0.78 and 2.95+/-1.14 percent injected dose per gram (%ID/g), respectively; tumor/muscle ratios were 5.57+/-0.82 and 7.69+/-2.18, respectively; the unidirectional influx rates (K(i)) were 0.013 and 0.018 ml/g per minute, respectively. CONCLUSION: Two novel [(18)F]- N(3)-substituted thymidine analogues have been synthesized in good yields, high purity and high specific activity. Preliminary in vivo studies demonstrated the efficacy of these [(18)F]- N(3) substituted thymidine analogues for PET imaging of tumors. PMID- 18678356 TI - In vitro evaluation of canine leukocytes radiolabeled in whole blood with (99m)Tc stannous colloid. AB - INTRODUCTION: Technetium-99m stannous colloid ((99m)TcSnC)-labeled leukocytes are used to investigate a variety of inflammatory diseases in human medicine. The present study investigates the in vitro behavior of canine leukocytes labeled in whole blood with (99m)TcSnC. METHODS: Blood samples from 10 healthy dogs were labeled with (99m)TcSnC using a standard procedure. The distribution of radioactivity among blood components (plasma, leukocyte layers and erythrocytes) was measured following separation of the radiolabeled samples across Histopaque density gradients. Phagocytic function of labeled and unlabeled leukocytes was estimated using zymosan particles. Labeling retention by leukocytes was determined at 1, 3, 4 and 7 h postlabeling. RESULTS: The mean+/-standard error percentage of radioactivity associated with plasma, erythrocyte and leukocyte fractions was 2.0+/-0.21%, 55.5+/-0.60% and 42.5+/-0.54%, respectively (the last comprising 70.2+/-0.83% in polymorphonuclear leukocytes and 29.8+/-0.83% in mononuclear leukocytes). Labeled canine leukocytes had a phagocytic activity of 91.3+/-0.28% (control, 91.7+/-0.26%). The radiolabeled canine leukocytes retained 94.1+/-0.30% of radioactivity at 7 h postlabeling. CONCLUSIONS: Radiolabeling of canine leukocytes in whole blood with (99m)TcSnC has minor adverse effect on their phagocytic function. The radiolabeled canine leukocytes retained a large percentage of radioactivity for at least 7 h postlabeling. PMID- 18678357 TI - Autoradiographic and small-animal PET comparisons between (18)F-FMISO, (18)F-FDG, (18)F-FLT and the hypoxic selective (64)Cu-ATSM in a rodent model of cancer. AB - INTRODUCTION: Copper(II)-diacetyl-bis(N(4)-methylthiosemicarbazone), or Cu-ATSM, a hypoxia imaging agent, has been shown to be predictive of response to traditional cancer therapies in patients with a wide range of tumors. It is known that the environment of the tumor results in a myriad of physiological consequences, including hypoxia, alterations in metabolism and proliferation. In an effort to better characterize the relationships between Cu-ATSM and other prominent radiopharmaceuticals, this current study was undertaken to compare the regional distribution of (64)Cu-ATSM with [(18)F]fluoromisonidazole ((18)F FMISO), [(18)F]fluoro-2-deoxy-d-glucose ((18)F-FDG) and [(18)F]fluorothymidine ((18)F-FLT) in 9L tumors. METHODS: Taking advantage of the different half-life of (18)F (t(1/2)=110 min) in comparison to (64)Cu (t(1/2)=12.7 h), we undertook a dual-tracer autoradiography study in 9L tumors. Four groups were examined: (a) (18)F-FMISO, 2 h postinjection (p.i.) and (64)Cu-ATSM, 10 min p.i.; (b) (18)F FMISO, 2 h p.i. and (64)Cu-ATSM, 24 h p.i.; (c) (18)F-FDG, 1 h p.i. and (64)Cu ATSM, 10 min p.i.; and (d) (18)F-FLT, 1 h p.i. and (64)Cu-ATSM, 10 min p.i. Small animal PET imaging was performed in 9L tumor-bearing rats with imaging on concurrent days comparing (64)Cu-ATSM with (18)F-FMISO and (18)F-FLT. RESULTS: It was shown that the regional distribution of (18)F-FMISO and (64)Cu-ATSM showed an excellent correlation when the (64)Cu-ATSM had been allowed to distribute for either 10 min (R(2)=.84) or 24 h (R(2)=.86). The regional comparisons between (64)Cu-ATSM (10 min) and (18)F-FDG (1 h) resulted in a very poor correlation (R(2)=.08) between the regional uptake of the two agents. The comparison between (18)F-FLT and (64)Cu-ATSM showed a strong relationship (R(2)=.83) between the two tracers. The small-animal PET images for the distribution comparisons between (64)Cu-ATSM and (18)F-FMISO and (18)F-FLT were in agreement with the data generated from the autoradiography studies. CONCLUSIONS: The data show that it is important to remember that a number of different metabolic situations can exist when considering the relationship between regions of high glucose uptake, proliferation and hypoxia. PMID- 18678358 TI - Validation of (68)Ge/(68)Ga generator processing by chemical purification for routine clinical application of (68)Ga-DOTATOC. AB - INTRODUCTION: Imaging of somatostatin receptor expressing tumours has been greatly enhanced by the use of (68)Ga-DOTATOC and PET/CT. METHODS: In this work, a purification method for the (68)Ge/(68)Ga generator eluate and a method to produce (68)Ga-DOTATOC suitable for clinical use were evaluated. The generator eluate was purified and concentrated on a cation-exchange cartridge in HCl/acetone media. The efficacy of this procedure in eliminating metal impurities from the (68)Ga solution was investigated by ICP-MS. The radiotracer quality was evaluated by radio-TLC, GC and gamma-ray spectrometry. RESULTS: (68)Ga-DOTATOC preparations (n=33) were carried out with a mean synthesis yield of 59.3+/-2.8% (not corrected for decay) and a batch activity ranging from 555 to 296 MBq. The radiochemical and radionuclidic purity were >98% and 99.9999%, respectively. With this purification process, >95% of the Fe(III), Zn(II) and Mn(II) were eliminated from the solution. CONCLUSIONS: (68)Ga-DOTATOC produced with this method can be efficiently used in nuclear medicine departments for PET evaluations. PMID- 18678359 TI - Automated synthesis of the generic peptide labelling agent N-succinimidyl 4 [(18)F]fluorobenzoate and application to (18)F-label the vasoactive transmitter urotensin-II as a ligand for positron emission tomography. AB - INTRODUCTION: The objectives of this work were to develop an automated production of N-succinimidyl 4-[(18)F]fluorobenzoate ([(18)F]-SFB) and to test whether the vasoactive peptide urotensin-II (U-II) could be labelled by conjugation with [(18)F]-SFB. METHODS: A TRACERlab MX(FDG) synthesizer including an HPLC unit was used. The MS Excel synthesis sequence and the standard disposable FDG cassette were modified to allow the synthesis of [(18)F]-SFB. U-II was subsequently conjugated with [(18)F]-SFB, and the resulting (18)F-labelled peptides were characterised using in vitro ligand binding assays. RESULTS: [(18)F]-SFB was successfully synthesised in the TRACERlab MX(FDG) in 44.3+/-2.5% (n=25) radiochemical yield in 98 min. [(18)F]-SFB (8-12 GBq) has been produced with specific activities in the range of 250-350 GBq/mumol and a radiochemical purity >95%. [(18)F]-SFB was subsequently used to label U-II. Two radiolabelled products, [(18)F]-(Glu(1))-U-II and [(18)F]-(Lys(8))-U-II, were formed in an isolated radiochemical yield from [(18)F]-SFB of 5.2+/-0.3% and 29.0+/-3.7%, respectively (n=7). Radioligand binding assays revealed that [(18)F]-(Glu(1))-U II had retained subnanomolar affinity. Binding to human skeletal muscle (n=3) was concentration dependent and saturable with K(d)=0.84+/-0.51 nM, B(max)=0.69+/ 0.14 fmol/mg protein and Hill slope (nH)=1.03+/-0.12. CONCLUSIONS: [(18)F]-SFB has been synthesised using the TRACERlab MX(FDG) module, allowing production of up to 8-12 GBq of [(18)F]-SFB with specific activities of 250-350 GBq/mumol. [(18)F]-SFB was used for the labelling of U-II. In vitro characterisation demonstrated that [(18)F]-(Glu(1))-U-II had retained desirable binding properties and may be suitable as a positron emission tomography radioligand for the imaging of the U-II receptor. PMID- 18678360 TI - Surviving sepsis campaign--the sequel. PMID- 18678361 TI - [Sympathetic response in primary hypertension]. AB - High blood pressure (BP) is a major cause of cardiovascular disease and primary hypertension is a frequent pathological condition. Sympathetic hyperactivity may be involved in primary hypertension. The purpose of this study was mainly to evaluate sympathetic activity when performing cardiovascular autonomic profile examination in patients with primary hypertension in comparison with normotensive subjects. PATIENTS AND METHODS: This prospective study included one group of hypertensive patients (n=120, mean age 54 years) compared with a control group (n=120, mean age 52 years) of normotensive subjects. Autonomic tests included deep-breathing (DB), hand-grip (HG) and echostress test (ES). Comparison tests between the two groups, similar in age, were expressed as mean+/-SE and made using the t Student test, p<0.05 was considered significant. RESULTS: Alpha adrenergic sympathetic response using ES method produced a BP response of 20,0%+/ 9,8 in hypertensive patients group and 15,2%+/-8,6 in the control group (p<0.001). Alpha-adrenergic sympathetic response using three minutes HG test was of 16,7%+/-7,5 in hypertensive patients group and 13,3%+/-6,5 in the control group (p<0.001). Vagal stimulation in hypertensive group after DB showed that electrocardiographic: ECG (EKG) waves R (RR) interval variation was of 30,2%+/ 8,1 meanwhile in the control group this RR variation was of 46,1%+/-21,1 p<0.001, and the one of HG of 15 seconds was 17,6%+/-10,2 versus 32,5%+/-12,7 p<0.001. CONCLUSION: Hypertensive patients had a significantly higher sympathetic response to central and peripheral stimulations and a significantly lower parasympathetic response when compared to normotensive controls. PMID- 18678362 TI - [Parapharyngeal tumors: diagnosis and treatment]. AB - OBJECTIVES: Parapharyngeal space tumors are rare and usually benign. Their treatment is surgical and many approaches have been described. We report our experience in managing these neoplasms. Two points are developed more fully: the correlation between imaging and surgical observation and the reliability of the cervical approach. MATERIAL AND METHODS: A retrospective review of primary parapharyngeal space tumors treated at the Rennes University Hospital between 1992 and 2004 is presented. Nine patients were included, all treated surgically with a cervical approach without parotidectomy. RESULTS: Eight MR imaging and seven CT scans were done. Two retrostyloid tumors and six prestyloid tumors, all independent of the deep lobe of the parotid gland, were found. They were benign in all cases (five salivary tumors and four schwannomas). Two patients treated for a pleomorphic adenoma presented recurrence and were reoperated successfully. The average follow-up was 66 months. None of the patients died. Definitive postoperative complications occurred only for neurogenic tumors. CONCLUSION: The cervical approach is safe and allows the surgical excision of retrostyloid and prestyloid tumors, independent of the deep lobe of the parotid gland. Large tumor (8cm in our study) excision can be done with few complications. An attentive study of preoperative imaging is necessary to choose the best surgical approach. PMID- 18678363 TI - In vivo osteopontin-induced macrophage accumulation is dependent on CD44 expression. AB - In order to assess the role of osteopontin (OPN) in leukocyte accumulation in inflammatory conditions, native OPN and its thrombin cleaved form (OPN+Thr) were studied in vivo using a rodent subcutaneous air pouch model (AP). Both forms of OPN-induced macrophage infiltration into the AP in wild-type mice. In animals lacking CD44, macrophage numbers were significantly reduced within the cavity, but cells still accumulated along the subcutaneous lining. In animals lacking endogenous OPN, no differences were found in exogenous OPN-induced macrophage accumulation, although macrophage exhibited increased alpha4 integrin expression. These studies reveal that both OPN and OPN+Thr attract macrophages in vivo through CD44. PMID- 18678364 TI - Three-dimensional arm movements at constant equi-affine speed. AB - It has long been acknowledged that planar hand drawing movements conform to a relationship between movement speed and shape, such that movement speed is inversely proportional to the curvature to the power of one-third. Previous literature has detailed potential explanations for the power law's existence as well as systematic deviations from it. However, the case of speed-shape relations for three-dimensional (3D) drawing movements has remained largely unstudied. In this paper we first derive a generalization of the planar power law to 3D movements, which is based on the principle that this power law implies motion at constant equi-affine speed. This generalization results in a 3D power law where speed is inversely related to the one-third power of the curvature multiplied by the one-sixth power of the torsion. Next, we present data from human 3D scribbling movements, and compare the obtained speed-shape relation to that predicted by the 3D power law. Our results indicate that the introduction of the torsion term into the 3D power law accounts for significantly more of the variance in speed-shape relations of the movement data and that the obtained exponents are very close to the predicted values. PMID- 18678366 TI - Elevated PCB levels in anglers and unsuspected transport of pollutants from aquatic food webs into human foods. PMID- 18678367 TI - Parental translocations and need for preimplantation genetic diagnosis? Distorting effects of ascertainment bias and the need for information-rich families. PMID- 18678368 TI - Antagonist/letrozole protocol in poor ovarian responders for intracytoplasmic sperm injection: a comparative study with the microdose flare-up protocol. AB - OBJECTIVE: To compare the effect of the GnRH antagonist/letrozole protocol (AL) with the microdose GnRH agonist flare-up (MF) protocol in poor ovarian responders for intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective, case-control study. SETTING: Anatolia Women and IVF Center. PATIENT(S): Eight hundred eighty five consecutive patients (1383 cycles) predicted to have or with a history of poor ovarian response who were undergoing ICSI were enrolled. The MF protocol was used in 673 patients (1026 cycles), and the AL protocol was used in the remaining 212 patients (357 cycles). INTERVENTION(S): Controlled ovarian hyperstimulation and ICSI. MAIN OUTCOME MEASUREMENT(S): Cycle cancellation rate, number of oocytes retrieved, fertilization rate, embryo quality, clinical pregnancy, and implantation rates. RESULT(S): The total gonadotropin consumption, duration of stimulation, E(2) level on the day of hCG administration, and number of oocytes retrieved were significantly lower with the AL protocol compared with the MF protocol. However, the fertilization rate and the rate of at least one top quality embryo transferred were higher with the AL compared with the MF protocol. The clinical pregnancy rates were comparable between the two groups. The implantation rates with the MF and AL protocols were 9.8% and 14.5%, respectively. CONCLUSION(S): The GnRH AL protocol is an effective protocol that may be used in poor ovarian responders for ICSI. PMID- 18678369 TI - Postpartum hemorrhage: new management? PMID- 18678371 TI - Viable pregnancies can occur after 24-hour incubation of ejaculated sperm before intracytoplasmic sperm injection. AB - This is the first report of viable pregnancies established with ICSI after 24 hour incubation of washed ejaculated sperm from men with poor sperm parameters. Because both pregnancy rates and viable pregnancy rates were significantly increased compared with 2- to 4-hour-incubated historical controls, 24-hour incubation before ICSI may be beneficial in selecting the most favorable ejaculated sperm. PMID- 18678372 TI - Basal metabolic rate is decreased in women with polycystic ovary syndrome and biochemical hyperandrogenemia and is associated with insulin resistance. AB - OBJECTIVE: To evaluate basal metabolic rate (BMR) in women with PCOS and to determine its association with insulin resistance (IR). DESIGN: Prospective assessment of BMR in women with PCOS. SETTING: Outpatient clinic of the Division of Reproductive Endocrinology. PATIENT(S): The study included 91 Greek women with PCOS and biochemical hyperandrogenemia, with mean age 24.03 +/- 0.55 years and mean body mass index (BMI) 26.67 +/- 0.69 kg/m(2), and 48 matched regularly menstruating women, with mean age 26.33 +/- 0.93 years and mean BMI 23.35 +/- 0.85 kg/m(2), as control subjects. INTERVENTION(S): Assessment of BMR by indirect calorimetry, IR by HOMA and QUICKI indices, fasting insulin, and fasting glucose/insulin ratio. MAIN OUTCOME MEASURE(S): Reduced BMR in PCOS with or without IR. RESULT(S): Adjusted BMR was 1,868 +/- 41 kcal/day in the control group, 1,445.57 +/- 76 in all PCOS women, 1,590 +/- 130 in PCOS women without IR and 1,116 +/- 106 in PCOS women with IR. Adjusted BMR showed a statistically significant difference between women with PCOS and control subjects, with lowest values in the group of PCOS women with IR, even after adjusting all groups for age and BMI. CONCLUSION(S): Women with PCOS, particularly those with IR, present a significantly decreased BMR. PMID- 18678374 TI - Parental translocations and the need for PGD? Distorting effects of ascertainment bias and the need for information-rich families. PMID- 18678375 TI - Kinetic chain of overarm throwing in terms of joint rotations revealed by induced acceleration analysis. AB - This study investigated how baseball players generate large angular velocity at each joint by coordinating the joint torque and velocity-dependent torque during overarm throwing. Using a four-segment model (i.e., trunk, upper arm, forearm, and hand) that has 13 degrees of freedom, we conducted the induced acceleration analysis to determine the accelerations induced by these torques by multiplying the inverse of the system inertia matrix to the torque vectors. We found that the proximal joint motions (i.e., trunk forward motion, trunk leftward rotation, and shoulder internal rotation) were mainly accelerated by the joint torques at their own joints, whereas the distal joint motions (i.e., elbow extension and wrist flexion) were mainly accelerated by the velocity-dependent torques. We further examined which segment motion is the source of the velocity-dependent torque acting on the elbow and wrist accelerations. The results showed that the angular velocities of the trunk and upper arm produced the velocity-dependent torque for initial elbow extension acceleration. As a result, the elbow joint angular velocity increased, and concurrently, the forearm angular velocity relative to the ground also increased. The forearm angular velocity subsequently accelerated the elbow extension and wrist flexion. It also accelerated the shoulder internal rotation during the short period around the ball-release time. These results indicate that baseball players accelerate the distal elbow and wrist joint rotations by utilizing the velocity-dependent torque that is originally produced by the proximal trunk and shoulder joint torques in the early phase. PMID- 18678376 TI - Children's knowledge of deceptive gaze cues and its relation to their actual lying behavior. AB - Eye gaze plays a pivotal role during communication. When interacting deceptively, it is commonly believed that the deceiver will break eye contact and look downward. We examined whether children's gaze behavior when lying is consistent with this belief. In our study, 7- to 15-year-olds and adults answered questions truthfully (Truth questions) or untruthfully (Lie questions) or answered questions that required thinking (Think questions). Younger participants (7- and 9-year-olds) broke eye contact significantly more when lying compared with other conditions. Also, their averted gaze when lying differed significantly from their gaze display in other conditions. In contrast, older participants did not differ in their durations of eye contact or averted gaze across conditions. Participants' knowledge about eye gaze and deception increased with age. This knowledge significantly predicted their actual gaze behavior when lying. These findings suggest that with increased age, participants became increasingly sophisticated in their use of display rule knowledge to conceal their deception. PMID- 18678377 TI - Central nervous system-directed effects of FTY720 (fingolimod). AB - FTY720, also known as fingolimod, is an orally administered sphingosine-1 phosphate (S1P) analogue that is under investigation as a therapy for both relapsing-remitting (RR) and progressive forms of multiple sclerosis (MS). The demonstrated beneficial effect of FTY720 on disease activity in RR-MS patients and in the animal model experimental autoimmune encephalomyelitis (EAE) is largely attributed to effects on the systemic immune system. In addition, unlike other current systemic immuno-modulators used in MS, the lipophilic nature of FTY720 allows it to cross the blood-brain barrier (BBB). Since S1P receptors are expressed on all cell types, FTY720 has the potential to exert effects directly on the BBB and on resident cells of the CNS. The latter include cells implicated in regulating immune reactivity within the CNS (astrocytes, microglia), those that are targeted by the disease process (oligodendrocytes, neurons), and those involved in repair (oligodendrocyte progenitor cells). In vitro studies document the dose-dependent effects of FTY720 on neural cell survival, differentiation, and cytoskeletal dynamics. Animal model studies, specifically EAE, indicate an overall neuroprotective effect of FTY720 mediated at least in part by its actions within the CNS. Ongoing studies will need to define the direct and indirect (via immune-modulation) effects of FTY720 on the CNS across the broad clinical spectrum of MS. PMID- 18678378 TI - Influence of statins treatment on survival in patients with amyotrophic lateral sclerosis. AB - BACKGROUND: Statins are increasingly recognized as causing muscle damage and, more rarely, peripheral neuropathy. A preliminary report that there are more cases of amyotrophic lateral sclerosis (ALS) among people treated with statins caused considerable concern. We considered the possibility that statins could affect survival in patients already diagnosed as having amyotropic lateral sclerosis who were taking statins for dyslipidemia. METHODS: We reviewed the clinical charts of 459 patients with ALS followed-up in our clinic between 1997 and 2007. Retrieved data included age, all administered medications, form of ALS at onset and survival. We compared the survival rates of patients taking any statins with that of patients not taking statins, while adjusting for other factors which influence disease progression, such as age, gender and ALS form at onset. RESULTS: 72 patients were on statins for dyslipidemia at disease onset. The doses ranged from 10-60 mg daily and varied throughout the disease course. As expected, the patients on statins were older than the non-treated ones (65.7+/-9 versus 57.5+/-13 years, respectively). After correcting for age, gender and disease form, there was no significant difference in survival between the groups. CONCLUSION: Our findings indicate that statins treatment for dyslipidemia in patients with ALS does not carry any survival risks. PMID- 18678379 TI - Serum vitamin B12 deficiency and hyperhomocystinemia: a reversible cause of acute chorea, cerebellar ataxia in an adult with cerebral ischemia. AB - Patients with vitamin B12 deficiency have protean neurological manifestations that are often insidious. Acute onset of cerebellar dysfunction and extrapyramidal manifestations like dystonia and chorea are rather uncommon in adults. We describe a patient who manifested with acute onset of language dysfunction, chorea and ataxia. There was no history of hypertension, diabetes or ischemic heart disease. He had low serum vitamin B12 and elevated serum homocystine levels. He improved dramatically following B12 replacement therapy. Our patient provides insight into the pathophysiological mechanism of this rare manifestation. Further the importance of considering vitamin B12 deficiency, in country like India, where vegetarian food practice is quite common, is being emphasized. PMID- 18678380 TI - Further observations in an area with an exceptionally low prevalence of Helicobacter pylori infection. PMID- 18678381 TI - Low validity of caretakers' reports on use of selected antimalarials and antibiotics in children with severe pneumonia at an urban hospital in Uganda. AB - Febrile children in low-income countries receive care from multiple sources, and caretakers' ability to report drug intake is crucial for appropriate prescription of drugs when reaching health facilities. This study describes and validates caretakers' reported use of sulfamethoxazole, chloroquine and sulfadoxine in their children. We performed a cross-sectional study in 139 children diagnosed with severe pneumonia at hospital in Kampala, Uganda. Caretakers were interviewed regarding treatments given prior to arrival at the hospital. Reported drug intake was compared to drug levels in blood sampled on filter paper, analyzed by HPLC methods. Caretakers under-reported intake of the studied drugs. Positive and negative predictive values were 67 and 64% for sulfamethoxazole, 69 and 52% for chloroquine and 85 and 62% for sulfadoxine. Many caretakers were unaware of what drug had been given to the child, and more so if treated outside the home (risk ratio 2.6, 95% CI 1.2-5.6). We conclude that caretakers' reports of drug intake have limited validity. Health workers need to improve counseling of caretakers during drug dispensing, especially for antibiotics. The roles and names of different drugs should be emphasized during counseling, and existing information systems such as immunization cards should be considered for record-keeping of treatment given. PMID- 18678382 TI - Denoising of quadrature ultrasound Doppler signal from bi-directional flow based on matching pursuit. AB - Denoising of Doppler signal is a preliminary and important step in medical ultrasound imaging. To denoise quadrature Doppler signal from bi-directional flow, we propose a novel method based on matching pursuit in this paper. The proposed method is an iterative decomposition algorithm which decomposes the original Doppler signal into a linear expansion of atoms in a time-frequency dictionary. The time-frequency dictionary is similar to Fourier transform domain and the atoms are similar to orthogonal bases in Fourier transform. In each step of the iteration, the atom which gives the largest inner product with the analyzed signal is selected from the dictionary, and the contribution of this atom is subtracted from the Doppler signal. This process is repeated on the residue until the SNR reaches the maximum. The linear expansion of the selected atoms is the denoised signal. Simulations were conducted on a simulation model with a sampling rate of 12.8 kHz. When the original SNRs are 0 dB, 2dB, 4 dB, 6 dB, 8 dB, 10 dB, the proposed method can improve the SNR for 7.9 dB, 7.8 dB, 7.5 dB, 7.3 dB, 7.05 dB, 6.8 dB respectively, reduce the root mean square error (RMSE) of the mean frequency waveform to 0.0441 kHz, 0.0303 kHz, 0.0245 kHz, 0.0215 kHz, 0.0161 kHz, 0.0125 kHz respectively, and suppress the RMSE of the spectral width waveform to 0.1774 kHz, 0.0591 kHz, 0.0486 kHz, 0.0170 kHz, 0.0145 kHz, 0.0117 kHz respectively. Preliminary in vivo evaluation was also carried out on a healthy 33-year-old male using B-K medical A/S 3535 ultrasound scanner, and the results showed that the proposed method can effectively enhance the Doppler spectrogram. PMID- 18678383 TI - Human T-cell leukemia virus type 1 Tax modulates interferon-alpha signal transduction through competitive usage of the coactivator CBP/p300. AB - We describe here Tax protein of human T-cell leukemia virus type 1 (HTLV-1) as an interferon (IFN)-alpha antagonist counteracting the transactivation function of IFN-stimulated gene factor 3 (ISGF3). Co-expression of Tax, but not the Tax mutant unable to bind to CBP, significantly inhibited the reporter gene expression directed by IFN-stimulated regulatory elements, despite that the formation of DNA-binding ISGF3 complex was unaffected. Gene activation induced by STAT2 transcription domain was also inhibited by expression of Tax. Furthermore, Tax-mediated transcriptional inhibition was reversed by overexpression of p300. These observations indicate that Tax interferes with IFN-alpha-induced JAK-STAT pathway by competition with STAT2 for CBP/p300 binding. Consistently, GST pull down assay showed that Tax dose-dependently inhibited binding of STAT2 to p300. This study suggests that Tax may prevent IFN-alpha from exerting its antiviral, antiproliferative and proapoptotic effects, thereby contributing to persistent viral infection and HTLV-1-associated oncogenesis. PMID- 18678384 TI - Proteolytic processing of polyproteins 1a and 1ab between non-structural proteins 10 and 11/12 of Coronavirus infectious bronchitis virus is dispensable for viral replication in cultured cells. AB - Coronavirus 3C-like proteinase (3CLpro) plays important roles in viral life cycle through extensive processing of the polyproteins 1a and 1ab into 12 mature, non structural proteins (nsp5-nsp16). Structural and biochemical studies have revealed that all confirmed 3CLpro cleavage sites have a conserved Gln residue at the P1 position, which is thought to be absolutely required for efficient cleavage. Recent studies on murine hepatitis virus (MHV) showed that processing of the 1a polyprotein at the position between nsp10-nsp11 is essential for viral replication. In this report, we investigated the requirement of processing at the equivalent position for replication of avian coronavirus infectious bronchitis virus (IBV), using an infectious cloning system. The results showed that mutation of the P1 Gln to Pro or deletion of the Gln residue in the nsp10-nsp11/12 site completely abolished the 3CLpro-mediated processing, but allowed production of infectious recombinant viruses with variable degrees of growth defect, suggesting that cleavage at the nsp10-nsp11/12 site of IBV is dispensable for viral replication in cultured cells. This study would pave a way for potential vaccine development by generation of attenuated IBV from field isolates through manipulation of the nsp10-nsp11/12 cleavage site. Similar approaches would be also applicable to other human and animal coronaviruses. PMID- 18678386 TI - Characterising humic acid fouling of nanofiltration membranes using bisphenol A as a molecular indicator. AB - Fouling of nanofiltration (NF) membranes by humic acids was investigated using bisphenol A (BPA) as an indicator chemical to differentiate between various mechanisms that may lead to a change in solute rejection. Three commercially available NF membranes were investigated and an accelerated fouling condition was achieved with a foulant mixture containing humic acids in an electrolyte matrix. The effects of membrane fouling on the rejection of BPA were interpreted with respect to the membrane pore sizes and the fouling characteristics. Results reported here indicate that calcium concentration in the feed solution could be a major factor governing the humic acid fouling process. Moreover, a critical concentration of calcium in the feed solution was observed, at which membrane fouling was most severe. Membrane fouling characteristics were observed by their influence on BPA rejection. Such influence could result in either an increase or decrease in rejection of BPA by the three different membranes depending on the rejection mechanisms involved. It is hypothesized that these mechanisms could occur simultaneously and that the effects of each might not be easily distinguished. However, it was observed that their relative contribution was largely dependent upon membrane pore size. Pore blocking, which resulted in a considerable improvement in rejection, was prominent for the more open pore size TFC-SR2 membrane. In contrast, the cake-enhanced concentration polarisation effect was more severe for the tighter NF-270 and NF-90 membranes. For hydrophobic solutes such as BPA, the formation of the fouling layer could also interfere with the solute-membrane interaction, and therefore, exert considerable influence on the separation process. PMID- 18678385 TI - Determinants of cyclophilin A-dependent TRIM5 alpha restriction against HIV-1. AB - TRIM5 alpha is a host protein that can bind to incoming retroviral capsid (CA) and inhibit retroviruses in a species-specific manner. The CA protein of HIV-1 also interacts with high affinity to the host protein cyclophilin A (CypA). This binding has been shown to positively affect some early stage of the viral life cycle in human cells. However, the CypA/CA interaction also renders HIV-1 more susceptible to rhesus TRIM5 alpha (rhTRIM5 alpha) restriction. We find that the ability of old world monkey TRIM5 alpha genes to restrict HIV-1 in a CypA dependent manner is widespread. On the other hand, we find that simian immunodeficiency viruses from tantalus monkeys (SIVagmTAN), is unlike HIV-1 in that CypA does not enhance the rhTRIM5 alpha restriction against the virus even though the CA of this virus, like HIV-1, does bind CypA. Mapping of the determinants for this phenotype by swapping regions on CA between SIVagmTAN and HIV-1 showed that when SIVagmTAN contains loops between helices 4/5 (4-5 loop) and 6/7 (6-7 loop) from HIV-1 CA, it becomes susceptible to the CypA-enhanced rhTRIM5 alpha restriction. Surprisingly, when SIVagmTAN contains either loop from HIV-1 CA, it gains sensitivity to TRIM5 alpha from species which originally have no effect on the wild-type virus. Moreover, we find that CypA/CA interaction occurs early after viral entry but the CypA-enhanced restriction mostly acts on the stage after reverse transcription. PMID- 18678387 TI - Diversity and antifungal susceptibility of yeasts from tropical freshwater environments in Southeastern Brazil. AB - Yeast communities were isolated from water and sediment samples of two unpolluted natural lakes, located inside Rio Doce State Park, and two rivers located outside of this Park in Southeastern Brazil. A total of 134 yeast isolates were obtained and identified as belonging to 36 species. The numbers of fecal coliforms and yeast species were higher in rivers than in lakes. The genus Candida had the highest number of species with the presence of opportunistic pathogens such as Candida krusei, C. tropicalis, C. guilliermondii and C. parapsilosis. Yeasts able to grow at 37 degrees C were tested in relation to their susceptibility to common used antifungal drugs. Yeast isolates (13%) were susceptible to ketoconazole, 79% to fluconazole, 31% to terbinafine and 78% of the strains were susceptible to amphotericin B. Seven isolates from different Candida species were resistant to all antifungals tested. The high number of fecal coliforms found in these aquatic environments and the presence of resistant yeast strains to common used antifungal drugs suggest that these environments can pose potential health risks for people utilizing the contaminated waters. PMID- 18678388 TI - Improvement of chromium biosorption by UV-HNO(2) cooperative mutagenesis in Candida utilis. AB - The present study focused on the improvement of chromium resistance and biosorption efficiency in Candida utilis CR-001 utilizing protoplast mutagenesis technology. Through ultraviolet (UV) radiation, HNO(2) treatment and chromium acclimatization, six preferred mutants of C. utilis CR-001 were screened out, namely, CRU132-26, CRC7-2, CRC2811-1, CRC2811-2, CRC2814-8 and CRY182-1. The removal efficiency of these mutants for 20mg/L Cr(VI) solutions were 85.6%, 95.2%, 87.0%, 82.5%, 94.7% and 82.7%, respectively, noticeably greater than that of the parent strain CR-001 (79.5%). Furthermore, CRC2811-1 exhibited outstanding application potential with high removal efficiency and low dosage over a wide range of pH. Cell surface and inner details of CRC2811-1 and its parent strain CR 001 were analyzed by scanning electron microscopy (SEM) and atomic force microscopy (AFM) in order to explore possible changes caused by inducement. The results showed that Cr-sorption of CR-001 mainly depended on intracellular accumulation, but for CRC2811-1, cell surface deposition was also involved in improving its chromium biosorption capacity. PMID- 18678389 TI - Effect of total solids on fecal coliform regrowth in anaerobically digested biosolids. AB - Fecal coliform (FC) concentrations in anaerobically digested biosolids can increase during centrifugal dewatering and afterwards in storage of dewatered cake. The immediate increase after centrifugation (reactivation) has been demonstrated to be the revitalization of fecal coliforms that had become non culturable. The increase during storage (regrowth) has been regarded as a subsequence of reactivated bacteria growing in a favorable environment. In this paper, however, regrowth is demonstrated without preceding reactivation, using intensive laboratory centrifugation to duplicate the levels of regrowth seen in full-scale centrifugation. Higher total solids (TS) levels of the dewatered biosolids lead to greater magnitudes of FC increase. The final TS level appears much more important than the level of shear imposed during centrifugation, based on comparison of different centrifugation/dilution procedures used to obtain similar TS levels. The greater TS levels also reduce methane production, suggesting that methanogens compete with, or inhibit, the fecal coliforms. The addition of bromoethanesulfonate as a methanogen-specific inhibitor decreased the production of methane gas, and also increased the number of fecal coliforms. PMID- 18678390 TI - Seasonal variations of monoterpene emissions from Pinus densiflora in East Asia. AB - The emission rates and compositions of monoterpene from Pinus densiflora were investigated in the Gumsung (GM) and Worak (WM) mountains. The standard emission rates (ERs: ERs is the monoterpene emission rate at standard temperature, 30 degrees C) from P. densiflora ranged from 0.817 to 1.704 (microgC/gdw-h). The ERs and beta-values of total monoterpene were measured at the two study sites (GM and WM). In the spring and summer, the ERs were the highest, while relatively low values (<0.058microgC/gdw-h) were measured in the autumn and winter. In GM and WM sites the beta-value obtained for the different seasons ranged from 0.047 to 0.179, with an average of 0.09. The major monoterpene compounds from P. densiflora were alpha-pinene, myrcene, beta-phellandrene, d-limonene and alpha terpinene. The fractional compositions of individual monoterpene compounds were significantly different between the two test sites in the summer and winter. The ERs of the older group (31-40 years) were higher than those in the younger group (21-30 years). However, the monoterpene compositions were similar between the two age groups. PMID- 18678391 TI - The application of novel coagulant reagent (polyaluminium silicate chloride) for the post-treatment of landfill leachates. AB - Relatively "old" (stabilized) landfill leachates are a special category of wastewaters, which are difficult to treat further, mainly due to their bio refractory organic content (humic substances). In this study, coagulation flocculation was examined as post-treatment method for the biologically pre treated stabilized leachates. The purpose was to examine the coagulation performance of alternative coagulant agents, i.e. the composite coagulant polyaluminium silicate chloride. Composite coagulants with different Al to Si molar ratio and different preparation methods were tested. Their efficiency was evaluated by monitoring from turbidity and phosphate content, other parameters strongly correlated with the presence of organic matter, such as UV absorbance at 254nm, COD and colour. The results suggest that the silica-based coagulants exhibit better coagulation performance, than the relevant conventional coagulant (alum) or simple pre-polymerized coagulants (PACl). Polyaluminium silicate chloride has greater tolerance against pH variation than alum or PACl, whereas this novel coagulant works better at pH values between 7 and 9. Coagulation flocculation has proved to be an efficient post-treatment method for the biologically pre-treated leachates, promoting the removal of the refractory humic substances, while the treatment efficiency of coagulation can be improved by the application of the new coagulant agent. PMID- 18678392 TI - Art and the Middle-to-Upper Paleolithic transition in Europe: comments on the archaeological arguments for an early Upper Paleolithic antiquity of the Grotte Chauvet art. AB - The spectacular art of the Grotte Chauvet stands out among all other examples of Aurignacian art, which are restricted to a handful of sites in other regions of western and Central Europe, which take the form of sophisticated carvings on organic materials and of simple engravings on rockshelter walls. Given its sophistication, Chauvet has understandably come to feature prominently in debates as to the nature of human symbolic origins, the behavioral capacities of Homo sapiens, the nature of the dispersal of modern humans across Europe, and the possibly contemporary extinction of Homo neanderthalensis. Significant objections to such an antiquity have, however, been made in recent years on the grounds of the style, themes, and technical practice of the art itself, and on the grounds of the AMS radiocarbon dating program that was first seen to suggest an early Upper Paleolithic age. To date, no attention has been paid to claims for an Aurignacian age on specifically archaeological grounds. Here, I undertake a critical examination of the archaeology of the cave and its wider region, as well as attempts to verify the antiquity of the art on the basis of comparison with well-dated Aurignacian art elsewhere. I conclude that none of the archaeological arguments withstand scrutiny and that many can be rejected as they are either incorrect or tautologous. By contrast, hypotheses that the art is of Gravettian Magdalenian age have not been successfully eliminated. The age of the art of the Grotte Chauvet should be seen as a scientific problem, not an established fact. While it may prove impossible to prove an Aurignacian age for some of the Chauvet art I suggest a set of expectations that would, in combination, strengthen the robusticity of the 'long chronology' argument. The onus is upon Chauvet long chronologists to do this, and until they do, we must conclude that the art of the Grotte Chauvet is not dated, and very possibly much younger than claimed. PMID- 18678393 TI - Early Man: a tribute to the late career of F. Clark Howell. PMID- 18678394 TI - A reappraisal of the terrestrial nitrogen cycle: what can we learn by extracting concepts from Gaia theory? AB - Although soil scientists and most environmental scientists are acutely aware of the interactions between the cycling of carbon and nitrogen, for conceptual convenience when portraying the nitrogen cycle in text books the N cycle tends to be considered in isolation from its interactions with the cycling of other elements and water, usually as a snap shot at the current time; the origins of dinitrogen are rarely considered, for example. The authors suggest that Lovelock's Gaia hypothesis provides a useful and stimulating framework for consideration of the terrestrial nitrogen cycle. If it is used, it suggests that urbanization and management of sewage, and intensive animal rearing are probably bigger global issues than nitrogen deposition from fossil fuel combustion, and that plant evolution may be driven by the requirement of locally sustainable and near optimal soil mineral N supply dynamics. This may, in turn, be partially regulating global carbon and oxygen cycles. It is suggested that pollutant N deposition may disrupt this essential natural plant and terrestrial ecosystem evolutionary process, causing biodiversity change. Interactions between the Earth and other bodies in the solar system, and possibly beyond, also need to be considered in the context of the global N cycle over geological time scales. This is because of direct potential impacts on the nitrogen content of the atmosphere, potential long-term impacts of past boloid collisions on plate tectonics and thus on global N cycling via subduction and volcanic emissions, and indirect effects upon C, O and water cycling that all may impact upon the N cycle in the long term. PMID- 18678395 TI - Potential problems with environmental sound barriers when used in mitigating surface transportation noise. AB - The public, increasingly well-informed about the problem of excessive noise, is taking actions for the development of new transport infrastructure projects and improvement of existing infrastructure. In addition, many countries have implemented mandatory Environmental Impact Assessment procedures. As a result, the construction of sound barriers has become a common measure, which can be used by an agency to mitigate potentially significant noise impacts. A sound barrier, eventually, will become part of the surrounding landscape and could be a cause of impact for ecosystems, the road users and those who live alongside the road. Basically, this article discusses these potential effects in the context of environmental assessment procedures. In addition, results of a pilot survey conducted at a residential area affected by the construction of a barrier are presented. Although most residents felt that sleeping conditions improved after the barrier was built, most important negative reactions are the loss of sunlight and visual impact. PMID- 18678396 TI - Effects of platinum from vehicle exhaust catalyst on carbon and nitrogen mineralization in soils. AB - There is strong evidence of continuously increasing contamination of soils with platinum group elements (PGE), in particular with platinum (Pt) from vehicle exhaust catalysts in roadside soils. However, knowledge about the effects of Pt contamination on soil processes is very limited. The objective of this study was to investigate whether the contamination of soils with Pt at realistic environmental levels leads to observable adverse effects on selected indicator parameters of the carbon and nitrogen turnover in soils. Incubation experiments with artificially contaminated soils and solutions containing dissolved organic matter (DOM) were carried out by the use of milled material from a Pt-containing vehicle exhaust catalyst. Interaction of the catalyst material with the soil resulted in a mobilization of Pt into the dissolved phase reaching up to 0.1% of the added Pt. The amount of Pt mobilization seemed to be mainly driven by the pH of the soil. Mineralization of carbon and nitrogen did not reveal any significant adverse effect of the Pt addition as compared to the control samples. Future studies dealing with Pt effects on soil processes should focus on environmental conditions favoring Pt mobilization, e.g. such as very low pH values or large concentrations of DOM. PMID- 18678397 TI - Lead shot contribution to blood lead of First Nations people: the use of lead isotopes to identify the source of exposure. AB - Although lead isotope ratios have been used to identify lead ammunition (lead shotshell pellets and bullets) as a source of exposure for First Nations people of Canada, the actual source of lead exposure needs to be further clarified. Whole blood samples for First Nations people of Ontario, Canada, were collected from participants prior to the traditional spring harvest of water birds, as well as post-harvest. Blood-lead levels and stable lead isotope ratios prior to, and after the harvest were determined by ICP-MS. Data were analyzed by paired t-tests and Wilcoxon Signed-Ranks tests. All participants consumed water birds harvested with lead shotshell during the period of study. For the group excluding six males who were potentially exposed to other sources of lead (as revealed through a questionnaire), paired t-tests and Wilcoxon Signed-Ranks tests showed consistent results: significant (p<0.05) increases in blood-lead concentrations and blood levels of (206)Pb/(204)Pb and (206)Pb/(207)Pb towards the mean values we previously reported for lead shotshell pellets; and a significant decrease in (208)Pb/(206)Pb values towards the mean for lead shotshell pellets. However, when we categorized the group further into a group that did not use firearms and did not eat any other traditional foods harvested with lead ammunition other than waterfowl, our predictions for (206)Pb/(204)Pb, (206)Pb/(207)Pb and (208)Pb/(206)Pb hold true, but there was not a significant increase in blood-lead level after the hunt. It appears that the activity of hunting (i.e., use of a shotgun) was also an important route of lead exposure. The banning of lead shotshell for all game hunting would eliminate a source of environmental lead for all people who use firearms and/or eat wild game. PMID- 18678398 TI - Reversal of vitamin K antagonist-associated coagulopathy: a survey of current practice. PMID- 18678399 TI - Novel chemotherapy approaches in chemoradiation protocols. AB - Locally advanced cervical carcinoma had been treated with radiation therapy until 1999, when five different large clinical trials showed an overall survival benefit when chemotherapy was administered concomitantly with radiotherapy. The chemotherapy agents used in these trials were cisplatin, cisplatin combined with fluorouracil or hydroxyurea. Weekly cisplatin (40 mg/m(2)) achieved the best responses, even when compared with the combination with fluorouracil. These results led the United States National Cancer Institute (NCI) to recommend platinum-based chemotherapy for the treatment of locally advanced cervical carcinoma. Other cytotoxic agents have been tried in combination with radiotherapy for the management of the disease, including carboplatin, paclitaxel, gemcitabine and even topotecan. Gemcitabine has shown promising results and the combination of paclitaxel and carboplatin has proved safe and effective. However, to date, there has been no agent or combination of agents to have shown superiority over weekly cisplatin. Biologic agents such as bevacizumab, cetuximab, sorafenib and erlotinib are currently being tried in different trials in combination with radiotherapy and cisplatin. Celecoxib, a COX 2 inhibitor was evaluated in an RTOG study in combination with cisplatin and flourouracil with radiation therapy with no apparent effect on DFS and poor rates of locoregional control. Chemoradiation is the current standard therapy in locally advanced cervical carcinoma. The integration of novel agents will be established by the ongoing clinical trials. PMID- 18678400 TI - SV40 early genes induce neoplastic properties in serous borderline ovarian tumor cells. AB - OBJECTIVE: Serous borderline ovarian tumors (SBOT) are slow growing, noninvasive ovarian epithelial neoplasms, which tend to recur as low-grade invasive carcinomas (LGC) with a much worse prognosis. We investigated the molecular basis of this progression. METHODS: We established cultures of three SBOTs and one LGC from tumor biopsies, and inactivated p53, Rb and PP2A in the cells with SV40 large T (LT) and small T (ST) antigen. They were examined for cadherins by immunofluorescence and immunoblotting, invasiveness in Boyden chambers, motility by scratch-wound healing assay, anchorage independence by growth in agarose, and protease activity by gelatin zymography, immunoassay and colorimetry. Cells were overexpressed with N-cadherin using an adenovirus. RESULTS: Inactivation of p53, Rb and PP2A by SV40 LT/ST antigen resulted in greatly enhanced growth potential, invasiveness, motility and anchorage independence, and in epithelio-mesenchymal transition, as indicated by morphology and substitution of N-cadherin for E cadherin. Overexpressed N-cadherin did not induce invasiveness of SBOT cells and there was no consistent change in protease activities, suggesting that these were not primary effectors of the enhanced neoplastic characteristics. Low passage LGC cells were more invasive than SBOT cells, but this difference disappeared with the introduction of LT/ST into the two cell types. CONCLUSION: Downregulation or inactivation of p53, Rb and/or PP2A plays a role in the progression from SBOT to invasive ovarian carcinomas. PMID- 18678401 TI - Oestrus cycle characteristics and prediction of ovulation in Catalonian jennies. AB - The Catalonian donkey breed is in danger of extinction, and much needs to be learned about the reproductive features of its females if breeding and conservation programmes are to be successful. This study reports the oestrous behaviour, oestrus cycle characteristics and dynamic ovarian events witnessed during 50 oestrous cycles (involving 106 ovulations) in 10 Catalonian jennies between March 2002 and January 2005. These jennies were teased, palpated transrectally and examined by ultrasound using a 5 MHz linear transducer-daily during oestrus and every other day during dioestrus. Predictors of ovulation were sought among the variables recorded. The most evident signs of oestrus were mouth clapping (the frequent vertical opening and closing of the mouth with ears depressed against the extended neck) and occasional urinating and winking of the vulval lips (homotypical behaviour). Interactions between jennies in oestrus were also recorded, including mounting, herding/chasing, the Flehmen response, and vocalization (heterotypical behaviour). Nine jennies ovulated regularly throughout the year; one had two anovulatory periods (54 and 35 days). The length of the oestrus cycle was 24.90 +/- 0.26 days, with oestrus itself lasting 5.64 +/ 0.20 days (mean +/- S.E.M.) and dioestrus 19.83 +/- 0.36 days. The incidence of single, double and triple ovulations was 55.66% (n=59), 42.45% (n=45) and 1.89% (n=2), respectively. No significant difference was seen in the number of ovulations involving the left and right ovaries (52.63% [n=70] compared to 47.37% [n=63] respectively; P>0.05). The mean interval between double ovulation was 1.44 +/- 3.98 days. The mean diameter of the preovulatory follicle at day -1 was 44.9 +/- 0.5 mm; the mean growth rate over the 5 days before ovulation was 3.7 mm/day. Data on preovulatory changes in oestrous behaviour, follicle size, follicle texture, the echographic appearance of the follicle and uterus, and uterine tone were subjected to stepwise logistic regression analysis to detect predictors of ovulation. The logit function showed the best predictors to be follicle size, follicular texture and oestrous behaviour. Certain combinations of these three variables allow the prediction of ovulation within 24 h with a probability of >75%. PMID- 18678402 TI - The Anterior Midline Field: coercion or decision making? AB - To study the neural bases of semantic composition in language processing without confounds from syntactic composition, recent magnetoencephalography (MEG) studies have investigated the processing of constructions that exhibit some type of syntax-semantics mismatch. The most studied case of such a mismatch is complement coercion; expressions such as the author began the book, where an entity-denoting noun phrase is coerced into an eventive meaning in order to match the semantic properties of the event-selecting verb (e.g., 'the author began reading/writing the book'). These expressions have been found to elicit increased activity in the Anterior Midline Field (AMF), an MEG component elicited at frontomedial sensors at approximately 400 ms after the onset of the coercing noun [Pylkkanen, L., & McElree, B. (2007). An MEG study of silent meaning. Journal of Cognitive Neuroscience, 19, 11]. Thus, the AMF constitutes a potential neural correlate of coercion. However, the AMF was generated in ventromedial prefrontal regions, which are heavily associated with decision-making. This raises the possibility that, instead of semantic processing, the AMF effect may have been related to the experimental task, which was a sensicality judgment. We tested this hypothesis by assessing the effect of coercion when subjects were simply reading for comprehension, without a decision-task. Additionally, we investigated coercion in an adjectival rather than a verbal environment to further generalize the findings. Our results show that an AMF effect of coercion is elicited without a decision-task and that the effect also extends to this novel syntactic environment. We conclude that in addition to its role in non-linguistic higher cognition, ventromedial prefrontal regions contribute to the resolution of syntax semantics mismatches in language processing. PMID- 18678403 TI - In vitro and in vivo suppression of cellular activity by guanidinoethyl disulfide released from hydrogel microspheres composed of partially oxidized hyaluronan and gelatin. AB - This paper describes the preparation of oxidized hyaluronan crosslinked gelatin microspheres for drug delivery. Microspheres were prepared by a modified water-in oil-emulsion crosslinking method, where three-dimensional crosslinked hydrogel microspheres formed in the absence of any extraneous crosslinker. SEM analyses of the microspheres showed rough surfaces in their dried state with an average diameter of 90 microm. Lyophilization of fully swollen microspheres revealed a highly porous structure. Guanidinoethyl disulfide (GED) was used as a model drug for incorporation into the microspheres; encapsulation of GED was confirmed by HPLC. There was an inverse correlation between the diameters of the microspheres with their GED loading. Macrophage was used as a model cell to evaluate the in vitro efficacy of GED release from the microspheres. The in vivo efficacy of the microspheres was further validated in a mouse full-thickness transcutaneous dermal wound model through suppression of cell infiltration. PMID- 18678404 TI - Surface antimicrobial activity and biocompatibility of incorporated polyethylenimine nanoparticles. AB - The antimicrobial effect and biocompatibility of insoluble cross-linked quaternary ammonium polyethylenimine (PEI) nanoparticles incorporated at 1 or 2%w/w in a resin composite were assayed. The antimicrobial effect against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli was tested using the direct contact test (DCT), agar diffusion test (ADT) and scanning electron microscopy (SEM). Biocompatibility was tested by assessing macrophage viability, and TNFalpha secretion. Samples incorporating 2%w/w nanoparticles inhibited the growth of all bacterial strains tested. Reducing the amount of the added nanoparticles to 1%w/w resulted in complete inhibition of S. aureus and E. faecalis, and decreased growth of S. epidermidis, P. aeruginosa and E. coli (p<0.0001). The DCT results were confirmed by SEM. However, ADT showed no inhibition halo in all test bacteria, indicating the antimicrobial nanoparticles are not diffusing into the agar milieu. Biocompatibility tests revealed macrophage viability, and TNFalpha secretion was not altered by the presence of the nanoparticles in the resin. Incorporation of PEI nanoparticles in a resin composite had a long lasing antimicrobial effect against a wide range of bacteria with no measured effect on biocompatibility. PMID- 18678405 TI - Cell adhesion and proliferation on hydrophilic dendritically modified surfaces. AB - Dendritically modified, or "dendronized" surfaces are generated by modification of a substrate with perfectly branched polymers, known as dendrimers. Here, such dendronized surfaces were prepared by initial chemisorption of poly(ethylene glycol)-mono-thiol (HS-PEG(650)-OH) onto gold-coated silicon wafers, followed by divergent synthesis of aliphatic polyester dendrons, generation 1-4, starting from the terminal PEG OH- group. The adhesion and proliferation of human corneal epithelial cells (HCEC) and mouse 3T3 fibroblasts (M-3T3) as model cells on these hydroxyl-terminated dendronized surfaces were investigated. In addition, the effect of covalently attaching PEG mono-methyl ether (PEG-OMe) chains (M(n)=2000 Da) to the peripheral hydroxyl groups of G1- and G2-dendronized surfaces on adhesion and proliferation of the same cell lines was studied. Little or no HCEC adhesion was noted on gold surfaces modified with PEG mono-thiol (HO-PEG-SH) in serum-free medium. These cells showed a greater affinity for the dendronized surfaces compared to the control Au surfaces at early incubation stages (1 day). At longer incubation times, HCEC proliferation increased exponentially on the dendronized surfaces. However, when G1- and G2-dendronized surfaces were modified with PEG-OMe chains, adhesion of both HCEC and M-3T3 cells was significantly reduced. Cell studies with M-3T3 fibroblasts, carried out in serum-containing medium, showed that cell attachment was diminished for the PEG-grafted Au surfaces compared to the control Au and G1-G4 dendronized surfaces. PMID- 18678406 TI - The effect of polypyrrole on arteriogenesis in an acute rat infarct model. AB - The conductive polymer polypyrrole was blended with alginate to investigate its potential in tissue engineering applications. This study showed that increasing the polypyrrole content altered the macroscopic structural morphology of the polymer blend scaffold, but did not alter the overall conductivity of the polymer blend, which was 10(-2)S/cm(2). Culturing of human umbilical vein endothelial cells on the polymer blend scaffolds showed that addition of polypyrrole mediated cell attachment to the polymer scaffold. However, cell proliferation was dependent on the polypyrrole content with 0.025% v/v polypyrrole giving the best results. Using an ischemia-reperfusion rat myocardial infarction model, local injection of 0.025% polypyrrole in alginate polymer blend into the infarct zone yielded significantly higher levels of arteriogenesis at 5 weeks post-treatment when compared with the saline control group and the alginate only treatment group. In addition, this alginate-polypyrrole polymer blend significantly enhanced infiltration of myofibroblasts into the infarct area when compared with the control group. The results of this study highlight the potential clinical benefit of using this alginate-polypyrrole polymer blend as an injectable scaffold to repair ischemic myocardium after myocardial infarction. PMID- 18678407 TI - Antioxidant effect of diphenyl diselenide on oxidative damage induced by smoke in rats: involvement of glutathione. AB - In the present study, the involvement of glutathione system in the restorative effect of diphenyl diselenide (PhSe)(2) on damage induced by cigarette smoke was investigated. Rat pups were progressively exposed to four, five, and six cigarettes for exposure periods of 15 min during their first, second, and third weeks of life. Thiobarbituric acid reactive species (TBARS) levels, components of the enzymatic antioxidant defenses (superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S transferase (GST) activities), and non-enzymatic antioxidant defenses (vitamin C and non-protein thiol (NPSH) levels) were examined in lungs of pups. The results demonstrated an increase in lipid peroxidation and the alteration in non enzymatic and enzymatic antioxidant defenses induced by cigarette smoke exposure in lung of pups. Administration of (PhSe)(2) (0.5mg/kg) restored TBARS levels and antioxidant defenses in lungs of rat pups exposed to cigarette smoke. (PhSe)(2) treatment increased NPSH levels and GST activity per se in lungs of rat pups. Together these results indicate that (PhSe)(2) restored oxidative damage induced by cigarette smoke exposure in lungs of rat pups. The glutathione system is involved in antioxidant effect of this compound. PMID- 18678408 TI - Women who kill: a comparison of the psychosocial background of female and male perpetrators. AB - The purpose of this study was to compare the psychosocial background of female and male perpetrators convicted of homicide in Sweden between 1995-2001. All women (n=43) who were convicted for lethal violence during the period and a corresponding number of randomly chosen men (n=43) were examined. In total, information about 86 individuals was collected retrospectively. Areas of interest were psychosocial variables during childhood and at the time of the current crime. Results showed that both female and male perpetrators were psychosocially encumbered already at an early age. Homicidal women had more severe childhood circumstances, but less aggressive childhood behaviour than did their male counterparts. At the time of the crime, women had a more ordered social situation, had more often been exposed to violence and searched for help than had the men. These gender differences suggest that specific actions are needed for preventing women's homicidal behaviour. PMID- 18678409 TI - Evaluation of background persistent organic pollutant levels in human from Taiwan: polychlorinated dibenzo-p-dioxins, dibenzofurans, and biphenyls. AB - This study was intended to determine the background levels of PCDD/PCDFs and PCBs in the Taiwanese population and to investigate factors potentially related to PCDD/PCDF and PCB levels. The levels of seventeen PCDD/PCDFs in the 251 serum samples collected from the general population in Taiwan ranged from 4.92 to 26.7 pg WHO(1998)-TEQ/g lipid (median: 11.5) and those of the twelve dioxin-like PCBs ranged between 1.74 and 21.6 pg WHO(1998)-TEQ/g lipid (median: 6.14). Five factors, age, gender, region of residence, dietary status, and smoking status, showed statistically significant association with the TEQ level of PCDD/PCDFs. The TEQ level of PCBs was statistically associated with age only, but not with the other four factors. The trends observed between age and the levels of PCDD/PCDFs and PCBs were not parallel in young subjects (<30 years old) and old subjects (>30 years old). The levels of PCDD/PCDFs and PCBs increased by 0.16 and 0.03 WHO(1998)-TEQ/g lipid per year for subjects above the age of 30, but there was no evidence of any association between age and the levels for subjects below the age of 30 years. These factors should be considered when investigating relationships between background serum levels of persistent organic pollutants and parameters associated with exposure sources or health outcomes. PMID- 18678410 TI - Occupational exposure of farm workers to pesticides: biochemical parameters and evaluation of genotoxicity. AB - To assess the effects of exposure to complex mixtures of pesticides in farm workers from two communities from Rio Grande do Sul, Brazil, we evaluated the activities of butyrylcholinesterase (BChE) and delta-aminolevulinic acid dehydratase (ALA-D) enzymes, hematological, lipid parameters, and genotoxicity using two tests to detect DNA damage, the Comet assay in peripheral blood leukocytes and the micronucleus (MN) test in oral mucosa cells. The use of personal protective equipment (PPE), age and smoke habits were considered in the analysis. There was a significant decrease in the BChE and ALA-D activities in farm workers (n=37) relative to the control group (n=20) (P< or =0.05 and P< or =0.001, respectively). The Comet assay in peripheral blood leukocytes showed that the Damage index and Damage frequency observed in the exposed group were significantly higher in relation to the controls (P< or =0.001, and P< or =0.05, respectively). No differences were detected regarding the hematological parameters, lipids profile, and MN frequencies. In addition, no significant differences were observed between younger (< or =38 years) and older subjects (>38 years), or between smokers and non-smokers within the groups, either by Comet assay or MN test. However, the use of PPE seems to be important in the prevention of contamination, as suggested by BChE levels and Comet assay results. PMID- 18678411 TI - Pollutants in particulate and gaseous fractions of ambient air interfere with multiple signaling pathways in vitro. AB - Traditionally, contamination of air has been evaluated primarily by chemical analyses of indicator contaminants and these studies have focused mainly on compounds associated with particulates. Some reports have shown that air contaminants can produce specific biological effects such as toxicity mediated by the aryl hydrocarbon receptor (AhR) or modulation of the endocrine system. This study assessed the dioxin-like toxicity, anti-/estrogenicity, anti-/androgenicity and anti-/retinoic activity of both the particulate and gas phase fractions of air in two regions with different types of pollution sources and a background locality situated in an agricultural area of Central Europe. The first region (A) is known to be significantly contaminated by organochlorine pesticides and chemical industry. The other region (B) has been polluted by historical releases of PCBs, but the major current sources of contamination are probably combustion sources from local traffic and heating. Samples of both particle and gas fractions produced dioxin-like (AhR-mediated) activity, anti-estrogenic and antiandrogenic effects, but none had any effect on retinoid signaling. AhR mediated activities were observed in all samples and the TEQ values were comparable in both fractions in region A, but significantly greater in the particulate fraction in region B. The greater AhR-mediated activity corresponded to a greater coincident antiestrogenicity of both phases in region B. Our study is the first report of antiestrogenicity and antiandrogenicity in ambient air. Anti-androgenicity was observed in the gas phase of all regions, while in the particulate phase only in one region due to the specific type of pollution in that area. Even though based on concentrations of individual compounds, except for the OCPs, the level of contamination of the two regions was similar, there were strong differences in responses in the bioassays between the two regions. Moreover, AhR-mediated activity and antiestrogenic potencies were greater in region B, where the pollution level according to the chemical analysis was similar or less than in the other region, which indicates the presence of other atmospheric pollutants with specific effects. The results document the advantage and utility of the simultaneous use of bioassays and chemical analysis in risk assessment of complex environmental samples. PMID- 18678412 TI - Intrafamilial outbreak of subcutaneous abscesses caused by PVL-positive methicillin-sensitive Staphylococcus aureus. PMID- 18678414 TI - Calcium channels, neuromuscular synaptic transmission and neurological diseases. AB - Voltage-dependent calcium channels are essential in neuronal signaling and synaptic transmission, and their functional alterations underlie numerous human disorders whether monogenic (e.g., ataxia, migraine, etc.) or autoimmune. We review recent work on Ca(V)2.1 or P/Q channelopathies, mostly using neuromuscular junction preparations, and focus specially on the functional hierarchy among the calcium channels recruited to mediate neurotransmitter release when Ca(V)2.1 channels are mutated or depleted. In either case, synaptic transmission is greatly compromised; evidently, none of the reported functional replacements with other calcium channels compensates fully. PMID- 18678413 TI - Hopelessness across phases of bipolar I or II disorder: a prospective study. AB - BACKGROUND: Hopelessness, a key risk factor for suicidal behaviour overall, has been studied little among bipolar disorder (BD) patients. For purposes of prevention, it is important to know whether it is predominantly a patient's permanent trait or merely reflects the highly variable illness states. We investigated the degree to which hopelessness is trait- or state-related during the course of BD. METHODS: The Jorvi Bipolar Study (JoBS) is a naturalistic prospective study representing psychiatric in- and outpatients with DSM-IV BD I and II. Repeated measurements with the Beck Hopelessness Scale of 188 patients at baseline, 6 months and 18 months were analysed using a linear regression model with general estimation equations. Factors covarying with hopelessness during follow-up were investigated. RESULTS: Levels of hopelessness varied markedly between illness phases, being highest in depressive and mixed phases, and lowest in euthymia, hypomania or mania. Hopelessness was independently associated with concurrent severity of depression (estimate 0.231, p<0.001), anxiety (0.105, p<0.001), fewer manic symptoms (-0.096, p=0.001) and comorbid personality disorder (1.741, p=0.001). However, the strongest predictor of degree of hopelessness during follow-up was previous hopelessness (0.403, p<0.001). LIMITATIONS: After baseline, relatively few patients had manic, hypomanic, mixed or depressive mixed phases. Hopelessness was measured at only three time-points. CONCLUSIONS: Level of hopelessness varies markedly between patients in different phases of BD, but is also, to a degree, a permanent feature. Among BD patients, hopelessness appears to be both a trait- and state-related characteristic. PMID- 18678415 TI - Inhibition of experimental autoimmune encephalomyelitis by a novel small molecular weight proinflammatory cytokine suppressing drug. AB - Experimental autoimmune encephalomyelitis (EAE) is a T cell-mediated, autoimmune disease of the central nervous system (CNS) that serves as a model for various cellular and molecular aspects of the human disease, multiple sclerosis (MS). Although EAE has long been considered a T cell-mediated disease, macrophages play a role in disease pathogenesis and are known to accumulate in the CNS under the control of chemokines. In the present report we demonstrate that mice induced to develop EAE were treated with a small molecular weight molecule that suppresses proinflammatory cytokine production which resulted in significantly decreased clinical EAE, CNS CCL2 expression and CNS macrophage accumulation. These results demonstrate the efficacy of a novel class of therapeutic molecules for CNS demyelinating disease. PMID- 18678416 TI - Who has inflamed my heart and makes my body burn? The Othello's dilemma in heart failure. PMID- 18678417 TI - Increased prevalence of migraine in Marfan syndrome. AB - OBJECTIVE: A high prevalence of migraine has been described in various forms of congenital heart disease, with and without shunt. In this study we investigated the prevalence of migraine in patients with Marfan syndrome (MFS). METHODS: All 457 adult patients with MFS from the participating centres and 194 controls received a validated questionnaire about headache. Migraine was diagnosed according to the International Headache Society criteria, by three independent neurologists, blinded to patient files. RESULTS: Response rate was 68% and 56% in Marfan patients and controls, respectively. Forty percent of the 309 responding MFS patients (mean age 40+/-14 years; 51% females) and 28% of the 102 controls (mean age 43+/-15 years; 58% females), suffered from migraine (p=0.03). The prevalence of migraine with aura (MA) was 22% in MFS patients and 14% in controls (p=0.06). We found MFS to be an independent risk factor for having overall migraine (OR 1.7; 95%CI 1.1-2.8), also after adjustment for age and gender (OR 1.9; 95%CI 1.1-3.1; p=0.02) and for MA after adjustment for gender (OR 2.0; 95%CI 1.1-3.7; p=0.04). In patient with MFS, previous aortic root surgery appeared to be an independent risk factor for having MA (OR 2.2; 95%CI; 1.2.-4.0, p=0.01) adjusted for gender. CONCLUSION: MFS is an independent risk factor for having overall migraine and MA. Moreover, we found that a history of aortic root surgery seems to be associated with an increased risk of MA. PMID- 18678418 TI - The development of left ventricular torsion and its clinical relevance. AB - Left ventricular torsion is a measurement derived from the twisting or wringing motion of the heart around its long axis. The calculation is made by measuring the magnitude of rotation at the apex of the heart, and subtracting the rotation at the base. Although the phenomenon of left ventricular twisting was first described in the 17th Century, it wasn't until the 1960s that the first invasive method of measurement was demonstrated. Silver tantalum clips were sutured into the epicardium during cardiac surgery and viewed using cineradiography. Non invasive torsion measurement has been subsequently developed, adopting Magnetic Resonance Imaging and 2D echocardiography. Interest in the changes of different components of torsion, during various cardiac disease states has developed with the advent of these non-invasive measurement techniques. This review article summarises the history of the development of torsion analysis and describes the known changes of torsion during different clinical circumstances. PMID- 18678419 TI - Circulating interleukin-18 concentrations and a loss-of-function P2X7 polymorphism in heart failure. AB - We examined if a loss-of-function polymorphism in the P2X(7) receptor (1513C) corresponded with circulating interleukin(IL)-18 concentrations in heart failure (HF) patients. IL-18 values were significantly elevated in HF subjects compared to healthy control subjects. No association was seen between the polymorphism and IL-18 concentrations in HF patients. In HF patients, IL-18 values had an inverse relationship with ejection fraction, mean arterial pressure and body mass index, while high IL-18 concentrations were associated with increased mortality. PMID- 18678420 TI - Chinese medical doctors' salaries. PMID- 18678421 TI - Left atrial compensatory function in subjects with early stage primary hypertension assessed by using left atrial volumetric emptying fraction acquired by transthoracic echocardiography. AB - PURPOSE: To evaluate left atrial (LA) function in subjects with early stage primary hypertension (HT) and without enlargement of LA, we used transthoracic echocardiogram and measured LA volumetric emptying fraction and compared the results with those in healthy volunteers. MATERIALS AND METHODS: 42 subjects with early stage primary HT (21 males, aged 61+/-12 years), within 1 year of HT diagnosis and starting initial treatment, were enrolled in the study. An additional inclusion criterion was normal sinus rhythm electrocardiogram without enlargement of LA. As the control group, 31 healthy volunteers with normal sinus rhythm electrocardiogram (13 male, aged 57+/-11 years) were enrolled. Maximum and minimum volume of LA (LAV max and min, respectively) were selected manually and visually in B mode images acquired from a four-chamber view from apex of left ventricle (LV) using a modified Simpson method, and the LA emptying fraction (LAEF) was calculated by the following formula: LAV min/LAV max x 100 (%). RESULTS: There were no significant differences in age, male:female ratio, end diastolic and end-systolic LV diameter, LA maximum volume, LV ejection fraction and E/A in mitral annulus between the two groups. However, we found that end diastolic inter-ventricular septum thickness and end-diastolic LV posterior wall thickness were significantly larger in the HT group in comparison with the control group (P<0.01), but the HT group did not fit the classical criteria of LV myocardial hypertrophy. The systolic and diastolic blood pressure, the end diastolic and end-systolic volume of LV, the total weight of LV myocardium and the LAEF were significantly higher in the HT group than in the control group (P<0.05). CONCLUSIONS: In subjects with early stage primary HT with normal sinus rhythm, in spite of a normal LAV, the LAEF may increase. This increase of LAEF may be regarded as one of the compensatory reactions against preload to the left side of the heart and precedes the occurrence of LA enlargement. PMID- 18678422 TI - Suitable solutions for reconstituting the ultrasound contrast agent "Levovist" used in contrast echocardiogram: in vitro and in vivo evaluation of the influence of osmotic pressure. AB - PURPOSE: To evaluate the influence of the osmotic pressure of solutions used for reconstituting the ultrasound contrast agent "Levovist" on the degree of video intensity of the enhancement and video intensity decay in contrast echocardiogram, we used 6 solutions with different osmotic pressures in both vitro and in vivo experiments. MATERIALS AND METHODS: In the in vitro experiments, Levovist was reconstituted with 6 kinds of solutions with different osmotic pressures (Lactate Ringer's solution, 2.5%, 5%, 7.5%, and 20% glucose and distilled water) and peak video intensities and video intensity decay were measured. In the in vivo experiments, contrast echocardiography was performed in 7 adult volunteers, following the intravenous injections of Levovist, previously reconstituted with one of 2 solutions (5% glucose or distilled water). RESULTS: In vitro, at peak time, Levovist reconstituted with either Lactate Ringer's solution, 2.5% glucose, 5% glucose, or distilled water had good peak video intensities. At 30 s after peak time, Levovist reconstituted with Lactate Ringer's solution had greater enhancement and less decay than the other 5 solutions (P<0.001). In vivo, at 180 heart beats after peak time, the video intensity decay with 5% glucose was greater than that with distilled water (150+/ 13 dB, 123+/-25 dB, respectively, P<0.05). CONCLUSION: In this study, among various (2.5-20%) glucose solutions, the stability of the microbubbles differed, depending on the degree of osmotic pressure of the respective solutions; 5% glucose was the best. However, overall, the most suitable solution for reconstituting Levovist, was Lactate Ringer's solution. These findings could lead to effective strategies for better contrast echocardiography using Levovist by changing the current solution of choice to Lactate Ringer's solution or 5% glucose. PMID- 18678423 TI - Free floating thrombus in the right atrium causing massive pulmonary embolism. AB - The movement of thrombi migrating from the veins of the lower limbs can give rise to pulmonary emboli within 24 hours. This is manifested as massive pulmonary embolism in 30% of cases, with a mortality rate of around 50%. Free-floating thrombi within the right cardiac cavities are rare, and the diagnosis is made mainly by transthoracic and transoesophageal echocardiography. Treatment includes surgery, invasive percutaneous embolectomy, thrombolysis and heparin administration. Here we report the case of an 80-years-old patient with massive pulmonary embolism caused by a free floating thrombus within the right atrium. Tenecteplase was administered with excellent results. PMID- 18678424 TI - External cardioversion of atrial fibrillation: the role of electrode position on cardioversion success. AB - The optimal method to perform external electrical cardioversion of atrial fibrillation has not been fully determined yet. In order to define the effects of different pad positions on cardioversion success rates, shock energy requirements and serum myocardial proteins levels we studied 62 patients with persistent atrial fibrillation who underwent elective external electrical cardioversion using a standardized step-up protocol of increasing energy levels. Electrode positions were randomly assigned as anterolateral (Group A) or anteroposterior (Group B). After all shocks were delivered there was no difference in the cardioversion success rate between the two groups. However, a significantly greater proportion of patients in group B were restored to sinus rhythm after the second shock of 300 J was delivered (p=0.005). Mean shock energy requirements and peak serum creatine kinase levels were lower for group B than for group A (p=0.049 and p=0.021 respectively). Troponin T serum levels were not increased after the cardioversion attempts in either group. We conclude that an anteroposterior electrode position is more effective in achieving restoration of sinus rhythm in lower energy shock levels compared to the anterolateral position. PMID- 18678425 TI - Paenibacillus tyraminigenes sp. nov. isolated from Myeolchi-jeotgal, a traditional Korean salted and fermented anchovy. AB - A bacterial strain H3029, a gram-positive, rod-shaped, oxidase-negative, endospore-forming bacterium that characteristically produces tyramine from tyrosine, was isolated from a Myeolchi-jeotgal, a traditional Korean salted and fermented anchovy (Engraulis japonicus). The H3029 strain showed a high ability to produce 4140 microg/ml of tyramine from the culture broth containing 5000 microg/ml tyrosine. On the other hand, the strain produced a relatively low level of other putrefactive amines, at 973 microg/ml of putrescine and 147 microg/ml of cadaverine from the media, with each 5000 microg/ml of ornithine hydrochloride and lysine hydrochloride. The H3029 strain produced no detectable level of histamine (detection limit of 4 microg/ml) from the media containing 5000 microg/ml of histidine hydrochloride. Meanwhile, tyramine, the main product of the strain, showed the antimicrobial activity at the level of over 1 mg/disk against Staphylococcus aureus by agar diffusion test, and the mutagenicity in Ames test at 0.1 mg/plate using Salmonella typhimurium TA98 and TA1535. On the basis of the polyphasic taxonomic study, the H3029(T) strain was assigned a novel species of the genus Paenibacillus as Paenibacillus tyraminigenes sp. nov. The type strain of which is strain H3029(T) (=KCTC 10694BP(T)). PMID- 18678426 TI - Diversity of virulence profiles of Shiga toxin-producing Escherichia coli serotypes in food-producing animals in Brazil. AB - The prevalence, serotypes and virulence profiles of Shiga toxin-producing Escherichia coli (STEC) were investigated in 205 healthy beef and dairy cattle, and 106 goats reared in the southeastern region of Minas Gerais State, Brazil. The prevalence of STEC was 57.5% (61/106) in goats, 39.2%, (40/102) in beef cattle and 17.5% (18/103) in dairy cattle. Among the 514 STEC isolates, 40 different serotypes were found and some of them were identified in a specific host. STEC isolates harboring stx1 corresponded to 15.6% (28/180), 26.7% (16/60) and 24.1% (66/274) in beef cattle, dairy cattle and goats, respectively. stx2 was found in 30% (54/180), 53.3% (32/60) and 34.7% (95/274) of beef and dairy cattle, and goats. stx1 plus stx2 sequences were harbored by 54.4% (98/180), 20% (12/60) and 41.2% (113/274) of beef cattle, dairy cattle and goats, respectively. The eae sequence was found in 15% (9/60) and 0.6% (1/180) of STEC isolates from dairy and beef cattle, respectively, and the toxB gene was found only in one O157:H7 strain isolated from beef cattle. Strains with the genetic profiles stx2 ehxA iha saa and stx1 stx2 ehxA iha saa were the most prevalent among STEC isolates from cattle. Profiles stx1 stx2 ehxA iha, stx2, and stx1 iha accounted for 75.5% (207 /274) of the STEC isolates from goats. While STEC strains carrying either stx2 alone or associated with stx1 were found more frequently in cattle, those harboring sequences stx1c and stx2d alone or associated with stx1c predominated in goats. Our data show a diversity of STEC strains in food-producing animals, most of them carrying genes linked to severe forms of human diseases. PMID- 18678428 TI - The detection of food soils and cells on stainless steel using industrial methods: UV illumination and ATP bioluminescence. AB - Open food contact surfaces were subjected to organic soiling to provide a source for transfer of microbial cells. Rapid industrial methods used for the detection of residual cells and soil e.g. ATP (adenosine triphosphate) bioluminescence and an ultraviolet (UV) light detection method were assessed for their ability to detect organic soils, or organic soil-cell mix on surfaces. A range of soils (complex [meat extract, fish extract, cottage cheese extract]; oils [cholesterol, fish oil, mixed fatty acids]; proteins [bovine serum albumin, fish peptones casein]; carbohydrates [glycogen, starch, lactose]); was used. Under UV, oily soils, mixed fatty acids, cholesterol and casein were detected at low concentrations, with detection levels ranging from 1% to 0.001% for different substances. Glycogen was the most difficult substance to detect at lower concentrations. Using UV wavelength bands (lambda) of 330-380 nm, 510-560 nm and 590-650 nm, wavelength bands of 330-380 nm, illuminated most of the soils well, whilst the wavelength band of 510-560 nm illuminated the fish extract, cholesterol and fatty acids; the 590-650 nm wavelength band illuminated the lactose. Soils at all concentrations were detected by the ATP bioluminescence method; the complex soils gave the highest readings. When complex soils were combined with Listeria monocytogenes Scott A or a non-pathogenic Escherichia coli O157:H7, ATP measurements increased by 1-2 logs. For UV illumination, the L. monocytogenes and cheese combination was the most intensely illuminated, with E. coli and meat the least. UV illumination is a simple well established method for detecting food soil, with little change in findings when microorganisms are included. Performance can be enhanced in certain circumstances by altering the wavelength. ATP bioluminescence is a proven system for hygienic assessment being especially useful in the presence of microorganisms rather than organic soil alone. PMID- 18678427 TI - Semantic annotation of Web data applied to risk in food. AB - A preliminary step to risk in food assessment is the gathering of experimental data. In the framework of the Sym'Previus project (http://www.symprevius.org), a complete data integration system has been designed, grouping data provided by industrial partners and data extracted from papers published in the main scientific journals of the domain. Those data have been classified by means of a predefined vocabulary, called ontology. Our aim is to complement the database with data extracted from the Web. In the framework of the WebContent project (www.webcontent.fr), we have designed a semi-automatic acquisition tool, called @WEB, which retrieves scientific documents from the Web. During the @WEB process, data tables are extracted from the documents and then annotated with the ontology. We focus on the data tables as they contain, in general, a synthesis of data published in the documents. In this paper, we explain how the columns of the data tables are automatically annotated with data types of the ontology and how the relations represented by the table are recognised. We also give the results of our experimentation to assess the quality of such an annotation. PMID- 18678429 TI - [French PMR and the action plan of the UEMS section of PMR]. PMID- 18678430 TI - A cardiac magnetic resonance imaging study of electrocardiographic Q waves in type 2 diabetes: the Fremantle Diabetes Study. AB - To investigate the evolution and significance of Q waves in type 2 diabetes, we studied 36 patients of mean (+/-S.D.) age 69.9+/-7.1 years from the longitudinal observational Fremantle Diabetes Study (FDS). All had (i) neither history/symptoms of coronary heart disease (CHD) nor pathological Q waves at FDS recruitment between 1993 and 1996, (ii) five consecutive annual assessments by FDS close-out in 2001, and (iii) contrast-enhanced cardiac magnetic resonance imaging in 2005. At this latter assessment, there were (i) 9 with no history of CHD or Q waves during follow-up (Group 1), (ii) 13 with Q waves on >/=1 electrocardiogram but no CHD history/symptoms (Group 2), and (iii) 14 with CHD history/symptoms irrespective of electrocardiographic status (Group 3). Of 20 episodes of new Q waves in 17 Group 2 or Group 3 patients during FDS follow-up, 17 (85%) resolved within 2 years. A myocardial infarction (MI) was detected by CMR in three patients (8.3%; one subendocardial in Groups 1 and 3, one non-full thickness in Group 3) but these did not correlate with electrocardiographic appearances. Q waves may have unreliable pathological significance in type 2 diabetes, including as a marker of silent MI. PMID- 18678431 TI - Transgenic expression of a mutated cyclin-dependent kinase 4 (CDK4/R24C) in pancreatic beta-cells prevents progression of diabetes in db/db mice. AB - In an attempt to rectify the hyperglycemic state in obese insulin resistant db/db mice, a transgenic line was generated (db/db-CDK4(R24C)) that expresses a constitutively active form of cyclin-dependent kinase 4 (CDK4/R24C) under the control of the insulin promoter. Compared with non-transgenic db/db littermates, adult db/db-CDK4(R24C) mice show near-complete glycemic normalization and improved plasma lipid concentrations, but are also more susceptible to weight gain and have significantly lower plasma adiponection levels. They have striking islet hypertrophy and beta-cell hyperplasia, and retain an insulin secretory response during the glucose tolerance test. We examined the expression of several key regulatory transcription factor genes involved in lipid and glucose metabolism in insulin target tissues of db/db-CDK4(R24C) as well as db/db mice, and found that the expression levels of members of the peroxisome proliferator activated receptor (PPAR) family are highly associated with metabolic alterations in a gene- and tissue-specific manner. We show for the first time that the Ppar delta in skeletal muscle and white adipose tissues is transcriptionally down regulated in db/db mice. The db/db-CDK4(R24C) mice present a novel model of leptin-resistant obesity with compensatory hyperinsulinemia and normalized blood glucose levels, and thus may be useful for future studies that aim to dissect relationships between insulin and leptin signaling. PMID- 18678432 TI - Is there discrimination in access to therapy for HCV patients? PMID- 18678433 TI - Molecular tools and hepatocellular carcinoma: Adding help or confusion in liver transplantation? PMID- 18678434 TI - Antiviral therapy after complete response to chemotherapy could be efficacious in HCV-positive non-Hodgkin's lymphoma. AB - BACKGROUND/AIMS: Prevalence of HCV infection in non-Hodgkin's lymphoma is high. The impact of antiviral therapy on the natural history of this subgroup of lymphomas after a successful chemotherapy regimen is still an argument of debate. METHODS: We retrospectively examined 343 chemotherapy-treated patients referred to our centre for five consecutive years. Clinical and histological characteristics, disease free-survival (DFS) and overall-survival (OS) were compared in HCV-positive (69/343) and HCV-negative (274/343) patients. Twenty five HCV-positive patients received antiviral treatment following chemotherapy discontinuation. Uni- and multivariate analyses were performed. RESULTS: 20% of lymphomas were HCV-positive. Indolent histology was prevalent in the HCV-positive group (p<0.05); no significant differences in OS or DFS were found between the two groups; in HCV-positive subjects, antiviral therapy, was associated with a longer DFS (p<0.05); none of the HCV-positive subjects who achieved a virological response experienced any lymphoma relapse; 29% of non responders did; at multivariate analysis, the independent factors related to a better clinical outcome were: indolent histology at the onset of lymphoma and antiviral therapy. CONCLUSIONS: Antiviral treatment in HCV-positive non-Hodgkin's lymphoma may be an important strategy to reinforce the results of a successful chemotherapy regimen; further studies are needed to validate this combined approach. PMID- 18678435 TI - Evaluation of Croatian model of polycentric health planning and decision making. AB - OBJECTIVE: To determine the progress in the development and implementation of health policies on a county level resulting from the learning-by-doing training provided through the County Public Health Capacity Building Program started in 2001 in Croatia. METHODS: Modular training using management tools, public health theory and practice, and SMDP's Healthy Plan-it tool, followed by the self evaluation of the progress made by county teams in health needs assessment and health policy development, implementation, and assurance. Fifteen county teams consisting of politicians, executive officers, public health professionals, and community members. RESULTS: Twelve of 15 county teams completed the program. The teams made progress in the evaluated areas, although to a different extent, which did not depend on the amount of time they had or the governance experience. The differences in improvement depended on the differences in the strength of political, executive, and professional components of the teams. Teams with a strong political and/or executive component, but weak public health professional and community components made major improvements in policy development and/or assurance function, but performed less well in the health needs assessment and constituency building. The reversed was also true. CONCLUSION: Learning-by-doing training program improved public health practices on a county level in Croatia. PMID- 18678436 TI - A new meshless approach for subject-specific strain prediction in long bones: Evaluation of accuracy. AB - BACKGROUND: The Finite Element Method is at present the method of choice for strain prediction in bones from Computed Tomography data. However, accurate methods rely on the correct topological representation of the bone surface, which requires a massive operator effort, thus restricting their applicability to clinical practice. Meshless methods, which do not rely on a pre-defined topological discretisation of the domain, might greatly improve the numerical process automation, but currently their application to biomechanics is negligible. METHODS: A meshless implementation of an innovative numerical approach based on a direct discrete formulation of physical laws, the Cell Method, was developed to predict strains in a cadaver femur from Computed Tomography data. The model accuracy was estimated by comparing the predicted strains with those experimentally measured on the same specimen in a previous study. As a reference, the results were compared to those obtained with a state of-the-art finite element model. FINDINGS: The Cell Method meshless model predicted strains highly correlated with the experimental measurements (R2=0.85) with a good global accuracy (RMSE=15.6%). The model performed slightly worse than the finite element one, but this was probably due to the need to sub-sample the original data, and the lower order of the interpolation used (linear vs parabolic). INTERPRETATION: Although there is surely room for improvement, the accuracy already obtained with this meshless implementation of the Cell Method makes it a good candidate for some clinical applications, especially considering the full automation of the method, which does not require any data pre processing. PMID- 18678437 TI - Sex, drugs, and HIV: rapid assessment of HIV risk behaviors among street-based drug using sex workers in Durban, South Africa. AB - South Africa is experiencing significant changes in patterns of illicit drug use, including increasing injection and non-injection drug use, and the use of drugs by persons engaged in sex work, both of which could further expand the HIV/AIDS epidemic. In 2005, a rapid ethnographic assessment was conducted in Durban, South Africa, to learn more about patterns of drug use and HIV risk behaviors among drug-using, street-based sex workers. Field teams recruited 52 current injection and non-injection drug users for key informant interviews and focus groups, and they conducted mapping and observation in identified high-risk neighborhoods. Key informants were offered free, voluntary counseling and HIV rapid testing. The results of the assessment indicate that in this population, drugs play an organizing role in patterns of daily activities, with sex work closely linked to the buying, selling, and using of drugs. Participants reported using multiple drugs including crack cocaine, heroin, Ecstasy and Mandrax, and their choices were based on their expectations about the functional role and behavioral and pharmacological properties of the drugs. The organization of sex work and patterns of drug use differ by gender, with males exercising more control over daily routines and drug and sexual transactions than females. Activities of female sex workers are subject to considerable control by individual pimps, many of whom also function as landlords and drug dealers. A strong hold over the overlapping economies of drugs and sex work by a few individuals extends to control of the physical and social settings in which sex is exchanged and drugs are sold and used as well as the terms under which sex work is carried out. The potential for accelerated HIV spread is considerable given the evidence of overlapping drug-using and sexual risk behaviors and the mixing patterns across drug and sexual risk networks. PMID- 18678438 TI - Subsequent ventricular fibrillation and survival in out-of-hospital cardiac arrests presenting with PEA or asystole. AB - BACKGROUND: The prognostic implications of conversion to ventricular fibrillation (VF) in out-of-hospital cardiac arrest (OHCA) patients with an initial non shockable rhythm are unclear. HYPOTHESIS: Among OHCA patients with an initial non shockable rhythm, survival is better in individuals who subsequently develop VF and are defibrillated. METHODS: DESIGN: Utstein style population-based cohort study. SUBJECTS: adults (age>or=18 years) with OHCA of presumed cardiac etiology and initial rhythm of pulseless electrical activity (PEA) or asystole treated by emergency medical services systems in Osaka, Japan from January 1, 2001 to December 31, 2005. Primary outcome measure was one-month neurologically favorable survival (CPCA/p.Arg113His. She had a progressive disease with development of tetraparesia in less than 6 years. Our observation confirms that ovarian failure in the context of a leukodystrophy warrants mutational analysis of the genes encoding the subunits of EIF2B. PMID- 18678443 TI - Growth inhibition in Chinese cabbage (Brassica rapa var. chinensis) growth exposed to di-n-butyl phthalate. AB - The toxicity and effects of di-n-butyl phthalate (DBP), an endocrine disruptor, on the growth of Chinese cabbage (Brassica rapa var. chinensis) were studied. Etiolation occurred on leaves of Chinese cabbage plant treated with 50mg/L of DBP for 42 d. DBP even below 1mg/L had a significant effect on the concentration of chlorophyll in Chinese cabbage and the biomass showed a severe decrease under treatment with more than 30 mg/L of DBP. At a concentration below 1mg/L of DBP, no significant difference in accumulation was found, but treatments with concentration exceeding 10, 30, 50 and 100mg/L all resulted in significant accumulation of DBP. Six protein spots extracted from leaf tissue of DBP-treated Chinese cabbage displaying a differential expression are shown in 2-DE maps. According to proteome level studies, three protein spots were found to increase and were identified, respectively, as acyl-[acyl-carrier-protein] desaturase (acyl-ACP desaturase), root phototropism protein 3 (RPT3) and ferredoxin-nitrite reductase (Fd-NiR). The other three protein spots were found to decrease and were identified respectively as dihydroflavonol-4-reductase (DFR), aminoacyl-tRNA synthetase (aaRS) and ATP synthase subunit beta. The key finding is that the other closely related plant, Bok choy (Brassica rapa subsp. chinensis), the subspecies of Chinese cabbage, respond differently to the same chemicals. PMID- 18678444 TI - Co-metabolism of 2,4-dichlorophenol and 4-Cl-m-cresol in the presence of glucose as an easily assimilated carbon source by Staphylococcus xylosus. AB - Comparison of the ability of Staphylococcus xylosus to degrade 2,4-dichlorophenol and 4-Cl-m-cresol in separate cultures is reported. Bacterial adaptation and the continuous presence of glucose, as a conventional carbon source, were found to stimulate the degrading efficiency of S. xylosus. 4-Cl-m-cresol exhibited higher substrate-induced toxicity with K(ig) value at 0.25 mM, comparing to 2,4 dichlorophenol (K(ig) value at 0.90 mM) at initial concentration ranging from 0.1 to 0.5 mM. Degradation rate of 4-Cl-m-cresol was found to decrease only, revealing lower value of inhibition degradation constant (K(i) at 0.019 mM) comparing to that of 2,4-dichlorophenol (K(i) at 0.41 mM). Both glucose and each one of the chloro-aromatic compounds tested were simultaneously consumed and an increase of chloride ions in the medium appeared, during the exponential phase of growth. The chloride ions increase was nearly stoichiometric in the presence of 2,4-dichlorophenol and one of its several intermediate products identified was 2 Cl-maleylacetic acid. In the case of 4-Cl-m-cresol, only one metabolic product was found and identified as 3-methyl-4-oxo-pentanoic acid. PMID- 18678445 TI - Prevenient dye-degradation mechanisms using UV/TiO2/carbon nanotubes process. AB - Photocatalysis research heavily emphasizes increasing photo-efficiency. This study presents the application of carbon nanotubes (CNTs) to increase the photocatalytic activity of TiO(2). It elucidates the effect of CNTs dose on the decolorization efficiency of aqueous azo dye, C.I. Reactive Red 2 (RR2), determines the effects of SO(4)(2-) formation and removal of total organic carbon (TOC), and measures the effects of various ultraviolet wavelengths. Scanning electron microscopy was used to elucidate the mixing phenomenon and the size of TiO(2) and CNTs; X-ray diffraction was used to determine crystallinity; a BET meter was used to measure surface area and a spectrophotometry was used to determine the decolorization of RR2. Experimental results indicated significant effects of photodegradation on the combination of TiO(2) with CNTs and electron transfer is higher for 410 nm irradiation than for 365 nm, revealing that solar light can be used. The electron transfer in the TiO(2)/CNTs composites reduced the electron/hole recombination and increased the photon efficiency and the prevenient dye-degradation mechanisms using UV/TiO(2)/CNTs were established. PMID- 18678446 TI - Alertness and feeding behaviors in ADHD: does the hypocretin/orexin system play a role? AB - Increasing evidence has suggested that patients with attention deficit/hyperactivity disorder (ADHD) may present with a deficit of alertness and sleep disturbances. Recent studies have also pointed out a previously underestimated association between ADHD and abnormal eating behaviors, including binge eating. Since sleep/alertness disturbances and eating disorders may significantly increase the functional impairment of ADHD, gaining insight into their pathophysiology as well as into their treatment is of relevance to provide a better clinical management of patients suffering from ADHD. The hypocretin/orexin system comprises two distinct peptides, located in the hypothalamus, which are involved in several homeostatic functions. In particular, it has been suggested that hypocretin/orexin neurons located in perifornical and dorsomedial hypothalamic nuclei increase arousal, whereas those located in the lateral hypothalamus are primarily implicated in reward processing, stimulating feeding and other reward seeking behaviors. Given the involvement of the hypocretin/orexin system in the control of alertness and reward seeking (including feeding), we hypothesize that hypocretin/orexin neurons located in perifornical and dorsomedial hypothalamic areas are hypoactivated, while those located in the lateral hypothalamus are overactivated in patients with ADHD. If confirmed by further neurophysiological, imaging, and genetics studies, our hypothesis may help us progress in the understanding of the complex pathophysiology of ADHD. This might set the basis for the study of novel molecules, acting on the hypocretin/orexin system, aimed at increasing wakefulness and reducing binge eating and other abnormal reward seeking behaviors in patients with ADHD. We also suggest future studies on the potential therapeutic role of other molecules which have a complex interplay with the hypocretin/orexin system, such as the histamine H(1) receptor agonists, the histamine H(3) receptor antagonists, and the neuropeptide Y receptor antagonists. All this body of research would provide a tremendous opportunity to improve the quality of life of patients with ADHD by means of pathophysiologically oriented treatment. PMID- 18678447 TI - Strain variation within Campylobacter species in fecal samples from dogs and cats. AB - To investigate the incidence of co-colonization of different strains of Campylobacter species present in canine and feline stool samples, isolates were recovered by culture from 40 samples from dogs (n=34) and cats (n=6). Animals were of different ages, with diarrhoea or without clinical signs. Three isolation procedures were used: two selective agars and a filtration method. In each stool sample, multiple colonies were identified to the species level by PCR, subsequently genotyped by Amplified Fragment Length Polymorphism (AFLP) and pattern similarities (451 isolates) were calculated to investigate their phylogenetic relationships. Genetic heterogeneity of strains in individual stool samples was detected within the species Campylobacter jejuni, C. upsaliensis and C. helveticus, though to a different degree in dogs and cats. In 3 of the 34 (9%) canine samples, more than one genotype of the same Campylobacter species was present, while strain variation was detected in four of the six feline samples. The results show that preferably, multiple colonies should be analyzed in molecular epidemiological and aetiological studies. PMID- 18678448 TI - Vestibular evoked myogenic potentials of undiagnosed dizziness. PMID- 18678449 TI - A MECP2 mutation in a highly conserved aminoacid causing mental retardation in a male. AB - MeCP2 is a protein that functions as a key factor in epigenetic transcriptional regulation. Mutations in MECP2 gene have been reported as being the major cause of Rett syndrome. These mutations may also cause a wide spectrum of neurological disorders in males. Here, we report the identification of the mutation p.P405L in a 19-year-old Brazilian male with mental retardation. This variant is localized in a highly conserved aminoacid from the carboxy terminal domain and may affect the protein function. Segregation analysis on the patient's mother revealed that this is a de novo mutation and it was not identified in the control sample. The programs SIFT, PolyPhen and A-GVGD considered that the p.P405L may be damaging. Despite the high frequency of non pathogenic variants that have been identified in this gene, our data lead us to consider the p.P405L a disease-causing mutation. PMID- 18678451 TI - [Rectosigmoid junction metastasis from melanoma: a case report]. PMID- 18678452 TI - Motion characterization of aortic wall and intimal flap by ECG-gated CT in patients with chronic B-dissection. AB - RATIONALE AND OBJECTIVES: To evaluate whether dynamic computed tomography (CT) imaging can provide functional vessel information in patients with chronic aortic dissection type Stanford-B (ADB). MATERIALS AND METHODS: In 32 patients, ECG gated CT-angiography images were obtained. Cross-sectional area change and wall distensibility were investigated by semiautomatic vessel area segmentation at the end of aortic arch. Significance of distensibility differences was tested with regard to the aortic diameter, and the oscillation of the intimal flap was analyzed. RESULTS: The aorta could be segmented successfully in all patients. These were separated into three subgroups: (A) 6 patients with an aortic diameter <4 cm and without a visible intimal flap, (B) 9 patients with an aortic diameter <4 cm, and (C) 17 individuals with an aortic diameter > or = 4 cm; (B) and (C) having a visible intimal flap. Differences in distensibility between the subgroups were not significant. Overall mean distensibility was D(tot)=(1.3+/ 0.6) x 10(-5) Pa(-1). Analysis of intimal flap oscillation showed a pulsatile short axis diameter decrease of the true lumen of up to 29%. CONCLUSION: Dynamic, ECG-gated CT-angiography can demonstrate pulsatile changes in aortic area and a highly variable motion of the intimal flap. Aortic distensibility appears independent of diameter or presence of a intimal flap. Follow-up studies may show correlation with possible complications. PMID- 18678453 TI - Vibrios dominate as culturable nitrogen-fixing bacteria of the Brazilian coral Mussismilia hispida. AB - Taxonomic characterization was performed on the putative N(2)-fixing microbiota associated with the coral species Mussismilia hispida, and with its sympatric species Palythoa caribaeorum, P. variabilis, and Zoanthus solanderi, off the coast of Sao Sebastiao (Sao Paulo State, Brazil). The 95 isolates belonged to the Gammaproteobacteria according to the 16S rDNA gene sequences. In order to identify the isolates unambiguously, pyrH gene sequencing was carried out. The majority of the isolates (n = 76) fell within the Vibrio core group, with the highest gene sequence similarity being towards Vibrio harveyi and Vibrio alginolyticus. Nineteen representative isolates belonging to V. harveyi (n = 7), V. alginolyticus (n = 8), V. campbellii (n = 3), and V. parahaemolyticus (n = 1) were capable of growing six successive times in nitrogen-free medium and some of them showed strong nitrogenase activity by means of the acetylene reduction assay (ARA). It was concluded that nitrogen fixation is a common phenotypic trait among Vibrio species of the core group. The fact that different Vibrio species can fix N(2) might explain why they are so abundant in the mucus of different coral species. PMID- 18678454 TI - Diversity of Lactobacillus sakei strains investigated by phenotypic and genotypic methods. AB - The diversity of 10 strains of Lactobacillus sakei, a commercially important species of lactobacilli, was characterized by studying food isolates. Growth characteristics varied among the strains when examined after growth in a complex medium and a defined medium with either glucose or ribose. A commercial starter culture strain showed the fastest growth rates and high biomass formation on all media, while two of the strains hardly grew on ribose. Based on acidification properties in a meat model, some of the strains had the ability to compete with the indigenous microbiota of the meat batter in addition to being fast acid producers. Carbohydrate-fermentation abilities revealed a relatively wide variation, clustering the strains into two phenotypic groups. The isolates were analyzed using different genetic fingerprinting techniques, demonstrating a distinction between two genetic groups, a grouping consistent with previous studies dealing with L. sakei strains. Comparative genome hybridization (CGH) was introduced for clustering the strains and the same division into two genetic groups was observed. Chromosomal sizes of the strains were estimated by pulsed field gel electrophoresis (PFGE) and were found to vary from 1884 to 2175 kb. The genetic groups did not correlate with the clustering obtained with carbohydrate fermenting abilities or with chromosomal sizes. PMID- 18678455 TI - Simultaneous determination of five main active constituents of Erigeron multiradiatus by HPLC-DAD-MS. AB - An HPLC-DAD-MS method was developed for simultaneous determination of the five major active constituents in Erigeron multiradiatus (Wall.) Benth, namely 6'-O cafferylerigeroside (1), scutellarin (2), apigenin-7-O-beta-d-glucuronide (3), apigenin (4) and kaempferol (5), respectively. They were identified by ESI-MS and comparisons with literature. A comprehensive validation of the method included tests of sensitivity, linearity, precision and accuracy. The linear regressions were acquired with r>0.999. The precision was evaluated by intra- and inter-day assays, and relative standard deviation (R.S.D.) values were reported within 2.7%. The recovery studies for the quantified compounds were observed in the range of 95.3-102.4% with R.S.D. values less than 2.3%. The overall procedure may be suitable for the qualitative and quantitative analyses of a large number of E. multiradiatus samples. Hierarchical clustering analysis based on the characteristics of the 5 investigated compound peaks in HPLC profiles showed that 18 samples were divided into 2 main clusters. The clusters corresponded to their content. The five constituents in E. multiradiatus are generally regarded as an index for the quality assessment of this herb. PMID- 18678456 TI - A liquid chromatography-mass spectrometry assay for simultaneous determination of two antimalarial thiazolium compounds in human and rat matrices. AB - A new class of antimalarial drugs targeting phospholipid metabolism of the malarial parasite is now in development. In the strategy of this development, two mono-thiazolium salts, T1 and T2, need to be monitored. A liquid chromatography mass spectrometry (LC-MS) method has been developed and validated according to FDA guidelines for simultaneous determination of T1 and T2 in plasma, whole blood and red blood cells (RBCs) from human and rat. The sample-pre-treatment procedure involved solid phase extraction after protein precipitation. Chromatography was carried out on a Zorbax eclipse XDB C8 column and mass spectrometric analysis was performed using an Agilent 1,100 quadrupole mass spectrometer working with an electrospray ionization source. LC-MS data were acquired in single ion monitoring mode at m/z 312, 326 and 227 for T1, T2 and the internal standard (T3), respectively. The drug/internal standard peak area ratios were linked via a quadratic relationship to concentrations (human and rat plasma: 2.25-900 microg/l; human blood and rat RBCs: 4.5-900 microg/kg). Precision was below 14.5% for T1 and below 13% for T2. Accuracy was 92.6-111% for T1 and 95.6-108% for T2. Extraction recoveries were >or=85% in plasma and >or=53% in blood and RBCs. For T1 and T2, the lower limits of quantitation were 2.25 microg/l in plasma, and 4.5 microg/kg in whole blood and RBCs. Stability tests under various conditions were also investigated. This highly specific and sensitive method was useful to analyse samples from pharmacokinetic studies carried out in rat and would also be useful in clinical trials at a later stage. PMID- 18678457 TI - Chemical fingerprint and quantitative analysis of Salvia plebeia R.Br. by high performance liquid chromatography. AB - To control the quality of Salvia plebeia R.Br., a simple and reliable method of high-performance liquid chromatography coupled with photodiode array detector (HPLC-DAD) was developed both for fingerprint analysis and quantitative determination of seven bioactive compounds, namely caffeic acid, luteolin-7 glucoside, nepetin-7-glucoside, homoplantaginin, luteolin, nepetin and hispidulin. In fingerprint analysis, twelve peaks were selected as characteristic peaks. In quantitative analysis, seven compounds showed good regression (R2>0.9995) within test ranges and the recovery of the method was in the range of 91.7-103.2%. The content ranges (mg/g) of seven compounds in the collected samples of S. plebeia were 0.80-1.67 (hispidulin), 2.18-5.75 (homoplantaginin), 0.52-1.22 (nepetin), 1.56-3.48 (nepetin-7-glucoside), 0.12-0.24 (luteolin), 0.97 2.22 (luteolin-7-glucoside) and 0.21-0.44 (caffeic acid), respectively. From the results obtained, the content of homoplantaginin was the highest. In addition, luteolin and luteolin-7-glucoside were isolated for the first time from S. plebeia. PMID- 18678458 TI - Determination of oxytocin in a dilute IV solution by LC-MS(n). AB - The most common drug prescribed to induce labor in the United States is oxytocin, a peptide hormone composed of nine amino acids. Oxytocin is often reconstituted in intravenous (IV) saline solutions at less than 0.05 units ml(-1) (125 ng ml( 1)) to be delivered at 1-4 drops per minute. Existing LC-UV methods for oxytocin do not have sufficient detection limits to quantitate and/or confirm oxytocin in IV solutions without sample concentration. A determinative and confirmatory method for oxytocin was developed using an LC-MS(n) ion trap instrument with an electrospray ionization (ESI) interface in positive ion mode. Separation was achieved on a C-18 column using an isocratic elution of water with 50% acetonitrile (v/v) and water with 0.05% formic acid (v/v) at a flow rate of 250 microl min(-1). Data was acquired from the selected ion monitoring (SIM) of the precursor ion (m/z 1007.3) and MS(2) scans from the collision induced dissociation of m/z 1007.3 at 30% collision energy. In this method, MS(2) full scans were utilized to obtain three structurally significant ions for the unambiguous identification of oxytocin. Calibration standards, prepared in de ionized water from 0.006 to 0.046 units ml(-1), were linear with an R(2) value of 0.9983. The methods LOD and LOQ were 0.00084 and 0.0029 units ml(-1) (2 and 7 ng ml(-1)), respectively. This LC-MS(n) method was used to determine the amount of oxytocin in a 0.04 units ml(-1) clinical sample that was prepared in 0.9% sodium chloride IV solution. PMID- 18678459 TI - Stability studies of anticancer agent bis(4-fluorobenzyl)trisulfide and synthesis of related substances. AB - Bis(4-fluorobenzyl)trisulfide, fluorapacin, has been extensively developed as a promising new anticancer drug candidate. Its degradation products were identified and verified by the newly synthesized compounds bis(4-fluorobenzyl)disulfide (A) and bis(4-fluorobenzyl)tetrasulfide (B) which were resulted from the disproportionation of fluorapacin under forced conditions. A stability-indicating HPLC method was used for the stability evaluation of active pharmaceutical ingredient (API) fluorapacin and finished pharmaceutical product (FPP) under various conditions. High recovery (99.57%) of API was found after three freeze thaw cycle processes of fluorapacin FPP. Susceptibility of fluorapacin to oxidative degradation was studied by treating fluorapacin and FPP in 30% hydrogen peroxide aqueous solution, and the result verified the oxidative stability of fluorapacin. However, treatment of this drug candidate under strong light (4500 Lx+/-500 Lx) for 10 days showed substantial effect on the recovery of fluorapacin, especially from fluorapacin FPP. Strong acid (1.0M, HCl) did not affect the recovery of fluorapacin while strong basic condition (1.0M, NaOH) accelerated the disproportionation of fluorapacin to its related substances A and B. The stability of fluorapacin in its aqueous media at a pH range of 2.0-10.0 for up to 6h was further investigated, and 4.0-8.0 was found to be the most stable pH range. Fluorapacin and FPP were exposed to the elevated temperatures of 40 and 60 degrees C for 10 days without obvious impact on their stability. The thermal stability of fluorapacin API and FPP under constant humidity with light protection was also thoroughly investigated under accelerated (40+/-2 degrees C, RH 75+/-5%, 6 months) and long-term (25+/-2 degrees C, RH 60+/-10%, 24 months) conditions. There was no significant change except minor color change of fluorapacin FPP. Therefore, fluorapacin has excellent stability as a potential drug candidate for further clinical development investigation. PMID- 18678460 TI - Quality assessment of unfractionated heparin using 1H nuclear magnetic resonance spectroscopy. AB - Due to problems, especially anaphylactoid reactions, raised by impure unfractionated heparin the quality assessment of heparin has to be reconsidered. Neither the USP nor the European Pharmacopoeia are able to guarantee the purity of heparin, i.e., the limitation of oversulfated chondroitin sulfate (OSCS) which was found to be the reason for the allergic adverse effects. In the first run the regulatory authorities ask for 1H NMR spectroscopic and capillary electrophoretic measurements in order to characterize the impurity profile of heparin. Using an optimized 1H NMR method the limit of detection for OSCS was found to be 0.1%. In addition, it is possible to reliably quantify both OSCS and dermatan sulfate (DS), the latter being an indicator of poor purification of the unfractionated heparin. Screening of more than 100 heparin samples collected from international markets revealed a high number of samples containing substantial amounts of DS and a number of samples containing OSCS in an amount higher than 0.1%. PMID- 18678461 TI - A comparison of two educational interventions for the cognitive empowerment of ambulatory orthopaedic surgery patients. AB - OBJECTIVE: The aim of this study was to compare the cognitive empowerment of ambulatory orthopaedic surgery patients when using Internet-based education (experiment) in contrast to face to face education conducted by a nurse (control). METHODS: Elective ambulatory orthopaedic surgery patients were randomized to either an experiment group (n = 72) receiving Internet-based education, or to a control group (n = 75) receiving face-to-face education with a nurse. The data were collected at three different time points: before the preoperative education session, after preoperative education and 2 weeks after the operation. Three structured instruments were used: the Knowledge Test, the Sufficiency of Knowledge and the Orthopaedic Patient Knowledge Instrument. RESULTS: Patients in both groups showed improvement in their knowledge. However, patients who received Internet-based education improved their knowledge level significantly more in the ethical (p = 0.005) and functional (p = 0.023) dimensions and also in total (p = 0.033) than those patients who underwent face to-face education with a nurse. In addition, patients in the experiment group had higher scores in sufficiency of knowledge in the experiential (p = 0.050) and financial (p = 0.048) dimensions and, moreover, their scores in sufficiency of knowledge in the ethical dimension improved significantly more (p = 0.008) during the study period than patients in the control group. CONCLUSION: Improvements in the patients' level and sufficiency of knowledge within both groups indicates an increase in patients' cognitive empowerment. PRACTICE IMPLICATIONS: Internet based education can be used in ambulatory orthopaedic surgery patient education for increasing patients' cognitive empowerment. PMID- 18678462 TI - Presence of neuropeptide Y in somatotrophs of cattle. AB - Neuropeptide Y (NPY), a 36-amino acid member of the pancreatic polypeptide family, was found to be present by immunohistochemistry in the bovine adenohypophysis. NPY mRNA expression was confirmed in the adenohypophysis by RT PCR. NPY immunoreactivity was present in about 38% of adenohypophyseal cells in the pars distalis. However, NPY immunoreactive cells (NPY-ir cells) were scarce in the zona tuberalis. Immunohistochemistry of NPY and specific hormones using mirror sections revealed that NPY was colocalized in GH immunoreactive cells. Over 90% of somatotrophs corresponded to NPY-ir cells. These results indicate that endogenous NPY is present in the bovine somatotroph and may act as an endocrine intercellular mediator in the adenohypophysis. PMID- 18678464 TI - [Is it necessary to revalue the risk of a gas embolism complicating an intervention with carbon dioxide insufflation? Answer elements]. PMID- 18678465 TI - [Osteoarticular manifestations of late congenital syphilis]. PMID- 18678466 TI - [PCR testing for Bordetella pertussis in household contacts as a diagnostic tool for atypical whooping cough in unvaccinated young infants]. AB - INTRODUCTION: False-negative findings of polymerase chain reaction (PCR) for genuine pertussis as well as the numerous atypical forms of whooping cough make it difficult to diagnose this disease in young babies. METHODS: For two years, real-time PCR was performed to test for Bordetella pertussis in 86 infants younger than 6 months hospitalized for apnea or paroxysmal and/or vomiting cough and in 205 of their household contacts, whether or not they coughed. RESULTS: Group 1 included 30 infants for whom PCR detected B. pertussis (25 of whom were also RSV+). PCR was also positive for at least one household contact in 25/30 families. This group included 16 babies with apnea and 12 who developed a whooping cough during follow-up. Group 2 comprised 12 infants whose PCR was negative while at least one household contact had positive results. Five of these infants had severe apnea and 6 developed a whooping cough. Group 3 included 44 infants (28 RSV +) for whom PCR was negative in the index case and in the household contacts: none developed a whooping cough during follow-up. Only 3 of the 54 positive household contacts had a paroxysmal cough or a typical whooping cough and 12 had no cough at all. CONCLUSION: Positive PCR in a household contact, symptomatic or not, is helpful for the diagnosis of atypical whooping cough in young infants. PMID- 18678467 TI - [Adult Henoch-Schonlein purpura]. AB - Henoch-Schonlein purpura (HSP) is a systemic IgA vasculitis affecting small vessels. HSP usually affect children whereas it is rare in adults (150 to 200 for 1) in which the disease is often more serious with more frequent and severe nephritis. Prevalence of adult PR is unknown and its annual incidence is 1 in 1 million. The dominant clinical features include cutaneous purpura, arthritis and gastrointestinal symptoms. Some times nephritis can add, typically as glomerulonephritis with IgA mesangial deposits. Pulmonary, cardiac, genital and neurological symptoms have also been observed. Although the cause is unknown, it is clear that IgA plays a pivotal role in the immunopathogenesis of HSP. Only symptomatic treatment is advised in case of self limited disease. Treatment of severe HSP, nephritis or gastrointestinal manifestations, is not established but some studies, which need to be confirmed, reported the benefit of corticosteroids combined with immunosuppressive drugs. Short term outcome depends on the severity of the gastro-intestinal manifestations. The long term prognosis is heavily dependent on the presence and severity of nephritis. Studies with prolonged follow-up show up to one third of adult patients reaching end stage renal failure. PMID- 18678468 TI - A fast nearest neighbor classifier based on self-organizing incremental neural network. AB - A fast prototype-based nearest neighbor classifier is introduced. The proposed Adjusted SOINN Classifier (ASC) is based on SOINN (self-organizing incremental neural network), it automatically learns the number of prototypes needed to determine the decision boundary, and learns new information without destroying old learned information. It is robust to noisy training data, and it realizes very fast classification. In the experiment, we use some artificial datasets and real-world datasets to illustrate ASC. We also compare ASC with other prototype based classifiers with regard to its classification error, compression ratio, and speed up ratio. The results show that ASC has the best performance and it is a very efficient classifier. PMID- 18678469 TI - An fMRI study of working memory in first-degree unaffected relatives of schizophrenia patients. AB - Identifying intermediate phenotypes of genetically complex psychiatric illnesses such as schizophrenia is important. First-degree relatives of persons with schizophrenia have increased genetic risk for the disorder and tend to show deficits on working memory (WM) tasks. An open question is the relationship between such behavioral endophenotypes and the corresponding brain activation patterns revealed during functional imaging. We measured task performance during a Sternberg WM task and used functional magnetic resonance imaging (fMRI) to assess whether 23 non-affected first-degree relatives showed altered performance and functional activation compared to 43 matched healthy controls. We predicted that a significant proportion of unaffected first-degree relatives would show either aberrant task performance and/or abnormal related fMRI blood oxygen level dependent (BOLD) patterns. While task performance in the relatives was not different than that of controls they were significantly slower in responding to probes., Schizophrenia relatives displayed reduced activation, most markedly in bilateral dorsolateral/ventrolateral (DLPFC/VLPFC) prefrontal and posterior parietal cortex when encoding stimuli and in bilateral DLPFC and parietal areas during response selection. Additionally, fMRI differences in both conditions were modulated by load, with a parametric increase in between-group differences with load in several key regions during encoding and an opposite effect during response selection. PMID- 18678471 TI - Tremendous ex vivo child skin absorption and permeation of eucalyptol. PMID- 18678472 TI - Effect of iontophoresis and penetration enhancers on transdermal absorption of metopimazine. AB - BACKGROUND: Metopimazine is an antiemetic drug already used by oral and rectal administration. It would be interesting to develop a new formulation for a transdermal administration. OBJECTIVE: The objective of this study was to determine the influence of iontophoresis on the metopimazine transdermal absorption and the possible synergistic enhancement with chemical enhancers. METHODS: Transdermal transport of metopimazine was studied in vitro in a Franz cell with pig skin according to the following protocol: 1h of iontophoresis followed by 7h of passive diffusion. Different current densities were applied: 0, 0.125, 0.25 and 0.5 mA/cm(2). Chemical enhancers used as solvent dilution were ethanol, propylene glycol and isopropyl myristate. Metopimazine was assayed by HPLC. Fourier transform infrared spectroscopy was used to determinate the interaction between chemical enhancers and stratum corneum. RESULTS: The iontophoresis has increased the percutaneous absorption of metopimazine and has decreased the lag time with 3.85+/-0.90 microg/(cm(2)h) and 1.9h for 0.5 mA/cm(2) and with 0.27+/-0.20 microg/(cm(2)h) and >8h for passive diffusion. Transdermal transport has been increased with current density and with isopropyl myristate and was not modified by ethanol or propylene glycol. CONCLUSION: Results indicated that iontophoresis is an effective method for transdermal administration of metopimazine. PMID- 18678470 TI - Abnormal expression of glutamate transporter and transporter interacting molecules in prefrontal cortex in elderly patients with schizophrenia. AB - Glutamate cycling is critically important for neurotransmission, and may be altered in schizophrenia. The excitatory amino acid transporters (EAATs) facilitate the reuptake of glutamate from the synaptic cleft and have a key role in glutamate cycling. We hypothesized that expression of the EAATs and the EAAT regulating proteins ARHGEF11, JWA, G-protein suppressor pathway 1 (GPS1), and KIAA0302 are altered in the brain in schizophrenia. To test this, we measured expression of EAAT1, EAAT2, EAAT3, and EAAT interacting proteins in postmortem tissue from the dorsolateral prefrontal and anterior cingulate cortex of patients with schizophrenia and a comparison group using in situ hybridization and Western blot analysis. We found increased EAAT1 transcripts and decreased protein expression, increased EAAT3 transcripts and protein, and elevated protein expression of both GPS1 and KIAA0302 protein. We did not find any changes in expression of EAAT2. These data indicate that proteins involved in glutamate reuptake and cycling are altered in the cortex in schizophrenia, and may provide potential targets for future treatment strategies. PMID- 18678473 TI - [Staphylococcus aureus isolated from a case of lymph node botryomycosis]. AB - Botryomycosis is an uncommon chronic bacterial infection, which usually involves the skin and the subcutaneous tissues. Visceral involvement often occurs in immunodepressed patients. We describe the case of a 4-year-old boy who presented a right inguinal inflammatory mass associated with pruritic papules without fever. Blood analyses showed a substantial inflammatory syndrome and blood hypereosinophilia. Histological examination of the right inguinal lymph node showed granulomatous adenitis with a Splendore-Hoeppli phenomenon surrounding Gram-positive bacteria, corresponding to Staphylococcus aureus after biopsy culture. The diagnosis of S. aureus lymph node botryomycosis was made and the child was successfully treated with a double dose of oxacillin over 3 weeks. PMID- 18678474 TI - Using support vector regression to model the correlation between the clinical metastases time and gene expression profile for breast cancer. AB - OBJECTIVE: Recently, the microarray analysis has been an important tool used for studying the cancer type, biological mechanism, and diagnostic biomarkers. There are several machine-learning methods being used to construct the prognostic model based on the microarray data sets. However, most of these previous studies were focused on the supervised classification for predicting the clinical type of patients. In this study, we investigate whether or not the expression level of some significant genes identified can be used to predict the clinical metastases time of patients. MATERIALS AND METHODS: We have used a regression method to remodel the data set of breast cancer published in 2002. Some significant genes were ranked and selected based on a wrapper method with 10-fold cross-validation procedure and the selected genes were used to fit the support vector regression (SVR) model. This method could model the relationship between the significant gene expression value and the clinical metastases time of breast cancer. RESULTS: 44 significant genes are selected for building the regression model and the corresponding cross-validated correlation coefficient obtained is 0.82 which is much superior to those reported previously by others using some different data sets. Moreover, there are two breast cancer related genes (the ligand 14 of the chemokine C-X-C motif (CXCL14) and estrogen receptor gene (ER)) selected in the gene set and one of them is never been included in the other data sets. CONCLUSION: In this report, we have shown that the expression level of some significant genes identified could strongly correlate with the clinical metastases time of breast cancer patients. The 44 selected genes may be used as a benchmark to evaluate the risk of recurrence of breast cancer. PMID- 18678475 TI - Drinking habits and health in Northern Italian and American men. AB - BACKGROUND AND AIM: The aim of this study was to investigate alcohol drinking habits in two male cohorts, one Italian and one American, and to investigate whether cardiovascular disease risk factors are related to different drinking patterns. Furthermore, socio-demographic characteristics were taken into account. METHODS: The Italian sample was drawn from the National Alpines Association. A dietary questionnaire was sent to the members of this association as an additional supplement to their monthly magazine. Eleven thousand one hundred and thirty-four men, 18-94 years, from Northern Italy were included in this analysis. The American sample is part of the Western New York Health Study (WNYHS) including 1927 male participants. RESULTS: In both populations, those who drank more than 4 drinks/day were the least educated and showed the highest percentage of current smokers; the highest prevalence of hypertension occurred in heavier drinkers and those who mostly drank without food. By contrast, lifetime abstainers exhibited the lowest percentage of hypertension and the highest level of serum cholesterol; in both populations the highest prevalence of diabetes was present in lighter drinkers. CONCLUSIONS: The current study shows that drinking habits are quite different in the two countries and are basically linked with socio-demographic and behavioral variables and support the notion that excess volume of alcohol consumed, and drinking without food, are associated with a higher risk of hypertension and hyperlipidaemia, particularly for Italians. PMID- 18678476 TI - Response to flavonoids as a factor influencing competitiveness and symbiotic activity of Rhizobium leguminosarum. AB - Flavonoids play a crucial role as signal molecules in promoting the formation of nodules by symbiotic bacteria commonly known as rhizobia. The early interaction between flavonoids and NodD regulatory protein activates nod gene transcription and the synthesis of Nod factor that initiates nodule primordium. In this study, we assessed response to flavonoids as factors influencing competitiveness of rhizobia and their symbiotic activity. Rhizobium leguminosarum nodule isolates belonging to three biovars, trifolii, viciae and phaseoli characterized earlier as competitive or uncompetitive relative to native rhizobia, were used. Investigating nodA promoter induction using plasmid lacZ fusion, we found that competitive strains more readily responded to a wide range of synthetic flavonoids and seed exudates in comparison to uncompetitive strains, albeit some exceptions were noticed. Of all the synthetic flavonoids and seed exudates studied, naringenin, hespertin and clover and vetch exudates were the most effective inducers of nodA promoter in competitive strains. Only one of the nine examined uncompetitive strains was highly induced by clover seed exudate. Subsequently, the effect of preinduction of R. leguminosarum bv. trifolii with clover exudate was assessed. Out of 18 pre-activated strains, nine strains (including competitive ones) increased clover wet mass of shoots and nodule number when used as inoculants. Our results demonstrate a plausible approach of isolating and characterizing flavonoid-responsive field isolates that could be further developed into relevant legume inoculants. PMID- 18678477 TI - Fatty acid-binding protein regulates LPS-induced TNF-alpha production in mast cells. AB - There has been increasing evidence for the involvement of fatty acid-binding proteins (FABPs) in the cytokine production of macrophages and dendritic cells probably through the control of cellular lipid metabolism and signal transduction. Since mast cells (MCs) are recently shown to be involved in immune response through modification of cytokine production, it is possible that some FABPs could also be involved in the immune response of MCs. In this study, we found that epidermal-type FABP (E-FABP) was expressed in murine bone marrow derived MCs (BMMCs). Using BMMCs from genetically E-FABP-null mutated mice, we demonstrated that E-FABP in BMMCs plays a key role in the production of TNF-alpha following lipopolysaccharide (LPS) stimulation. In the in vivo septic peritonitis model (cecal ligation and puncture model), E-FABP-null mice showed a significantly increased mortality compared to wild-type mice. However, no significant difference in antigen-induced cytokine production was observed between wild-type and E-FABP-null BMMCs, and systemic anaphylaxis was equally induced in vivo in both wild-type and E-FABP-null mice. These results suggest that E-FABP is specifically involved in the LPS-induced cytokine production of MCs, and could play a role in the host-defense against bacterial infection, possibly through regulation of TNF-alpha production. PMID- 18678478 TI - Peripheral endothelial dysfunction in patients with pulmonary arterial hypertension. AB - BACKGROUND: Pulmonary endothelium plays an important role in the mechanism of pulmonary arterial hypertension (PAH). However, there is only a few data regarding the systemic endothelium in this syndrome. This study focused on the systemic endothelial involvement in PAH. METHODS: Endothelial function was evaluated in 54 patients with idiopathic (n=28), scleroderma-associated (n=10), chronic thromboembolic (n=7), or Eisenmenger (n=9) PAH and 21 controls (13 healthy; eight scleroderma and normal pulmonary pressure). All underwent clinical evaluation, pulmonary assessment, echocardiography, and pulmonary cardiac stress test. Endothelial function was evaluated by measuring the forearm blood flow dilatation response to brachial arterial occlusion by a non-invasive plethysmograph, yielding a peripheral arterial tone (PAT) ratio. RESULTS: The PAT ratio was significantly lower (p<0.05) than healthy controls in all patients except the Eisenmenger group (control: 2.20+/-0.25; idiopathic 1.84+/-0.51; scleroderma 1.66+/-0.66; thromboembolic 1.89+/-0.32; Eisenmenger 2.17+/-0.62). The impaired hyperemic response significantly correlated with disease severity, as measured by NYHA classification (r=-0.210, p=0.035), pulmonary pressure (r= 0.228, p=0.035), 6 min walking distance (r=0.215, p=0.047), and oxygen desaturation on effort (r=0.207, p=0.038). Mean systolic pulmonary pressure among patients was 54-99 mmHg. CONCLUSION: A systemic component of endothelial dysfunction might be involved in idiopathic, scleroderma-associated and chronic thromboembolic PAH that is correlated with disease severity. PMID- 18678479 TI - Feasibility study of a soil-based rubberized CLSM. AB - The development of beneficial uses of recycled scrap tires is always in great demand around the world. The disposal of on-site surplus excavated soil and the production of standard engineering aggregates have also been facing increasing environmental and ecological challenges in congested islands, such as Taiwan. This paper presents an experimental study using recycled crumb rubber and native silty sand to produce a lightweight, soil-based, rubberized controlled low strength material (CLSM) for a bridge approach repair. To assess the technical feasibility of this material, the effects of weight ratios of cement-to-water (C/W) and water-to-solid (W/S), and of rubber content on the engineering properties for different mixtures were investigated. The presented test results include flowability, unit weight, strength, settlement potential, and bearing capacity. Based on the findings, we conclude that a soil-based rubberized CLSM with 40% sand by weight and an optimal design ratio of 0.7 for C/W and 0.35 for W/S can be used for the proposed bridge approach repair. Such a mixture has demonstrated acceptable flowability, strength, and bearing capacity. Its lower unit weight, negligible compressibility, and hydrocollapse potential also help ensure that detrimental settlement is unlikely to occur. The results illustrate a novel scheme of CLSM production, and suggest a beneficial alternative for the reduction of scrap tires as well as conservation of resources and environment. PMID- 18678480 TI - Monitoring household waste recycling centres performance using mean bin weight analyses. AB - This paper describes a modelling approach used to investigate the significance of key factors (vehicle type, compaction type, site design, temporal effects) in influencing the variability in observed nett amenity bin weights produced by household waste recycling centres (HWRCs). This new method can help to quickly identify sites that are producing significantly lighter bins, enabling detailed back-end analyses to be efficiently targeted and best practice in HWRC operation identified. Tested on weigh ticket data from nine HWRCs across West Sussex, UK, the model suggests that compaction technique, vehicle type, month and site design explained 76% of the variability in the observed nett amenity weights. For each factor, a weighting coefficient was calculated to generate a predicted nett weight for each bin transaction and three sites were subsequently identified as having similar characteristics but returned significantly different mean nett bin weights. Waste and site audits were then conducted at the three sites to try and determine the possible sources of the remaining variability. Significant differences were identified in the proportions of contained waste (bagged), wood, and dry recyclables entering the amenity waste stream, particularly at one site where significantly less contaminated waste and dry recyclables were observed. PMID- 18678481 TI - A methodology for rapid detection of Salmonella typhimurium using label-free electrochemical impedance spectroscopy. AB - A pathogen detection methodology based on Bayesian decision theory has been developed for rapid and reliable detection of Salmonella typhimurium. The methodology exploits principles from statistical signal processing along with impedance spectroscopy in order to analytically determine the existence of pathogens in the target solution. The proposed technique is validated using a cost-effective and portable immunosensor. This device uses label-free, electrochemical impedance spectroscopy for pathogen detection and has been demonstrated to reliably detect pre-infectious levels of pathogen in sample solutions. The detection process does not entail any pathogen enrichment procedures. The results using the proposed technique indicate a detection time of approximately 6min (5min for data acquisition, 1min for analysis) for pathogen concentrations in the order of 500CFU/ml. The detection methodology presented here has demonstrated high accuracy and can be generalized for the detection of other pathogens with healthcare, food, and environmental implications. Furthermore, the technique has a low computational complexity and uses a minimal data-set (only 30 data-samples) for data analysis. Hence, it is ideal for use in hand-held pathogen detectors. PMID- 18678482 TI - Thermoresponsive protein adsorption of poly(N-isopropylacrylamide)-modified streptavidin on polydimethylsiloxane microchannel surfaces. AB - The control of protein adsorption on microchannel surfaces is important for biosensors. In this study, we demonstrated protein adsorption method that is controlled through temperature change, i.e., thermoresponsive protein adsorption, on polydimethylsiloxane (PDMS) microchannel surfaces using a thermoresponsive polymer, poly(N-isopropylacrylamide) (PNIPAAm). To provide general protein adsorption control method, we adopted biotin-streptavidin chemistry and synthesized streptavidin covalently modified with PNIPAAm (PNIPAAm-StAv). Modification of streptavidin, a hydrophilic protein, with PNIPAAm induced successful thermoresponsive adsorption on a PDMS microchannel surfaces: PNIPAAm StAv adsorbed at 37 degrees C and desorbed at 10 degrees C on the surfaces. We also demonstrated the thermoresponsive adsorption of biotinylated immunoglobulin G (IgG-b) using PNIPAAm-StAv. Conjugation of IgG-b with PNIPAAm-StAv induced successful thermoresponsive IgG-b adsorption on PDMS. Modification of PDMS surfaces with PNIPAAm reduced physical adsorption of the partially hydrophobic IgG-b on the surface and contributed to the high-contrast thermoresponsive adsorption of IgG-b: less than 1% of the IgG-b adsorbed at 37 degrees C was detected after the PNIPAAm-PDMS surface was washed at 10 degrees C. The controllable adsorption of this system is expected to be applied to the regeneration of biosensor chips and to on-chip protein manipulation. PMID- 18678483 TI - Application of the self-organizing map as a prediction tool for an integrated constructed wetland agroecosystem treating agricultural runoff. AB - A self-organizing map (SOM) model was applied as a prediction tool for the performance of an integrated constructed wetland (ICW) agroecosystem treating agricultural runoff to protect receiving watercourses. By utilizing the SOM model, the time-consuming to measure expensive biochemical oxygen demand outflow concentrations were predicted well by other inexpensive variables, which were quicker and easier to measure. Correct predictions for the outflow biochemical oxygen demand concentrations were between 89% and 100%. This novel approach allows for the real time control of the outflow water quality of the ICW and potentially also of other treatment system applications. Moreover, the missing values and outliers from the large but incomplete ICW data set were replaced accurately by most likely values determined by the SOM model. This was important because the proportions of unusable entries for chemical oxygen demand, suspended solids and biochemical oxygen demand were very high: 41%, 54%, and 61%, respectively. PMID- 18678484 TI - A novel 5-[1,3,4-oxadiazol-2-yl]-N-aryl-4,6-pyrimidine diamine having dual EGFR/HER2 kinase activity: design, synthesis, and biological activity. AB - A novel 5-[1,3,4-oxadiazol-2-yl]-N-aryl-4,6-pyrimidine diamine was synthesized and found to have potent dual EGFR/HER2 kinase inhibitory activity. The structure based drug design of this molecule as well as the kinase and cellular inhibition of HER2 kinase dependent cell lines will be discussed. PMID- 18678485 TI - Studies on the porcine liver esterase-catalyzed hydrolysis of pentaacetyl catechin and epicatechin: application to the synthesis of novel dimers and trimers. AB - Porcine liver esterase-catalyzed hydrolysis of 3,5,7,3',4'-pentaacetylated catechin was studied. The selectivity of the enzyme in hydrolyzing the acetate moiety is time dependent. Careful control of the duration of hydrolysis makes it possible to isolate the differentially protected catechins. Similar result was also obtained in the epicatechin series. These results are important for elaboration of epicatechin or catechin into different derivatives with defined regiochemistry. These include novel dimeric and trimeric architectures. PMID- 18678487 TI - Synthesis of triazole-oxazolidinones via a one-pot reaction and evaluation of their antimicrobial activity. AB - C-5-substituted triazole-oxazolidinones were synthesized using a bromide catalyzed cycloaddition between aryl isocyanates and epibromohydrin followed by a three-component Huisgen cycloaddition. The library of compounds was screened for antibacterial activity against Mycobacterium smegmatis ATCC 14468, Bacillus subtilis ATCC 6633, and Enterococcus faecalis ATCC 29212. Notably, the 3-(4 acetyl-phenyl)-5-(1H-1,2,3-triazol-1-yl)methyl)-oxazolidin-2-one (18) showed an MIC of 1 microg/mL against M. smegmatis ATCC 14468, fourfold lower than the MIC measured for isoniazid. PMID- 18678486 TI - Structure-activity relationship study on a novel series of cyclopentane containing macrocyclic inhibitors of the hepatitis C virus NS3/4A protease leading to the discovery of TMC435350. AB - SAR analysis performed with a limited set of cyclopentane-containing macrocycles led to the identification of N-[17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8 methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo [13.3.0.0(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide (TMC435350, 32c) as a potent inhibitor of HCV NS3/4A protease (K(i)=0.36nM) and viral replication (replicon EC(50)=7.8nM). TMC435350 also displayed low in vitro clearance and high permeability, which were confirmed by in vivo pharmacokinetic studies. TMC435350 is currently being evaluated in the clinics. PMID- 18678488 TI - 8-Biarylchromen-4-one inhibitors of the DNA-dependent protein kinase (DNA-PK). AB - The synthesis and biological evaluation of libraries of 8-biarylchromen-4-ones enabled the elucidation of structure-activity relationships for inhibition of the DNA-dependent protein kinase (DNA-PK), with 8-(3-(thiophen-2-yl)phenyl)chromen-4 one and 8-(3-(thiophen-3-yl)phenyl)chromen-4-one being especially potent inhibitors. PMID- 18678489 TI - Discovery of a potent and selective aurora kinase inhibitor. AB - This communication describes the discovery of a novel series of Aurora kinase inhibitors. Key SAR and critical binding elements are discussed. Some of the more advanced analogues potently inhibit cellular proliferation and induce phenotypes consistent with Aurora kinase inhibition. In particular, compound 21 (SNS-314) is a potent and selective Aurora kinase inhibitor that exhibits significant activity in pre-clinical in vivo tumor models. PMID- 18678490 TI - Determination of the optimal locations of surface-mounted markers on the tibial segment. AB - This study aims to determine optimal locations on the lower limbs for skin mounted markers representing the tibial segment in three-dimensional (3D) gait analysis. It was predicted that markers located on the anterior tibial crest and malleoli would be least susceptible to soft tissue movement. Ten retro-reflective markers were attached to each tibial segment for 20 participants. Participants performed 10 walking trials and two different range-of-movement tasks (knee flexion/extension and ankle plantarflexion/dorsiflexion). The results showed a subset of four markers with inter-marker pair distances on the tibia have less than 1.6 mm variation (standard deviation (S.D.)) during walking. Minimal variation was also found in isolated ROM tasks, where marker pairs showed variability of less than 2.2 mm. Other marker locations, the femoral epicondyles and the tibial tuberosity varied up to 4 mm during walking and up to 11 mm during the isolated ROM tasks. The four marker locations that are optimal for defining the tibia are the proximal anterior tibial crest, the distal anterior tibial crest, the lateral malleolus and the medial malleolus. PMID- 18678491 TI - Synthesis and evaluation of curcumin analogues as potential thioredoxin reductase inhibitors. AB - Series of curcumin derivatives were synthesized; the inhibitory activities on thioredoxin reductase (TrxR) of all analogues were evaluated by DTNB assay in vitro. It is found that most of the analogues can inhibit TrxR in the low micromolar range; Structure-activity relationship analysis reveals that analogues with furan moiety have excellent inhibitory effect on TrxR in an irreversible manner, indicating that the furan moiety may serve as a possible pharmacophore during the interaction of curcumin analogues with TrxR. The effect of selected curcuminoids on growth of different TrxR overexpressed cancer cell lines was also investigated and discussed. PMID- 18678492 TI - Design, synthesis and in vitro antitumor activity of 4-aminoquinoline and 4 aminoquinazoline derivatives targeting EGFR tyrosine kinase. AB - Two series of new 6-alkoxy-4-substituted-aminoquinazolines (2-4f) and their bioisoteric quinoline congeners (5-7c) were designed and synthesized. Virtual screening was carried out through docking the designed compounds into the ATP binding site of epidermal growth factor receptor (EGFR) to predict if these compounds have analogous binding mode to the EGFR inhibitors. The newly synthesized compounds were tested in vitro on human breast carcinoma cell line (MCF-7) in which EGFR is highly expressed. Most of the tested compounds exploited potent antitumor activity with IC(50) values in the nanomolar range in particular compound 3b which displayed the highest activity among the tested compounds with IC(50) equal to 0.13 nmol. PMID- 18678493 TI - Allele-specific inhibition of divergent protein tyrosine phosphatases with a single small molecule. AB - A central challenge of chemical biology is the development of small-molecule tools for controlling protein activity in a target-specific manner. Such tools are particularly useful if they can be systematically applied to the members of large protein families. Here we report that protein tyrosine phosphatases can be systematically 'sensitized' to target-specific inhibition by a cell-permeable small molecule, Fluorescein Arsenical Hairpin Binder (FlAsH), which does not inhibit any wild-type PTP investigated to date. We show that insertion of a FlAsH binding peptide at a conserved position in the PTP catalytic-domain's WPD loop confers novel FlAsH sensitivity upon divergent PTPs. The position of the sensitizing insertion is readily identifiable from primary-sequence alignments, and we have generated FlAsH-sensitive mutants for seven different classical PTPs from six distinct subfamilies of receptor and non-receptor PTPs, including one phosphatase (PTP-PEST) whose three-dimensional catalytic-domain structure is not known. In all cases, FlAsH-mediated PTP inhibition was target specific and potent, with inhibition constants for the seven sensitized PTPs ranging from 17 to 370 nM. Our results suggest that a substantial fraction of the PTP superfamily will be likewise sensitizable to allele-specific inhibition; FlAsH-based PTP targeting thus potentially provides a rapid, general means for selectively targeting PTP activity in cell-culture- or model-organism-based signaling studies. PMID- 18678494 TI - Novel synthesis and pharmacological evaluation as alpha2-adrenoceptor ligands of O-phenylisouronium salts. AB - The synthesis of nine new mono- and bis-O-phenylisouronium compounds (2, 6b-10b and 12b-14b) and their Boc-protected isourea precursors (2a, 6a-10a and 12a-14a) is described. The carbodiimide 4, which was formed, had been suggested as the reactive intermediate species and driving force of the reaction. All final substrates were tested as potential alpha(2)-ARs ligands in human brain tissue by means of radioligand binding experiments and were compared to the potential antidepressant 1, as well as other related guanidine containing derivatives. PMID- 18678495 TI - Structure-activity relationships of new inhibitors of breast cancer resistance protein (ABCG2). AB - At the end of the last century tariquidar (XR9576) was synthesized, pharmacologically investigated, and classified as a promising 3rd generation P glycoprotein (P-gp) modulator. Following the discovery of BCRP in 1998 an increasing number of substances were studied in relation to their potency to interact with this transporter. Recently it has been shown that XR9576 inhibits both P-gp and BCRP transport function similarly to GF120918 (elacridar). This observation prompted us to investigate 5 XR compounds and 25 structurally related derivatives synthesized in our laboratory for their BCRP inhibitory effect. The biological activity data were determined by our new Hoechst 33342 assay that has been transferred from P-gp to BCRP overexpressing cells. 3D-QSAR models (CoMFA and CoMSIA) were generated and validated by the leave-many-out method and the scrambling stability test. The best models yielded an internal predictive squared correlation coefficient higher than 0.8 and contained steric, electrostatic, hydrophobic, and hydrogen bond donor fields. To our knowledge, this is the first 3D-QSAR analysis of BCRP inhibitors. Additionally the biological activity data determined in P-gp overexpressing cells on one side and BCRP overexpressing cells on the other side were compared to identify selective and non-selective inhibitors of P-gp and BCRP. The results may help to get a better insight which structural elements are necessary to direct the interaction of these compounds with P-gp and/or BCRP. PMID- 18678496 TI - TX-2152: a conformationally rigid and electron-rich diyne analogue of FTY720 with in vivo antiangiogenic activity. AB - We designed FTY720 analogues with conformationally rigid and electron-rich acetylenic chains as antiangiogenic agents (the monoyne 1: TX-2148, the diyne 2: TX-2152, the triyne 3: TX-2256). Molecular orbital (MO) calculations of our designed acetylenic analogues and FTY720 showed that the localization of the lowest unoccupied MO and the highest occupied MO increased from phenyl ring to acetylenic chain compared with that of FTY720. These acetylenic analogues were synthesized from p-hydroxyphenylethanol as a starting material. The construction of the acetylenic chain was carried out by an iterative strategy using a Sonogashira cross-coupling reaction and desilylative bromination in two steps. The corresponding overall yields of the monoyne 1, the diyne 2, and the triyne 3 were 27% (11 steps), 13% (13 steps), and 10% (15 steps). The in vivo antiangiogenic activities of these acetylenic analogues and FTY720 were evaluated by the chick embryo chorioallantoic membrane (CAM) assay and compared to the activities of the known antiangiogenic agent TNP-470. The diyne 2 showed more potent antiangiogenic activity (90% inhibition) than FTY720 (77% inhibition) and other acetylenic analogues (the monoyne 1: 42% inhibition, the triyne 3: 60% inhibition), and TNP-470 (82% inhibition) at a dose of 10 microg/CAM, without showing toxicity. The diyne 2 also had potent inhibitory activity at a dose of 5 and 2.5 microg/CAM. These results indicate that the flexibility of C8 alkyl chain of FTY720 is not required for its antiangiogenic activity. We suggest that the diyne 2 (TX-2152) may be a promising candidate as an antiangiogenic agent for antineoplastic drug discovery. PMID- 18678497 TI - Structure-activity relationship studies on a novel class of antiproliferative agents derived from Lavendustin A. Part I: Ring A modifications. AB - The potent antiproliferative agent SDZ LAP 977, which has shown efficacy in a clinical proof of concept study in actinic keratosis patients, has been previously demonstrated to block the cell cycle in mitosis. In the present study, we further explored the mode of action: SDZ LAP 977 binds to the "colchicine binding site" on tubulin and, thus, inhibits tubulin polymerization in vitro. Moreover, we established structure-activity relationships for the effect of modifications in the 2,5-dimethoxyphenyl moiety ("ring A") of the molecule on in vitro antiproliferative activity. PMID- 18678498 TI - Antifungal activity of synthetic di(hetero)arylamines based on the benzo[b]thiophene moiety. AB - The antifungal activity of several di(hetero)arylamine derivatives of the benzo[b]thiophene system was evaluated against clinically relevant Candida, Aspergillus, and dermatophyte species by a broth macrodilution test based on CLSI (formerly NCCLS) guidelines. The most active compound showed a broad spectrum of activity (against all tested fungal strains, including fluconazole-resistant fungi), with particularly low MICs for dermatophytes. Results from the inhibition of the dimorphic transition in Candida albicans and flow cytometry studies further confirmed their biological activity. With this study it was possible to establish some structure-activity relationships (SARs). The hydroxy groups proved to be essential for the activity in the aryl derivatives. Furthermore, the spectrum of activity in the pyridine derivatives was broadened by the absence of the ester group on position 2 of the benzo[b]thiophene system. PMID- 18678499 TI - Chemical synthesis of scyllo-inosamine and catabolism studies in Sinorhizobium meliloti. AB - Rhizopines such as scyllo-inosamine (SIA) and L-3-O-methyl-scyllo-inosamine (3-O MSI) play an intricate role as nutritional mediators during the establishment of the symbiotic relationship between legumes and rhizobia. The mechanism of action is not well understood. One challenge is the availability of rhizopines, which occur in only minute amounts in plant nodules. We herewith report an efficient synthesis of scyllo-inosamine and its biochemical activity in specific bacteria. SIA was prepared in 7 steps and 32% overall yield from readily available myo inositol. The chemically synthesized SIA was tested to determine whether it can serve as sole carbon and nitrogen source for Sinorhizobium meliloti wild-type strain L5-30 and for strains carrying mutations in the rhizopine degradation (moc) genes. The analysis of the phenotype of the mutant strains revealed that the moc genes previously shown to be essential for the breakdown of the rhizopines isolated from root nodules are also essential for the utilization of the chemically synthesized SIA. PMID- 18678500 TI - Synthesis and cellular effects of cycloterpenals: cyclohexadienal-based activators of neurite outgrowth. AB - An unusual class of diterpenoid natural products, 'cycloterpenals' (with a central cyclohexadienal core), that arise in nature by condensation of retinoids and other isoprenes, have been isolated from a variety of organisms including marine sponges as well as from the human eye. A milk whey protein has also demonstrated the formation of a cycloterpenal derived from beta ionylidineacetaldehyde. Here, we generate a synthetic library of these molecules where we detail reaction conditions required to effect cross condensation of alpha,beta-unsaturated aldehydes as opposed to homodimerization. The ability of this class of molecules to activate neurite outgrowth activity is reported. PMID- 18678501 TI - 1,3,4-Oxadiazole-3(2H)-carboxamide derivatives as potential novel class of monoamine oxidase (MAO) inhibitors: synthesis, evaluation, and role of urea moiety. AB - A new series of 1,3,4-oxadiazole-3(2H)-carboxamide derivatives have been synthesized by direct heterocyclization reaction of substituted benzoylisocyanate with various aroylhydrazones as novel monoamine oxidase inhibitors (MAOIs). The target molecules have been identified on the basis of satisfactory analytical and spectra (IR, (1)H NMR, (13)C NMR, and HR-MS) data. The newly synthesized compounds were evaluated for their MAO inhibitory activity by kynuramine fluorimetric assay method. The preliminary results showed that most of the compounds have moderate inhibitory activities toward MAO at the concentration of 10(-5)-10(-3)M. This work may provide a novel class of lead compounds with potential MAO inhibitions for further optimization. PMID- 18678502 TI - Identification of SVM-based classification model, synthesis and evaluation of prenylated flavonoids as vasorelaxant agents. AB - Support vector machine (SVM) was applied to predict vasorelaxation effect of different structural molecules. A good classification model had been established, and the accuracy in prediction for the training, test, and overall datasets was 93.0%, 82.6%, and 89.5%, respectively. Furthermore, the model was used to predict the activity of a series of prenylated flavonoids. According to the estimated result, eleven molecules 1-11 were selected and synthesized. Their vasodilatory activities were determined experimentally in rat aorta rings that were pretreated with phenylephrine (PE). Structure-activity relationship (SAR) analysis revealed that flavanone derivatives showed the most potent activities, while flavone and chalcone derivatives exhibited medium activities. PMID- 18678503 TI - Lethal events in V79 cells irradiated by low-energy protons and correlations with distribution patterns of energy deposition, radical concentration and DNA damage. AB - Published survival data of V79 cells irradiated by 0.5-5.0 MeV (7-40 keV/microm) protons have been analyzed with a detailed radiobiological model to estimate the per-track yields of lethal lesions. Their correlations with distribution patterns of deposited energy, radical concentrations and with the yields of specific classes of DNA damage have been studied. The observed correlations indicate a potential interpretation of DNA damage lethal for the cell and the initial physical and chemical processes leading to such damage. PMID- 18678504 TI - Esophagorespiratory fistulas of tumorous origin. Non-operative management of 264 cases in a 20-year period. AB - OBJECTIVE: Esophagorespiratory fistulas developing from malignant tumors have serious complications by maintaining continuous airway contamination. The objective was to reveal the incidence, causes and characteristics of fistula formation and to examine the possibilities and efficiency of the treatment. METHODS: In a single-center study between 1984 and 2004, the data of 2113 patients with tumorous esophageal stenosis were analyzed. Esophagorespiratory fistulas were detected in 264 cases (12.5%). Successful esophageal intubation, stent correction or replacement was performed in 188 cases, while there was one lethal complication. Twenty-seven patients had an intervention for nutritional support: 25 gastrostomies, 1 jejunostomy and 1 percutaneous endoscopic gastrostomy. RESULTS: The mean survival period of all patients was 2.8 months; patients with implanted endoprosthesis 3.4 months; with nutritional support 1.1 months and with only supportive therapy 1.3 months, respectively. The differences between the endoprosthesis implanted group and the other two groups were significant (p<0.001). CONCLUSIONS: By sealing the fistula, a successful endoscopic esophageal intubation ends the severe respiratory contamination and the inability to swallow, improving the quality of life and survival period. After the procedure, it is the malignant tumor and not the fistula that determines the future of the patient. PMID- 18678505 TI - Fractured prosthetic valve leaflet. PMID- 18678506 TI - Heart allograft transplanted twice. AB - We present the case of a man who underwent successful heart transplantation with an allograft that was obtained from a donor who had already received heart transplantation. PMID- 18678507 TI - Attenuation of receptor-dependent and -independent vasoconstriction in the human radial artery. AB - BACKGROUND: Vasodilator strategies used to treat bypass grafts in the operating theatre, such as nitrates, phosphodiesterase inhibitors and calcium channel antagonists have a broad but short-lived effect against a variety of vasoconstrictor stimuli. Treatments that react irreversibly with proteins modulating vasoconstriction have the advantage that their effects can last well into the postoperative period. In addition systemic effects are avoided as the treatment is localised to the treated graft. This study investigated the use of two clinically applied drugs; fluphenazine (SKF7171A, HCl), an irreversible calmodulin antagonist and minoxidil sulphate, an irreversible potassium channel opener. Treatments were tested against receptor and non-receptor-mediated contraction in the human radial artery. METHOD: Isometric tension was measured in response to angiotensin II, KCl and vasopressin in 108 radial artery rings (taken from 31 patients undergoing coronary artery bypass grafting). Control responses were compared with rings pretreated with fluphenazine or minoxidil sulphate. Vasopressin responses were also compared in the presence of glyceryl trinitrate or the reversible Rho kinase inhibitor Y27632. RESULTS: Fluphenazine pretreatment significantly suppressed vasoconstriction to all agonists tested. Maximal responses to angiotensin II, vasopressin and KCl were reduced by 42+/-19%, 35+/ 8% and 48+/-15% respectively, without any measurable effect on the EC(50). Minoxidil sulphate showed no discernable effect. Vasopressin-induced contraction was also reduced by high levels of glyceryl trinitrate (220 microM; 50 microg/ml) or 10 microM Y27632. CONCLUSIONS: The irreversible calmodulin antagonist fluphenazine has potential to be developed as an inhibitor of contraction in arterial graft vessels. The involvement of Rho kinase indicates that other vasoconstrictors and surgical stress can sensitize radial artery to vasopressin induced contraction. Strategies targeting this pathway also have future potential. PMID- 18678508 TI - Chest wall reconstruction with two types of biodegradable polymer prostheses in dogs. AB - OBJECTIVE: Currently, the choice of chest wall prosthesis remains a challenging problem for thoracic and reconstructive surgeons. The purpose of this study is to investigate the feasibility of newly developed biodegradable prostheses. METHODS: Two types of chest wall prostheses made from degradable polymer, collagen coated polydioxanone (CCP) mesh and chitin fiber reinforced polycaprolactone (CFRP) strut, were developed and studied. Adult mongrel dogs were subjected to extensive resection and reconstruction of anterior-lateral chest wall, CCP mesh was used in six dogs, the combination of CCP mesh and CFRP strut was used in four dogs, and polypropylene (PP) mesh in two dogs, as contrast. RESULTS: With good integration with tissue, CCP meshes maintained strength in the chest wall for more than 8 weeks and were completely resorbed within 24 weeks, and satisfactory short-term and long-term chest wall stabilization was achieved. The combined use of CCP mesh with CFRP strut provided a firmer chest wall in the early postoperative course. A mild wound infection developed in one animal with CCP mesh but resolved without sequelae, and no added complications were observed with the additional use of CFRP strut. CONCLUSIONS: Our experimental study shows that the CCP mesh and CFRP prosthesis were favorable for chest wall repair. The advantages of biodegradable copolymer give them promise as an excellent addition to the available reconstructive techniques currently in use. PMID- 18678509 TI - Hybrid procedures can reduce the risk of congenital cardiovascular surgery. AB - BACKGROUND: Minimally invasive operations and percutaneous interventions are well accepted options in the treatment of congenital heart defects. However, percutaneous interventions may be associated with an increased risk due to limited vascular access or a very tortuous catheter course. In these cases, combining operative and interventional approaches with direct puncture of the heart or the great vessels may facilitate implantation of even large devices. Furthermore, in some situations, cardiopulmonary bypass or circulatory arrest can be omitted when doing a hybrid procedure. PATIENTS: Between January 2000 and April 2007 17 patients were operated in a hybrid fashion. Age ranged from 14 days to 45 years. Operative procedures consisted of implantation of an atrial septal defect occluder via direct puncture of the right atrium (n=4), closure of a ventricular septal defect via direct puncture of the right ventricle (n=1), implantation of isthmus stents via the ascending aorta (n=5), redilation of an isthmus stent (n=1), redilation of a ductal stent (n=1), angioplasty of a pulmonary artery stenosis (n=1), interventional occlusion of an intrahepatic porto-caval shunt (n=1), stent implantation into the right pulmonary artery (n=1) and into the right ventricular outflow tract (n=1) under direct vision as well as atrioseptoplasty combined with a bilateral pulmonary artery banding in one newborn with a single ventricle and very low birth weight (n=1). RESULTS: The planned intervention could be performed in all cases under the assistance of intraoperative fluoroscopy, transesophageal or epicardial echocardiography, or under direct vision. In all cases, the primary hemodynamic objectives were achieved. CONCLUSION: In selected patients, the combination of a surgical procedure and a percutaneous intervention may help to reduce both operative and interventional risks. This concept may enable new treatment options, especially in patients with complex congenital heart defects or complex vascular situations. PMID- 18678510 TI - Modified Bentall operation. PMID- 18678511 TI - Outcomes following non-valved autologous reconstruction of the right ventricular outflow tract in neonates and infants. AB - OBJECTIVE: Controversy surrounds the optimal method of establishing right ventricle to pulmonary artery continuity in neonates and infants with congenital heart disease. We reviewed our experience with non-valved autologous reconstruction of the right ventricular outflow tract to determine mid-term outcome and risk factors for reintervention. METHODS: Between 1998 and 2006, 34 consecutive patients underwent non-valved autologous right ventricular outflow tract reconstruction. The need for postoperative catheter-based intervention or reoperation was assessed using relevant patient and procedure-related variables. RESULTS: Diagnoses included tetralogy of Fallot with anomalous coronary (n=3), tetralogy of Fallot with pulmonary atresia (n=10), truncus arteriosus communis (n=15), and other (n=6). Median age at surgery was 5 days (1-270 days). Twenty six (76%) patients were neonates. Median weight was 3.1kg (1.8-7.3kg). At a median follow-up of 43 months (1-90 months), 15 (50%) patients underwent reoperation and 7 (23%) underwent catheter-based intervention, with a total of 16 (53%) undergoing either reoperation or catheter-based intervention. Kaplan-Meier freedom from reintervention at 6 months, 1 year, 3 years, and 5 years was 67%, 47%, 47%, and 35% for truncus arteriosus versus 87%, 82%, 68%, and 65% for diagnoses other than truncus arteriosus (p=0.05). CONCLUSIONS: Mid-term outcome following non-valved autologous reconstruction of the right ventricular outflow tract is satisfactory and constitutes a sound alternative to the use of small diameter conduits in neonates and infants. In our hands, this strategy favors certain anatomic subtypes. Non-truncus patients have significantly lower rates of reintervention. Technical details associated with the anatomical reconstruction of the posterior autologous pathway may play an important role in outcomes. PMID- 18678512 TI - Cardiac silhouette following a mitral valve replacement, tricuspid annuloplasty and atrioplasty of both atria. PMID- 18678513 TI - Chromatographic NMR in NMR solvents. AB - Recently, it was demonstrated that pseudo-chromatographic NMR experiments could be performed using typical chromatographic solids and solvents. This first setup yielded improved separation of the spectral components of the NMR spectra of mixtures using PFG self-diffusion measurements. The method (dubbed Chromatographic NMR) was successively shown to possess, in favorable cases, superior resolving power on non-functionalized silica, compared to its LC counterpart. To further investigate the applicability of the method, we studied here the feasibility of Chromatographic NMR in common deuterated solvents. Two examples are provided, using deuterated chloroform and water, for homologous compounds soluble in these solvents, namely aromatic molecules and alcohols, respectively. PMID- 18678514 TI - Spatial and temporal mapping of water content across Nafion membranes under wetting and drying conditions. AB - Water transport and water management are fundamental to polymer electrolyte membrane fuel cell operation. Accurate measurements of water content within and across the Nafion layer are required to elucidate water transport behavior and validate existing numerical models. We report here a direct measurement of water content profiles across a Nafion layer under wetting and drying conditions, using a novel magnetic resonance imaging methodology developed for this purpose. This method, multi-echo double half k-space spin echo single point imaging, based on a pure phase encode spin echo, is designed for high resolution 1D depth imaging of thin film samples. The method generates high resolution (<8 microm) depth images with an SNR greater than 20, in an image acquisition time of less than 2 min. The high temporal resolution permits water content measurements in the transient states of wetting and drying, in addition to the steady state. PMID- 18678515 TI - Design of peptidomimetic inhibitors of aspartic protease of HIV-1 containing -Phe Psi Pro- core and displaying favourable ADME-related properties. AB - Aspartic protease (PR) of HIV-1 virus represents a valid therapeutic target for the design of antiviral agents suitable for treatment of AIDS. We have designed peptidomimetic PR inhibitors containing a novel dihydroxyethylenediamine -Phe Psi[CHOH-CHOH]-Pro- core using molecular modelling approach that predicts the inhibitory potencies (IC(50)(pre)) in terms of computed relative enzyme-inhibitor complexation Gibbs free energies (Delta Delta G(comp)). The modelling approach considers not only the enzyme-inhibitor interactions, but includes also the solvent and entropic effects affecting the enzyme inhibition. The objectives of this study were to optimize the number and type of flanking residues that occupy the S(3), S(2) and S(2'), S(3') positions in the PR binding pocket and to select potent lead candidates, which display also favourable ADME-related properties. The structure-based design was combined with a synthetic strategy used to prepare a training set of 10 analogues sharing the -Phe Psi Pro- core. This strategy couples stereochemical control with full flexibility in the choice of the flanking residues and in vitro activity assays. A QSAR model correlating calculated Delta Delta G(comp) with the measured IC(50)(exp) values for the training set was prepared and confirmed that our computational approach can serve for reliable prediction of PR inhibitory potencies of peptidomimetics. The appropriate choice of the flanking residues allowed us to design virtual lead compounds, such as FP14, FP23 and FP76, with reduced molecular weight, predicted inhibitory potencies in the picomolar range, promising ADME profiles and a potential to escape drug resistance due to favourable interactions with the PR backbone. PMID- 18678516 TI - Relationship between GH response and glycemic fluctuations in the glucagon stimulation test. AB - OBJECTIVE: Verifying the association between glycemic fluctuation and GH response in the glucagon stimulation test. Basal evaluation of growth hormone (GH) has poor diagnostic accuracy due to its pulsatile secretion. GH-stimulation tests are used for an adequate evaluation of somatotrophic axis. Various stimuli can be employed, among them glucagon, which has an elusive mechanism of action. Since hypoglycemia reportedly occurs during the test, investigation of its role as a stimulus to GH release is granted. DESIGN: Retrospective analysis of glucagon stimulated GH tests performed in 128 children (36.7% female; age 12.4+3.3 years), at Fleury Functional Tests Facility from July 2000 to 2006. GH and blood glucose (BG) curves, IGF-1, and IGFBP-3 have been assessed. Positive GH response was defined by a peak GH value >or=3.3 microg/L. Normal IGF-1 levels were defined as those between 2.5th and 97.5th percentiles for age and gender. RESULTS: Hypoglycemia under 2.2 mmol/L did not occur during the test. BG decrease occurred with lower magnitude and was not associated to GH response. Comparison between patients with negative and positive GH response showed, respectively, BG nadir 3.74 vs. 3.62 mmol/L, glucose AUC 23.3 vs. 22.4, and glycemic decrease (below 3.3 mmol/L) 19% vs. 35.5% (with P non-significant for all comparisons). CONCLUSION: Hypoglycemia was not seen after glucagon stimulation and decrease in BG occurred above levels physiologically expected to stimulate GH release, being apparently not associated to GH response. PMID- 18678517 TI - A missense variant in desmoglein-2 predisposes to dilated cardiomyopathy. AB - Familial Dilated Cardiomyopathy (FDCM) is caused by mutations in genes encoding myocardial force transduction proteins. Desmoglein-2 (DSG2) and Desmocollin-2 (DSC2) provide cellular adhesion and force transduction by cell-to-cell anchorage. To test whether perturbations of DSG2 or DSC2 exhibit a pathogenic impact on DCM pathogenesis, we sequenced both genes in 73 patients with FDCM and assessed prevalence of missense variations in matched control cohorts. We detected two missense variations in DSG2 (V55M and V919G) which were absent in 360 control alleles. Surprisingly, both variants were previously reported in patients with arrhythmogenic right ventricular cardiomyopathy. Yet, in the present study only the DSG2-V55M variant showed segregation with DCM in a family pedigree. Subsequent, analysis of 538 patients with idiopathic DCM and 617 consecutive control individuals resulted in identification of thirteen DSG2-V55M carriers with DCM, whereas only three control subjects harbored the variant. DSG2 immunostaining revealed pale structures of the intercalated disc in myocardium of one unique homozygous DSG2-V55M carrier. Furthermore, myocardial desmosomal structures were significantly shortened when compared to DCM myocardium negative for DSG2-V55M. Thus, our study identified the DSG2-V55M polymorphism as a novel risk variant for DCM associated with shortened desmosomes of the cardiac intercalated disc. PMID- 18678518 TI - Viral hepatitis in a multi-ethnic neighborhood in the Netherlands: results of a community-based study in a low prevalence country. AB - OBJECTIVES: The prevalence of viral hepatitis varies worldwide. Although the prevalence of hepatitis A virus (HAV) and hepatitis B virus (HBV) infection is generally low in Western countries, pockets of higher prevalence may exist in areas with large immigrant populations. The aim of this study was to obtain further information on the prevalence of viral hepatitis in a multi-ethnic area in the Netherlands. METHODS: We conducted a community-based study in a multi ethnic neighborhood in the city of Rotterdam, the Netherlands, including both native Dutch and migrant participants, who were tested for serological markers of hepatitis A, hepatitis B, and hepatitis C infection. RESULTS: Markers for hepatitis A infection were present in 68% of participants. The prevalence of hepatitis B core antibodies (anti-HBc), a marker for previous or current infection, was 20% (58/284). Prevalence of hepatitis A and B varied by age group and ethnicity. Two respondents (0.7%) had chronic HBV infection. The prevalence of hepatitis C was 1.1% (3/271). High levels of isolated anti-HBc were found. CONCLUSIONS: We found a high prevalence of (previous) viral hepatitis infections. This confirms previous observations in ethnic subgroups from a national general population study and illustrates the high burden of viral hepatitis in areas with large immigrant populations. PMID- 18678519 TI - High rate of complications associated with extrafocal kirschner wire pinning for distal radius fractures. AB - Kirschner wire (K-wire) fixation is a common technique aimed at treating unstable distal radius fractures. We report our experience with extrafocal K-wire fixation in the treatment of unstable distal radius fractures in 34 patients (AO classification: 10 A2, 5 A3, 14 C1, and 5 C2). The patients' median age was 63 years (range, 16-87 years). K-wires remained in place for a median duration period of 42 days, and a splint was applied during 44 days in median. The median follow-up was 2.5 years (range, 1.3-5 years). At the final follow-up examination, all patients were assessed clinically and bilateral radiographs were taken. The median final range of motion was 69 degrees /64 degrees for extension/flexion, 88 degrees/82 degrees for pronation/supination and 36 degrees/21 degrees for ulnar/radial deviation. Grip strength was 81% of the contralateral side. Using the Gartland-Werley score, 12 patients (35%) had excellent results, 17 (50%) good, and five patients (15%) had fair results. The median final radiographic dorsal tilt was 4.5 degrees (range, 0-14 degrees) and 79% of patients had a loss of volar reduction (median 8 degrees). Radial inclination was in median 20 degrees (range, 10-32 degrees) with 82% of the patients with loss of reduction (median 5 degrees). Radial shortening was in median 0.9 mm (range, -2 to +5 mm) and 62% of patients had median loss of radial length of 1.1mm. Radial shortening was the major source of pain due to ulnar impingement. We found a high complication rate, with 15 patients (44%) suffering from at least one of the 30 reported complications. We did not find any direct correlation between radiographic results and mobility. Although good clinical outcome may be obtained, K-wire fixation cannot be routinely recommended. PMID- 18678520 TI - Hemiballism may not be due to isolated injury to the subthalamic nucleus. PMID- 18678522 TI - Comes a time. AB - The circadian clock is a selfsustaining oscillator with an endogenous period of approximately 24 hours. The Arabidopsis clock is composed of a set of interlocking negative feedback loops entailing transcriptional, post transcriptional, and post-translational, particularly regulated proteolysis, control. Clock control of the transcriptome is widespread; up to 90% of the transcriptome cycles in at least one condition in seedlings exposed to a variety of environmental cycles. Clock control extends to the metabolome, though diurnal oscillations in enzyme activities and metabolites are less dramatic than oscillations in cognate transcripts. Metabolites, including organic nitrogen intermediates, feed back to modulate clock function, consistent with the view of the circadian clock as a key integrator of metabolic signals to coordinate metabolism and physiology with the environment. PMID- 18678521 TI - A non-parametric vessel detection method for complex vascular structures. AB - Modern medical imaging techniques enable the acquisition of in vivo high resolution images of the vascular system. Most common methods for the detection of vessels in these images, such as multiscale Hessian-based operators and matched filters, rely on the assumption that at each voxel there is a single cylinder. Such an assumption is clearly violated at the multitude of branching points that are easily observed in all, but the most focused vascular image studies. In this paper, we propose a novel method for detecting vessels in medical images that relaxes this single cylinder assumption. We directly exploit local neighborhood intensities and extract characteristics of the local intensity profile (in a spherical polar coordinate system) which we term as the polar neighborhood intensity profile. We present a new method to capture the common properties shared by polar neighborhood intensity profiles for all the types of vascular points belonging to the vascular system. The new method enables us to detect vessels even near complex extreme points, including branching points. Our method demonstrates improved performance over standard methods on both 2D synthetic images and 3D animal and clinical vascular images, particularly close to vessel branching regions. PMID- 18678523 TI - EPR, mass, electronic, IR spectroscopic and thermal studies of bimetallic copper(II) complexes with tetradentate ligand, 1,4-diformyl piperazine bis(carbohydrazone). AB - The synthesis of novel bimetallic Cu(II) complexes with general stoichiometry [Cu(2)(H(2)L)X(2)(H(2)O)(2)], [Cu(2)(H(2)L)(CH(3)COO)(2)] and [Cu(2)(H(2)L)SO(4)(H(2)O)(2)] (where H(2)L=dideprotonated ligand and X=NO(3)(-) and Cl(-)) derived from tetradentate ligand obtained by the condensation of 1,4 diformyl piperazine with carbohydrazide has been discussed. The complexes were characterized by elemental analyses, molar conductance measurements, magnetic susceptibility measurements, IR, mass, UV, EPR spectral studies and thermogravimetric analyses. The value of magnetic moments indicates that the complexes are paramagnetic and show the antiferromagnetic interaction between the two metal centres. The complexes possess the square planar coordination environment. The values of covalency measurements, i.e., in-plane sigma-bonding alpha(2), in-plane pi-bonding beta(2) and orbital reduction factor k indicate the covalent nature of complexes. PMID- 18678524 TI - Cyanobacterial chlorophyll as a sensitizer for colloidal TiO2. AB - Chlorophyll has been extracted from cyanobacteria. The adsorption of chlorophyll on the surface of colloidal TiO(2) through electrostatic interaction was observed. The apparent association constant (K(app)) of chlorophyll-TiO(2) obtained from absorption spectra is 3.78x10(4)M(-1). The K(app) value of chlorophyll-TiO(2) as determined from fluorescence spectra is 1.81x10(4)M(-1), which matches well with that determined from the absorption spectra changes. These data indicate that there is an interaction between chlorophyll and colloidal TiO(2) nanoparticle surface. The dynamics of photoinduced electron transfer from chlorophyll to the conduction band of colloidal TiO(2) nanoparticle has been observed and the mechanism of electron transfer has been confirmed by the calculation of free energy change (DeltaG(et)) by applying Rehm-Weller equation as well as energy level diagram. Lifetime measurements gave the rate constant (k(et)) for electron injection from the excited state chlorophyll into the conduction band of TiO(2) is 4.2x10(8)s(-1). PMID- 18678525 TI - Merocyanine solvatochromic dyes in the study of synergistic effects in mixtures of chloroform with hydrogen-bond accepting solvents. AB - The molar transition energy (E(T)) polarity values for the solvatochromic probes 2,6-diphenyl-4-(2,4,6-triphenylpyridinium)phenolate (1), 4[(1-methyl-4-(1H) pyridinylidene)-ethylidene]-2,5-cyclohexadien-1-one (2), and 4-[4 (dimethylamino)styryl]-1-methylpyridinium iodide (3) were collected in binary mixtures comprising chloroform and a hydrogen-bond accepting (HBA) solvent [dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), acetone or acetonitrile], aiming to investigate the ability of the chlorinated component to act as hydrogen-bond donating (HBD) solvent. Plots of E(T) as a function of X(2), the mole fraction of chloroform, were obtained and the data were analysed to investigate the preferential solvation (PS) of each probe in terms of both solute-solvent and solvent-solvent interactions. For dyes 1 and 2 a strong synergistic behavior was observed for all mixtures studied, indicating that the dyes are preferentially solvated by complexes formed through hydrogen bonding between chloroform and the HBA component in the mixtures. A study of 1 in deuterated chloroform with an HBA component (DMF and DMA) demonstrated that while almost no differences occur with the DMF mixtures, the presence of deuterated chloroform in its mixtures with DMA increases the synergistic effect, suggesting that it interacts more strongly with DMA, making its mixtures more polar. These data were successfully fitted to a model based on solvent-exchange equilibria. The features of the mixtures with dye 3 revealed a very different profile in comparison with the other two dyes, which suggests that in mixtures containing chloroform, the microenvironment of the dye seems to be important in determining the contribution of the structure resonances responsible for the stability of the dye. PMID- 18678526 TI - Clinical trials update from Heart Rhythm 2008 and Heart Failure 2008: ATHENA, URGENT, INH study, HEART and CK-1827452. AB - This article provides information and a commentary on trials relevant to the pathophysiology, prevention and treatment of heart failure, presented at the Heart Rhythm Society meeting in San Francisco, USA and the Heart Failure Association meeting of the European Society of Cardiology which was held in Milan, Italy in June 2008. Unpublished reports should be considered as preliminary data, as analyses may change in the final publication. The ATHENA study showed that dronedarone reduced the incidence of the composite outcome of cardiovascular hospitalisation or death, in patients with atrial fibrillation or flutter, 29% of whom had a history of heart failure, compared with placebo. The URGENT study demonstrated that treatment of acute heart failure with standard therapy, including intravenous diuretics and nitrates, leads to a rapid resolution of breathlessness in the sitting position but that orthopnoea often persists. The INH study showed that a disease management programme could reduce mortality compared to usual care but not hospitalisation rates. The HEART study failed to recruit its planned number of patients, although it is the largest randomised trial of revascularisation in heart failure reported to date. At a median follow-up of 5 years no difference in mortality was observed but the study lacked power to provide a conclusive result. The selective myosin activator CK 1827452 produced a concentration dependent increase in systolic ejection time, stroke volume and fractional shortening in patients with heart failure compared to placebo. PMID- 18678527 TI - Knowlesi malaria: newly emergent and of public health importance? AB - Several questions on public health impact have arisen from the discovery of a large focus of the simian malaria parasite, Plasmodium knowlesi, in the human population. P. knowlesi malaria is not newly emergent and was overlooked until molecular tools to distinguish between P. knowlesi and the morphologically similar Plasmodium malariae became available. Knowlesi malaria is a zoonosis that is widely distributed in Southeast Asia and can be fatal. Information on knowlesi malaria should be included in medical and public health guidelines to encourage the accurate diagnosis and treatment of patients, and monitor the incidence and distribution of cases. A complete emergence of P. knowlesi into the human population could be overwhelming and, although challenging, the prevention of this situation deserves serious consideration. PMID- 18678528 TI - Vaccination of dogs against Echinococcus granulosus: a means to control hydatid disease? AB - Hydatid disease continues to be a substantial cause of morbidity and mortality worldwide. Elimination is difficult with current control options, but reducing egg production by Echinococcus granulosus (Eg) in canines might help to reduce transmission in areas where the parasite is endemic. Recently obtained data using recombinant protein-based and live attenuated Salmonella vaccines are preliminary but encouraging and auger well for the future development of an effective dog vaccine against Eg, although much additional work is required before this becomes a reality. New approaches to control and for the prevention of hydatidosis have been described recently, and important additional gains should be expected if the efficacy of the dog vaccines is confirmed and leads to their incorporation into future control options. PMID- 18678529 TI - Mosquitocidal vaccines: a neglected addition to malaria and dengue control strategies. AB - The transmission of vector-borne diseases is dependent upon the ability of the vector to survive for longer than the period of development of the pathogen within the vector. One means of reducing mosquito lifespan, and thereby reducing their capacity to transmit diseases, is to target mosquitoes with vaccines. Here, the principle behind mosquitocidal vaccines is described, their potential impact in malaria and dengue control is modeled and the current research that could make these vaccines a reality is reviewed. Mosquito genome data, combined with modern molecular techniques, can be exploited to overcome the limited advances in this field. Given the large potential benefit to vector-borne disease control, research into the development of mosquitocidal vaccines deserves a high profile. PMID- 18678530 TI - Management of vesicoureteral reflux without indwelling catheter and drain, using trigonoplasty technique. AB - PURPOSE: Gil-Vernet trigonoplasty is a simple, fast and effective technique for treating primary VUR, and is the only technique used to manage unilateral reflux that does not result in contralateral new reflux, due to its inherent bilateral nature. We have tried to further simplify postoperative management of the procedure by eliminating the use of an indwelling urethral catheter and drain. PATIENTS AND METHODS: In a prospective study during a 15-month period, 65 children with 103 refluxing units (56 girls, nine boys), aged 1-15 years, underwent the designed surgery. RESULTS: Preoperative voiding cystourethrogram revealed bilateral reflux in 38 and unilateral reflux in 27 children. Of 130 renal units, 103 were associated with reflux grade I, II, III, IV and V in 8.73%, 24.27%, 35.92%, 30.9% and 0.9% units, respectively. Of 103 refluxing units, 97 units had stopped refluxing on cystogram 3 months after surgery, giving the overall success rate of 94.1% for all grades of reflux included in the study. Only 7.6% of the children needed urethral catheterization after surgery. No ureteral obstruction occurred. Two patients developed urinary extravasation, both managed conservatively. No patient needed reoperation. The procedure was done on an outpatient basis in 40% of cases, while 60% of the children were hospitalized with a mean admission time of 3.4 days. CONCLUSION: The Gil-Vernet trigonoplasty technique is simple, safe and effective. Our study further simplified postoperative management by avoiding an indwelling catheter and drain. Also, we have shown that this can be done as an outpatient procedure. PMID- 18678531 TI - Obesity and skill attainment in early childhood. AB - This paper investigates the association between obesity and skill attainment in early childhood (aged 2-3 years). Data from the German Socio-Economic Panel Study are used to estimate models of developmental functioning in four critical areas (verbal skills, activities of daily living, motor skills, and social skills) as a function of various measures of weight (including body mass index and obesity) controlling for a rich set of child, parent, and family characteristics. The findings indicate that, among boys, obesity is associated with reduced verbal skills, social skills, motor skills, and activities of daily living. Among girls, obesity is associated with reduced verbal skills. Further investigations show that the correlations exist even for those preschool children who spend no time in day care, which implies that the correlations cannot be due solely to discrimination by teachers, classmates, or day care providers. PMID- 18678532 TI - Shape-memory NiTi foams produced by replication of NaCl space-holders. AB - NiTi foams were created with a structure (32-36% open pores 70-400 microm in size) and mechanical properties (4-25 GPa stiffness, >1000 MPa compressive strength, >42% compressive ductility, and shape-memory strains up to 4%) useful for bone implant applications. A mixture of NiTi and NaCl powders was hot isostatically pressed at 950 and 1065 degrees C and the NaCl phase was then dissolved in water. The resulting NiTi foams show interconnected pores that replicate the shape and size of the NaCl powders, indicating that NiTi powders densified significantly before NaCl melted at 801 degrees C. Densifying NiTi or other metal powders above the melting point of the space-holder permits the use of NaCl, with the following advantages compared with higher-melting, solid space holders such as oxides and fluorides used to date: (i) no temperature limit for densification; (ii) lower cost; (iii) greater flexibility in powder (and thus pore) shape; (iv) faster dissolution; (v) reduced metal corrosion during dissolution; (vi) lower toxicity if space-holder residues remain in the foam. PMID- 18678533 TI - Performance of vertebral cancellous bone augmented with compliant PMMA under dynamic loads. AB - Increased fracture risk has been reported for the adjacent vertebral bodies after vertebroplasty. This increase has been partly attributed to the high Young's modulus of commonly used polymethylmethacrylate (PMMA). Therefore, a compliant bone cement of PMMA with a bulk modulus closer to the apparent modulus of cancellous bone has been produced. This compliant bone cement was achieved by introducing pores in the cement. Due to the reduced failure strength of that porous PMMA cement, cancellous bone augmented with such cement could deteriorate under dynamic loading. The aim of the present study was to assess the potential of acute failure, particle generation and mechanical properties of cancellous bone augmented with this compliant cement in comparison to regular cement. For this purpose, vertebral biopsies were augmented with porous- and regular PMMA bone cement, submitted to dynamic tests and compression to failure. Changes in Young's modulus and height due to dynamic loading were determined. Afterwards, yield strength and Young's modulus were determined by compressive tests to failure and compared to the individual composite materials. No failure occurred and no particle generation could be observed during dynamical testing for both groups. Height loss was significantly higher for the porous cement composite (0.53+/-0.21%) in comparison to the biopsies augmented with regular cement (0.16+/-0.1%). Young's modulus of biopsies augmented with porous PMMA was comparable to cancellous bone or porous cement alone (200-700 MPa). The yield strength of those biopsies (21.1+/-4.1 MPa) was around two times higher than for porous cement alone (11.6+/-3.3 MPa). PMID- 18678534 TI - Posttraumatic compensatory lipodystrophy. PMID- 18678535 TI - Gunshot injury in west Dublin over a 5-year period: a plastic surgery perspective. PMID- 18678536 TI - Clinical study of treatment for recalcitrant ingrown toenail by partial distal phalanx removal. AB - The aim of this study was to identify putative aetiological factors of recalcitrant ingrown toenail and then to introduce a new surgical technique for its treatment on the basis of this identification. We found that many of our patients had an upturned morphology on the distal phalanx beneath the recalcitrant ingrown toenail. We thereby designed a new operative technique to treat this problem. From October 1997 to May 2006, 31 patients (38 toes), who were operated on using the new technique, were assigned to the experimental group. Another 38 patients, who were randomly selected from the population without an ingrown toenail, were assigned to the control group. Briefly, the operation is performed as follows: make an elliptical skin incision distal to the hyponychium. Remove the wedge of tissue through incision, together with the periosteum on the lateral side of the distal phalanx. Expose and then transect the distal part of the distal phalanx. Twenty-nine patients (36 toes) were included in the follow up which varied in length from 8 to 29 months. None of them had recurrent symptoms. In conclusion, an upturned abnormality of the distal phalanx may contribute, at least partly, to the formation of recalcitrant ingrown toenail. The partial distal phalanx removal could be considered as an effective technique in recalcitrant ingrown toenail therapy. PMID- 18678537 TI - The prefabricated pedicled anterolateral thigh flap for reconstruction of a full thickness defect of the urethra. AB - The ideal reconstructive method for a urethral defect should provide durable, stable coverage, a patent tube passage for voiding and ejaculation, and a natural aesthetic contour, while simultaneously minimising morbidity of both the recipient and donor sites. Since the first report of the anterolateral thigh flap in 1984, this has become one of the most commonly used flaps for the reconstruction of various soft tissue defects. In this report we present our experience with a prefabricated anterolateral thigh flap used for a full thickness defect of the urethra. The flap was used in a 40-year-old male patient with a defect located on the proximal penile region, including full-thickness urethra and ventral penile skin measuring 2.5x4 cm after delay and prefabrication. To the best of our knowledge this is the only report in the literature to date of an anterolateral thigh flap being used for a urethra defect. The authors conclude that with its evident structural and cosmetic advantages, the anterolateral thigh flap can be considered an excellent and ideal flap option for selected defects of the urethra. PMID- 18678538 TI - Primary care diabetes in Norway. AB - Usually patients with type 1 diabetes are treated by endocrinologists or specialists in internal medicine after the age of 16-18 years, and patients with type 2 diabetes in primary care. Quality of care has improved. A National Diabetes Strategy was launched in 2006. New national clinical guidelines will soon be published. PMID- 18678539 TI - [Lymphoid neogenesis and lymphangiogenesis: two newcomers in the pathophysiology of chronic rejection]. AB - Chronic rejection is one of the main causes of late allograft failure and no therapy is currently available to prevent efficiently its development. Improving the comprehension of the mechanisms involved in the pathophysiology of chronic rejection is a mandatory step to propose innovative therapies that would prolong grafts' survival. Using the rat aortic interposition model of chronic vascular rejection, we have demonstrated that the intragraft inflammatory infiltrate progressively organized itself into a functional ectopic lymphoid tissue (tertiary lymphoid organ) supporting the local synthesis of alloantibody. Thus, during chronic rejection the graft is at the same time the target and the site of elaboration of the humoral allo-immune response. This hypothesis has been confirmed in the clinical setting by the analysis of human grafts (kidneys, hearts and lungs) removed for terminal failure due to chronic rejection. This lymphoid neogenesis process, previously identified in other chronic inflammatory diseases, occurs with a strikingly high frequency in chronically rejected grafts, suggesting that an additional mechanism synergizes to initiate the development of tertiary lymphoid organs during chronic rejection. We propose that the defective lymphatic drainage of chronically rejected organs triggers lymphoid neogenesis and we discuss the complex crosstalk between lymphoid neogenesis and lymphangiogenesis that takes place during chronic rejection. PMID- 18678540 TI - [Patients on dialysis and the psychological aging process]. AB - In reference to psychopathology, the authors highlight several indicators of the aging dialysed patient's psychological functioning. The aim of this work is to extract some indicators in order to help these patients to experience a pleasant aging, in spite of a difficult disease state, constraining care and the presence of several side effects. PMID- 18678541 TI - Besides increasing surveillance and waiting for an effective vaccine to emerge in the future, what else can be done to save the lives of HFMD victims? PMID- 18678542 TI - Is eradication of Helicobacter pylori the feasible way to prevent gastric cancer? New evidence and progress, but still a long way to go. AB - Epidemiological, animal and biological studies provide compelling evidence for the role of Helicobacter pylori infection in gastric carcinogenesis. The finding that H. pylori-induced chronic atrophic gastritis is the major cause of gastric cancer suggests that eradication of the bacterium may prevent this malignancy. Computer-simulation studies have confirmed the cost-effectiveness of eradication in high-risk subjects; however, unresolved issues complicate active testing for and treatment of H. pylori infection among asymptomatic carriers. Concerns include the enormous costs for developing countries to implement strategies, the inconclusiveness of data from randomized controlled studies, the potential induction of antimicrobial resistance, and the uncertain effect of eliminating this organism on the spectrum of modern disease. Although current evidence is insufficient to recommend universal testing and treatment, it is possible to identify highly susceptible individuals who are most likely to benefit from treatment. Novel biomarkers for predicting risk are under extensive investigation, including genetic, epigenetic and proteinomic factors. The emerging evidence suggests that treatment of H. pylori infection in asymptomatic carriers may decrease the burden of gastric cancer. However, confirmation of long term benefits remains a long way off. PMID- 18678543 TI - Effects of sodium azide, barium ion, d-amphetamine and procaine on inward rectifying potassium channel 6.2 expressed in Xenopus oocytes. AB - BACKGROUND/PURPOSE: Inward rectifying potassium channel 6.2 (Kir6.2DelataC26 channel) is closely related to ATP-sensitive potassium channels. Whether sodium azide, barium ion, d-amphetamine or procaine acts directly on the Kir6.2DeltaC26 channel remains unclear. We studied the effects of these compounds on Kir6.2DeltaC26 channel expressed in Xenopus oocytes. METHODS: The coding sequence of a truncated form of mouse Kir6.2 (GenBank accession number NP_034732.1), Kir6.2(1-364) (i.e. Kir6.2DeltaC26), was subcloned into the pET20b(+) vector. Plasmid containing the correct T7 promoter-Kir6.2(1-364) cDNA fragment [Kir6.2/pET20b(+)] was then subject to NotI digestion to generate the templates for in vitro run-off transcriptions. The channel was expressed in Xenopus laevis oocytes. Two-electrode voltage clamping was used to measure the effects of sodium azide, barium ion, d-amphetamine and procaine on Kir6.2DeltaC26 channel current. RESULTS: Sodium azide activated and barium ion and d-amphetamine inhibited the Kir6.2DeltaC26 channel. Procaine did not have any significant effect on the Kir6.2DeltaC26 channel. CONCLUSION: Kir6.2DeltaC26 channel expressed in Xenopus oocytes can be used as a pharmacological tool for the study of inward rectifying potassium channels. PMID- 18678544 TI - Cytotoxicity analysis of strontium ranelate on cultured human periodontal ligament fibroblasts: a preliminary report. AB - BACKGROUND/PURPOSE: The aim of this study was to analyze the cytotoxicity of strontium ranelate (SR) on human periodontal ligament fibroblasts (PDL cells) in vitro. METHODS: PDL cells were obtained from healthy human third molars and cultured in Dulbeccos Modified Eagles Medium. The experimental groups were: G1, cultures treated with fresh medium (control); and G2, G3, G4 and G5: treated with SR at 20, 10, 5 and 2.5 mg/mL, respectively. The experimental times were 1, 6, 12 and 24 hours (short-term) for viability, and 2, 4, 6 and 8 days (long-term) for cell survival. The cells were counted using a hemocytometer. Data were then analyzed by one-way ANOVA and Tukey's tests (p < 0.05). RESULTS: Cultures treated with the highest SR concentrations (G2 and G3) had significantly lower cell viability and cell numbers (p < 0.05) than those in G1, G4 and G5. SR at 2.5 mg/mL was non-cytotoxic to PDL cells. CONCLUSION: SR was non-toxic at appropriate concentrations. Preclinical tests are needed to further assess its safety and effectiveness for tooth resorption prior to clinical use. PMID- 18678545 TI - Is hepatectomy beneficial in the treatment of multinodular hepatocellular carcinoma? AB - BACKGROUND/PURPOSE: Hepatectomy remains the standard treatment for primary hepatocellular carcinoma (HCC). However, its role in the treatment of multinodular HCC (MNHCC) is unknown. METHODS: The study consisted of 599 patients undergoing curative hepatic resection for HCC between October 1990 and June 2006, in which 112 patients had MNHCC (tumor number > or = 2). The type of MNHCC was classified into: A, nodules involving one or two adjoining segments; B, large tumor with satellite nodules involving three or more segments; C, three or fewer nodules that are scattered in remote segments; and D, more than three separate tumors. Univariate and multivariate analyses were used to identify the prognostic factors related to postoperative survival. During the same period of time, and from our database of 178 patients with pathologically proven MNHCC who were undergoing nonsurgical multidisciplinary therapy, 48 patients with serum albumin level > or = 3.5 g/dL, total bilirubin < 2 mg/dL, tumor number < or = 3, and tumor size < or = 5 cm were compared with 38 patients with the same condition treated with hepatectomy, in which 16 received one-block resection and 22 underwent multiple-site resection. RESULTS: The overall 1-, 3- and 5-year survival rates for patients with single-tumor HCC and MNHCC were 88.0%, 69.2% and 58.4%, and 86.1%, 55.5% and 29.9%, respectively (p < 0.001). Alpha-fetoprotein > 400 ng/mL, total tumor size > 5 cm, largest tumor size > 5 cm, total tumor number > 3, microvascular invasion, non-A type MNHCC and multiple-site resection were poor prognostic factors for MNHCC in the hepatectomy group. Multivariate analysis revealed that only multiple-site hepatic resection was an independent adverse factor related to postoperative survival. In addition, patients who underwent one block resection had significantly better survival compared with the nonsurgical group (p = 0.0016), but the multiple-site resection subgroup did not. CONCLUSION: The prognosis of MNHCC is poor in comparison with that of single-nodular HCC. Hepatectomy is the treatment of choice if the tumors can be removed by one-block resection and liver function reserve is acceptable. PMID- 18678546 TI - Retinal nerve fiber layer thickness measured by optical coherence tomography in non-glaucomatous Taiwanese. AB - BACKGROUND/PURPOSE: Assessment of the peripapillary retinal nerve fiber layer (RNFL) is essential for neuroretinal diseases, especially for early prediction of glaucomatous damage. The purpose of this study was to measure RNFL thickness by Stratus optical coherence tomography (OCT) in normal Taiwanese subjects according to age group. METHODS: The thickness of the RNFL around the disc of normal subjects was obtained, after pupil dilation, by means of 3.4 mm diameter fast mode circle OCT scan. Data for one randomly selected eye of each subject were used for statistical analysis. Differences in RNFL thickness were determined by ANOVA. RESULTS: A total of 162 participants were evaluated: 61 male and 101 female, with mean age of 41.3 +/- 20 years (range, 6-74 years). Mean RNFL thickness was 108.7 +/- 9.4 micrometers (range, 85.5-133.7 micrometers). The RNFL was thickest in the inferior (135.8 +/- 16.3 micrometers) and superior (133.9 +/- 18.0 micrometers) quadrants, followed by the nasal (82.6 +/- 16.0 micrometers) and temporal (82.4 +/- 17.8 micrometers) quadrants (F = 551.9, p < 0.001). CONCLUSION: RNFL thickness was determined by OCT for a normal Taiwanese population aged 6-74 years. The normative data from this study may offer valuable information to enable comparisons of ocular diseases involving RNFL across different populations. PMID- 18678547 TI - Quality of care for lung cancer in Taiwan: a pattern of care based on core measures in the Taiwan Cancer Database registry. AB - BACKGROUND/PURPOSE: To investigate the quality of care (QOC) for lung cancer in Taiwan, as measured by pattern of care (POC) variation. METHODS: Based on core measures in the Taiwan Cancer Database (TCDB) registry, QOC for lung cancer was measured as variation in POC for 16 selected core measures for different hospital characteristics. Statistical significance in variation was evaluated by the Chi squared test. RESULTS: Among the 26 participating hospitals (one excluded as an outlier), 6624 cases of lung cancer were reported in 2004. Among the 16 core measures (6 in the diagnostic and 10 in the therapeutic domain), no significant variation in POC was noted in 12 in northern and non-northern hospitals. However, significant variation in POC was noted for most (5/6) of the core measures in the diagnostic domain for other hospital characteristics (large vs. small, medical center vs. regional hospital, public vs. private). Increasing utilization of tissue diagnosis, diagnostic computed tomography (CT), and CT or magnetic resonance imaging for staging advanced non-small cell lung cancer was noted in the four participating hospitals from 2002 to 2004. CONCLUSION: It is very likely that significant variation in QOC for lung cancer in Taiwan exists among different types but not locations of hospitals, at least in the diagnostic domain. The introduction of internal benchmarking (TCDB and core measures) was associated with some changes, at least in some diagnostic domains, which may lead to improvement in QOC for lung cancer in Taiwan. PMID- 18678548 TI - Factors related to perceived needs of primary caregivers of patients with schizophrenia. AB - BACKGROUND/PURPOSE: Schizophrenia is a chronic mental illness, and sufferers are usually dependent on family, primary caregivers in particular. The present study was designed to assess the perceived needs of caregivers so that adequate services can be provided for them in the community. METHODS: A total of 177 primary caregivers were interviewed with the structured burden-and-need schedules to determine their perceived needs, and the related clinical and demographic factors. Fourteen perceived needs were identified and classified into different need clusters using the generalized association plots. A multiple regression of logistic model was adopted to explore the relationships between the related factors and perceived needs. RESULTS: Four clusters of perceived needs were identified, which included assistant patient care (77.6%), access to relevant information (66.1%), societal support (68.2%), and burden release (27.2%). These needs were significantly related to number of admissions, duration of illness, relationship between caregiver and patient, and education level of the caregiver. CONCLUSION: Four clusters of caregivers' perceived needs were identified and found to be related to psychopathologic and demographic factors. These data are of value in designing appropriate community psychiatric programs to improve the quality of care and enhance the capacity of primary caregivers to care for patients. PMID- 18678549 TI - Longitudinal changes in bone mineral density of healthy elderly men in southern Taiwan. AB - BACKGROUND/PURPOSE: Longitudinal data on bone decline for Chinese elderly people are sparse, especially for the healthy aged male. We report the longitudinal change in bone mineral density (BMD) at the femoral neck, great trochanter and Ward's triangle in healthy older Taiwanese men. METHODS: A prospective cohort study was conducted. We screened 1500 subjects aged > or = 65 years. One hundred and seventy men were eligible for hip evaluation, and 167 had hip BMD measured. Two years later, 142 men completed follow-up BMD measurement. Linear regression was performed between aging and bone loss. Paired t test was used to determine changes in BMD between the intervals. RESULTS: In the initial study, subjects showed significant bone loss through aging by linear regression at all three sites (p < 0.001). Two years later, there was a significant decrease in BMD at all three sites (p < 0.001). For the age cohort, all the age groups showed a significant decrease in BMD at the three study sites (p < 0.05), except those aged > or = 75 years at Ward's triangle (p = 0.667) and the great trochanter (p = 0.1). There was a peak loss of BMD in men aged 65-69 years, as high as 5.57% annually at Ward's triangle. CONCLUSION: BMD was negatively related to aging in healthy men. The loss of BMD in the 65-69 years age group was faster at Ward's triangle than at other sites. Bone loss in Chinese men is of concern because it is greater than in Caucasian men. PMID- 18678550 TI - Hemorrhagic bullae caused by Bacteroides fragilis in a patient with intra abdominal infection. AB - Skin manifestations due to intra-abdominal infection are uncommon but could be a warning sign of severe infection. We report a 58-year-old uremic female who had acute cholecystitis and pneumatosis intestinalis. She developed periumbilical hemorrhagic bullae and finally had a fatal outcome with medical therapy. Severe intra-abdominal infection such as pneumatosis intestinalis should be suspected when periumbilical bullae increase in size. PMID- 18678551 TI - Denture mis-swallowing in the sliding esophageal hiatal hernia mimics esophageal perforation. AB - Mis-swallowing of a foreign body in the esophagus coexisting with sliding hernia might be misdiagnosed as esophageal perforation with mediastinal abscess. We report an 89-year-old woman, bedridden for a long period in a nursing home after a previous cerebrovascular accident, who was sent to our emergency department in a state of sepsis because she had swallowed a radio-opaque partial denture. The retention of the denture as an esophageal foreign body was complicated with mediastinitis and bilateral pleural effusion. The inability of the patient to give a reliable clinical history delayed the diagnosis. This report highlights the difficulty in precisely locating a partial denture because of conflicting radiologic findings and the coexistence of esophageal sliding hernia, all of which led to a misdiagnosis of possible esophageal perforation. A right posterolateral thoracotomy with gastrostomy was performed to remove the lower esophageal foreign body after esophagoscopy failed. The surgical finding of a coincidental sliding esophageal hiatal hernia correlated well with the clinical presentation. Managing such a complicated esophageal foreign body in this elderly patient was challenging. PMID- 18678552 TI - Total laparoscopic repair for abdominal aortic aneurysm. AB - Surgical resection and grafting have long been the standard treatment for abdominal aortic aneurysm and provide an excellent long-term outcome. However, there is tremendous impact on patients due to the surgical invasiveness. Endovascular aortic repair using stent graft was introduced in 1991. After refinement of the techniques and technology, endovascular aortic repair was approved by most health authorities and is associated with less periprocedural morbidities. In between these two extremes, some surgeons endeavored to create an alternative and perform less invasive surgeries. Hand-assisted laparoscopic aortic surgery and laparoscopic-assisted aortic surgery were introduced in 1996. In 2001, total laparoscopic abdominal aortic aneurysm resection with tube graft interposition was first performed in Canada. Till now, only a few vascular units in North America and Europe perform these delicate techniques. We report our first case of total laparoscopic abdominal aortic aneurysm repair. Laparoscopic aortic surgery provides better visualization of the aneurysm neck, less bowel manipulation and avoidance of hypothermia. The minimal invasiveness could translate to better perioperative outcome. To our knowledge, this is also the first case report in Asia. The detailed techniques are described. PMID- 18678553 TI - Laminitis: drink deep, or taste not the Pierian Spring. PMID- 18678554 TI - In vitro and in vivo modulation of the equine immune response by parapoxvirus ovis. AB - REASON FOR PERFORMING STUDY: While immune modulators are used routinely in equine medicine, their mechanism of action is not always known. OBJECTIVES: To determine the effect of a commercial preparation of inactivated parapoxvirus ovis (Orf virus; PPVO) on cytokine gene expression by equine peripheral blood mononuclear cells (PBMC) both in vitro and in vivo. METHODS: PBMC were prepared from 6 mixed breed yearlings and cultured in vitro with PPVO with or without Concanavalin A (Con A) for 24 h. Effects on the expression of IFNalpha, IFNbeta IFNgamma, TNFalpha and IL-18 were analysed by real time quantitative PCR (RT-PCR). In addition, 12 yearling horses were treated with PPVO and whole blood RNA samples were prepared at regular intervals to assess effects on in vivo cytokine gene expression. Six of those yearlings were later treated with saline and served as treatment controls. Nine additional yearlings were injected intradermally with a single dose and their injection sites biopsied at 24 and 48 h for cytokine expression. RESULTS: In vitro culture of PBMC with PPVO led to a significant increase in IFNalpha and IFNbeta gene expression compared to mock-stimulated cultures. In addition, expression of IFNgamma and TNFalpha was significantly higher in PBMC stimulated with PPVO and Con A, than those stimulated with Con A alone. No changes were observed in IL-18 gene expression in vitro. Treatment of horses with a 3-dose regimen of PPVO resulted in elevation of IFNgamma gene expression, which was detected 24 h after the first dose and declined thereafter. Intradermal inoculation led to increased expression of IFNgamma along with IFNbeta, IL-15 and IL-18. CONCLUSIONS: Together these results indicate that PPVO stimulated IFNgamma production both in vitro and in vivo. Increased cytokine expression could account for its immunomodulatory activity. POTENTIAL RELEVANCE: The absence of adverse reactions and clear indications of increased expression of cytokine gene expression supports previous clinical uses for this immune modulator in those situations when increased expression of IFNgamma is warranted. PMID- 18678555 TI - Trust models in ubiquitous computing. AB - We recapture some of the arguments for trust-based technologies in ubiquitous computing, followed by a brief survey of some of the models of trust that have been introduced in this respect. Based on this, we argue for the need of more formal and foundational trust models. PMID- 18678556 TI - Living in a ubiquitous world. PMID- 18678557 TI - A second life for eHealth: prospects for the use of 3-D virtual worlds in clinical psychology. AB - The aim of the present paper is to describe the role played by three-dimensional (3-D) virtual worlds in eHealth applications, addressing some potential advantages and issues related to the use of this emerging medium in clinical practice. Due to the enormous diffusion of the World Wide Web (WWW), telepsychology, and telehealth in general, have become accepted and validated methods for the treatment of many different health care concerns. The introduction of the Web 2.0 has facilitated the development of new forms of collaborative interaction between multiple users based on 3-D virtual worlds. This paper describes the development and implementation of a form of tailored immersive e-therapy called p-health whose key factor is interreality, that is, the creation of a hybrid augmented experience merging physical and virtual worlds. We suggest that compared with conventional telehealth applications such as emails, chat, and videoconferences, the interaction between real and 3-D virtual worlds may convey greater feelings of presence, facilitate the clinical communication process, positively influence group processes and cohesiveness in group-based therapies, and foster higher levels of interpersonal trust between therapists and patients. However, challenges related to the potentially addictive nature of such virtual worlds and questions related to privacy and personal safety will also be discussed. PMID- 18678558 TI - The effects of gestational age and growth restriction on compensatory kidney growth. AB - BACKGROUND: Low birth weight is associated with altered renal development, adult onset hypertension and renal disease. The aim of this prospective longitudinal study was to estimate the renal growth during the first 2 years of life in small for-gestational age (SGA) infants of varied gestational age (GA) and with differing degrees of growth retardation (GR) at birth. Material and methods. The study included 466 children: SGA, n = 243, and appropriate-for-gestational age (AGA), n = 223, classified according to GA into three groups (28-34, 34-36 and >36 weeks, respectively). The SGA children were also classified according to the degree of GR: birth weight <3rd percentile, and birth weight 3-10th percentiles. Serial renal ultrasonography (US) for kidney length (KL) measurement was performed at the ages of 36 and 40 weeks corrected age and 3, 6, 12 and 24 months of chronological age. The ratios of KL(3) to crown to heel length (CHL), body weight (BW) and body surface area (BSA) were used as estimators of relative kidney length (RKL). RESULTS: A total of 1898 measurements were performed. In the full-term and near-term SGA infants (GA >36 weeks), RKL was similar to or even higher than that in AGA controls (P < 0.05 at 12 and 24 months). In two groups of preterm infants (GA 34-36, 28-34 weeks), RKL was lower than in AGA controls either after the first 6 months (GA 34-36 group, P < 0.05) or throughout the study period (GA 28-34 group, P < 0.05). The absolute KL was more severely affected in the preterm babies (GA <36 weeks) with BW <3rd percentile than in those of GA 3rd-10th percentile. CONCLUSION: While in full-term and near-term SGA infants RKL is similar to or even higher than that of AGA infants, in smaller preterm babies (<36 weeks of GA) the RKL is impaired up to the second year of life. PMID- 18678560 TI - From registry data collection to international comparisons: examples of haemodialysis duration and frequency. AB - BACKGROUND: The purpose of this study was to investigate haemodialysis (HD) dose practice patterns in different European countries in the light of the European Best Practice Guidelines (EBPG) and to study the associations of patient characteristics and country with weekly dialysis duration. METHODS: Renal registries in Europe were asked to contribute to the study with individual patient data on weekly HD duration, number of HD sessions a week and last measured Kt/V. Additional items were age, sex, date of first renal replacement therapy (RRT), dry weight, height, HD modality, HD technique, diabetes status and vascular access type. Multivariate logistic regression was used to study the probability of receiving HD for <12 h per week. RESULTS: Seven registries contributed data on 26 136 patients on HD on 31 December 2005. Eighty-three percent of the patients received HD for at least 12 h per week as recommended by the EBPG (range 49.0-97.3% across countries). Multivariate analysis showed significant differences across countries concerning the risk of receiving <12 h. Other risk factors included age (older), sex (female), BMI (low) and duration of RRT (shorter). Diabetes was associated with longer total HD duration. CONCLUSION: This study shows a great international variability in weekly HD duration and some discrepancies between current practices and the EBPG. It also points out the difficulty of obtaining and comparing Kt/V values under current registry practices. PMID- 18678559 TI - Evidence for involvement of nonesterified fatty acid-induced protonophoric uncoupling during mitochondrial dysfunction caused by hypoxia and reoxygenation. AB - BACKGROUND: Proximal tubules subjected to hypoxia in vitro under conditions relevant to ischaemia in vivo develop an energetic deficit that is not corrected even after full reoxygenation. We have provided evidence that accumulation of nonesterified fatty acids (NEFA) is the primary reason for this energetic deficit. In this study, we have further investigated the mechanism for the NEFA induced energetic deficit. METHODS: Mitochondrial membrane potential (Deltapsi) was measured in digitonin-permeabilized, freshly isolated proximal tubules by safranin O uptake. Addition of the potassium/proton exchanger nigericin enables the determination of the mitochondrial proton motive force (Deltap) and the proton gradient (DeltapH). ATP was measured luminometrically and NEFA colorimetrically. RESULTS: Tubule ATP content was depleted after hypoxia and recovered incompletely, even after full reoxygenation. Mitochondrial safranin O uptake was decreased in proximal tubules after hypoxia and reoxygenation (H/R). This decrease was attenuated by delipidated bovine serum albumin (dBSA) or citrate. Addition of nigericin increased safranin O uptake of mitochondria in normoxic proximal tubules, but not in proximal tubules after H/R. Addition of dBSA restored the effect of nigericin to increase mitochondrial safranin O uptake. Addition of the NEFA oleate had the same impact on mitochondrial safranin O uptake as subjecting proximal tubules to H/R. CONCLUSION: The mechanism of the NEFA-induced energetic deficit in freshly isolated rat proximal tubules induced by H/R is characterized by impaired ATP production after full reoxygenation, impaired recovery of Deltapsi and Deltap, abrogation of DeltapH and sensitivity to citrate, consistent with involvement of the tricarboxylate carrier. The data support the concept that protonophoric uncoupling by NEFA movement on anion carriers plays a critical role in proximal tubule mitochochondrial dysfunction after H/R. PMID- 18678561 TI - Label-retaining cells and tubular regeneration in postischaemic kidney. AB - BACKGROUND: In this study, we have examined rat kidneys after ischaemic injury (35 min) with regard to the dynamics of S3 tubule regeneration. METHODS: One day before ischaemia, each rat received four successive i.p. injections of BrdU (5 bromo-2'-deoxyuridine: 80 mg/kg) at 2 h intervals. Groups of experimental animals (n = 4) were killed every 2 h during the first 24 h post-ischaemia as well as 2, 3, 7 and 14 days post-ischaemia. Renal sections were processed to characterize by immunohistochemistry the distribution and phenotype of BrdU-positive cells. RESULTS: Renal regeneration after ischaemia was associated with a typical sequence of transient events: (1) absence of immunostaining during the first 8 h after reperfusion; (2) between 8 and 16 h, detection of a small population of BrdU-positive cells (CD44(+), vimentin(+), CD45(-)) restricted to the lumen of blood vessels characterized by the endothelial expression of selectin E; (3) between 16 and 24 h, progressive decrease of labelled cells in renal capillaries and a concomitant increase in the interstitial compartment; (4) after 1 day, labelled cells disappeared progressively from peritubular interstitium and were mainly observed in regenerating S3 tubules, and (5) after 3 days numerous positive cells were only present in regenerated tubules. CONCLUSIONS: Our data suggest that positive cells (BrdU(+), CD44(+), vimentin(+) and CD45(-)) observed in kidney tubules after ischaemia could originate from an extrarenal source and reach the renal parenchyma via blood vessels. We postulate that these immature cells migrate to injured tubules, proliferate and finally differentiate into mature epithelial cells leading to the replacement of a majority (>80%) of altered S3 cells. PMID- 18678562 TI - Quantitative serum proteomics from surface plasmon resonance imaging. AB - The detection and quantification of specific proteins in complex mixtures is a major challenge for proteomics. For example, the development of disease-related biomarker panels will require fast and efficient methods for obtaining multiparameter protein profiles. We established a high throughput, label-free method for analyzing serum using surface plasmon resonance imaging of antibody microarrays. Microarrays were fabricated using standard pin spotting on bare gold substrates, and samples were applied for binding analysis using a camera-based surface plasmon resonance system. We validated the system by measuring the concentrations of four serum proteins using part of a 792-feature microarray. Transferrin concentrations were measured to be 2.1 mg/ml in human serum and 1.2 mg/ml in murine serum, which closely matched ELISA determinations of 2.6 and 1.2 mg/ml, respectively. In agreement with expected values, human and mouse albumin levels were measured to be 24.3 and 23.6 mg/ml, respectively. The lower limits of detection for the four measurements ranged from 14 to 58 ng/ml or 175 to 755 pm. Where purified target proteins are not available for calibration, the microarrays can be used for relative protein quantification. We used the antibody microarray to compare the serum protein profiles from three liver cancer patients and three non-liver cancer patients. Hierarchical clustering of the serum protein levels clearly distinguished two distinct profiles. Thirty-nine significant protein changes were detected (p < 0.05), 10 of which have been observed previously in serum. alpha-Fetoprotein, a known liver cancer marker, was observed to increase. These results demonstrate the feasibility of this high throughput approach for both absolute and relative protein expression profiling. PMID- 18678563 TI - Soluble amyloid-beta peptides potently disrupt hippocampal synaptic plasticity in the absence of cerebrovascular dysfunction in vivo. AB - Long before the onset of clinical Alzheimer's disease non-fibrillar, soluble assembly states of amyloid-beta (Abeta) peptides are believed to cause cognitive problems by disrupting synaptic function in the absence of significant neurodegeneration. Since many of the risk factors for Alzheimer's disease are vascular, impairment of cerebral blood flow by soluble Abeta has been proposed to be critical in triggering these early changes. However, it is not known if soluble Abeta can affect cerebrovascular function at the concentrations required to cause inhibition of synaptic plasticity mechanisms believed to underlie the early cognitive deficits of Alzheimer's disease. Here we developed a new method to simultaneously assess the ability of soluble Abeta to impair plasticity at synapses and to affect resting and activity-dependent local blood flow in the rat hippocampus in vivo. Intracerebroventricular injection of soluble synthetic Abeta(40) dimers rapidly inhibited plasticity of excitatory synaptic transmission at doses (10-42 pmol) comparable to natural Abeta, but failed to affect vascular function measured using laser-Doppler flowmetry (LDF). Like wild-type Abeta(40), the more vasculotropic Abeta produced by people with familial hemorrhagic stroke of the Dutch type (Abeta(40)E22Q), impaired hippocampal plasticity without causing a significant change in local blood flow. Furthermore, neither resting nor activation-evoked hippocampal perfusion was affected by soluble Abeta(42), even at a concentration that markedly (25%) reduced baseline synaptic transmission. These findings demonstrate that the putative synaptotoxic soluble Abeta species of early Alzheimer's disease cause synaptic dysfunction in the absence of detectible changes in local blood flow. This strongly indicates that early cognitive deficits can be caused by soluble Abeta independently of deleterious effects on cerebrovascular dynamics. PMID- 18678564 TI - Selective neuronal loss in rescued penumbra relates to initial hypoperfusion. AB - Selective neuronal loss (SNL) in the rescued penumbra could account for suboptimal clinical recovery despite effective early reperfusion. Previous studies of SNL used single-photon emission tomography (SPECT), did not account for potential volume loss secondary to collapse of the infarct cavity, and failed to show a relationship with initial hypoperfusion. Here, we obtained acute-stage computerized tomography (CT) perfusion and follow-up quantitative (11)C flumazenil (FMZ)-PET to map SNL in the non-infarcted tissue and assess its relationship with acute-stage hypoperfusion. We prospectively recruited seven patients with evidence of (i) acute (<6 h) extensive middle cerebral artery territory ischaemia based on clinical deficit (National Institutes of Health stroke scale, NIHSS score range: 8-23) and CT Perfusion (CTp) findings and (ii) early recanalization (spontaneous or following thrombolysis) based on spectacular clinical recovery (DeltaNIHSS > or =6 at 24 h), good clinical outcome (NIHSS < or =5) and small final infarct (6/7 subcortical) on late-stage MRI. Ten age-matched controls were also studied. FMZ image analysis took into account potential post stroke volume loss. Across patients, clusters of significantly reduced FMZ binding were more prevalent and extensive in the non-infarcted middle cerebral artery cortical areas than in the non-affected hemisphere (P = 0.028, Wilcoxon sign rank test). Voxel-based between-group comparisons revealed several large clusters of significantly reduced FMZ binding in the affected peri-insular, superior temporal and prefrontal cortices (FDR P < 0.05), as compared with no cluster on the unaffected side. Finally, comparing CTp and PET data revealed a significant negative correlation between FMZ binding and initial hypoperfusion. Applying correction for volume loss did not substantially alter the significance of these results. Although based on a small patient sample sometimes studied late after the index stroke, and as such preliminary, our results establish the presence and distribution of FMZ binding loss in ultimately non-infarcted brain areas after stroke. In addition, the data suggest that this binding loss is proportional to initial hypoperfusion, in keeping with the hypothesis that the rescued penumbra is affected by SNL. Although its clinical counterparts remain uncertain, it is tempting to speculate that peri-infarct SNL could represent a new therapeutic target. PMID- 18678565 TI - Lateropulsion, pushing and verticality perception in hemisphere stroke: a causal relationship? AB - The relationships between perception of verticality by different sensory modalities, lateropulsion and pushing behaviour and lesion location were investigated in 86 patients with a first stroke. Participants sat restrained in a drum-like framework facing along the axis of rotation. They gave estimates of their subjective postural vertical by signalling the point of feeling upright during slow drum rotation which tilted them rightwards-leftwards. The subjective visual vertical was indicated by setting a line to upright on a computer screen. The haptic vertical was assessed in darkness by manually setting a rod to the upright. Normal estimates ranged from -2.5 degrees to 2.5 degrees for visual vertical and postural vertical, and from -4.5 degrees to 4.5 degrees for haptic vertical. Of six patients with brainstem stroke and ipsilesional lateropulsion only one had an abnormal ipsilesional postural vertical tilt (6 degrees ); six had an ipsilesional visual vertical tilt (13 +/-.4 degrees ); two had ipsilesional haptic vertical tilts of 6 degrees . In 80 patients with a hemisphere stroke (35 with contralesional lateropulsion including 6 'pushers'), 34 had an abnormal contralesional postural vertical tilt (average -8.5 +/- 4.7 degrees ), 44 had contralesional visual vertical tilts (average -7 +/- 3.2 degrees ) and 26 patients had contralesional haptic vertical tilts (-7.8 +/- 2.8 degrees ); none had ipsilesional haptic vertical or postural vertical tilts. Twenty-one (26%) showed no tilt of any modality, 41 (52%) one or two abnormal modality(ies) and 18 (22%) a transmodal contralesional tilt (i.e. PV + VV + HV). Postural vertical was more tilted in right than in left hemisphere strokes and specifically biased by damage to neural circuits centred around the primary somatosensory cortex and thalamus. This shows that thalamo-parietal projections have a functional role in the processing of the somaesthetic graviceptive information. Tilts of the postural vertical were more closely related to postural disorders than tilts of the visual vertical. All patients with a transmodal tilt showed a severe lateropulsion and 17/18 a right hemisphere stroke. This indicates that the right hemisphere plays a key role in the elaboration of an internal model of verticality, and in the control of body orientation with respect to gravity. Patients with a 'pushing' behaviour showed a transmodal tilt of verticality perception and a severe postural vertical tilt. We suggest that pushing is a postural behaviour that leads patients to align their erect posture with an erroneous reference of verticality. PMID- 18678566 TI - Deceptively amorphous macroscopic appearance of the brain conceals untold complexities of regional specification. PMID- 18678567 TI - Influence of spontaneous pursed lips breathing on walking endurance and oxygen saturation in patients with moderate to severe chronic obstructive pulmonary disease. AB - OBJECTIVE: To evaluate how spontaneously used pursed lips breathing influences walking endurance, oxygen saturation and dyspnoea in patients with moderate to severe chronic obstructive pulmonary disease. DESIGN: A randomized open-label, cross-over study of chronic obstructive pulmonary disease patients participating in a rehabilitation programme. SETTING: Outpatient pulmonary rehabilitation centre at a university hospital. SUBJECTS: Thirty-two patients with moderate to severe chronic obstructive pulmonary disease. INTERVENTION: All patients performed two endurance shuttle walking tests in random order. During endurance shuttle walking test I a mouthpiece was used in order to prevent spontaneous pursed lips breathing. During endurance shuttle walking test II spontaneous pursed lips breathing was used freely. Heart rate, oxygen saturation and the patients' estimated dyspnoea and leg fatigue on a Borg Category Ratio 10 scale were recorded before, directly after, and 5 and 10 minutes after the tests. RESULTS: When spontaneous pursed lips breathing was used the patients walked on an average for 37 seconds (16%) longer (P<0.01) than when pursed lips breathing was prevented. The patients desaturated considerably during both walking tests but the average drop in oxygen saturation was 1.2% less when spontaneous pursed lips breathing was employed. There were no significant differences in rated degree of dyspnoea or leg fatigue with or without pursed lips breathing. CONCLUSION: Spontaneous pursed lips breathing can be a useful technique to increase walking endurance and reduce oxygen desaturation during walking in patients with moderate to severe chronic obstructive pulmonary disease. PMID- 18678568 TI - Functional thumb orthosis for type I and II boutonniere deformity on the dominant hand in patients with rheumatoid arthritis: a randomized controlled study. AB - OBJECTIVE: To evaluate the effectiveness of a functional thumb orthosis on the dominant hand of patients with rheumatoid arthritis and boutonniere thumb. METHODS: Forty patients with rheumatoid arthritis and boutonniere deformity of the thumb were randomly distributed into two groups. The intervention group used the orthosis daily and the control group used the orthosis only during the evaluation. Participants were evaluated at baseline as well as after 45 and 90 days. Assessments were preformed using the O'Connor Dexterity Test, Jamar dynamometer, pinch gauge, goniometry and the Health Assessment Questionnaire. A visual analogue scale was used to assess thumb pain in the metacarpophalangeal joint. RESULTS: Patients in the intervention group experienced a statistically significant reduction in pain. The thumb orthosis did not disrupt grip and pinch strength, function, Health Assessment Questionnaire score or dexterity in either group. CONCLUSION: The use of thumb orthosis for type I and type II boutonniere deformities was effective in relieving pain. PMID- 18678569 TI - Electrical stimulation of the upper extremity in stroke: cyclic versus EMG triggered stimulation. AB - OBJECTIVE: To compare the effect of cyclic and electromyography (EMG)-triggered electrical stimulation on motor impairment and function of the affected upper extremity in chronic stroke. DESIGN: Randomized controlled trial. SETTING: Outpatient clinic of a rehabilitation centre. SUBJECTS AND INTERVENTION: Twenty two subjects in the chronic stage after stroke were randomly assigned to receive either cyclic (n=11) or EMG-triggered electrical stimulation (n=11) of the wrist and finger extensor muscles for a six-week period. OUTCOME MEASURES: The primary outcome measure was the Action Research Arm test (0-57 points) to assess arm function. Grip strength, Fugl-Meyer Motor Assessment and Motricity Index were secondary outcome measures. Assessments were made at the start of the treatment and after 4, 6 and 12 weeks. RESULTS: Both groups improved on the Action Research Arm test. The group receiving cyclic stimulation improved by 2.3 points, and the group receiving EMG-triggered stimulation improved by 4.2 points. The difference in functional gain was not statistically significant. Differences in gain on the secondary outcome measures were not significant either. CONCLUSION: The present study did not detect a significant difference between EMG-triggered and cyclic electrical stimulation with respect to improvement of motor function of the affected arm in chronic stroke. PMID- 18678570 TI - The effect of early customized vestibular rehabilitation on balance after acoustic neuroma resection. AB - OBJECTIVE: To compare the effect of early customized vestibular rehabilitation with general instructions in patients after acoustic neuroma surgery. SETTING: Tertiary referral centre. SUBJECTS: Fifty-three patients after surgery. DESIGN: A prospective assessor-blinded, randomized controlled trial. INTERVENTIONS: After stratification for age (<50 years; >or=50 years), patients were randomly allocated into groups receiving general instructions or customized vestibular rehabilitation protocols for 12 weeks. OUTCOME MEASURES: Standing Balance Sum, Timed Up and Go test, Tandem Gait and Dynamic Gait Index. Balance performance was assessed preoperatively, at discharge (one week after surgery), three, six, nine, 12, 26 and 52 weeks after surgery. RESULTS AND DISCUSSION: All subjects clearly improved within the first six weeks after surgery. However, older subjects receiving vestibular rehabilitation performed significantly (P<0.05) better on standing balance, Timed Up and Go test and Tandem Gait, when compared with the older group that received only general instructions. This effect persisted up to 12 weeks and also became apparent on the Dynamic Gait Index. In addition, the older vestibular rehabilitation group had better balance scores at 12 weeks when compared with their original preoperative scores (P<0.05). This pattern remained even up to one year after surgery (P<0.05). CONCLUSION: In patients over 50, early vestibular rehabilitation facilitates recovery of postural control after acoustic neuroma surgery. Customized vestibular rehabilitation should be given in addition to general instructions that stress the need of exposure to movement. Retention of the early beneficial effects even one year after surgery stresses the importance of customized vestibular rehabilitation. PMID- 18678571 TI - A phase II exploratory cluster randomized controlled trial of a group mobility training and staff education intervention to promote urinary continence in UK care homes. AB - OBJECTIVES: To assess feasibility, acceptability and potential efficacy of group exercise and staff education intervention to promote continence in older people residing in care homes. To establish measures and information to inform a larger trial. DESIGN: Phase II pilot exploratory cluster randomized controlled trial. SETTING: Six purposely selected care homes in the West Midlands, UK. SUBJECTS: Thirty-four care home residents (mean age 86, 29 female), 23 with cognitive impairments. INTERVENTION: Physiotherapy-led group exercise and staff continence and mobility facilitation training. MAIN OUTCOME MEASURES: Reported continence status, Rivermead Mobility Index. Feasibility was assessed by uptake and compliance, and acceptability by verbal feedback. A staff knowledge questionnaire was used. RESULTS: Thirty-three residents, cluster sizes from 3 to 7. The number of residents agreeing with the statement 'Do you ever leak any urine when you don't mean to?' in the intervention group decreased from 12/17 at baseline to 7/17 at six weeks in the intervention group and increased from 9/16 at baseline to 9/15 at six weeks. The Rivermead Mobility Index scores were better in the intervention group (n=17; baseline: 6.1, six weeks: 6.2) compared with controls (n=16; baseline: 5.9, six weeks: 4.75). The intervention was feasible, well received and had good compliance. CONCLUSIONS: Group mobility training and staff education to promote continence is feasible and acceptable for use with care home residents, including those with cognitive impairment. PMID- 18678572 TI - A randomized controlled trial evaluating the effect of a support and education programme for spouses of people affected by stroke. AB - OBJECTIVE: To determine whether a nurse-led support and education programme for spouses of patients affected by stroke improved the psychological health of the spouses. DESIGN: A longitudinal, open, randomized controlled trial. SAMPLE: One hundred spouses of stroke patients were randomly assigned to either an intervention or a control group. SETTING: The study was conducted in a hospital setting. INTERVENTION: The intervention consisted of six group meetings during six months, with a follow-up after further six months. Comparison between the intervention and the control groups was made at baseline, after six and 12 months using analysis with repeated measures. MAIN MEASURES: The Comprehensive Psychopathological Rating Scale--Self-Affective for psychological health. RESULTS: No significant difference was found between the intervention and control groups concerning overall psychological health. However, a subanalysis revealed that those who participated more frequently in the group meetings (five or six times) had significantly stronger psychological health (P<0.05). Knowledge about stroke increased over time in both groups, but participants in the intervention group learned more (P=0.041). CONCLUSION: Encouraging participation in the group meetings of a support programme might have a positive effect on psychological health. PMID- 18678573 TI - A cognitive behavioural group therapy for patients diagnosed with mild cognitive impairment and their significant others: feasibility and preliminary results. AB - OBJECTIVE: To evaluate the feasibility and present preliminary results of a cognitive behavioural group therapy for patients with mild cognitive impairment and their significant others. DESIGN: One group pretest-posttest design. SUBJECTS: Twenty-two patients with mild cognitive impairment and their significant others, running in four group programmes. INTERVENTION: The main goal of the cognitive behavioural group therapy was to strengthen adaptive behaviour in 10 weekly 2-hour sessions. MAIN MEASURES: Distress and mood: The RAND-36, Geriatric Depression Scale--short form; Acceptance and helplessness: Subscales Acceptance and Helplessness from the Illness Cognition Questionnaire; Marital satisfaction: Maudsley Marital Questionnaire; Alertness to memory failure and behaviour changes: Informant Questionnaire on Cognitive Decline in the Elderly and the Revised Memory and Behaviour Problems Checklist Burden. The burden of caregiving reported by the significant others: Sense of competence Questionnaire and Behaviour Problems Checklist Burden, Hindrance subscale. RESULTS: No changes were found on distress and mood measures in both patients and their significant others. Patients showed a significant increased level of acceptance (P<0.05) and a trend for an increased marital satisfaction (P<0.1). The significant others reported an increased awareness of memory and behavioural problems (P<0.05). Attendance was high, indicating a high motivation for this intervention. CONCLUSION: Preliminary results show evidence for positive changes after a cognitive behavioural group therapy for patients with mild cognitive impairment and their significant others. In addition, the developed programme is applicable and feasible. The programme's effectiveness should be studied further, with an estimated sample size of 70 couples in a controlled study design. PMID- 18678574 TI - Comparison study of chest physiotherapy home training programmes on respiratory functions in patients with muscular dystrophy. AB - OBJECTIVE: To compare the effects of home training programmes, threshold inspiratory muscle training and breathing exercise on spirometry and maximal pressures in patients with muscular dystrophy. DESIGN: Prospective blinded 12 week study. SETTINGS: Cardiopulmonary department of university hospital. SUBJECTS: Twenty-three patients with muscular dystrophy (17 patients with limb girdle muscular dystrophy and 6 patients with Becker muscular dystrophy) assigned to the threshold inspiratory muscle training and breathing exercise groups with alternate allocation. METHODS: Spirometry, maximal inspiratory pressure (PImax) and maximal expiratory pressure (PEmax) were measured before and after training. In the threshold inspiratory muscle training group threshold pressure load was determined as equal to 30% of weekly PImax measurement. In the breathing exercise group, patients performed deep and forceful diaphragmatic and segmental exercises. All patients performed exercises at home and once a week at hospital under supervision. RESULTS: The improvement of PImax in the threshold inspiratory muscle training group was more significant than the improvement observed in the breathing exercise group (P=0.05). PEmax increased significantly only in the breathing exercise group (P=0.01). Spirometry results did not change significantly in both groups after the training. CONCLUSIONS: We conclude that respiratory muscle strength is enhanced by training in the patients with muscular dystrophy who are ambulatory, but inspiratory and/or expiratory training effect is specific to the trained muscles. The techniques that improve the strength of respiratory muscles should be included in the physiotherapy management of patients with muscular dystrophy. PMID- 18678575 TI - Cultural adaptation and validation of the Persian version of the Disabilities of the Arm, Shoulder and Hand (DASH) outcome measure. AB - OBJECTIVE: To translate and validate the Persian version of the Disabilities of the Arm, Shoulder and Hand (DASH) outcome measure (Persian DASH). DESIGN: Cultural translation and psychometric testing. SETTING: Outpatient departments of orthopaedics surgery, primary care settings, rehabilitation medicine and physical therapy. SUBJECTS: Two hundred and seventy-one consecutive Persian-speaking patients with upper extremity disorders including subacromial impingement syndrome, rotator cuff disease, epicondylitis, ulnar nerve entrapment, bursitis, instability, carpal tunnel syndrome, tenosynovitis and adhesive capsulitis. METHODS: The translation and cultural adaptation of the original questionnaire was carried out in accordance with published guidelines. The participants were asked to complete a questionnaire booklet including the Persian DASH, the Short Form General Health Survey (SF-36) and a visual analogue scale (VAS) of pain. In addition, 31 randomly selected patients were asked to complete the questionnaire 48 hours later for the second time. RESULTS: Cronbach's alpha coefficient for the Persian DASH was 0.96. The Persian DASH showed excellent test-retest reliability with intraclass correlation coefficient equal to 0.82 (P<0.01). The correlation between the Persian DASH and the functional scales of the Iranian SF-36 showed desirable results indicating a good convergent validity (Pearson's coefficients ranged from -0.25 to -0.72; P<0.001). The correlation between the Persian DASH and the visual analogue scale was 0.52 (P<0.01). CONCLUSIONS: The Persian DASH is a reliable and valid instrument to measure functional status in Persian-speaking patients with upper extremity disorders in Iran. It is simple and easy to use and now can be applied in clinical settings and future outcome studies in Iran and other Persian-speaking communities. PMID- 18678576 TI - Somatosensory impairment after stroke: frequency of different deficits and their recovery. AB - OBJECTIVE: To investigate the frequency of somatosensory impairment in stroke patients within different somatosensory modalities and different body areas, and their recovery. DESIGN: Prospective observational study. SETTING: Two stroke rehabilitation units. SUBJECTS: Seventy patients with a first stroke (36 men, 34 women; average age, 71, SD 10.00 years; average time since stroke onset, 15 days) were assessed on admission and two, four and six months after stroke. INTERVENTIONS: Not applicable. MAIN MEASURE: Nottingham Sensory Assessment. RESULTS: Somatosensory impairment was common after stroke; 7-53% had impaired tactile sensations, 31-89% impaired stereognosis, and 34-64% impaired proprioception. When comparing somatosensory modalities within body areas the kappa values were low (kappa values<0.54). Recovery occurred over time, though not significantly in lower limb tactile sensations. Stroke severity was the main factor influencing initial somatosensory impairment, but accounted for a small amount of the variance (21-41%). Initial somatosensory impairment was significantly related to somatosensory ability at six months, accounting for 46 71% of the variance. CONCLUSIONS: Proprioception and stereognosis were more frequently impaired than tactile sensations. The different somatosensory modalities showed only slight agreement between impairment within the same body areas, suggesting that the modalities are independent of each other and all should be assessed. High agreements were found between different body areas for each somatosensory modality. Somatosensory impairment was associated with stroke severity, however low variance indicated other factors were involved. PMID- 18678577 TI - Expression of cathepsin K and matrix metalloproteinase 1 indicate persistent osteodestructive activity in long-standing ankylosing spondylitis. AB - BACKGROUND: Ankylosing spondylitis (AS) is a common, largely genetically determined, rheumatic disease that is characterised by spinal inflammation and new bone formation. However, the exact pathogenesis and pathology are still not clear. OBJECTIVE: To analyse tissue obtained at spinal surgery by immunohistochemistry and compare the specimen of patients with AS to those with degenerative disc disease (DDD). METHODS: Bony and soft tissue specimens of 30 patients with AS and 20 with DDD were obtained during spinal osteotomy from different anatomic regions including articular and spinous processes, interspinous ligaments and intervertebral disks. Immunohistolochemistry was performed with established markers for cathepsin K, matrix metalloproteinase (MMP)1, MMP3 and receptor activator for nuclear factor kappaB (RANK) ligand. RESULTS: Cathepsin K and MMP1-positive cells were only observed in AS specimens. Cathepsin K-positive multinucleated cells were detected at articular processes adjacent to fibrous tissues. MMP1 was expressed in smaller mononuclear cells attached to bone. Invasion of bone by MMP1 cells was seen at entheseal sites. In the intervertebral disks, most mononuclear cells were cathepsin K-positive. Isolated cells expressing these matrix-degrading enzymes found in DDD never showed signs of invasion. No differences were found for MMP3 between AS and DDD. Clear expression of RANK ligand was only detected in one patient with AS. CONCLUSIONS: Cathepsin K is strongly expressed in different regions of the spine in AS. Cathepsin K was mainly expressed by mononuclear cells, fibroblast-like cells and cells attached to bone and at sites of bone remodelling, suggestive of high osteoclastic activity. This supports the role of persistent inflammation in the pathogenesis of AS. How these changes relate to osteoproliferation remains to be determined. PMID- 18678578 TI - Local delivery of a recombinant adenoassociated vector containing a tumour necrosis factor alpha antagonist gene in inflammatory arthritis: a phase 1 dose escalation safety and tolerability study. AB - OBJECTIVE: To examine the safety and tolerability of a single intra-articular injection of rAAV2-TNFR:Fc, an adenoassociated virus serotype 2 vector containing the cDNA for the human tumour necrosis factor-immunoglobulin Fc fusion gene (tgAAC94), in subjects with inflammatory arthritis. METHODS: In a double-blind, placebo-controlled, phase 1, dose-escalation study, 15 subjects with inflammatory arthritis (14 with rheumatoid arthritis and 1 with ankylosing spondylitis) not receiving tumour necrosis factor alpha (TNFalpha) inhibitors with persistent moderate (grade 2) or severe (grade 3) swelling in a target joint due to inflammatory arthritis received a single intra-articular injection of rAAV2 TNFR:Fc at 1 x 10(10) (n = 5) or 1 x 10(11) (n = 6) DNase resistant particles per ml joint volume or placebo (n = 4) into a knee (n = 14) or ankle (n = 1). Safety was assessed through adverse event monitoring. As a secondary objective, changes in injected joint tenderness and swelling scores, each measured on a four-point scale, were evaluated. RESULTS: Intra-articular injections of rAAV2-TNFR:Fc were well tolerated with no major safety issues. One event, mild knee pruritus, was considered probably related. Synovial fluid TNFR:Fc protein was not detected (nor expected) at the doses used. At 12 weeks after injection, a two-point decrease in swelling was noted in 2/11 and 2/4 subjects injected with rAAV2-TNFR:Fc and placebo, respectively. CONCLUSION: A single dose of intra-articular rAAV2-TNFR:Fc appears to be safe and well tolerated in subjects without concurrent systemic TNFalpha antagonist use. It is thus feasible to proceed with larger trials to further test the safety and efficacy of local TNFR:Fc gene transfer as a therapeutic modality for patients with inflammatory arthritis. PMID- 18678579 TI - The cyclic GMP-dependent protein kinase II gene associates with gout disease: identified by genome-wide analysis and case-control study. AB - OBJECTIVE: To identify the position of a gout susceptibility gene. METHODS: A genome-wide scan was performed using 382 random polymorphic microsatellite markers spread across 22 autosomes in a Taiwanese family with gout to screen for the gout susceptibility genetic marker. Its association with gout by 33 single nucleotide polymorphisms (SNP) in 148 matched case-control subjects was confirmed. The family with gout comprised eight patients with gout and 10 gout free subjects; case-control subjects were 74 male patients with gout and 74 healthy controls matched by age. RESULTS: Analysis of the genome-wide scan results by a non-parametric linkage method found that chromosome 4q21 contains a locus significantly linked with gout (D4S3243 at 81 289 553 bp; p = 0.004; LOD score = 5.13). In SNP genotyping analysis at the neighbourhood regions of marker D4S3243 for the case-control subjects, the polymorphisms rs7688672 and rs6837293, located on the cGMP-dependent protein kinase II (cGK II) gene, were found to relate significantly to gout disease in a recessive model after adjustment of hyperuricaemia (OR = 2.89, 95% CI 1.19 to 7.02 and OR = 2.72, 95% CI 1.13 to 6.54, respectively). CONCLUSIONS: This study suggests that the cGK II gene on chromosome 4q21 is most likely to harbour gout disease independently of hyperuricaemia and is inherited recessively. PMID- 18678580 TI - What pain assessment guidelines tell us and what they may miss. PMID- 18678581 TI - The stress-reducing effects of art in pediatric health care: art preferences of healthy children and hospitalized children. AB - Art is assumed to possess therapeutic benefits of healing for children, as part of patient-focused design in health care. Since the psychological and physiological well-being of children in health care settings is extremely important in contributing to the healing process, it is vitally important to identify what type of art supports stress reduction. Based on adult studies, nature art was anticipated to be the most preferred and to have stress-reducing effects on pediatric patients. Nature art refers to art images dominated by natural vegetation, flowers or water. The objective of this study was to investigate what type of art image children prefer, and what type of art image has potentially stress-reducing effects on children in hospitals. This study used a three-phase, multi-method approach with children aged 5-17 years: a focus group study (129 participants), a randomized study (48 participants), and a quasi experimental study design (48 participants). Findings were evaluated from three phases. PMID- 18678582 TI - Fear in children with cancer: observations at an outpatient visit. AB - The aim of the study was to describe interactions within the family and between them and professionals on a routine visit at a paediatric oncology outpatient clinic where the visiting child was likely to be fearful. Observations were performed. Data were analysed by qualitative content analysis. The behaviours most frequently observed as expressing fear were being quiet, withdrawn or providing detailed descriptions of experiences. Within the theme ;Recognition of the fear', an attentive attitude to the fear was traced; fear was confirmed and cooperation was seen. Although many efforts were made to meet the fear, this was not always successful. Within the theme ;Lack of attention to the fear', the fear was not in focus due to parental worries and concerns about the child's health, and organizational disturbances. The results can serve as a basis for collegial reflections of how to handle fear in children with cancer. PMID- 18678583 TI - Victimization of youth with type-1 diabetes by teachers: relations with adherence and metabolic control. AB - This study examined rates of teacher victimization and relationships between teacher victimization, adherence and metabolic control in 167 young people with type-1 diabetes. Questions that assessed diabetes-related victimization from teachers were administered to young people with type-1 diabetes. Metabolic control was measured with HbA1c values and diabetes self-management was assessed using the parent version of the Diabetes Self-Management Profile (DSMP). The results indicated that teacher victimization was associated significantly and negatively with the DSMP Diet Subscale (p = .03), DSMP Insulin Adherence Subscale (p = .00) and overall adherence (p = .01) for younger children (aged 8-11 years), but not for older youth (12-17 years). The implications and recommendations for practitioners are discussed. PMID- 18678584 TI - Cardiovascular disease risk in adolescent smokers: evidence of a 'smoker lifestyle'. AB - This study assessed the prevalence of traditional cardiovascular disease risk factors in 119 adolescent smokers, compared to an age-matched sample of non smokers. Participants were assessed for body mass index (BMI), relative body fat, systolic/diastolic/mean arterial blood pressure, total serum cholesterol, aerobic fitness, family (parents or siblings) smoking habits and history of cardiovascular disease. The results showed that BMI and relative body fat were significantly increased, while aerobic fitness was significantly decreased in smokers. Male smokers further demonstrated increased diastolic blood pressure and mean arterial pressure. Smokers also showed increased prevalence for family smoking habits and cardiovascular disease history. Prevalence rates for all other cardiovascular disease risk factors were increased in smokers, but the differences between groups did not reach statistical significance. It is concluded that the studied adolescent smokers demonstrated a higher risk for developing cardiovascular disease later in life, compared to their non-smoking counterparts. PMID- 18678585 TI - Physical activity levels, exercise attitudes, self-perceptions and BMI type of 11 to 12-year-old children. AB - This study examined elementary school pupils with different body mass index (BMI) as to attitudes towards exercise, self-perception, lesson satisfaction in physical education and participation in physical activity. Seven hundred and seventy-five pupils participated in this study, aged 11-12 years. The study was conducted through questionnaires. Students were divided according to their BMI: normal, overweight and obese. Two-way analysis of variance was used, with gender and BMI type as independent variables. The results revealed that when compared to students with a normal BMI, the obese and overweight students had lower scores in lesson satisfaction, negative views of their body and reduced levels of physical activity. Also, the results showed that the obese and overweight students adopted more sedentary daily habits, such as many hours of TV watching and PC usage. The results of this study imply the need for necessary school interventions in order to encourage healthier behaviours and habits. PMID- 18678586 TI - Non-puerperal induced lactation: an infant feeding option in paediatric HIV/AIDS in tropical Africa. AB - A major problem in the management of infants exposed to HIV is the issue of feeding, which stems from the need to avoid transmission of the virus via breast milk. Other important issues in the nutrition of infants exposed to the virus include severe maternal illness, which makes suckling extremely difficult, and feeding orphans. Wet nursing is one of the recommended steps in addressing the feeding problems of such infants but for reasons of sociocultural disapproval, it appears not to be popular in traditional African settings. Non-puerperal induced lactation or re-lactation of a close relation, usually a grandmother, which hitherto has been used to rehabilitate severely malnourished motherless infants, may be equally useful. The procedure of re-lactation and the limitations of the method are highlighted. Also, the need to employ information, education and communication in improving the sociocultural acceptability of this veritable infant feeding method in tropical Africa is discussed. PMID- 18678587 TI - Model-based prediction of sequence alignment quality. AB - MOTIVATION: Multiple sequence alignment (MSA) is an essential prerequisite for many sequence analysis methods and valuable tool itself for describing relationships between protein sequences. Since the success of the sequence analysis is highly dependent on the reliability of alignments, measures for assessing the quality of alignments are highly requisite. RESULTS: We present a statistical model-based alignment quality score. Unlike other quality scores, it does not require several parallel alignments for the same set of sequences or additional structural information. Our quality score is based on measuring the conservation level of reference alignments in Homstrad. Reference sequences were realigned with the Mafft, Muscle and Probcons alignment programs, and a sum-of pairs (SP) score was used to measure the quality of the realignments. Statistical modelling of the SP score as a function of conservation level and other alignment characteristics makes it possible to predict the SP score for any global MSA. The predicted SP scores are highly correlated with the correct SP scores, when tested on the Homstrad and SABmark databases. The results are comparable to that of multiple overlap score (MOS) and better than those of normalized mean distance (NorMD) and normalized iRMSD (NiRMSD) alignment quality criteria. Furthermore, the predicted SP score is able to detect alignments with badly aligned or unrelated sequences. AVAILABILITY: The method is freely available at http://www.mtt.fi/AlignmentQuality/. PMID- 18678588 TI - Powerful fusion: PSI-BLAST and consensus sequences. AB - MOTIVATION: A typical PSI-BLAST search consists of iterative scanning and alignment of a large sequence database during which a scoring profile is progressively built and refined. Such a profile can also be stored and used to search against a different database of sequences. Using it to search against a database of consensus rather than native sequences is a simple add-on that boosts performance surprisingly well. The improvement comes at a price: we hypothesized that random alignment score statistics would differ between native and consensus sequences. Thus PSI-BLAST-based profile searches against consensus sequences might incorrectly estimate statistical significance of alignment scores. In addition, iterative searches against consensus databases may fail. Here, we addressed these challenges in an attempt to harness the full power of the combination of PSI-BLAST and consensus sequences. RESULTS: We studied alignment score statistics for various types of consensus sequences. In general, the score distribution parameters of profile-based consensus sequence alignments differed significantly from those derived for the native sequences. PSI-BLAST partially compensated for the parameter variation. We have identified a protocol for building specialized consensus sequences that significantly improved search sensitivity and preserved score distribution parameters. As a result, PSI-BLAST profiles can be used to search specialized consensus sequences without sacrificing estimates of statistical significance. We also provided results indicating that iterative PSI-BLAST searches against consensus sequences could work very well. Overall, we showed how a very popular and effective method could be used to identify significantly more relevant similarities among protein sequences. AVAILABILITY: http://www.rostlab.org/services/consensus/. PMID- 18678589 TI - VistaClara: an expression browser plug-in for Cytoscape. AB - SUMMARY: VistaClara is a plug-in for Cytoscape which provides a more flexible means to visualize gene and protein expression within a network context. An extended attribute browser is provided in the form of a graphical and interactive permutation matrix that resembles the heat map displays popular in gene expression analysis. This extended browser permits a variety of display options and interactions not currently available in Cytoscape. AVAILABILITY: http://chianti.ucsd.edu/cyto_web/plugins/index.php. PMID- 18678590 TI - Phylocom: software for the analysis of phylogenetic community structure and trait evolution. AB - MOTIVATION: The increasing availability of phylogenetic and trait data for communities of co-occurring species has created a need for software that integrates ecological and evolutionary analyses. Capabilities: Phylocom calculates numerous metrics of phylogenetic community structure and trait similarity within communities. Hypothesis testing is implemented using several null models. Within the same framework, it measures phylogenetic signal and correlated evolution for species traits. A range of utility functions allow community and phylogenetic data manipulation, tree and trait generation, and integration into scientific workflows. AVAILABILITY: Open source at: http://phylodiversity.net/phylocom/. PMID- 18678591 TI - Monitoring of controlled drugs in England is still lacking in some aspects, commission finds. PMID- 18678592 TI - George Godber at 100. PMID- 18678593 TI - Is early intervention in the major psychiatric disorders justified? Yes. PMID- 18678594 TI - Is early intervention in the major psychiatric disorders justified? No. PMID- 18678595 TI - Violence between intimate partners: working with the whole family. PMID- 18678596 TI - Central pontine and extrapontine myelinolysis after alcohol withdrawal. AB - Central pontine myelinolysis (CPM) and extrapontine myelinolysis (EPM) are well recognized syndromes that are related to various conditions such as rapid correction of hyponatremia and chronic alcoholism. We report a very case of a patient with dysarthria, dysphagia and psychiatric symptoms including abnormal behavior starting after alcohol withdrawal, with radiological evidence of CPM and EPM. There was little improvement in the dysarthria or psychiatric symptoms in the first month. PMID- 18678597 TI - Eya1 gene dosage critically affects the development of sensory epithelia in the mammalian inner ear. AB - Haploinsufficiency of the transcription co-activator EYA1 causes branchio-oto renal syndrome, congenital birth defects that account for as many as 2% of profoundly deaf children; however, the underlying cause for its dosage requirement and its specific role in sensory cell development of the inner ear are unknown. Here, an allelic series of Eya1 were generated to study the basis of Eya1 dosage requirements for sensory organ development. Our results show different threshold requirements for the level of Eya1 in different regions of the inner ear. Short and disorganized hair cell sterocilia was observed in wild type/null heterozygous or hypomorphic/hypomorphic homozygous cochleae. Patterning and gene-marker analyses indicate that in Eya1 hypomorphic/null heterozygous mice, a reduction of Eya1 expression to 21% of normal level causes an absence of cochlear and vestibular sensory formation. Eya1 is initially expressed in the progenitors throughout the epithelium of all six sensory regions, and later on during sensory cell differentiation, its expression becomes restricted to the differentiating hair cells. We provide genetic evidence that Eya1 activity, in a concentration-dependent manner, plays a key role in the regulation of genes known to be important for sensory development. Furthermore, we show that Eya1 co localizes with Sox2 in the sensory progenitors and both proteins physically interact. Together, our results indicate that Eya1 appears to be upstream of very early events during the sensory organ development, hair cell differentiation and inner-ear patterning. These results also provide a molecular mechanism for understanding how hypomorphic levels of EYA1 cause inner-ear defects in humans. PMID- 18678598 TI - Transcript and in silico analysis of CLN3 in juvenile neuronal ceroid lipofuscinosis and associated mouse models. AB - Juvenile neuronal ceroid lipofuscinoses (JNCL), commonly known as Batten disease, is a progressive neurodegenerative disorder of childhood characterized by blindness, seizures, motor and cognitive decline, leading to death in early adulthood. Mutations within the CLN3 gene, which encodes a putative lysosomal protein of unknown function, are the underlying cause of JNCL. Over 85% of JNCL patients harbor a 1 kb deletion that is predicted to result in a truncated CLN3 protein and is presumed to be a null mutation. A recent study by Kitzmuller et al. (1) suggested that the 1 kb deletion-associated truncated protein may have partial function, and proposed that JNCL is a mutation-specific disease. In addition, the validity of the original and most widely utilized JNCL mouse model, the Cln3(Deltaex1-6) mouse, as a true null mutant was questioned. We report a substantial decrease in the transcript level of the truncated CLN3 gene product in cells from 1 kb deletion patients. We contend that the truncated CLN3 protein is unlikely to be expressed in JNCL patients since cellular quality control mechanisms at the RNA and protein levels are likely to degrade the mutant transcript and polypeptides. Moreover, we present analysis identifying the expressed transcripts present in Cln3(Deltaex1-6) mouse brain. From the analysis of expressed Cln3(Deltaex1-6) mouse transcripts, combined with in silico prediction of the expected consequences of the Cln3(Deltaex1-6) mutation on these transcripts, we argue that aberrant Cln3 proteins are unlikely to be expressed in this disease model. Taken together our results indicate that the most common mutation associated with JNCL results in a loss of functional CLN3, that the Cln3(Deltaex1-6) mouse harbors a null Cln3 allele, and that it therefore represents a valid model for this disease. PMID- 18678599 TI - A novel deletion in the GTPase domain of OPA1 causes defects in mitochondrial morphology and distribution, but not in function. AB - Autosomal dominant optic atrophy (ADOA), the commonest cause of inherited optic atrophy, is caused by mutations in the ubiquitously expressed gene optic atrophy 1 (OPA1), involved in fusion and biogenesis of the inner membrane of mitochondria. Bioenergetic failure, mitochondrial network abnormalities and increased apoptosis have all been proposed as possible causal factors. However, their relative contribution to pathogenesis as well as the prominent susceptibility of the retinal ganglion cell (RGC) in this disease remains uncertain. Here we identify a novel deletion of OPA1 gene in the GTPase domain in three patients affected by ADOA. Muscle biopsy of the patients showed neurogenic atrophy and abnormal morphology and distribution of mitochondria. Confocal microscopy revealed increased mitochondrial fragmentation in fibroblasts as well as in myotubes, where mitochondria were also unevenly distributed, with clustered organelles alternating with areas where mitochondria were sparse. These abnormalities were not associated with altered bioenergetics or increased susceptibility to pro-apoptotic stimuli. Therefore, changes in mitochondrial shape and distribution can be independent of other reported effects of OPA1 mutations, and therefore may be the primary cause of the disease. The arrangement of mitochondria in RGCs, which degenerate in ADOA, may be exquisitely sensitive to disturbance, and this may lead to bioenergetic crisis and/or induction of apoptosis. Our results highlight the importance of mitochondrial dynamics in the disease per se, and point to the loss of the fine positioning of mitochondria in the axons of RGCs as a possible explanation for their predominant degeneration in ADOA. PMID- 18678600 TI - Dynamic variation in allele-specific gene expression of Paraoxonase-1 in murine and human tissues. AB - Differential allelic expression has been shown to be common in mice, humans and maize, and variability in the expression of polymorphic alleles has been associated with human disease. Here, we describe the differential expression pattern of Paraoxonase-1, a gene involved in lipid metabolism and implicated in the formation of atherosclerotic lesions. We measured the expression of the murine Paraoxonase-1 gene (Pon1) in livers at different stages of embryonic development using F1 hybrid crosses and quantified the transcriptional level of both parental alleles. Using human foetal tissues, we analysed the expression of the human orthologue (PON1) and found monoallelic or preferential allelic expression in 6/7 and 4/4 samples from liver and pancreas, respectively. We observed that Pon1 does not show a parent-of-origin preference in its allelic expression, but has dramatic variations in allele-specific expression occurring throughout development. This study has important repercussions in the analysis of haplotypes at disease loci, since it implies that the expression of polymorphic alleles can be unequal and dynamic. PMID- 18678601 TI - Goitrous autoimmune thyroiditis in a pediatric population: a longitudinal study. AB - BACKGROUND: Patients with autoimmune thyroiditis can present with thyroid function that varies from euthyroidism to frank hypothyroidism or occasionally hyperthyroidism. Although there is a risk of progression from the euthyroid or subclinical hypothyroid state to frank hypothyroidism, the rate of progression is not known. OBJECTIVES: Subjects with diffuse goiter and autoimmune thyroiditis were followed up to observe the rate of deterioration in thyroid function from euthyroid and subclinical hypothyroid states to hypothyroidism. METHODS: Patients who presented with goiter and autoimmune thyroiditis were grouped as those with euthyroidism, subclinical hypothyroidism, and overt hypothyroidism on the basis of levels of thyroxine and thyrotropin at presentation. Patients were followed up for a minimum duration of 24 months with periodic monitoring of thyroid function. RESULTS: Ninety-eight consecutive subjects (aged of 8-18 years) with a diagnosis of autoimmune thyroiditis and diffuse goiter were studied. At presentation, in 24 subjects (24.5%) thyroid function was normal (euthyroidism), 32 (32.6%) had subclinical hypothyroidism, and the remaining 42 subjects (42.9%) had hypothyroidism. All of the subjects with hypothyroid were maintained euthyroid on thyroxine during follow-up. Hypothyroidism developed in 3 of 24 patients with euthyroidism and in 4 of 32 patients with subclinical hypothyroidism. CONCLUSIONS: Subjects with goitrous autoimmune thyroiditis need periodic monitoring of thyroid function. Development of thyroid dysfunction is insidious and may not be accompanied by symptoms and clinical signs. In pediatric and adolescent age groups it is imperative to correct thyroid dysfunction to achieve optimal growth and development. PMID- 18678602 TI - Facial nerve palsy complicating Kawasaki disease. AB - The diagnosis of Kawasaki disease, the most common cause of pediatric acquired heart disease, is difficult and often delayed for children whose age falls outside the typical range of 6 months to 5 years, especially in those with incomplete Kawasaki disease and atypical features. Delayed diagnosis is associated with an increased incidence of coronary artery pathology. Here we describe 2 cases of lower motor neuron facial nerve palsy complicating Kawasaki disease. In both cases the diagnosis of Kawasaki disease was not made acutely, and both patients developed extensive coronary artery lesions. These cases highlight the importance of considering Kawasaki disease in children with unexplained prolonged fever at any age, particularly those without full diagnostic criteria and with unusual features. PMID- 18678603 TI - Gastric volvulus in infants and children. AB - INTRODUCTION: Gastric volvulus is an important cause of nonbilious emesis that must be recognized early to ensure a good outcome. We reviewed 7 cases from our institution, Children's Healthcare of Atlanta (Egleston campus). In addition, we reviewed all gastric volvulus cases in children published to date in the English literature to draw general conclusions about the presentation and treatment of this unusual disease. METHODS: An electronic literature search was performed to find all published cases of pediatric gastric volvulus. The care of all children from January 2002 to December 2007 who were treated for gastric volvulus was also reviewed. RESULTS: There have been 581 cases of gastric volvulus in children published in English between 1929 and 2007. Of these, 252 were acute and 329 were chronic cases. The most common presentation of acute gastric volvulus is in a child <5 years old with nonbilious emesis, epigastric distention, and abdominal pain. Acute gastric volvulus is often associated with deformities of adjacent organs. Definitive diagnosis is made with upper gastrointestinal studies, and definitive therapy requires repair of associated defects and anterior fixation of the stomach to the abdominal wall. The most common presentation of chronic volvulus is in an infant <1 year old with emesis, epigastric distention, feeding difficulties, and growth failure. Treatment may be medical or surgical depending on the underlying etiology of the volvulus. CONCLUSIONS: Acute gastric volvulus is a potentially life-threatening occurrence with a good outcome when treated in a timely fashion. Chronic volvulus may be more difficult to recognize. The common features of acute and chronic gastric volvulus described in this review should assist pediatric health care providers in promptly diagnosing and treating this disease. PMID- 18678604 TI - Quantification of carnitine and acylcarnitines in biological matrices by HPLC electrospray ionization-mass spectrometry. AB - BACKGROUND: Analysis of carnitine and acylcarnitines by tandem mass spectrometry (MS/MS) has limitations. First, preparation of butyl esters partially hydrolyzes acylcarnitines. Second, isobaric nonacylcarnitine compounds yield false-positive results in acylcarnitine tests. Third, acylcarnitine constitutional isomers cannot be distinguished. METHODS: Carnitine and acylcarnitines were isolated by ion-exchange solid-phase extraction, derivatized with pentafluorophenacyl trifluoromethanesulfonate, separated by HPLC, and detected with an ion trap mass spectrometer. Carnitine was quantified with d(3)-carnitine as the internal standard. Acylcarnitines were quantified with 42 synthesized calibrators. The internal standards used were d(6)-acetyl-, d(3)-propionyl-, undecanoyl-, undecanedioyl-, and heptadecanoylcarnitine. RESULTS: Example recoveries [mean (SD)] were 69.4% (3.9%) for total carnitine, 83.1% (5.9%) for free carnitine, 102.2% (9.8%) for acetylcarnitine, and 107.2% (8.9%) for palmitoylcarnitine. Example imprecision results [mean (SD)] within runs (n = 6) and between runs (n = 18) were, respectively: total carnitine, 58.0 (0.9) and 57.4 (1.7) micromol/L; free carnitine, 44.6 (1.5) and 44.3 (1.2) micromol/L; acetylcarnitine, 7.74 (0.51) and 7.85 (0.69) micromol/L; and palmitoylcarnitine, 0.12 (0.01) and 0.11 (0.02) micromol/L. Standard-addition slopes and linear regression coefficients were 1.00 and 0.9998, respectively, for total carnitine added to plasma, 0.99 and 0.9997 for free carnitine added to plasma, 1.04 and 0.9972 for octanoylcarnitine added to skeletal muscle, and 1.05 and 0.9913 for palmitoylcarnitine added to skeletal muscle. Reference intervals for plasma, urine, and skeletal muscle are provided. CONCLUSIONS: This method for analysis of carnitine and acylcarnitines overcomes the observed limitations of MS/MS methods. PMID- 18678605 TI - Chronic calorie restriction attenuates experimental autoimmune encephalomyelitis. AB - Calorie restriction (CR) prevents many age-associated diseases and prolongs the lifespan. CR induces multiple metabolic and physiologic modifications, including anti-inflammatory, antioxidant, and neuroprotective effects that may be beneficial in multiple sclerosis (MS). The present studies sought to determine whether CR or increased calorie intake alters the course of experimental autoimmune encephalomyelitis (EAE), the leading animal model for MS. SJL and C57BL/6 mice were subjected to 40% CR beginning at 5 weeks of age. After 5 weeks of CR, EAE was induced by immunizing with proteolipid protein in SJL mice and with myelin oligodendrocyte glycoprotein in C57BL/6 mice. Clinical, histologic, and immunologic features of EAE were compared with mice fed ad libitum and to SJL mice fed a high-fat, high-calorie diet. CR ameliorated clinical EAE in both mouse strains with less severe inflammation, demyelination, and axon injury. No suppression of immune function was observed. A high-calorie diet did not alter the EAE course. CR was associated with increased plasma levels of corticosterone and adiponectin and reduced concentrations of IL-6 and leptin. The CR-induced hormonal, metabolic, and cytokine changes observed in our studies suggest a combined anti-inflammatory and neuroprotective effect. CR with adequate nutrition and careful medical monitoring should be explored as a potential treatment for MS. PMID- 18678607 TI - Insights into mechanisms behind arteriogenesis: what does the future hold? AB - Arteriogenesis, the enlargement of collateral vessels, seems a promising new target to improve blood flow to ischemic regions in patients suffering from cardiovascular conditions. With the growing knowledge of the mechanisms involved in arteriogenesis and the factors that influence the process, an increasing number of clinical trials are being performed to stimulate arteriogenesis, providing more insight in therapeutic opportunities for arteriogenesis. The expression of growth factors and the cooperation of surrounding and infiltrating cells seem to be essential in orchestrating the complex processes during arteriogenesis. In this review, we will discuss the regulating mechanisms of arteriogenesis, including the role of growth factors and different cell types and their implementation in a clinical setting. Furthermore, individual differences in the arteriogenic response will be considered, in light of the effect this will have on the success of therapeutic strategies to improve blood flow to ischemic tissue. PMID- 18678606 TI - Regulation of STAT pathways and IRF1 during human dendritic cell maturation by TNF-alpha and PGE2. AB - Maturation of dendritic cells (DCs) by TLR ligands induces expression of IFN-beta and autocrine activation of IFN-inducible Stat1-dependent genes important for DC function. In this study, we analyzed the regulation of STAT signaling during maturation of human DCs by TNF-alpha and PGE2, which induced maturation of human DCs comparably with LPS but did not induce detectable IFN-beta production or Stat1 tyrosine phosphorylation. Consistent with these results, TNF-alpha and PGE2 did not induce Stat1 DNA binding to a standard Stat1-binding oligonucleotide. Instead, TNF-alpha and PGE2 increased Stat1 serine phosphorylation and Stat4 tyrosine phosphorylation and activated expression of the NF-kappaB and Stat1 target gene IFN regulatory factor 1 (IRF1), which contributes to IFN responses. TNF-alpha and PGE2 induced a complex that bound an oligonucleotide derived from the IRF1 promoter that contains a STAT-binding sequence embedded in a larger palindromic sequence, and this complex was recognized by Stat1 antibodies. These results suggest that TNF-alpha and PGE2 activate STAT-mediated components of human DC maturation by alternative pathways to the IFN-beta-mediated autocrine loop used by TLRs. PMID- 18678608 TI - Neonatal diabetes with end-stage nephropathy: pancreas transplantation decision. AB - OBJECTIVE: To describe the diagnosis of a patient with neonatal diabetes who had been misdiagnosed with type 1 diabetes and referred to our hospital for pancreas and kidney transplantation because of end-stage renal disease. RESEARCH DESIGN AND METHODS: A diagnosis of neonatal diabetes was made after a molecular genetic study revealed a mutation in exon 34 of the ABCC8 gene. Pancreas transplantation was ill-advised. RESULTS: The patient was switched from insulin to glibenclamide 4 months after kidney transplantation, confirming that pancreas transplantation would not have been a good decision. CONCLUSIONS: This is the first report of a patient with neonatal diabetes who developed diabetic nephropathy that progressed to end-stage renal disease. This report illustrates that careful endocrinological evaluation, including molecular genetic studies, if necessary, is mandatory before a decision to perform a pancreas transplant is made. PMID- 18678609 TI - Financial and clinical impact of team-based treatment for medicaid enrollees with diabetes in a federally qualified health center. AB - OBJECTIVE: The purpose of this study was to determine whether multidisciplinary team-based care guided by the chronic care model can reduce medical payments and improve quality for Medicaid enrollees with diabetes. RESEARCH DESIGN AND METHODS: This study was a difference-in-differences analysis comparing Medicaid patients with diabetes who received team-based care versus those who did not. Team-based care was provided to patients treated at CareSouth, a multisite rural federally qualified community health center located in South Carolina. Control patients were matched to team care patients using propensity score techniques. Financial outcomes compared Medicaid (and Medicare for dually eligible patients) payments 1 year before and after intervention. Trends over time in levels of A1C, BMI, and systolic blood pressure (SBP) were analyzed for intervention patients during the postintervention period. RESULTS: Although average claims payments increased for both the CareSouth patients and control patients, there were no statistically significant differences in total payments between the two groups. In the intervention group, patients with A1C >9 at baseline experienced an average reduction of 0.75 mg/dl per year (95% CI 0.50-0.99), patients with BMI >30 at baseline had an average reduction of 2.3 points per year (95% CI 0.99 3.58), and patients with SBP >140 mmHg at baseline had an average reduction of 2.2 mmHg per year (95% CI 0.44-3.88). CONCLUSIONS: Team-based care following the chronic care model has the potential to improve quality without increasing payments. Short-term savings were not evident and should not be assumed when designing programs. PMID- 18678611 TI - SERENADE: the Study Evaluating Rimonabant Efficacy in Drug-naive Diabetic Patients: effects of monotherapy with rimonabant, the first selective CB1 receptor antagonist, on glycemic control, body weight, and lipid profile in drug naive type 2 diabetes. AB - OBJECTIVE: The purpose of this study was to assess the glucose-lowering efficacy and safety of rimonabant monotherapy in drug-naive type 2 diabetic patients. RESEARCH DESIGN AND METHODS: The Study Evaluating Rimonabant Efficacy in Drug Naive Diabetic Patients (SERENADE) was a 6-month, randomized, double-blind, placebo-controlled trial of 20 mg/day rimonabant in drug-naive patients with type 2 diabetes (A1C 7-10%). The primary end point was A1C change from baseline; secondary end points included body weight, waist circumference, and lipid profile changes. RESULTS: A total of 281 patients were randomly assigned; 278 were exposed to treatment, and 236 (84.9%) completed the study. Baseline A1C (7.9%) was reduced by -0.8% with rimonabant versus -0.3% with placebo (Delta A1C -0.51%; P = 0.0002), with a larger rimonabant effect in patients with baseline A1C >or=8.5% (Delta A1C -1.25%; P = 0.0009). Weight loss from baseline was -6.7 kg with rimonabant versus -2.8 kg with placebo (Delta weight -3.8 kg; P < 0.0001). Rimonabant induced improvements from baseline in waist circumference (-6 vs. -2 cm; P < 0.0001), fasting plasma glucose (-0.9 vs. -0.1 mmol/l; P = 0.0012), triglycerides (-16.3 vs. +4.4%; P = 0.0031), and HDL cholesterol (+10.1 vs. +3.2%; P < 0.0001). Adverse events of interest that occurred more frequently with rimonabant versus placebo were dizziness (10.9 vs. 2.1%), nausea (8.7 vs. 3.6%), anxiety (5.8 vs. 3.6%), depressed mood (5.8 vs. 0.7%), and paresthesia (2.9 vs. 1.4%). CONCLUSIONS: Rimonabant monotherapy resulted in meaningful improvements in glycemic control, body weight, and lipid profile in drug-naive type 2 diabetic patients. Further ongoing studies will better establish the benefit-to-risk profile of rimonabant and define its place in type 2 diabetes management. PMID- 18678610 TI - Efficacy and safety of Technosphere inhaled insulin compared with Technosphere powder placebo in insulin-naive type 2 diabetes suboptimally controlled with oral agents. AB - OBJECTIVE: This double-blind, placebo-controlled, randomized, multicenter, parallel-group study compared the efficacy, safety, and tolerability of Technosphere insulin with Technosphere powder as placebo in insulin-naive type 2 diabetic patients whose diabetes was suboptimally controlled with oral antidiabetic agents. RESEARCH DESIGN AND METHODS: Patients (n = 126) were randomly assigned to 12 weeks of therapy with Technosphere insulin or Technosphere powder after lifestyle education on nutrition, exercise, and instructions on inhaler use. The primary efficacy outcome was change in A1C from baseline to study end, and the secondary efficacy outcome was area under the curve for postprandial glucose levels during a meal test at treatment weeks 4, 8, and 12. RESULTS: A1C reduction from a mean baseline of 7.9% was greater with Technosphere insulin than with Technosphere powder (-0.72 vs. -0.30%; P = 0.003). Postprandial glucose excursions were reduced by 56% with Technosphere insulin compared with baseline, and maximal postprandial glucose levels were reduced by 43% compared with Technosphere powder. Incidences of hypoglycemia, hyperglycemia, cough, and other adverse events were low in both groups. Body weight was unchanged in both groups. CONCLUSIONS: Technosphere insulin was well tolerated and demonstrated significant improvement in glycemic control with clinically meaningful reductions in A1C levels and postprandial glucose concentrations after 12 weeks of treatment. PMID- 18678612 TI - Lack of FFAR1/GPR40 does not protect mice from high-fat diet-induced metabolic disease. AB - OBJECTIVE: FFAR1/GPR40 is a G-protein-coupled receptor expressed predominantly in pancreatic islets mediating free fatty acid-induced insulin secretion. However, the physiological role of FFAR1 remains controversial. It was previously reported that FFAR1 knockout (Ffar1(-/-)) mice were resistant to high-fat diet-induced hyperinuslinemia, hyperglycemia, hypertriglyceridemia, and hepatic steatosis. A more recent report suggested that although FFAR1 was necessary for fatty acid induced insulin secretion in vivo, deletion of FFAR1 did not protect pancreatic islets against fatty acid-induced islet dysfunction. This study is designed to investigate FFAR1 function in vivo using a third line of independently generated Ffar1(-/-) mice in the C57BL/6 background. RESEARCH DESIGN AND METHODS: We used CL-316,243, a beta3 adrenergic receptor agonist, to acutely elevate blood free fatty acids and to study its effect on insulin secretion in vivo. Ffar1(+/+) (wild-type) and Ffar1(-/-) (knockout) mice were placed on two distinct high-fat diets to study their response to diet-induced obesity. RESULTS: Insulin secretion was reduced by approximately 50% in Ffar1(-/-) mice, confirming that FFAR1 contributes significantly to fatty acid stimulation of insulin secretion in vivo. However, Ffar1(+/+) and Ffar1(-/-) mice had similar weight, adiposity, and hyperinsulinemia on high-fat diets, and Ffar1(-/-) mice showed no improvement in glucose or insulin tolerance tests. In addition, high-fat diet induced comparable levels of lipid accumulation in livers of Ffar1(+/+) and Ffar1(-/-) mice. CONCLUSIONS: FFAR1 is required for normal insulin secretion in response to fatty acids; however, Ffar1(-/-) mice are not protected from high-fat diet-induced insulin resistance or hepatic steatosis. PMID- 18678613 TI - Oxidative stress regulates adipocyte apolipoprotein e and suppresses its expression in obesity. AB - OBJECTIVE: Endogenous expression of apolipoprotein E (apoE) has a significant impact on adipocyte lipid metabolism and is markedly suppressed in obesity. Adipose tissue oxidant stress is emerging as an important mediator of adipocyte dysfunction. These studies were undertaken to evaluate the role of oxidant stress for regulation of adipocyte apoE. RESEARCH DESIGN AND METHODS: ApoE gene and protein expression in 3T3-L1 adipocytes or mature adipocytes and adipose tissue from C57/BL6 mice was evaluated after induction of oxidant stress. The response of adipose tissue and adipocytes from obese compared with lean mice to antioxidants was also assessed. RESULTS: Oxidant stress in 3T3-L1 cells or adipocytes and adipose tissue from lean mice significantly reduced apoE mRNA and protein level. Inclusion of an antioxidant eliminated this reduction. Oxidant stress was accompanied by activation of the nuclear factor-kappaB (NF-kappaB) transcription complex, and its effect on apoE was eliminated by an NF-kappaB activation inhibitor. Treatment of freshly isolated adipose tissue or mature adipocytes from obese mice with antioxidant increased apoE expression but had no effect on cells or tissue from lean mice. Incubation of freshly isolated adipocytes from lean mice with stromovascular cells from obese mice significantly suppressed adipocyte apoE compared with incubation with stromovascular cells from lean mice, but this suppression was reversed by inclusion of antioxidant or a neutralizing antibody to tumor necrosis factor-alpha. CONCLUSIONS: Oxidant stress significantly modulates adipose tissue and adipocyte apoE expression. Furthermore, oxidant stress contributes to suppression of adipocyte apoE in obesity. This suppression depends on interaction between adipose tissue stromovascular cells and adipocytes. PMID- 18678614 TI - Common missense variant in the glucokinase regulatory protein gene is associated with increased plasma triglyceride and C-reactive protein but lower fasting glucose concentrations. AB - OBJECTIVE: Using the genome-wide association approach, we recently identified the glucokinase regulatory protein gene (GCKR, rs780094) region as a novel quantitative trait locus for plasma triglyceride concentration in Europeans. Here, we sought to study the association of GCKR variants with metabolic phenotypes, including measures of glucose homeostasis, to evaluate the GCKR locus in samples of non-European ancestry and to fine- map across the associated genomic interval. RESEARCH DESIGN AND METHODS: We performed association studies in 12 independent cohorts comprising >45,000 individuals representing several ancestral groups (whites from Northern and Southern Europe, whites from the U.S., African Americans from the U.S., Hispanics of Caribbean origin, and Chinese, Malays, and Asian Indians from Singapore). We conducted genetic fine-mapping across the approximately 417-kb region of linkage disequilibrium spanning GCKR and 16 other genes on chromosome 2p23 by imputing untyped HapMap single nucleotide polymorphisms (SNPs) and genotyping 104 SNPs across the associated genomic interval. RESULTS: We provide comprehensive evidence that GCKR rs780094 is associated with opposite effects on fasting plasma triglyceride (P(meta) = 3 x 10(-56)) and glucose (P(meta) = 1 x 10(-13)) concentrations. In addition, we confirmed recent reports that the same SNP is associated with C-reactive protein (CRP) level (P = 5 x 10(-5)). Both fine-mapping approaches revealed a common missense GCKR variant (rs1260326, Pro446Leu, 34% frequency, r(2) = 0.93 with rs780094) as the strongest association signal in the region. CONCLUSIONS: These findings point to a molecular mechanism in humans by which higher triglycerides and CRP can be coupled with lower plasma glucose concentrations and position GCKR in central pathways regulating both hepatic triglyceride and glucose metabolism. PMID- 18678615 TI - Persistence of pre-diabetes in overweight and obese Hispanic children: association with progressive insulin resistance, poor beta-cell function, and increasing visceral fat. AB - OBJECTIVE: To examine changes in risk factors in overweight and obese Hispanic children at high risk of developing type 2 diabetes. RESEARCH DESIGN AND METHODS: We recruited 128 overweight/obese Hispanic children with a family history of type 2 diabetes primarily from clinics in East Los Angeles. Children were evaluated annually for 4 years with an oral glucose tolerance test, applying American Diabetes Association criteria to define diabetes and pre-diabetes. Insulin sensitivity (S(i)), acute insulin response (AIR) to glucose, and beta-cell function (BCF) were determined from frequently sampled intravenous glucose tolerance tests, and total body fat by dual-energy X-ray absorptiometry and intra abdominal and subcutaneous abdominal adipose tissue (IAAT and SAAT) by magnetic resonance imaging were assessed in years 1, 2, and 4. RESULTS: No subjects developed type 2 diabetes, 40% never had pre-diabetes, 47% had intermittent pre diabetes with no clear pattern over time, and 13% had persistent pre-diabetes. At baseline, those with persistent pre-diabetes had lower BCF and higher IAAT. In repeated measures, S(i) deteriorated regardless of pre-diabetes, and there was a significant effect of pre-diabetes on AIR (42% lower in pre-diabetes; P = 0.01) and disposition index (34% lower in pre-diabetes; P = 0.021) and a significant interaction of pre-diabetes and time on IAAT (greater increase over time in those with pre-diabetes; P = 0.034). CONCLUSIONS: In this group of Hispanic children at high risk of type 2 diabetes, 1) pre-diabetes is highly variable from year to year; 2) the prevalence of persistent pre-diabetes over 3 years is 13%; and 3) children with persistent pre-diabetes have lower BCF, due to a lower AIR, and increasing visceral fat over time. PMID- 18678616 TI - Lower intrinsic ADP-stimulated mitochondrial respiration underlies in vivo mitochondrial dysfunction in muscle of male type 2 diabetic patients. AB - OBJECTIVE: A lower in vivo mitochondrial function has been reported in both type 2 diabetic patients and first-degree relatives of type 2 diabetic patients. The nature of this reduction is unknown. Here, we tested the hypothesis that a lower intrinsic mitochondrial respiratory capacity may underlie lower in vivo mitochondrial function observed in diabetic patients. RESEARCH DESIGN AND METHODS: Ten overweight diabetic patients, 12 first-degree relatives, and 16 control subjects, all men, matched for age and BMI, participated in this study. Insulin sensitivity was measured with a hyperinsulinemic-euglycemic clamp. Ex vivo intrinsic mitochondrial respiratory capacity was determined in permeabilized skinned muscle fibers using high-resolution respirometry and normalized for mitochondrial content. In vivo mitochondrial function was determined by measuring phosphocreatine recovery half-time after exercise using (31)P-magnetic resonance spectroscopy. RESULTS: Insulin-stimulated glucose disposal was lower in diabetic patients compared with control subjects (11.2 +/- 2.8 vs. 28.9 +/- 3.7 micromol x kg(-1) fat-free mass x min(-1), respectively; P = 0.003), with intermediate values for first-degree relatives (22.1 +/- 3.4 micromol x kg(-1) fat-free mass x min(-1)). In vivo mitochondrial function was 25% lower in diabetic patients (P = 0.034) and 23% lower in first-degree relatives, but the latter did not reach statistical significance (P = 0.08). Interestingly, ADP-stimulated basal respiration was 35% lower in diabetic patients (P = 0.031), and fluoro-carbonyl cyanide phenylhydrazone-driven maximal mitochondrial respiratory capacity was 31% lower in diabetic patients (P = 0.05) compared with control subjects with intermediate values for first-degree relatives. CONCLUSIONS: A reduced basal ADP stimulated and maximal mitochondrial respiratory capacity underlies the reduction in in vivo mitochondrial function, independent of mitochondrial content. A reduced capacity at both the level of the electron transport chain and phosphorylation system underlies this impaired mitochondrial capacity. PMID- 18678617 TI - Type 1 diabetic akita mouse hearts are insulin sensitive but manifest structurally abnormal mitochondria that remain coupled despite increased uncoupling protein 3. AB - OBJECTIVE: Fatty acid-induced mitochondrial uncoupling and oxidative stress have been proposed to reduce cardiac efficiency and contribute to cardiac dysfunction in type 2 diabetes. We hypothesized that mitochondrial uncoupling may also contribute to reduced cardiac efficiency and contractile dysfunction in the type 1 diabetic Akita mouse model (Akita). RESEARCH DESIGN AND METHODS: Cardiac function and substrate utilization were determined in isolated working hearts and in vivo function by echocardiography. Mitochondrial function and coupling were determined in saponin-permeabilized fibers, and proton leak kinetics was determined in isolated mitochondria. Hydrogen peroxide production and aconitase activity were measured in isolated mitochondria, and total reactive oxygen species (ROS) were measured in heart homogenates. RESULTS: Resting cardiac function was normal in Akita mice, and myocardial insulin sensitivity was preserved. Although Akita hearts oxidized more fatty acids, myocardial O(2) consumption was not increased, and cardiac efficiency was not reduced. ADP stimulated mitochondrial oxygen consumption and ATP synthesis were decreased, and mitochondria showed grossly abnormal morphology in Akita. There was no evidence of oxidative stress, and despite a twofold increase in uncoupling protein 3 (UCP3) content, ATP-to-O ratios and proton leak kinetics were unchanged, even after perfusion of Akita hearts with 1 mmol/l palmitate. CONCLUSIONS: Insulin deficient Akita hearts do not exhibit fatty acid-induced mitochondrial uncoupling, indicating important differences in the basis for mitochondrial dysfunction between insulin-responsive type 1 versus insulin-resistant type 2 diabetic hearts. Increased UCP3 levels do not automatically increase mitochondrial uncoupling in the heart, which supports the hypothesis that fatty acid-induced mitochondrial uncoupling as exists in type 2 diabetic hearts requires a concomitant increase in ROS generation. PMID- 18678619 TI - Nonsteroidal anti-inflammatory drugs induced endothelial apoptosis by perturbing peroxisome proliferator-activated receptor-delta transcriptional pathway. AB - Recent studies have shown that use of nonsteroidal anti-inflammatory drugs (NSAIDs) is associated with an increased risk of myocardial infarction. To explore whether NSAIDs may induce endothelial apoptosis and thereby enhance atherothrombosis, we treated human umbilical vein endothelial cells (HUVECs) with sulindac sulfide (SUL), indomethacin (IND), aspirin (ASA), or sodium salicylate (NaS), and we analyzed apoptosis. SUL and/or IND significantly increased annexin V-positive cells, cleaved poly(ADP-ribose) polymerase (PARP) and caspase-3. ASA and NaS at 1 mM did not induce PARP cleavage or caspase-3 and at 5 mM, ASA but not NaS increased apoptosis. Because peroxisome proliferator-activated receptor delta-mediated 14-3-3epsilon up-regulation was reported to play a crucial role in protecting against apoptosis, we determined whether NSAIDs suppress this transcriptional pathway. SUL, IND, and ASA (5 mM) suppressed PPARdelta and 14-3-3 proteins in a manner parallel to PARP cleavage. Neither ASA nor NaS at 1 mM interfered with PPARdelta or 14-3-3epsilon expression. SUL inhibited PPARdelta promoter activity, which correlated with 14-3-3epsilon promoter suppression. Suppression of 14-3-3epsilon was associated with increased Bad translocation to mitochondria. Neither carbaprostacylin nor 4-(3-(2-propyl-3-hydroxy-4-acetyl) phenoxy)propyloxyphenoxy acetic acid (L-165041) prevented HUVECs from SUL-induced apoptosis. Because of suppression of ectopic PPARdelta by sulindac, adenoviral PPARdelta transduction failed to restore 14-3-3epsilon or prevent PPAR cleavage. Our findings suggest that NSAIDs, but not aspirin (<1 mM) induce endothelial apoptosis via suppression of PPARdelta-mediated 14-3-3epsilon expression. PMID- 18678618 TI - Comprehensive association study of type 2 diabetes and related quantitative traits with 222 candidate genes. AB - OBJECTIVE: Type 2 diabetes is a common complex disorder with environmental and genetic components. We used a candidate gene-based approach to identify single nucleotide polymorphism (SNP) variants in 222 candidate genes that influence susceptibility to type 2 diabetes. RESEARCH DESIGN AND METHODS: In a case-control study of 1,161 type 2 diabetic subjects and 1,174 control Finns who are normal glucose tolerant, we genotyped 3,531 tagSNPs and annotation-based SNPs and imputed an additional 7,498 SNPs, providing 99.9% coverage of common HapMap variants in the 222 candidate genes. Selected SNPs were genotyped in an additional 1,211 type 2 diabetic case subjects and 1,259 control subjects who are normal glucose tolerant, also from Finland. RESULTS: Using SNP- and gene-based analysis methods, we replicated previously reported SNP-type 2 diabetes associations in PPARG, KCNJ11, and SLC2A2; identified significant SNPs in genes with previously reported associations (ENPP1 [rs2021966, P = 0.00026] and NRF1 [rs1882095, P = 0.00096]); and implicated novel genes, including RAPGEF1 (rs4740283, P = 0.00013) and TP53 (rs1042522, Arg72Pro, P = 0.00086), in type 2 diabetes susceptibility. CONCLUSIONS: Our study provides an effective gene-based approach to association study design and analysis. One or more of the newly implicated genes may contribute to type 2 diabetes pathogenesis. Analysis of additional samples will be necessary to determine their effect on susceptibility. PMID- 18678620 TI - Muscarinic modulation of synaptic transmission via endocannabinoid signalling in the rat midbrain periaqueductal gray. AB - The midbrain periaqueductal gray (PAG) is involved in organizing behavioral responses to threat, stress, and pain. These PAG functions are modulated by cholinergic agents. In the present study, we examined the cholinergic modulation of synaptic transmission in the PAG using whole-cell voltage-clamp recordings from rat midbrain slices. We found that the cholinergic agonist carbachol reduced the amplitude of evoked inhibitory and excitatory postsynaptic currents (IPSCs and EPSCs, respectively) in all PAG neurons, and this was abolished by the muscarinic receptor antagonist atropine. Carbachol increased the paired pulse ratio of evoked IPSCs and EPSCs, and it reduced the rate, but not the amplitude of spontaneous miniature IPSCs. The carbachol inhibition of evoked IPSCs was mimicked by the acetylcholinesterase inhibitor physostigmine and was reduced by the M1 and M1/M3 muscarinic receptor antagonists pirenzepine and 4 diphenylacetoxy-N-methylpiperidine, but not by the M2 and M4 antagonists gallamine and PD-102807 (3,6a,11,14-tetrahydro-9-methoxy-2-methyl-(12H)-isoquino [1,2-b]pyrrolo[3,2-f][1,3]benzoxazine-1-carboxylic acid, ethyl ester). The carbachol inhibition of evoked IPSCs was reduced by the cannabinoid CB(1) receptor antagonist AM251 (1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-1 piperidinyl-1H-pyrazole-3-carboxamide) and the diacylglycerol (DAG) lipase inhibitor tetrahydrolipstatin, and it was abolished in the presence of both AM251 and gallamine. The carbachol inhibition of evoked EPSCs was also reduced in the combined presence of gallamine and AM251. These results indicate that M1 induced inhibition of GABAergic transmission within the PAG is mediated via endocannabinoids, which are produced via the phospholipase C/DAG lipase pathway and activate presynaptic cannabinoid CB(1) receptors. Thus, presynaptic muscarinic modulation of PAG function is mediated indirectly by M1 receptor induced endocannabinoid signaling and directly by M2 receptors. PMID- 18678621 TI - Positive allosteric modulation of the alpha7 nicotinic acetylcholine receptor: ligand interactions with distinct binding sites and evidence for a prominent role of the M2-M3 segment. AB - The alpha7 nicotinic acetylcholine receptor (nAChR), a homopentameric, rapidly activating and desensitizing ligand-gated ion channel with relatively high degree of calcium permeability, is expressed in the mammalian central nervous system, including regions associated with cognitive processing. Selective agonists targeting the alpha7 nAChR have shown efficacy in animal models of cognitive dysfunction. Use of positive allosteric modulators selective for the alpha7 receptor is another strategy that is envisaged in the design of active compounds aiming at improving attention and cognitive dysfunction. The recent discovery of novel positive allosteric modulators such as 1-(5-chloro-2-hydroxyphenyl)-3-(2 chloro-5-trifluoromethylphenyl)urea (NS-1738) and 1-(5-chloro-2,4 dimethoxyphenyl)-3-(5-methylisoxazol-3-yl)urea (PNU-120596) that are selective for the alpha7 nAChRs but display significant phenotypic differences in their profile of allosteric modulation, suggests that these molecules may act at different sites on the receptor. Taking advantage of the possibility to obtain functional receptors by the fusion of proteins domains from the alpha7 and the 5 HT(3) receptor, we examined the structural determinants required for positive allosteric modulation. This strategy revealed that the extracellular N-terminal domain of alpha7 plays a critical role in allosteric modulation by NS-1738. In addition, alpha7-5HT(3) chimeras harboring the M2-M3 segment showed that spontaneous activity in response to NS-1738, which confirmed the critical contribution of this small extracellular segment in the receptor gating. In contrast to NS-1738, positive allosteric modulation by PNU-120596 could not be restored in the alpha7-5HT(3) chimeras but was selectively observed in the reverse 5HT(3)-alpha7 chimera. All together, these data illustrate the existence of distinct allosteric binding sites with specificity of different profiles of allosteric modulators and open new possibilities to investigate the alpha7 receptor function. PMID- 18678622 TI - The major limitation to exercise performance in COPD is inadequate energy supply to the respiratory and locomotor muscles. PMID- 18678623 TI - The major limitation to exercise performance in COPD is lower limb muscle dysfunction. PMID- 18678624 TI - The major limitation to exercise performance in COPD is dynamic hyperinflation. PMID- 18678625 TI - The major limitation to exercise performance in COPD is inadequate energy supply to the respiratory and locomotor muscles vs. lower limb muscle dysfunction vs. dynamic hyperinflation. The real cause of exercise limitation in COPD. PMID- 18678626 TI - The major limitation to exercise performance in COPD is inadequate energy supply to the respiratory and locomotor muscles vs. lower limb muscle dysfunction vs. dynamic hyperinflation. Interpretation of exercise intolerance in COPD requires an integrated, multisystemic approach. PMID- 18678627 TI - The major limitation to exercise performance in COPD is inadequate energy supply to the respiratory and locomotor muscles vs. lower limb muscle dysfunction vs. dynamic hyperinflation. Might an imbalance between energy supplies and demands curtail incremental exercise in Gold stage I COPD patients? PMID- 18678628 TI - The major limitation to exercise performance in COPD is inadequate energy supply to the respiratory and locomotor muscles vs. lower limb muscle dysfunction vs. dynamic hyperinflation. Defining 'dynamic hyperinflation'. PMID- 18678629 TI - Last Word on Point:Counterpoint: The major limitation to exercise performance in COPD is 1) inadequate energy supply to the respiratory and locomotor muscles, 2) lower limb muscle dysfunction, 3) dynamic hyperinflation. PMID- 18678630 TI - Last Word on Point:Counterpoint: The major limitation to exercise performance in COPD is 1) inadequate energy supply to the respiratory and locomotor muscles, 2) lower limb muscle dysfunction, 3) dynamic hyperinflation. PMID- 18678631 TI - Last Word on Point:Counterpoint: The major limitation to exercise performance in COPD is 1) inadequate energy supply to the respiratory and locomotor muscles, 2) lower limb muscle dysfunction, 3) dynamic hyperinflation. PMID- 18678632 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? PMID- 18678633 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? PMID- 18678634 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? PMID- 18678635 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? PMID- 18678636 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? PMID- 18678637 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? PMID- 18678638 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? PMID- 18678639 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? Enhanced vascular function: a rationale for exercise and marker of cardiovascular health. PMID- 18678640 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? Habitual exercise and vascular function. PMID- 18678641 TI - Commentary on Viewpoint: Exercise and cardiovascular risk reduction: time to update the rationale for exercise? Considering the role of sex in modulating direct effects of exercise on the vasculature. PMID- 18678642 TI - Nitric oxide inhibits Kv4.3 and human cardiac transient outward potassium current (Ito1). AB - AIMS: Chronic atrial fibrillation (CAF) is characterized by a shortening of the plateau phase of the action potentials (AP) and a decrease in the bioavailability of nitric oxide (NO). In this study, we analysed the effects of NO on Kv4.3 (I(Kv4.3)) and on human transient outward K(+) (I(to1)) currents as well as the signalling pathways responsible for them. We also analysed the expression of NO synthase 3 (NOS3) in patients with CAF. METHODS AND RESULTS: I(Kv4.3) and I(to1) currents were recorded in Chinese hamster ovary cells and in human atrial and mouse ventricular dissociated myocytes using the whole-cell patch clamp. The expression of NOS3 was analysed by western blotting. AP were recorded using conventional microelectrode techniques in mouse atrial preparations. NO and NO donors inhibited I(Kv4.3) and human I(to1) in a concentration- and voltage dependent manner (IC(50) for NO: 375.0 +/- 48 nM) as a consequence of the activation of adenylate cyclase and the subsequent activation of the cAMP dependent protein kinase and the serine-threonine phosphatase 2A. The density of the I(to1) recorded in ventricular myocytes from wild-type (WT) and NOS3 deficient mice (NOS3(-/-)) was not significantly different. Furthermore, the duration of atrial AP repolarization in WT and NOS3(-/-) mice was not different. The increase in NO levels to 200 nM prolonged the plateau phase of the mouse atrial AP and lengthened the AP duration measured at 20 and 50% of repolarization of the human atrial CAF-remodelled AP as determined using a mathematical model. However, the expression of NOS3 was not modified in left atrial appendages from CAF patients. CONCLUSION: Our results suggested that the increase in the atrial NO bioavailability could partially restore the duration of the plateau phase of CAF-remodelled AP by inhibiting the I(to1) as a result of the activation of non canonical enzymatic pathways. PMID- 18678643 TI - Fate of connexin43 in cardiac tissue harbouring a disease-linked connexin43 mutant. AB - AIMS: More than 40 mutations in the GJA1 gene encoding connexin43 (Cx43) have been linked to oculodentodigital dysplasia (ODDD), a pleiotropic, autosomal dominant disorder. We hypothesized that even with a significant reduction in the levels of Cx43 in a mutant mouse model of ODDD (Gja1(Jrt/+)) harbouring a G60S mutation (Cx43(G60S)), cardiomyocyte function may only be moderately compromised given that a majority of mutant mice typically survive. METHODS AND RESULTS: Western blotting and quantitative reverse transcriptase-polymerase chain reaction in conjunction with immunofluorescence were used to assess the expression and localization of Cx43 in hearts and cultured cardiomyocytes from wild-type and Gja1(Jrt/+) mice. Dye-coupling and dual whole cell patch-clamp recordings were also used to assess the gap junction channel status in cultured cardiomyocytes from wild-type and mutant mice. Cardiac tissue from adult Gja1(Jrt/+) mice revealed a 60-80% reduction in Cx43 protein with a preferential loss of the highly phosphorylated forms of Cx43. Compensation via the up-regulation of Cx40 or Cx45 was not observed. Immunofluorescent analysis of cultured cardiomyocytes revealed a trafficking defect, with a decrease in Cx43 plaques and a large population of Cx43 being retained in the Golgi apparatus. However, cultured cardiomyocytes from mutant mice remained beating with a 50% decrease in coupling conductance. CONCLUSION: These results suggest that the Cx43(G60S) mutant impairs normal trafficking and function of co-expressed Cx43 with no dramatic effect on cardiomyocyte function, suggesting that Cx43 is biosynthesized in excess of an essential need. PMID- 18678644 TI - Epigenetic regulation of retrotransposons within the nucleolus of Drosophila. AB - R2 retrotransposable elements exclusively insert into a conserved region of the tandemly organized 28S rRNA genes. Despite inactivating a subset of these genes, R2 elements have persisted in the ribosomal DNA (rDNA) loci of insects for hundreds of millions of years. Controlling R2 proliferation was addressed in this study using lines of Drosophila simulans previously shown to have either active or inactive R2 retrotransposition. Lines with active retrotransposition were shown to have high R2 transcript levels, which nuclear run-on transcription experiments revealed were due to increased transcription of R2-inserted genes. Crosses between R2 active and inactive lines indicated that an important component of this transcriptional control is linked to or near the rDNA locus, with the R2 transcription level of the inactive parent being dominant. Pulsed field gel analysis suggested that the R2 active and inactive states were determined by R2 distribution within the locus. Molecular and cytological analyses further suggested that the entire rDNA locus from the active line can be silenced in favor of the locus from the inactive line. This silencing of entire rDNA loci represents an example of the large-scale epigenetic control of transposable elements and shares features with the nucleolar dominance frequently seen in interspecies hybrids. PMID- 18678645 TI - CSIG inhibits PTEN translation in replicative senescence. AB - Using a suppressive subtractive hybridization system, we identified CSIG (cellular senescence-inhibited gene protein; RSL1D1) that was abundant in young human diploid fibroblast cells but declined upon replicative senescence. Overexpression or knockdown of CSIG did not influence p21(Cip1) and p16(INK4a) expressions. Instead, CSIG negatively regulated PTEN and p27(Kip1) expressions, in turn promoting cell proliferation. In PTEN-silenced HEK 293 cells and PTEN deficient human glioblastoma U87MG cells, the effect of CSIG on p27(Kip1) expression and cell division was abolished, suggesting that PTEN was required for the role of CSIG on p27(Kip1) regulation and cell cycle progression. Investigation into the underlying mechanism revealed that the regulation of PTEN by CSIG was achieved through a translational suppression mechanism. Further study showed that CSIG interacted with PTEN mRNA in the 5' untranslated region (UTR) and that knockdown of CSIG led to increased luciferase activity of a PTEN 5' UTR luciferase reporter. Moreover, overexpression of CSIG significantly delayed the progression of replicative senescence, while knockdown of CSIG expression accelerated replicative senescence. Knockdown of PTEN diminished the effect of CSIG on cellular senescence. Our findings indicate that CSIG acts as a novel regulatory component of replicative senescence, which requires PTEN as a mediator and involves in a translational regulatory mechanism. PMID- 18678646 TI - A gene signature-based approach identifies mTOR as a regulator of p73. AB - Although genomic technologies have advanced the characterization of gene regulatory networks downstream of transcription factors, the identification of pathways upstream of these transcription factors has been more challenging. In this study we present a gene signature-based approach for connecting signaling pathways to transcription factors, as exemplified by p73. We generated a p73 gene signature by integrating whole-genome chromatin immunoprecipitation and expression profiling. The p73 signature was linked to corresponding signatures produced by drug candidates, using the in silico Connectivity Map resource, to identify drugs that would induce p73 activity. Of the pharmaceutical agents identified, there was enrichment for direct or indirect inhibitors of mammalian Target of Rapamycin (mTOR) signaling. Treatment of both primary cells and cancer cell lines with rapamycin, metformin, and pyrvinium resulted in an increase in p73 levels, as did RNA interference-mediated knockdown of mTOR. Further, a subset of genes associated with insulin response or autophagy exhibited mTOR-mediated, p73-dependent expression. Thus, downstream gene signatures can be used to identify upstream regulators of transcription factor activity, and in doing so, we identified a new link between mTOR, p73, and p73-regulated genes associated with autophagy and metabolic pathways. PMID- 18678647 TI - Noncanonical E2 variant-independent function of UBC13 in promoting checkpoint protein assembly. AB - The E2 ubiquitin-conjugating enzyme UBC13 plays pivotal roles in diverse biological processes. Recent studies have elucidated that UBC13, in concert with the E3 ubiquitin ligase RNF8, propagates the DNA damage signal via a ubiquitylation-dependent signaling pathway. However, mechanistically how UBC13 mediates its role in promoting checkpoint protein assembly and its genetic requirement for E2 variants remain elusive. Here we provide evidence to support the idea that the E3 ubiquitin ligase complex RNF8-UBC13 functions independently of E2 variants and is sufficient in facilitating ubiquitin conjugations and accumulation of DNA damage mediator 53BP1 at DNA breaks. The RNF8 RING domain serves as the molecular platform to anchor UBC13 at the damaged chromatin, where localized ubiquitylation events allow sustained accumulation of checkpoint proteins. Intriguingly, we found that only a group of RING domains derived from E3 ubiquitin ligases, which have been shown to interact with UBC13, enabled UBC13 mediated FK2 and 53BP1 focus formation at DNA breaks. We propose that the RNF8 RING domain selects and loads a subset of UBC13 molecules, distinct from those that exist as heterodimers, onto sites of double-strand breaks, which facilitates the amplification of DNA damage signals. PMID- 18678648 TI - The T body, a new cytoplasmic RNA granule in Saccharomyces cerevisiae. AB - A large share of mRNA processing and packaging events occurs cotranscriptionally. To explore the hypothesis that transcription defects may affect mRNA fate, we analyzed poly(A)(+) RNA distribution in Saccharomyces cerevisiae strains harboring mutations in Rpb1p, the largest subunit of RNA polymerase II. In certain rpb1 mutants, a poly(A)(+) RNA granule, distinct from any known structure, strongly accumulated in a confined space of the cytoplasm. RNA and protein expressed from Ty1 retrovirus-like elements colocalized with this new granule, which we have consequently named the T body. A visual screen revealed that the deletion of most genes with proposed functions in Ty1 biology unexpectedly does not alter T-body levels. In contrast, the deletion of genes encoding the Mediator transcription initiation factor subunits Srb2p and Srb5p as well as the Ty1 transcriptional regulator Spt21p greatly enhances T-body formation. Our data disclose a new cellular body putatively involved in the assembly of Ty1 particles and suggest that the cytoplasmic fate of mRNA can be affected by transcription initiation events. PMID- 18678649 TI - Sprouty2-mediated inhibition of fibroblast growth factor signaling is modulated by the protein kinase DYRK1A. AB - Raf-MEK-extracellular signal-regulated kinase (Erk) signaling initiated by growth factor-engaged receptor tyrosine kinases (RTKs) is modulated by an intricate network of positive and negative feedback loops which determine the specificity and spatiotemporal characteristics of the intracellular signal. Well-known antagonists of RTK signaling are the Sprouty proteins. The activity of Sprouty proteins is modulated by phosphorylation. However, little is known about the kinases responsible for these posttranslational modifications. We identify DYRK1A as one of the protein kinases of Sprouty2. We show that DYRK1A interacts with and regulates the phosphorylation status of Sprouty2. Moreover, we identify Thr75 on Sprouty2 as a DYRK1A phosphorylation site in vitro and in vivo. This site is functional, since its mutation enhanced the repressive function of Sprouty2 on fibroblast growth factor (FGF)-induced Erk signaling. Further supporting the idea of a functional interaction, DYRK1A and Sprouty2 are present in protein complexes in mouse brain, where their expression overlaps in several structures. Moreover, both proteins copurify with the synaptic plasma membrane fraction of a crude synaptosomal preparation and colocalize in growth cones, pointing to a role in nerve terminals. Our results suggest, therefore, that DYRK1A positively regulates FGF-mitogen-activated protein kinase signaling by phosphorylation-dependent impairment of the inhibitory activity of Sprouty2. PMID- 18678651 TI - Apoptosis and autophagy induction in mammalian cells by small interfering RNA knockdown of mRNA capping enzymes. AB - Addition of a 5' cap to RNA polymerase II transcripts, the first step of pre-mRNA processing in eukaryotes from yeasts to mammals, is catalyzed by the sequential action of RNA triphosphatase, guanylyltransferase, and (guanine-N 7)methyltransferase. The effects of knockdown of these capping enzymes in mammalian cells were investigated using T7 RNA polymerase-synthesized small interfering RNA and also a lentivirus-based inducible, short hairpin RNA system. Decreasing either guanylyltransferase or methyltransferase resulted in caspase-3 activation and elevated terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining characteristic of apoptosis. Induction of apoptosis was independent of p53 tumor suppressor but dependent on BAK or BAX. In addition, levels of the BH3 family member Bim increased, while Mcl-1 and Bik levels remained unchanged during apoptosis. In contrast to capping enzyme knockdown, apoptosis induced by cycloheximide inhibition of protein synthesis required BAK but not BAX. Both Bim and Mcl-1 levels decreased in cycloheximide induced apoptosis while Bik levels were unchanged, suggesting that apoptosis in siRNA-treated cells is not a direct consequence of loss of mRNA translation. siRNA-treated BAK(-/-) BAX(-/-) double-knockout mouse embryonic fibroblasts failed to activate capase-3 or increase TUNEL staining but instead exhibited autophagy, as demonstrated by proteolytic processing of microtubule-associated protein 1 light chain 3 (LC3) and translocation of transfected green fluorescent protein-LC3 from the nucleus to punctate cytoplasmic structures. PMID- 18678650 TI - Posttranscriptional regulation of chicken ccn2 gene expression by nucleophosmin/B23 during chondrocyte differentiation. AB - CCN2/CTGF is a multifunctional factor that plays a crucial role in the growth and differentiation of chondrocytes. The chicken ccn2 gene is regulated not only at the transcriptional level but also by the interaction between a posttranscriptional element in the 3' untranslated region (3'-UTR) and a cofactor. In the present study, we identified a nucleophosmin (NPM) (also called B23) as this cofactor. Binding of NPM to the element was confirmed, and subsequent analysis revealed a significant correlation between the decrease in cytosolic NPM and the increased stability of the ccn2 mRNA during chondrocyte differentiation in vivo. Furthermore, recombinant chicken NPM enhanced the degradation of chimeric RNAs containing the posttranscriptional cis elements in a chicken embryonic fibroblast extract in vitro. It is noteworthy that the RNA destabilization effect by NPM was far more prominent in the cytosolic extract of chondrocytes than in that of fibroblasts, representing a chondrocyte-specific action of NPM. Stimulation by growth factors to promote differentiation changed the subcellular distribution of NPM in chondrocytes, which followed the expected patterns from the resultant change in the ccn2 mRNA stability. Therefore, the present study reveals a novel aspect of NPM as a key player in the posttranscriptional regulation of ccn2 mRNA during the differentiation of chondrocytes. PMID- 18678652 TI - Crystal structure of human Edc3 and its functional implications. AB - Edc3 is an enhancer of decapping and serves as a scaffold that aggregates mRNA ribonucleoproteins together for P-body formation. Edc3 forms a network of interactions with the components of the mRNA decapping machinery and has a modular domain architecture consisting of an N-terminal Lsm domain, a central FDF domain, and a C-terminal YjeF-N domain. We have determined the crystal structure of the N-terminally truncated human Edc3 at a resolution of 2.2 A. The structure reveals that the YjeF-N domain of Edc3 possesses a divergent Rossmann fold topology that forms a dimer, which is supported by sedimentation velocity and sedimentation equilibrium analysis in solution. The dimerization interface of Edc3 is highly conserved in eukaryotes despite the overall low sequence homology across species. Structure-based site-directed mutagenesis revealed dimerization is required for efficient RNA binding, P-body formation, and likely for regulating the yeast Rps28B mRNA as well, suggesting that the dimeric form of Edc3 is a structural and functional unit in mRNA degradation. PMID- 18678653 TI - Direct interaction between DNA methyltransferase DIM-2 and HP1 is required for DNA methylation in Neurospora crassa. AB - DNA methylation is involved in gene silencing and genomic stability in mammals, plants, and fungi. Genetics studies of Neurospora crassa have revealed that a DNA methyltransferase (DIM-2), a histone H3K9 methyltransferase (DIM-5), and heterochromatin protein 1 (HP1) are required for DNA methylation. We explored the interrelationships of these components of the methylation machinery. A yeast two hybrid screen revealed that HP1 interacts with DIM-2. We confirmed the interaction in vivo and demonstrated that it involves a pair of PXVXL-related motifs in the N-terminal region of DIM-2 and the chromo shadow domain of HP1. Both regions are essential for proper DNA methylation. We also determined that DIM-2 and HP1 form a stable complex independently of the trimethylation of histone H3K9, although the association of DIM-2 with its substrate sequences depends on trimethyl-H3K9. The DIM-2/HP1 complex does not include DIM-5. We conclude that DNA methylation in Neurospora is largely or exclusively the result of a unidirectional pathway in which DIM-5 methylates histone H3K9 and then the DIM-2/HP1 complex recognizes the resulting trimethyl-H3K9 mark via the chromo domain of HP1. PMID- 18678654 TI - Lack of evidence for "Acinetobacter septicus" as a species different from Acinetobacter ursingii? PMID- 18678655 TI - Volume of blood cultured and APACHE II score: an intriguing diptych for diagnosis of bloodstream infections? PMID- 18678656 TI - Moraxella catarrhalis binding to host cellular receptors is mediated by sequence specific determinants not conserved among all UspA1 protein variants. AB - The Moraxella catarrhalis ubiquitous surface proteins (UspAs) are autotransporter molecules reported to interact with a variety of different host proteins and to affect processes ranging from serum resistance to cellular adhesion. The role of UspA1 as an adhesin has been confirmed with a number of different human cell types and is mediated by binding to eukaryotic proteins including carcinoembryonic antigen-related cellular adhesion molecules (CEACAMs), fibronectin, and laminin. A distinct difference in the ability of prototypical M. catarrhalis strains to adhere to CEACAM-expressing cell lines prompted us to perform strain-specific structure-function analyses of UspA1 proteins. In this study, we characterized CEACAM binding by a diverse set of UspA1 proteins and showed that 3 out of 10 UspA1 proteins were incapable of binding CEACAM. This difference resulted from the absence of a distinct CEACAM binding motif in nonadhering strains. Our sequence analysis also revealed a single M. catarrhalis isolate that lacked the fibronectin-binding motif and was defective in adherence to Chang conjunctival epithelial cells. These results clearly demonstrate that UspA1-associated adhesive functions are not universally conserved. Instead, UspA1 proteins must be considered as variants with the potential to confer both different cell tropisms and host cell responses. PMID- 18678657 TI - Two-partner secretion systems of Neisseria meningitidis associated with invasive clonal complexes. AB - The two-partner secretion (TPS) pathway is widespread among gram-negative bacteria and facilitates the secretion of very large and often virulence-related proteins. TPS systems consist of a secreted TpsA protein and a TpsB protein involved in TpsA transport across the outer membrane. Sequenced Neisseria meningitidis genomes contain up to five TpsA- and two TpsB-encoding genes. Here, we investigated the distribution of TPS-related open reading frames in a collection of disease isolates. Three distinct TPS systems were identified among meningococci. System 1 was ubiquitous, while systems 2 and 3 were significantly more prevalent among isolates of hyperinvasive clonal complexes than among isolates of poorly invasive clonal complexes. In laboratory cultures, systems 1 and 2 were expressed. However, several sera from patients recovering from disseminated meningococcal disease recognized the TpsAs of systems 2 and 3, indicating the expression of these systems during infection. Furthermore, we showed that the major secreted TpsAs of systems 1 and 2 depend on their cognate TpsBs for transport across the outer membrane and that the system 1 TpsAs undergo processing. Together, our data indicate that TPS systems may contribute to the virulence of N. meningitidis. PMID- 18678658 TI - Serial isolates of persistent Haemophilus influenzae in patients with chronic obstructive pulmonary disease express diminishing quantities of the HMW1 and HMW2 adhesins. AB - In patients with chronic obstructive pulmonary disease (COPD), the lower respiratory tract is commonly colonized by bacterial pathogens, including nontypeable Haemophilus influenzae. The H. influenzae HMW1 and HMW2 adhesins are homologous proteins that promote bacterial adherence to respiratory epithelium and are the predominant targets of the host immune response. These adhesins undergo graded phase variation, controlled by the numbers of 7-bp repeats upstream of the HMW1 and HMW2 structural genes (hmw1A and hmw2A, respectively). In this study, we examined the levels of HMW1 and HMW2 expressed by H. influenzae isolates collected serially from patients with COPD. We found that expression of HMW1 and HMW2 in a given strain decreased over time in a majority of patients, reflecting progressive increases in the numbers of 7-bp repeats and associated with high serum titers of HMW1/HMW2-specific antibodies. We speculate that the presence of high titers of antibodies against the HMW1 and HMW2 adhesins and other immune factors in the lower respiratory tracts of patients with COPD may result in gradual selection for bacteria with reduced levels of HMW1 and HMW2. PMID- 18678659 TI - Modular arrangement of allelic variants explains the divergence in Moraxella catarrhalis UspA protein function. AB - Ubiquitous surface protein A molecules (UspAs) of Moraxella catarrhalis are large, nonfimbrial, autotransporter proteins that can be visualized as a "fuzzy" layer on the bacterial surface by transmission electron microscopy. Previous studies attributed a wide array of functions and binding activities to the closely related UspA1, UspA2, and/or UspA2H protein, yet the molecular and phylogenetic relationships among these activities remain largely unexplored. To address this issue, we determined the nucleotide sequence of the uspA1 genes from a variety of independent M. catarrhalis isolates and compared the deduced amino acid sequences to those of the previously characterized UspA1, UspA2, and UspA2H proteins. Rather than being conserved proteins, we observed a striking divergence of individual UspA1, UspA2, and UspA2H proteins resulting from the modular assortment of unrelated "cassettes" of peptide sequence. The exchange of certain variant cassettes correlates with strain-specific differences in UspA protein function and confers differing phenotypes upon these mucosal surface pathogens. PMID- 18678660 TI - Toll-like receptor 2 recognition of the microsporidia Encephalitozoon spp. induces nuclear translocation of NF-kappaB and subsequent inflammatory responses. AB - Microsporidia are obligate intracellular parasites that are ubiquitous in nature and have been recognized as causing an important emerging disease among immunocompromised individuals. Limited knowledge exists about the immune response against these organisms, and virtually nothing is known about the receptors involved in host recognition. Toll-like receptors (TLR) are pattern recognition receptors that bind to specific molecules found on pathogens and signal a variety of inflammatory responses. In this study, we show that both Encephalitozoon cuniculi and Encephalitozoon intestinalis are preferentially recognized by TLR2 and not by TLR4 in primary human macrophages. This is the first demonstration of host receptor recognition of any microsporidian species. TLR2 ligation is known to activate NF-kappaB, resulting in inflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha) and interleukin-8 (IL-8). We found that the infection of primary human macrophages leads to the nuclear translocation of NF kappaB in as early as 1 h and the subsequent production of TNF-alpha and IL-8. To verify the direct role of TLR2 parasite recognition in the production of these cytokines, the receptor was knocked down in primary human macrophages using small interfering RNA. This knockdown resulted in decreases in both the nuclear translocation of NF-kappaB and the levels of TNF-alpha and IL-8 after challenge with spores. Taken together, these experiments directly link the initial inflammatory response induced by Encephalitozoon spp. to TLR2 stimulation in human macrophages. PMID- 18678661 TI - Enumeration of cytotoxic CD8 T cells ex vivo during the response to Listeria monocytogenes infection. AB - Cytotoxicity is a key effector function of CD8 T cells. However, what proportion of antigen-specific CD8 T cells in vivo exert cytotoxic activity during a functional CD8 T-cell response to infection still remains unknown. We used the Lysispot assay to directly enumerate cytotoxic CD8 T cells from the spleen ex vivo during the immune response to infection with the intracellular bacterium Listeria monocytogenes. We demonstrate that not all antigen-responsive gamma interferon (IFN-gamma)-secreting T cells display cytotoxic activity. Most CD8 T cells detected at early time points of the response were cytotoxic. This percentage continuously declined during both the expansion and contraction phases to about 50% at the peak and to <10% of IFN-gamma-producing cells in the memory phase. As described for clonal expansion, this elaboration of a program of differentiation after an initial stimulus was not affected by antigen or CD4 help but, like proliferation, could be influenced by later reinfection. These data indicate that cytotoxic effector function during the response to infection is regulated independently from IFN-gamma secretion or expansion or contraction of the overall CD8 T-cell response. PMID- 18678662 TI - Tumor suppressor Foxo3a is involved in the regulation of lipopolysaccharide induced interleukin-8 in intestinal HT-29 cells. AB - Enteric bacteria and their products play an important role in intestinal inflammation; however, the complete mechanisms are not elucidated yet. Tumor suppressor Foxo3a regulates gene expression in the nucleus, and its translocation to the cytosol leads to inactivation. Proximally, Foxo3a is regulated by different pathways including the phosphoinositide 3-kinase (PI3K) pathway. The aim of this study was to determine the effect of bacterial infection on Foxo3a in intestinal epithelial cells and to examine the contribution of Foxo3a in intestinal inflammation. Bacterial lipopolysaccharide (LPS) and infection with mouse pathogen Citrobacter rodentium induce translocation of the nuclear Foxo3a into the cytosol, where it degrades in human HT-29 and mouse CMT-93 cells. In colonic epithelia of healthy mice, Foxo3a is localized in the epithelia at the bottom of the crypts in both the nucleus and the cytosol, while in C. rodentium infected colon Foxo3a is expressed along the crypts and located mainly in the cytosol, suggesting its inactivation. LPS utilized the PI3K pathway to inhibit Foxo3a. Additionally, inhibition of PI3K attenuated LPS-induced proinflammatory interleukin-8 (IL-8). LPS-induced IL-8 is increased in HT-29 cells with silenced Foxo3a. Moreover, in HT-29 cells with silenced Foxo3a, the amount of IkappaBalpha, an NF-kappaB inhibitor, is decreased. In conclusion, LPS and bacterial infection inactivate Foxo3a in intestinal epithelia via the PI3K pathway and inactivated Foxo3a leads to the upregulation of IL-8 by suppressing inhibitory IkappaBalpha. PMID- 18678663 TI - Antibiotic-induced perturbations of the intestinal microbiota alter host susceptibility to enteric infection. AB - Intestinal microbiota comprises microbial communities that reside in the gastrointestinal tract and are critical to normal host physiology. Understanding the microbiota's role in host response to invading pathogens will further advance our knowledge of host-microbe interactions. Salmonella enterica serovar Typhimurium was used as a model enteric pathogen to investigate the effect of intestinal microbiota perturbation on host susceptibility to infection. Antibiotics were used to perturb the intestinal microbiota. C57BL/6 mice were treated with clinically relevant doses of streptomycin and vancomycin in drinking water for 2 days, followed by oral infection with Salmonella enterica serovar Typhimurium. Alterations in microbiota composition and numbers were evaluated by fluorescent in situ hybridization, differential plating, and Sybr green staining. Antibiotics had a dose-dependent effect on intestinal microbiota composition. The chosen antibiotic regimen did not significantly alter the total numbers of intestinal bacteria but altered the microbiota composition. Greater preinfection perturbations in the microbiota resulted in increased mouse susceptibility to Salmonella serovar Typhimurium intestinal colonization, greater postinfection alterations in the microbiota, and more severe intestinal pathology. These results suggest that antibiotic treatment alters the balance of the microbial community, which predisposes the host to Salmonella serovar Typhimurium infection, demonstrating the importance of a healthy microbiota in host response to enteric pathogens. PMID- 18678664 TI - Enhanced innate immune responsiveness to pulmonary Cryptococcus neoformans infection is associated with resistance to progressive infection. AB - Genetically regulated mechanisms of host defense against Cryptococcus neoformans infection are not well understood. In this study, pulmonary infection with the moderately virulent C. neoformans strain 24067 was used to compare the host resistance phenotype of C57BL/6J with that of inbred mouse strain SJL/J. At 7 days or later after infection, C57BL/6J mice exhibited a significantly greater fungal burden in the lungs than SJL/J mice. Characterization of the pulmonary innate immune response at 3 h after cryptococcal infection revealed that resistant SJL/J mice exhibited significantly higher neutrophilia, with elevated levels of inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) and keratinocyte-derived chemokine (KC)/CXCL1 in the airways, as well as increased whole-lung mRNA expression of chemokines KC/CXCL1, MIP-1alpha/CCL3, MIP 1beta/CCL4, MIP-2/CXCL2, and MCP-1/CCL2 and cytokines interleukin 1beta (IL 1beta) and IL-1Ra. At 7 and 14 days after infection, SJL/J mice maintained significantly higher levels of TNF-alpha and KC/CXCL1 in the airways and exhibited a Th1 response characterized by elevated levels of lung gamma interferon (IFN-gamma) and IL-12/IL-23p40, while C57BL/6J mice exhibited Th2 immunity as defined by eosinophilia and IL-4 production. Alveolar and resident peritoneal macrophages from SJL/J mice also secreted significantly greater amounts of TNF-alpha and KC/CXCL1 following in vitro stimulation with C. neoformans. Intracellular signaling analysis demonstrated that TNF-alpha and KC/CXCL1 production was regulated by NF-kappaB and phosphatidylinositol 3 kinase in both strains; however, SJL/J macrophages exhibited heightened and prolonged activation in response to C. neoformans infection compared to that of C57BL/6J. Taken together, these data demonstrate that an enhanced innate immune response against pulmonary C. neoformans infection in SJL/J mice is associated with natural resistance to progressive infection. PMID- 18678665 TI - Haemophilus ducreyi LspA proteins are tyrosine phosphorylated by macrophage encoded protein tyrosine kinases. AB - The LspA proteins (LspA1 and LspA2) of Haemophilus ducreyi are necessary for this pathogen to inhibit the phagocytic activity of macrophage cell lines, an event that can be correlated with a reduction in the level of active Src family protein tyrosine kinases (PTKs) in these eukaryotic cells. During studies investigating this inhibitory mechanism, it was discovered that the LspA proteins themselves were tyrosine phosphorylated after wild-type H. ducreyi cells were incubated with macrophages. LspA proteins in cell-free concentrated H. ducreyi culture supernatant fluid could also be tyrosine phosphorylated by macrophages. This ability to tyrosine phosphorylate the LspA proteins was not limited to immune cell lineages but could be accomplished by both HeLa and COS-7 cells. Kinase inhibitor studies with macrophages demonstrated that the Src family PTKs were required for this tyrosine phosphorylation activity. In silico methods and site directed mutagenesis were used to identify EPIYG and EPVYA motifs in LspA1 that contained tyrosines that were targets for phosphorylation. A total of four tyrosines could be phosphorylated in LspA1, with LspA2 containing eight predicted tyrosine phosphorylation motifs. Purified LspA1 fusion proteins containing either the EPIYG or EPVYA motifs were shown to be phosphorylated by purified Src PTK in vitro. Macrophage lysates could also tyrosine phosphorylate the LspA proteins and an LspA1 fusion protein via a mechanism that was dependent on the presence of both divalent cations and ATP. Several motifs known to interact with or otherwise affect eukaryotic kinases were identified in the LspA proteins. PMID- 18678666 TI - TrxR, a new CovR-repressed response regulator that activates the Mga virulence regulon in group A Streptococcus. AB - Coordinate regulation of virulence factors by the group A streptococcus (GAS) Streptococcus pyogenes is important in this pathogen's ability to cause disease. To further elucidate the regulatory network in this human pathogen, the CovR repressed two-component system (TCS) trxSR was chosen for further analysis based on its homology to a virulence-related TCS in Streptococcus pneumoniae. In a murine skin infection model, an insertion mutation in the response regulator gene, trxR, led to a significant reduction in lesion size, lesion severity, and lethality. Curing the trxR mutation restored virulence comparable to the wild type strain. The trxSR operon was defined in vivo, and CovR was found to directly repress its promoter in vitro. DNA microarray analysis established that TrxR activates transcription of Mga-regulated virulence genes, which may explain the virulence attenuation of the trxR mutant. This regulation appears to occur by activation of the mga promoter, Pmga, as demonstrated by analysis of a luciferase reporter fusion. Complementation of the trxR mutant with trxR on a plasmid restored expression of Mga regulon genes and restored virulence in the mouse model to wild-type levels. TrxR is the first TCS shown to regulate Mga expression. Because it is CovR repressed, TrxR defines a new pathway by which CovR can influence Mga to affect pathogenesis in the GAS. PMID- 18678667 TI - Identification of the smallest structure capable of evoking opsonophagocytic antibodies against Streptococcus pneumoniae type 14. AB - Synthetic overlapping oligosaccharide fragments of Streptococcus pneumoniae serotype 14 capsular polysaccharide (Pn14PS), [6)-[beta-D-Galp-(1-->4)-]beta-D GlcpNAc-(1-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp-(1-->](n), were conjugated to CRM(197) protein and injected into mice to determine the smallest immunogenic structure. The resulting antibodies were then tested for Pn14PS specificity and for their capacity to promote the phagocytosis of S. pneumoniae type 14 bacteria. Earlier studies have reported that the oligosaccharide corresponding to one structural repeating unit of Pn14PS, i.e., Gal-Glc-(Gal-)GlcNAc, induces a specific antibody response to Pn14PS. The broader study described here, which evaluated 16 oligosaccharides, showed that the branched trisaccharide element Glc (Gal-)GlcNAc is essential in inducing Pn14PS-specific antibodies and that the neighboring galactose unit at the nonreducing end contributes clearly to the immunogenicity of the epitope. Only the oligosaccharide conjugates that produce antibodies recognizing Pn14PS were capable of promoting the phagocytosis of S. pneumoniae type 14. In conclusion, the branched tetrasaccharide Gal-Glc-(Gal )GlcNAc may be a serious candidate for a synthetic oligosaccharide conjugate vaccine against infections caused by S. pneumoniae type 14. PMID- 18678668 TI - Binding of the Streptococcus gordonii DL1 surface protein Hsa to the host cell membrane glycoproteins CD11b, CD43, and CD50. AB - Infective endocarditis is frequently attributed to oral streptococci. The mechanisms of pathogenesis, however, are not well understood, although interaction between streptococci and phagocytes are thought to be very important. A highly expressed surface component of Streptococcus gordonii, Hsa, which has sialic acid-binding activity, contributes to infective endocarditis in vivo. In the present study, we found that S. gordonii DL1 binds to HL-60 cells differentiated into monocytes, granulocytes, and macrophages. Using a glutathione S-transferase (GST) fusion to the NR2 domain, which is the sialic acid-binding region of Hsa, we confirmed that the Hsa NR2 domain also binds to differentiated HL-60 cells. To identify which sialoglycoproteins on the surface of differentiated HL-60 cells are receptors for Hsa, intrinsic membrane proteins were assessed by bacterial overlay and far-Western blotting. S. gordonii DL1 adhered to 100- to 150-kDa proteins, a reaction that was abolished by neuraminidase treatment. These sialoglycoproteins were identified as CD11b, CD43, and CD50 by GST pull-down assay and immunoprecipitation with each specific monoclonal antibody. These data suggest that S. gordonii DL1 Hsa specifically binds to three glycoproteins as receptors and that this interaction may be the initial bacterial binding step in infective endocarditis by oral streptococci. PMID- 18678669 TI - Amylase-binding protein B of Streptococcus gordonii is an extracellular dipeptidyl-peptidase. AB - The oral commensal bacterium Streptococcus gordonii interacts with salivary amylase via two amylase-binding proteins, AbpA and AbpB. Based on sequence analysis, the 20-kDa AbpA protein is unique to S. gordonii, whereas the 82-kDa AbpB protein appears to share sequence homology with other bacterial dipeptidases. The aim of this study was to verify the peptidase activity of AbpB and further explore its potential functions. The abpB gene was cloned, and histidine-tagged AbpB (His-AbpB) was expressed in Escherichia coli and purified. Its amylase-binding activity was verified in an amylase ligand binding assay, and its cross-reactivity was verified with an anti-AbpB antibody. Both recombinant His-AbpB and partially purified native AbpB displayed dipeptidase activity and degraded human type VI collagen and fibrinogen, but not salivary amylase. Salivary amylase precipitates not only AbpA and AbpB but also glucosyltransferase G (Gtf-G) from S. gordonii supernatants. Since Streptococcus mutans also releases Gtf enzymes that could also be involved in multispecies plaque interactions, the effect of S. gordonii AbpB on S. mutans Gtf-B activity was also tested. Salivary amylase and/or His-AbpB caused a 1.4- to 2-fold increase of S. mutans Gtf-B sucrase activity and a 3- to 6-fold increase in transferase activity. An enzyme linked immunosorbent assay verified the interaction of His-AbpB and amylase with Gtf-B. In summary, AbpB demonstrates proteolytic activity and interacts with and modulates Gtf activity. These activities may help explain the crucial role AbpB appears to play in S. gordonii oral colonization. PMID- 18678670 TI - Cryptococcus neoformans enters the endolysosomal pathway of dendritic cells and is killed by lysosomal components. AB - Cryptococcus neoformans is an opportunistic fungal pathogen that primarily causes disease in immunocompromised individuals. Dendritic cells (DCs) can phagocytose C. neoformans, present cryptococcal antigen, and kill C. neoformans. However, early events following C. neoformans phagocytosis by DCs are not well defined. We hypothesized that C. neoformans traffics to the endosome and the lysosome following phagocytosis by DCs and is eventually killed in the lysosome. Murine bone marrow-derived DCs (BMDCs) or human monocyte-derived DCs (HDCs) were incubated with live, encapsulated C. neoformans yeast cells and opsonizing antibody. Following incubation, DCs were intracellularly stained with antibodies against EEA1 (endosome) and LAMP-1 (late endosome/lysosome). As assessed by confocal microscopy, C. neoformans trafficked to endosomal compartments of DCs within 10 min and to lysosomal compartments within 30 min postincubation. For HDCs, the studies were repeated using complement-sufficient autologous plasma for the opsonization of C. neoformans. These data showed results similar to those for antibody opsonization, with C. neoformans localized to endosomes within 20 min and to lysosomes within 60 min postincubation. Additionally, the results of live real-time imaging studies demonstrated that C. neoformans entered lysosomal compartments within 20 min following the initiation of phagocytosis. The results of scanning and transmission electron microscopy demonstrated conventional zipper phagocytosis of C. neoformans by DCs. Finally, lysosomal extracts were purified from BMDCs and incubated with C. neoformans to determine their potential to kill C. neoformans. The extracts killed C. neoformans in a dose-dependent manner. This study shows that C. neoformans enters into endosomal and lysosomal pathways following DC phagocytosis and can be killed by lysosomal components. PMID- 18678671 TI - Toxoplasma gondii rhoptry discharge correlates with activation of the early growth response 2 host cell transcription factor. AB - Toxoplasma gondii is a ubiquitous apicomplexan parasite that can cause severe disease in fetuses and immune-compromised patients. Rhoptries, micronemes, and dense granules, which are secretory organelles unique to Toxoplasma and other apicomplexan parasites, play critical roles in parasite growth and virulence. To understand how these organelles modulate infected host cells, we sought to identify host cell transcription factors triggered by their release. Early growth response 2 (EGR2) is a host cell transcription factor that is rapidly upregulated and activated in Toxoplasma-infected cells but not in cells infected with the closely related apicomplexan parasite Neospora caninum. EGR2 upregulation occurred only when live parasites were in direct contact with the host cell and not from exposure to cell extracts that contain dense granule or micronemal proteins. When microneme-mediated attachment was blocked by pretreating parasites with a calcium chelator, EGR2 expression was significantly reduced. In contrast, when host cells were infected with parasites in the presence of cytochalasin D, which allows rhoptry secretion but prevents parasite invasion, EGR2 was activated. Finally, we demonstrate that Toxoplasma activation of host p38 mitogen activated protein kinase is necessary but not sufficient for EGR2 activation. Collectively, these data indicate that EGR2 is specifically upregulated by a parasite-derived secreted factor that is most likely a resident rhoptry protein. PMID- 18678672 TI - Immunization with Vibrio cholerae outer membrane vesicles induces protective immunity in mice. AB - The gram-negative bacterium Vibrio cholerae releases outer membrane vesicles (OMVs) during growth. In this study, we immunized female mice by the intranasal, intragastric, or intraperitoneal route with purified OMVs derived from V. cholerae. Independent of the route of immunization, mice induced specific, high titer immune responses of similar levels against a variety of antigens present in the OMVs. After the last immunization, the half-maximum total immunoglobulin titer was stable over a 3-month period, indicating that the immune response was long lasting. The induction of specific isotypes, however, was dependent on the immunization route. Immunoglobulin A, for example, was induced to a significant level only by mucosal immunization, with the intranasal route generating the highest titers. We challenged the offspring of immunized female mice with V. cholerae via the oral route in two consecutive periods, approximately 30 and 95 days after the last immunization. Regardless of the route of immunization, the offspring was protected against colonization with V. cholerae in both challenge periods. Our results show that mucosal immunizations via both routes with OMVs derived from V. cholerae induce long-term protective immune responses against this gastrointestinal pathogen. These findings may contribute to the development of "nonliving," OMV-based vaccines against V. cholerae and other enteric pathogens, using the oral or intranasal route of immunization. PMID- 18678674 TI - The anthrax vaccine adsorbed vaccine generates protective antigen (PA)-Specific CD4+ T cells with a phenotype distinct from that of naive PA T cells. AB - Cellular immune responses against protective antigen (PA) of Bacillus anthracis in subjects that received the anthrax vaccine adsorbed (AVA) vaccine were examined. Multiple CD4(+) T-cell epitopes within PA were identified by using tetramer-guided epitope mapping. PA-reactive CD4(+) T cells with a CD45RA(-) phenotype were also detected by direct ex vivo staining of peripheral blood mononuclear cells (PBMC) with PA-specific tetramers. Surprisingly, PA-specific T cells were also detected in PBMC of nonvaccinees after a single cycle of in vitro PA stimulation. However, PA-reactive CD4(+) T cells in nonvaccinees occurred at lower frequencies than those in vaccinees. The majority of PA-reactive T cells from nonvaccinees were CD45RA(+) and exhibited a Th0/Th1 cytokine profile. In contrast, phenotyping and cytokine profile analyses of PA-reactive CD4(+) T cells from vaccinees indicated that vaccination leads to commitment of PA-reactive T cells to a Th2 lineage, including generation of PA-specific, pre-Th2 central memory T cells. These results demonstrate that the current AVA vaccine is effective in skewing the development of PA CD4(+) T cells to the Th2 lineage. The data also demonstrated the feasibility of using class II tetramers to analyze CD4(+) cell responses and lineage development after vaccination. PMID- 18678673 TI - Characterization of chromosomal regions conserved in Yersinia pseudotuberculosis and lost by Yersinia pestis. AB - The transformation of the enteropathogenic bacterium Yersinia pseudotuberculosis into the plague bacillus, Yersinia pestis, has been accompanied by extensive genetic loss. This study focused on chromosomal regions conserved in Y. pseudotuberculosis and lost during its transformation into Y. pestis. An extensive PCR screening of 78 strains of the two species identified five regions (R1 to R5) and four open reading frames (ORFs; orf1 to orf4) that were conserved in Y. pseudotuberculosis and absent from Y. pestis. Their conservation in Y. pseudotuberculosis suggests a positive selective pressure and a role during the life cycle of this species. Attempts to delete two ORFs (orf3 and orf4) from the chromosome of strain IP32953 were unsuccessful, indicating that they are essential for its viability. The seven remaining loci were individually deleted from the IP32953 chromosome, and the ability of each mutant to grow in vitro and to kill mice upon intragastric infection was evaluated. Four loci (orf1, R2, R4, and R5) were not required for optimal growth or virulence of Y. pseudotuberculosis. In contrast, orf2, encoding a putative pseudouridylate synthase involved in RNA stability, was necessary for the optimal growth of IP32953 at 37 degrees C in a chemically defined medium (M63S). Deletion of R1, a region predicted to encode the methionine salvage pathway, altered the mutant pathogenicity, suggesting that the availability of free methionine is severely restricted in vivo. R3, a region composed mostly of genes of unknown functions, was necessary for both optimal growth of Y. pseudotuberculosis at 37 degrees C in M63S and for virulence. Therefore, despite their loss in Y. pestis, five of the nine Y. pseudotuberculosis-specific chromosomal loci studied play a role in the survival, growth, or virulence of this species. PMID- 18678675 TI - Cortactin recruitment by enterohemorrhagic Escherichia coli O157:H7 during infection in vitro and ex vivo. AB - Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is an important human pathogen that colonizes the gut mucosa via attaching and effacing (A/E) lesions; A/E lesion formation in vivo and ex vivo is dependent on the type III secretion system (T3SS) effector Tir. Infection of cultured cells by EHEC leads to induction of localized actin polymerization, which is dependent on Tir and a second T3SS effector protein, TccP, also known as EspF(U). Recently, cortactin was shown to bind both the N terminus of Tir and TccP via its SH3 domain and to play a role in EHEC-triggered actin polymerization in vitro. In this study, we investigated the recruitment of cortactin to the site of EHEC adhesion during infection of in vitro-cultured cells and mucosal surfaces ex vivo (using human terminal ileal in vitro organ cultures [IVOC]). We have shown that cortactin is recruited to the site of EHEC adhesion in vitro downstream of TccP and N-WASP. Deletion of the entire N terminus of Tir or replacing the N-terminal polyproline region with alanines did not abrogate actin polymerization or cortactin recruitment. In contrast, recruitment of cortactin to the site of EHEC adhesion in IVOC is TccP independent. These results imply that cortactin is recruited to the site of EHEC adhesion in vitro and ex vivo by different mechanisms and suggest that cortactin might have a role during EHEC infection of mucosal surfaces. PMID- 18678676 TI - An orphan comes of age. PMID- 18678677 TI - State mental health policy: Overview and goals of the Mississippi Youth Programs Around the Clock (MYPAC). AB - This column briefly outlines the Mississippi Youth Programs Around the Clock (MYPAC) Demonstration Project. MYPAC is a federally funded effort to examine community-based alternatives to residential placement for youths with severe emotional disturbance. The project is one of ten similarly structured grants awarded nationally and is among the first to reach implementation. Because such a large amount of funding has been dedicated to such a substantial cross-section of national public mental health services ($66,000,000 in Mississippi alone), it is important to make the field aware of developments in these projects as they occur. PMID- 18678679 TI - Personal accounts: Rodolph H. Turcotte, M.D.: another perspective. PMID- 18678678 TI - Economic grand rounds: Financial burden of medical care and risk of forgoing care among Europeans with depressive symptoms. AB - Little is known about the financial burden of individuals with depressive symptoms. This study explored that burden, using data from the Survey of Health, Ageing, and Retirement in Europe. To assess the association between depressive symptoms and the individuals' financial burden for medical care and whether they forwent medical care because of costs, logistic regressions were performed that adjusted for age, gender, marital status, education, and chronic diseases. A total of 16,696 noninstitutionalized individuals aged 50-79 years were included in the study. Individuals with depressive symptoms and those without such symptoms bore a similar financial burden. However, individuals with depressive symptoms were at increased risk of forgoing care because of costs, which may worsen their health and financial situation. PMID- 18678680 TI - Access to medical care among persons with psychotic and major affective disorders. AB - OBJECTIVE: People with serious mental illness have higher mortality rates than the general population, and this difference is not explained by such causes as suicide or accidents. This study therefore examined access and barriers to medical care among persons with serious mental illness. METHODS: Using a nationally representative sample, the authors examined access and barriers to medical care among individuals reporting psychotic and mood disorders. The National Health Interview Survey (NHIS) and NHIS-Disability Component for 1994 and 1995 were merged to provide a sample of 156,475 people over age 18. Individuals with psychotic disorders, bipolar disorder, or major depression were compared with persons without mental disorders on the following outcomes: having a primary care physician, being unable to get needed medical care, being unable to get a needed prescription medication, and delaying medical care because of cost. RESULTS: Persons with psychotic disorders (odds ratio [OR]=.55, 95% confidence interval [CI]=.44-.69) and bipolar disorder (OR=.74, CI=.56-.98) had significantly reduced odds of having a primary care physician compared with people without mental disorders. For any barriers to care, persons with psychotic disorders, bipolar disorder, or major depressive disorder had greatly increased odds (ORs=2.5-7.0) of reporting difficulties in accessing care. CONCLUSIONS: Persons with psychotic disorders and bipolar disorder reported markedly more difficulty in obtaining a primary care physician and greater barriers to care than the general population. Interventions are needed to improve provision of primary medical care to this population. PMID- 18678681 TI - Adequacy of medical treatment among older persons with schizophrenia. AB - OBJECTIVE: This study used Krause's model of illness behavior in later life to examine factors associated with adequacy of medical treatment in a racially diverse sample of older patients (age 55 and over) with schizophrenia. METHODS: In a cross-sectional community study, 119 persons were identified from a sample of 198 persons with schizophrenia age 55 or older who developed the disorder before age 45; for comparison, 57 persons were identified from a group of 113 without the disorder. The groups were compared for treatment of one or more of four common medical disorders. Thirteen independent variables were inserted into Krause's model for the comparisons. A dependent variable was created to indicate whether persons received medication treatment for all of their medical disorders (high-treatment group). RESULTS: Significant differences were found between the schizophrenia and control groups in the proportion receiving treatment for the four physical conditions (.61 versus .83, respectively, p=.004). By logistic regression, four of 13 variables for the schizophrenia group were significantly associated (p<.01) with being in the high-treatment group: absence of depression, lower frequency of medical visits, fewer positive symptoms, and more negative symptoms. CONCLUSIONS: Although older patients with schizophrenia and their age peers had similar levels of physical disorders and rates of health visits, the former received less adequate treatment. More aggressive management of comorbid depression and positive symptoms of schizophrenia may help in improving the health of this population. PMID- 18678682 TI - The impact of integrating mental and general health services on mental health's share of total health care spending in Alberta. AB - OBJECTIVE: In April 2003 the Alberta government integrated specialized mental health services, formerly organized independently, with the health regions, which are responsible for general health services. The objective of this article is to determine whether the transfer was associated with an increase or decrease in the share of resources in the region allocated to mental health care relative to total spending for health care. METHODS: The measure of the share for mental health care is the total costs for mental health care resources as a percentage of total health care spending. Resources and spending examined were those that were actually or potentially under the regions' control. Annual costs for mental health services in the province were obtained for a seven-year period (fiscal year [FY] 2000 through FY 2006) from provincial utilization records for all residents in the province. Unit costs were assigned to each visit. The trend in the share measure was plotted for each year. RESULTS: The share for mental health care increased overall from FY 2000 (7.6%) to FY 2003 (8.2%), but returned to pre FY 2003 levels in the three years after the transfer (7.6%). CONCLUSIONS: Despite concerns expressed before the transfer by federal and provincial reports over the level of expenditures devoted to mental health care, the integration of mental health services with other health services did not result in an increase of the share for mental health care. PMID- 18678683 TI - Family physicians' experiences with community mental health centers: a multilevel analysis. AB - OBJECTIVE: The measurement of professionals' views of quality is a key component in the evaluation of health care delivery; however, few studies have assessed the feasibility of using such evaluations as quality indicators. The effects of individual and organizational factors on family physicians' experiences with community mental health centers were assessed by responses to a self-completed questionnaire and publicly available data. METHODS: Questionnaires were mailed in spring 2006 to family physicians in Norway for their evaluation of 71 community mental health centers; 2,120 (61%) practitioners responded. They evaluated centers on five scales: competence, discharge letter, contact with and help from the center in emergency situations, guidance from the center to practitioners, and workforce situation. Multilevel regression analyses were used to divide the variance between the two levels and to analyze associations with the five scales for individual- and center-level variables. RESULTS: Center characteristics accounted for a substantial part of the variation; the intraclass correlation coefficient ranged from 10% for discharge letter to 23% for workforce situation. General impression of psychiatric services, knowledge of the center, negative episodes with the center, gender, and the center's location were significantly related to the scales. Explained variance at the individual level ranged from 24% for emergency situations to 38% for competence. Explained variance at the center level ranged from 40% for emergency situations to 70% for competence. CONCLUSIONS: A substantial part of the variation at both levels was explained for all scales. Family physicians' evaluation of community mental health centers had substantial variation at the center level, supporting the use of such evaluations as quality indicators. PMID- 18678684 TI - Telephone-delivered, interpersonal psychotherapy for HIV-infected rural persons with depression: a pilot trial. AB - OBJECTIVE: Rural areas account for approximately 6% of AIDS cases in the United States. Many HIV-infected persons in rural areas live with elevated levels of psychiatric distress, suicidal ideation, and loneliness. This pilot study tested whether brief interpersonal psychotherapy delivered via telephone could reduce psychiatric distress among persons living with HIV-AIDS in rural areas in the United States. METHODS: Seventy-nine participants were assigned randomly to a usual care control condition or to a six-session, telephone-delivered, interpersonal psychotherapy intervention (hereafter referred to as the teletherapy group); participants in the teletherapy group continued to receive standard services available to them in the community. Participants completed self administered surveys pre- and postintervention that assessed depressive and psychiatric symptoms, perceptions of loneliness, and social support. RESULTS: Participants in the teletherapy group evidenced greater reductions in depressive symptoms and in overall levels of psychiatric distress, compared with those in the control group. Nearly one-third of teletherapy participants reported clinically meaningful reductions in psychiatric distress from pre- to postintervention. CONCLUSIONS: The telephone-delivered interpersonal therapy intervention showed potential to reduce depressive and psychiatric symptoms among HIV-infected persons in rural areas. On the basis of these encouraging findings, additional research examining this intervention with this clinical population is warranted. PMID- 18678685 TI - Employment outcomes of persons with a mental disorder and comorbid chronic pain. AB - OBJECTIVE: This study examined the independent and interactive effects of common mental disorders and chronic pain conditions on employment and work outcomes among individuals under 65 years old. METHODS: Cross-sectional data were analyzed from the second wave of Healthcare for Communities (HCC2), a household telephone survey of U.S. civilian adults conducted in 2000-2001 (N=5,328). Common mental disorders were assessed by using the short-form version of the World Health Organization's Composite International Diagnostic Interview. Chronic pain conditions and employment outcomes were identified by self-report. Logistic and linear regression analyses were used to provide estimates for work impairment on the basis of the presence of a mental disorder or a chronic pain condition or both. RESULTS: The interaction between presence of a mental disorder and presence of a chronic pain condition was significantly associated with no work for pay in the past 12 months (odds ratio=2.3, 95% confidence interval=1.2-4.2) and number of days of work missed in the past month because of health (regression coefficient=1.47, SE=.59). In stratified analyses this effect persisted for women but not for men. The presence of a mental disorder and the presence of a chronic pain condition were each independently associated with limitations in work and any work missed in the past 30 days because of health, although the interaction was not significant. CONCLUSIONS: Mental disorders and chronic pain are each associated with work disability. Mental disorders are more strongly associated with some work disability outcomes when they are accompanied by chronic pain, especially among women. PMID- 18678686 TI - Key factors for implementing supported employment. AB - OBJECTIVE: This study examined strategies and barriers for implementing supported employment in routine mental health services. METHODS: Qualitative and fidelity data from a two-year period (2002-2004) were examined for nine sites participating in the National Evidence-Based Practices Project. RESULTS: At baseline, none of the sites were providing high-fidelity supported employment. However, by the two-year follow-up, eight of the nine sites successfully implemented high-fidelity programs. Three factors, leadership, mastery, and attitudes, were identified as strongly influencing the implementation (both positively and negatively) across the nine sites. CONCLUSIONS: The findings indicate the need for strong leadership on both the administrative and program levels, an in-depth understanding of the nature and level of training and consultation needed for program leaders and employment specialists, and the value of hiring staff with clinical or business skills. The study also demonstrated that employing staff who doubt and challenge the evidence-based model slows down the implementation process, suggesting the critical role of hiring staff who believe in recovery and supported employment principles. PMID- 18678688 TI - Influence of family involvement and substance use on sustained utilization of services for schizophrenia. AB - OBJECTIVE: This observational study assessed the influence of family support and substance abuse on patterns of service use by individuals with schizophrenia. METHODS: Polychotomous logistic regression was used to analyze an existing database for 258 individuals with schizophrenia who were between the ages of 18 and 67 and were recruited from public mental health care settings. Analyses determined the extent to which two consumer-identified factors, family support and substance abuse status, influenced patterns of outpatient service use (regular, irregular, and infrequent) for schizophrenia. RESULTS: After the analysis adjusted for insight into illness, cognitive functioning, rural or urban residence, and gender, comorbid substance abuse and the interaction between substance abuse status and family support were significantly associated with patterns of service use. Comorbid substance abuse predicted irregular or infrequent patterns of service use over time. Stratified analyses indicated that weekly family support substantially reduced the adverse impact of substance abuse status on consumers' patterns of service use, especially for those living in rural areas. CONCLUSIONS: This study provides evidence that ongoing family support is associated with substantial reductions in the adverse impact of substance abuse on consumers' patterns of service use, especially for consumers living in rural areas. If confirmed in other populations, study findings suggest that reinforcing services and support for family members who provide informal care helps to sustain involvement in care by the especially vulnerable population of individuals with a dual diagnosis of schizophrenia and substance abuse. PMID- 18678687 TI - Service utilization differences for Axis I psychiatric and substance use disorders between white and black adults. AB - OBJECTIVE: Although studies have shown disparities between black and white populations in service utilization for mental disorders, little information exists on whether such disparities apply equally across disorders. The objective of this study was to examine racial differences in lifetime prevalence of service utilization for mood and anxiety disorders and for alcohol and drug use disorders, with controls for predisposing, enabling, and need-for-service variables unequally distributed between racial-ethnic groups. METHODS: Data were from a face-to-face epidemiologic survey of 32,752 non-Hispanic white or black adults ages 18 and older residing in households and group quarters in the United States. Main outcome measures were treatment for mood, anxiety, and alcohol and drug use disorders. RESULTS: White adults were consistently more likely than black adults to have had treatment for mood disorders (odds ratio [OR]=2.16, 95% confidence interval [CI]=1.80-2.59) and anxiety disorders (OR=1.77, 95% CI=1.43 2.19) after adjustment for predisposing and enabling factors and need for service (severity of disorder). In contrast no evidence of lower service utilization for treatment of alcohol use disorders emerged among black respondents (OR=.87, 95% CI=.69-1.10). Moreover, white respondents with drug use disorders were significantly less likely than black respondents to receive treatment for a drug problem (OR=.64, 95% CI=.47-.88). CONCLUSIONS: Differences in treatment between black and white adults depended on the specific disorder and type of treatment considered. Prevention and intervention strategies should address disorder specific disparities in services received. PMID- 18678689 TI - Use of buprenorphine for addiction treatment: perspectives of addiction specialists and general psychiatrists. AB - OBJECTIVE: In 2002 buprenorphine (Suboxone or Subutex) was approved by the U.S. Food and Drug Administration for office-based treatment of opioid addiction. The goal of office-based pharmacotherapy is to bring more opiate-dependent people into treatment and to have more physicians address this problem. This study examined prescribing practices for buprenorphine, including facilitators and barriers, and the organizational settings that facilitate its being incorporated into treatment. METHODS: Addiction specialists and other psychiatrists in four market areas were surveyed by mail and Internet in fall 2005 to examine prescribing practices for buprenorphine. Respondents included 271 addiction specialists (72% response rate) and 224 psychiatrists who were not listed as addiction specialists but who had patients with addictions in their practice (57% response rate). RESULTS: Three years after approval of buprenorphine for office based addiction treatment, nearly 90% of addiction specialists had been approved to prescribe it and two-thirds treated patients with buprenorphine. However, fewer than 10% of non-addiction specialist psychiatrists prescribed it. Regression-adjusted factors predicting prescribing of buprenorphine included support of training and use of buprenorphine by the physician's main affiliated organization, less time in general psychiatry compared with addictions treatment, more time in group practice rather than solo, ten or more opiate-dependent patients, belief that drugs play a large role in addiction treatment, and patient demand. CONCLUSIONS: Office-based pharmacotherapy offers a promising path to improved access to addictions treatment, but prescribing has expanded little beyond the addiction specialist community. PMID- 18678690 TI - Medical services for clients in community mental health centers: results from a national survey. AB - OBJECTIVE: This study provides national data on community mental health centers' (CMHCs') capacity to screen for and address their clients' general medical conditions. METHODS: A survey was distributed to members of the National Council for Community Behavioral Healthcare, the oldest and largest association of CMHCs. RESULTS: Among the 181 CMHCs responding to the survey, more than two-thirds reported having protocols or procedures to screen for common medical problems (hypertension, obesity, dyslipidemia, and diabetes). However, only one-half could provide treatment or referral for those conditions, and less than one-third could provide general medical services on site. Barriers to providing general medical services included problems in reimbursement, workforce limitations, physical plant constraints (for example, lack of available space or equipment), and lack of options for referrals to local community medical providers. CONCLUSIONS: Although most CMHCs had the capacity to screen for common medical conditions, they reported a variety of barriers to providing medical care for those problems either on site or via referral. PMID- 18678691 TI - Perceived barriers to medical care and mental health care among veterans with serious mental illness. AB - OBJECTIVES: This study examined perceived barriers to mental health care and medical care and the relationship between demographic and clinical characteristics and perceived barriers among veterans with serious mental illness. METHODS: Veterans diagnosed as having serious mental illnesses, hospitalized for psychiatric concerns, and at risk for treatment dropout (N=136) completed an interview as part of a larger study of a critical time intervention. RESULTS: Many participants perceived barriers to accessing mental health care (67%) and medical care (60%). Personal factors were cited most often as barriers; overall, however, personal barriers were more likely to be perceived to impede mental health care (56%) than medical care (43%). Psychiatric symptoms were associated with greater perceived barriers to mental health care and medical care. CONCLUSIONS: Veterans with serious mental illness at risk of treatment dropout perceived barriers to mental health care and medical services. Strategies to overcome barriers are needed and should target illness-related factors that may impede service use. PMID- 18678692 TI - Psychiatric disease burden profiles among veterans with epilepsy: the association with health services utilization. AB - OBJECTIVE: This brief report describes patterns of psychiatric comorbidities among patients with epilepsy and their relationship with health care utilization. METHODS: The study identified psychiatric comorbid conditions in a cohort of veterans identified as having epilepsy in fiscal year 1999. From these diagnoses, nine psychiatric disease burden profiles were created. Logistic regression examined variation in emergency, neurology, and primary care for groups having different profiles and compared them with those with only epilepsy. RESULTS: Of the 23,752 individuals identified, 48% had comorbid psychiatric conditions; most had multiple psychiatric diagnoses. Compared with patients with epilepsy only, those with comorbid psychiatric conditions were more likely to have emergency care and high primary care utilization; those with serious mental illness (psychotic disorders) were less likely to receive neurology care. CONCLUSIONS: Multiple co-occurring psychiatric diseases are common among patients with epilepsy. Addressing the mental health and medical needs of these patients, particularly those with serious mental illness, represents a challenge for health organizations. PMID- 18678693 TI - Use of cancer-screening services among persons with serious mental illness in Sacramento County. AB - OBJECTIVE: This study examined the use of breast, cervical, colorectal, and prostate cancer-screening services by persons with serious mental illness enrolled in the Sacramento County Mental Health clinics. METHODS: Of 387 outpatients approached from January 2005 to May 2007, 229 were interviewed. RESULTS: Whereas 97% of the women had received cervical cancer screening at least once in their lifetime, more than 50% of eligible persons over age 50 had never received colorectal cancer screening. Recent use of screening services was highest for cervical cancer (69% had had a Pap test in the past three years) and lowest for colorectal cancer (12% had had a fecal occult blood stool test in the past year or a flexible sigmoidoscopy or colonoscopy in the past five years). CONCLUSIONS: Among persons with serious mental illness, lifetime screening of cervical cancer was higher than for breast, prostate, and colorectal cancers. Receipt of routine, timely cancer screening was low, especially for colorectal cancer. PMID- 18678694 TI - HIV prevention in the clubhouse. PMID- 18678695 TI - The Family Member Provider Outreach program. PMID- 18678698 TI - A question of disclosure. PMID- 18678699 TI - ACT for people with intellectual disabilities and mental health problems. PMID- 18678700 TI - Occupational exposures and the risk of COPD: dusty trades revisited. AB - BACKGROUND: The contribution of occupational exposures to chronic obstructive pulmonary disease (COPD) and, in particular, their potential interaction with cigarette smoking remains underappreciated. METHODS: Data from the FLOW study of 1202 subjects with COPD (of which 742 had disease classified as stage II or above by Global Obstructive Lung Disease (GOLD) criteria) and 302 referent subjects matched by age, sex and race recruited from a large managed care organisation were analysed. Occupational exposures were assessed using two methods: self reported exposure to vapours, gas, dust or fumes on the longest held job (VGDF) and a job exposure matrix (JEM) for probability of exposure based on occupation. Multivariate analysis was used to control for age, sex, race and smoking history. The odds ratio (OR) and adjusted population attributable fraction (PAF) associated with occupational exposure were calculated. RESULTS: VGDF exposure was associated with an increased risk of COPD (OR 2.11; 95% CI 1.59 to 2.82) and a PAF of 31% (95% CI 22% to 39%). The risk associated with high probability of workplace exposure by JEM was similar (OR 2.27; 95% CI 1.46 to 3.52), although the PAF was lower (13%; 95% CI 8% to 18%). These estimates were not substantively different when the analysis was limited to COPD GOLD stage II or above. Joint exposure to both smoking and occupational factors markedly increased the risk of COPD (OR 14.1; 95% CI 9.33 to 21.2). CONCLUSIONS: Workplace exposures are strongly associated with an increased risk of COPD. On a population level, prevention of both smoking and occupational exposure, and especially both together, is needed to prevent the global burden of disease. PMID- 18678701 TI - Effect of maternal asthma, exacerbations and asthma medication use on congenital malformations in offspring: a UK population-based study. AB - BACKGROUND: Clinical advice to pregnant women with asthma is to maintain optimal therapeutic management; however, potential adverse effects of asthma treatments on fetal development remain uncertain. A study was undertaken to assess the association between maternal asthma and gestational exposure to asthma medications with risk of congenital malformation in offspring. METHODS: A matched case-control study was performed using The Health Improvement Network primary care database. Children with malformations were matched to control children on birth year, general practice and singleton or twin delivery. RESULTS: 5124 cases of liveborn children with major congenital malformations and 30,053 controls were included in the study. The risk of any malformation in children born to women with asthma was marginally higher than that in children born to women without asthma (adjusted OR 1.10, 95% CI 1.01 to 1.20). However, no association was present in children born to mothers receiving asthma treatment in the year before or during pregnancy (OR 1.06, 95% CI 0.94 to 1.20). In assessing teratogenicity of medications, no increased risk of malformation was found with gestational exposures to short- or long-acting beta agonists, inhaled corticosteroids, oral corticosteroids, other bronchodilators or cromones. These findings were similar for each of 11 system-specific malformation groups, except for an increase in musculo-skeletal system malformation associated with cromone exposure. CONCLUSIONS: Gestational exposure to commonly used asthma medications was found to be safe overall, although a moderate teratogenic risk of cromones cannot be excluded. There was some evidence of a small increased risk of congenital malformation in children born to women with asthma, but this was not explained by gestational exposure to asthma drugs. PMID- 18678702 TI - Pulmonary tumours in the Netherlands: focus on temporal trends in histology and stage and on rare tumours. AB - BACKGROUND: Recent temporal trends in histology and stage of pulmonary tumours in the Netherlands were studied. The incidence of rare pulmonary tumours was determined. METHODS: All tumours originating from the trachea, bronchus and lung recorded in the Netherlands Cancer Registry were included. Based on ICD-O morphology codes, five major subgroups were constructed: squamous carcinoma (SC), adenocarcinoma (AC), large cell (undifferentiated) carcinoma (LC), small cell lung cancer (SCLC) and other (including uncommon tumours). RESULTS: Between 1989 and 2003, 134,894 tumours were diagnosed. In men the age-adjusted incidence of SC and SCLC decreased, AC remained stable and LC increased. In women the incidence of all subgroups increased. Since 1996, a stage shift was observed with fewer patients in stage I and more patients in stage IV at diagnosis. This stage shift occurred equally in SC, AC and LC. In SC, fewer patients presented with stage IV disease than in AC and LC (25% vs 44% and 49% in 2003, respectively). The incidence of adenosquamous carcinoma decreased from 0.6 to 0.29/100 000 (p<0.001). The incidence of carcinoid tumours, sarcomatoid carcinomas and primary pulmonary sarcomas remained stable (0.44, 0.17 and 0.08/100 000, respectively). CONCLUSION: The incidence of smoking-related tumours decreased in men (especially SC and SCLC) and increased in women (all subgroups). More patients presented with stage IV disease. The incidence of non-smoking-related uncommon tumours remained constant. PMID- 18678703 TI - Ventilation heterogeneity in children with well controlled asthma with normal spirometry indicates residual airways disease. AB - BACKGROUND: In adults with asthma, ventilation heterogeneity, independent of inflammation, has been hypothesised to be associated with airway remodelling. Bronchial biopsy in preschool children with wheeze demonstrates early structural changes. Ventilation heterogeneity is sensitive to airway disease in other paediatric respiratory conditions such as cystic fibrosis, so may be sensitive to early airway disease in asthma. An observational study was performed in which it was hypothesised that ventilation heterogeneity (lung clearance index (LCI) and phase III slope indices (S(cond) and S(acin))) were more sensitive than conventional measurements (forced expiratory volume in 1 s (FEV(1)) and exhaled nitric oxide (Feno)) for detecting residual airways disease in children with well controlled asthma. METHODS: In 31 children with asthma of mean age 10.6 years (range 5-15), FEV(1), LCI, S(cond) and S(acin) were measured at two separate visits, before and after blinded salbutamol or placebo, with Feno measured once. 29 healthy volunteers of mean age 11.2 years (range 5-16) completed measurements at one visit only. RESULTS: Baseline mean (SD) LCI was significantly higher in children with asthma than in controls (6.69 (0.91) vs 6.24 (0.47), p = 0.02). There were no significant differences in FEV(1) or median Feno. Following salbutamol there was a small significant change in mean (SD) FEV(1) (from -1.26 (1.25) to -0.93 (0.23), p = 0.03) but not in LCI, S(cond) or S(acin). Importantly, LCI remained significantly higher after bronchodilator in children with asthma than in controls (6.64 (0.69), p = 0.01). CONCLUSION: This study identifies the presence of residual ventilation heterogeneity in children with well controlled asthma and normal FEV(1). The role of LCI in measuring early airway disease in children with asthma requires further exploration, possibly as a surrogate of structural remodelling. PMID- 18678704 TI - Associations of wheezing phenotypes in the first 6 years of life with atopy, lung function and airway responsiveness in mid-childhood. AB - BACKGROUND: Patterns of wheezing during early childhood may indicate differences in aetiology and prognosis of respiratory illnesses. Improved characterisation of wheezing phenotypes could lead to the identification of environmental influences on the development of asthma and airway diseases in predisposed individuals. METHODS: Data collected on wheezing at seven time points from birth to 7 years from 6265 children in a longitudinal birth cohort (the ALSPAC study) were analysed. Latent class analysis was used to assign phenotypes based on patterns of wheezing. Measures of atopy, airway function (forced expiratory volume in 1 s (FEV(1)), mid forced expiratory flow (FEF(25-75))) and bronchial responsiveness were made at 7-9 years of age. RESULTS: Six phenotypes were identified. The strongest associations with atopy and airway responsiveness were found for intermediate onset (18 months) wheezing (OR for atopy 8.36, 95% CI 5.2 to 13.4; mean difference in dose response to methacholine 1.76, 95% CI 1.41 to 2.12 %FEV(1) per mumol, compared with infrequent/never wheeze phenotype). Late onset wheezing (after 42 months) was also associated with atopy (OR 6.6, 95% CI 4.7 to 9.4) and airway responsiveness (mean difference 1.61, 95% CI 1.37 to 1.85 %FEV(1) per mumol). Transient and prolonged early wheeze were not associated with atopy but were weakly associated with increased airway responsiveness and persistent wheeze had intermediate associations with these outcomes. CONCLUSIONS: The wheezing phenotypes most strongly associated with atopy and airway responsiveness were characterised by onset after age 18 months. This has potential implications for the timing of environmental influences on the initiation of atopic wheezing in early childhood. PMID- 18678705 TI - Reciprocal interaction with G-actin and tropomyosin is essential for aquaporin-2 trafficking. AB - Trafficking of water channel aquaporin-2 (AQP2) to the apical membrane and its vasopressin and protein kinase A (PKA)-dependent regulation in renal collecting ducts is critical for body water homeostasis. We previously identified an AQP2 binding protein complex including actin and tropomyosin-5b (TM5b). We show that dynamic interactions between AQP2 and the actin cytoskeleton are critical for initiating AQP2 apical targeting. Specific binding of AQP2 to G-actin in reconstituted liposomes is negatively regulated by PKA phosphorylation. Dual color fluorescence cross-correlation spectroscopy reveals local AQP2 interaction with G-actin in live epithelial cells at single-molecule resolution. Cyclic adenosine monophosphate signaling and AQP2 phosphorylation release AQP2 from G actin. In turn, AQP2 phosphorylation increases its affinity to TM5b, resulting in reduction of TM5b bound to F-actin, subsequently inducing F-actin destabilization. RNA interference-mediated knockdown and overexpression of TM5b confirm its inhibitory role in apical trafficking of AQP2. These findings indicate a novel mechanism of channel protein trafficking, in which the channel protein itself critically regulates local actin reorganization to initiate its movement. PMID- 18678706 TI - A Cdo-Bnip-2-Cdc42 signaling pathway regulates p38alpha/beta MAPK activity and myogenic differentiation. AB - The p38alpha/beta mitogen-activated protein kinase (MAPK) pathway promotes skeletal myogenesis, but the mechanisms by which it is activated during this process are unclear. During myoblast differentiation, the promyogenic cell surface receptor Cdo binds to the p38alpha/beta pathway scaffold protein JLP and, via JLP, p38alpha/beta itself. We report that Cdo also interacts with Bnip-2, a protein that binds the small guanosine triphosphatase (GTPase) Cdc42 and a negative regulator of Cdc42, Cdc42 GTPase-activating protein (GAP). Moreover, Bnip-2 and JLP are brought together through mutual interaction with Cdo. Gain- and loss-of-function experiments with myoblasts indicate that the Cdo-Bnip-2 interaction stimulates Cdc42 activity, which in turn promotes p38alpha/beta activity and cell differentiation. These results reveal a previously unknown linkage between a cell surface receptor and downstream modulation of Cdc42 activity. Furthermore, interaction with multiple scaffold-type proteins is a distinctive mode of cell surface receptor signaling and provides one mechanism for specificity of p38alpha/beta activation during cell differentiation. PMID- 18678707 TI - Microtubule cross-linking triggers the directional motility of kinesin-5. AB - Although assembly of the mitotic spindle is known to be a precisely controlled process, regulation of the key motor proteins involved remains poorly understood. In eukaryotes, homotetrameric kinesin-5 motors are required for bipolar spindle formation. Eg5, the vertebrate kinesin-5, has two modes of motion: an adenosine triphosphate (ATP)-dependent directional mode and a diffusive mode that does not require ATP hydrolysis. We use single-molecule experiments to examine how the switching between these modes is controlled. We find that Eg5 diffuses along individual microtubules without detectable directional bias at close to physiological ionic strength. Eg5's motility becomes directional when bound between two microtubules. Such activation through binding cargo, which, for Eg5, is a second microtubule, is analogous to known mechanisms for other kinesins. In the spindle, this might allow Eg5 to diffuse on single microtubules without hydrolyzing ATP until the motor is activated by binding to another microtubule. This mechanism would increase energy and filament cross-linking efficiency. PMID- 18678708 TI - Bat3 deficiency accelerates the degradation of Hsp70-2/HspA2 during spermatogenesis. AB - Meiosis is critical for sexual reproduction. During meiosis, the dynamics and integrity of homologous chromosomes are tightly regulated. The genetic and molecular mechanisms governing these processes in vivo, however, remain largely unknown. In this study, we demonstrate that Bat3/Scythe is essential for survival and maintenance of male germ cells (GCs). Targeted inactivation of Bat3/Scythe in mice results in widespread apoptosis of meiotic male GCs and complete male infertility. Pachytene spermatocytes exhibit abnormal assembly and disassembly of synaptonemal complexes as demonstrated by abnormal SYCP3 staining and sustained gamma-H2AX and Rad51/replication protein A foci. Further investigation revealed that a testis-specific protein, Hsp70-2/HspA2, is absent in Bat3-deficient male GCs at any stage of spermatogenesis; however, Hsp70-2 transcripts are expressed at normal levels. We found that Bat3 deficiency induces polyubiquitylation and subsequent degradation of Hsp70-2. Inhibition of proteasomal degradation restores Hsp70-2 protein levels. Our findings identify Bat3 as a critical regulator of Hsp70-2 in spermatogenesis, thereby providing a possible molecular target in idiopathic male infertility. PMID- 18678709 TI - Ku80 removal from DNA through double strand break-induced ubiquitylation. AB - The Ku70/Ku80 heterodimer, or Ku, is the central component of the nonhomologous end joining (NHEJ) pathway of double strand break (DSB) repair. Because Ku forms a ring through which the DSB threads, it likely becomes topologically attached to DNA during repair. The mechanism for its removal was unknown. Using a method to identify proteins recruited to DSBs in Xenopus laevis egg extract, we show that DSB-containing DNAs accumulate members of the Skp1-Cul1-F-box complex and K48 linked polyubiquitylated proteins in addition to known repair proteins. We demonstrate that Ku80 is degraded in response to DSBs in a ubiquitin-mediated manner. Strikingly, K48-linked polyubiquitylation, but not proteasomal degradation, is required for the efficient removal of Ku80 from DNA. This removal is DNA length dependent, as Ku80 is retained on duplex oligonucleotides. Finally, NHEJ completion and removal of Ku80 from DNA are independent from one another. We propose that DSB-induced ubiquitylation of Ku80 provides a mechanism to efficiently eliminate Ku from DNA for pre- and postrepair processes. PMID- 18678710 TI - FGF-23-Klotho signaling stimulates proliferation and prevents vitamin D-induced apoptosis. AB - Fibroblast growth factor 23 (FGF-23) and Klotho are secretory proteins that regulate mineral-ion metabolism. Fgf-23(-/-) or Klotho(-/-) knockout mice exhibit several pathophysiological processes consistent with premature aging including severe atrophy of tissues. We show that the signal transduction pathways initiated by FGF-23-Klotho prevent tissue atrophy by stimulating proliferation and preventing apoptosis caused by excessive systemic vitamin D. Because serum levels of active vitamin D are greatly increased upon genetic ablation of Fgf-23 or Klotho, we find that these molecules have a dual role in suppression of apoptotic actions of vitamin D through both negative regulation of 1alpha hydroxylase expression and phosphoinositide-3 kinase-dependent inhibition of caspase activity. These data provide new insights into the physiological roles of FGF-23 and Klotho. PMID- 18678711 TI - Dynamic partitioning of mitotic kinesin-5 cross-linkers between microtubule-bound and freely diffusing states. AB - The dynamic behavior of homotetrameric kinesin-5 during mitosis is poorly understood. Kinesin-5 may function only by binding, cross-linking, and sliding adjacent spindle microtubules (MTs), or, alternatively, it may bind to a stable "spindle matrix" to generate mitotic movements. We created transgenic Drosophila melanogaster expressing fluorescent kinesin-5, KLP61F-GFP, in a klp61f mutant background, where it rescues mitosis and viability. KLP61F-GFP localizes to interpolar MT bundles, half spindles, and asters, and is enriched around spindle poles. In fluorescence recovery after photobleaching experiments, KLP61F-GFP displays dynamic mobility similar to tubulin, which is inconsistent with a substantial static pool of kinesin-5. The data conform to a reaction-diffusion model in which most KLP61F is bound to spindle MTs, with the remainder diffusing freely. KLP61F appears to transiently bind MTs, moving short distances along them before detaching. Thus, kinesin-5 motors can function by cross-linking and sliding adjacent spindle MTs without the need for a static spindle matrix. PMID- 18678712 TI - Activation of mitotic kinesin by microtubule bundling. AB - Kinesin-5 family members cross-link and slide parallel microtubules of opposite polarity, an activity that is essential for the formation of a bipolar spindle during mitosis. In this issue, Kapitein et al. (Kapitein, L.C., B.H. Kwok, J.S. Weinger, C.F. Schmidt, T.M. Kapoor, and E.J.G. Peterman. 2008. J. Cell Biol. 182:421-428) demonstrate that microtubule cross-linking triggers the conversion of kinesin-5 motility from a diffusive mode to a directional mode, initiating antiparallel microtubule sliding. PMID- 18678714 TI - Healthcare system error: beyond apology. PMID- 18678716 TI - Making a blood transfusion safe: vigilance and audit. PMID- 18678717 TI - Quality of dying--how are we doing? PMID- 18678718 TI - Changes in practice and organisation surrounding blood transfusion in NHS trusts in England 1995-2005. AB - BACKGROUND: Between 1996 and 2005 in the UK, the Serious Hazards of Transfusion (SHOT) scheme has reported 105 deaths and 296 patients developing major morbidity due to transfusion. Accurate patient identification and monitoring of patients during blood transfusion are vital in ensuring patient safety, and national guidelines have been in place since 1999. There have been numerous initiatives in the UK in recent years promoting safe and appropriate use of blood and this paper reports the results of the 2005 National Comparative Audit of transfusion practice, and compares this audit with previous audits and survey results. METHODS: The 2005 audit consisted of two parts looking at organisational factors and bedside practice. To enable comparison with previous audits and surveys the 2005 data have been limited to English NHS sites (217 sites for organisational, 211 sites for bedside practice). RESULTS: Hospital transfusion committees were well established by 2003 though hospital transfusion teams have lagged behind. 86% of hospitals reported having established teams by 2005 although only 52% reported having all essential elements of the team in place. Only 38% reported having trained at least half of their nurses in blood transfusion. Bedside practice has improved, although in 2005, 6% of patients receiving a blood transfusion had no identification wristband in place, and in 9% of those who did, the details were incomplete. Observation of vital signs during transfusions has also improved, although in 2005, 13% of patients receiving a transfusion had had no observations recorded. CONCLUSION: This paper document the progress that has been made in the UK in establishing an effective infrastructure for the support of safe transfusion practice, and the measurable improvements in bedside transfusion practice. There remain, however, many areas of poor practice, and the improvements have not been seen across all hospitals. It is still too early to say whether progress made is being translated into a reduction in serious transfusion errors at the bedside. Further progress needs to be made. PMID- 18678719 TI - Quality of dying in a New Zealand teaching hospital. AB - OBJECTIVE: Studies suggest that there is a need to improve the way we deliver care at the end of life. Based on recommendations from end-of-life experts, metrics were identified to measure the quality of dying in Dunedin Hospital. DESIGN: A retrospective observational study was performed to assess the care provided to patients who died in the hospital in 2003. SETTING: Dunedin Hospital is a 350-bed tertiary care teaching hospital located in the South Island of New Zealand. SUBJECTS AND METHOD: Medical records of 200 consecutive decedents were reviewed to evaluate communication, interventions, and symptom control during their terminal hospitalisation. RESULTS: Mean hospital length-of-stay was 8 days; 38 patients (19%) died following an ICU admission. There was documentation of end of-life discussion with either the patient or the family in 164 patients (82%). 74% had a DNR order. Pain status was documented in 140 patients (70%); 134 of these patients were pain-free. CONCLUSION: Overall, the results suggest that the ideals in end-of-life care pertaining to pain control, communication and avoidance of unnecessary interventions were achieved in a majority of the decedents during the study period. The socialised healthcare system, the availability of resources, societal expectations and a lack of a litigious environment are theorised to positively influence end-of-life care delivery in New Zealand. PMID- 18678720 TI - Peer support: healthcare professionals supporting each other after adverse medical events. AB - The patient safety movement in healthcare is beginning to openly acknowledge the need to support the human side of adverse medical events in conjunction with evidence-based improvement initiatives. While medical literature has sporadically reported on the emotional impact of adverse events on healthcare professionals, little has been documented on the implementation of support services following these events. This article describes an adverse medical event where open communication and apology catalysed the development and implementation of a structured peer support service for care providers at the Brigham and Women's Hospital following adverse events. The Peer Support Service bypasses the stigmas that limit the utilisation of formal support services and offers care providers a safe environment to share the emotional impact of adverse events while serving as a foundation for open communication and a renewal of compassion in the workplace. As the breadth of stressors impacting healthcare professionals is revealed, the Peer Support Service is being recognised as a vital hospital-wide service. It also appears to offer an important leap forward in the critical areas of patient safety and quality of care. PMID- 18678721 TI - Detection of adverse events in surgical patients using the Trigger Tool approach. AB - BACKGROUND: Most studies of healthcare complications identify surgery as a major contributor to the overall burden of complicated care that leads to injury or death. Indeed, surgical adverse events account for one-half to three-quarters of all adverse events in these studies. Despite the intensive current focus on improving medical quality and safety, only a minority of quality improvement efforts are focused on surgery. This study reports on the development and testing of a Trigger Tool to detect adverse events among patients undergoing inpatient surgery. METHODS: Rather than relying on traditional voluntary reporting for safety outcome measures such as incident reports, surgical peer review, or morbidity and mortality conferences, the Institute for Healthcare Improvement (IHI) has employed a new method for the detection of surgical adverse events (SAEs). This approach, commonly referred to as the "Trigger Tool", identifies adverse events using a form of retrospective record review that has been developed and implemented in many areas of care. RESULTS: During a 12-month IHI Perioperative Safety Collaborative, 11 hospitals voluntarily submitted data from surgical inpatient record reviews. In 854 patients, 138 SAEs were detected in 125 records for a rate of 16 SAEs per 100 patients or 14.6% of patients; 61 (44%) of these events contributed to increased length of stay or readmission and 12 (8.7%) events required life-saving intervention or resulted in permanent harm or death. Hospital review teams reported verbally that most of the events identified during the Trigger Tool review process had not been detected or reported via any other existing mechanism. CONCLUSIONS: The IHI Surgical Trigger Tool may offer a practical, easy-to-use approach to detecting safety problems in patients undergoing surgery; it can be the basis not only for estimating the frequency of adverse events in an organisation, but also determining the impact of interventions that focus on reducing adverse events in surgical patients. PMID- 18678722 TI - Analysis of 23 364 patient-generated, physician-reviewed malpractice claims from a non-tort, blame-free, national patient insurance system: lessons learned from Sweden. AB - CONTEXT: In Sweden, patient malpractice claims are handled administratively and compensated if an independent physician review confirms patient injury resulting from medical error. Full access to all malpractice claims and hospital discharge data for the country provided a unique opportunity to assess the validity of patient claims as indicators of medical error and patient injury. OBJECTIVE: To determine: (1) the percentage of patient malpractice claims validated by independent physician review, (2) actual malpractice claims rates (claims frequency / clinical volume) and (3) differences between Swedish and other national malpractice claims rates. Design, setting and material: Swedish national malpractice claims and hospital discharge data were combined, and malpractice claims rates were determined by county, hospital, hospital department, surgical procedure, patient age and sex and compared with published studies on medical error and malpractice. RESULTS: From 1997 to 2004, there were 23 364 inpatient malpractice claims filed by Swedish patients treated at hospitals reporting 11 514 798 discharges. The overall claims rate, 0.20%, was stable over the period of study and was similar to that found in other tort and administrative compensation systems. Over this 8-year period, 49.5% (range 47.0-52.6%) of filed claims were judged valid and eligible for compensation. Claims rates varied significantly across hospitals; surgical specialties accounted for 46% of discharges, but 88% of claims. There were also large differences in claims rates for procedures. CONCLUSIONS: Patient-generated malpractice claims, as collected in the Swedish malpractice insurance system and adjusted for clinical volumes, have a high validity, as assessed by standardised physician review, and provide unique new information on malpractice risks, preventable medical errors and patient injuries. Systematic collection and analysis of patient-generated quality of care complaints should be encouraged, regardless of the malpractice compensation system in use. PMID- 18678723 TI - Impact of NICE guidance on rates of haemorrhage after tonsillectomy: an evaluation of guidance issued during an ongoing national tonsillectomy audit. AB - BACKGROUND: The National Institute for Health and Clinical Excellence (NICE) issued guidance on surgical techniques for tonsillectomy during a national audit of surgical practice and postoperative complications. OBJECTIVE: To assess the impact of the guidance on tonsillectomy practice and outcomes. DESIGN: An interrupted time-series analysis of routinely collected Hospital Episodes Statistics data, and an analysis of longitudinal trends in surgical technique using data from the National Prospective Tonsillectomy Audit. PARTICIPANTS: Patients undergoing tonsillectomy in English NHS hospitals between January 2002 and December 2004. MAIN OUTCOME MEASURE: Postoperative haemorrhage within 28 days. RESULTS: The rate of haemorrhage increased by 0.5% per year from 2002, reaching 6.4% when the guidance was published. After publication, the rate of haemorrhage fell immediately to 5.7% (difference 0.7%: 95% CI -1.3% to 0.0%) and the rate of increase appeared to have stopped. Data from the National Prospective Tonsillectomy Audit showed that the fall coincided with a shift in surgical techniques, which was consistent with the guidance. CONCLUSION: NICE guidance influenced surgical tonsillectomy technique and in turn produced an immediate fall in postoperative haemorrhage. The ongoing national audit and strong support from the surgical specialist association may have aided its implementation. PMID- 18678724 TI - Implementation of proven interventions in general medical inpatients: development and evaluation of a new quality indicator for drug therapy. AB - BACKGROUND: Among adult general medical inpatients, there are numerous interventions whose benefits outweigh their risks. However, there are no published reports describing the overall use of such proven interventions in this population. OBJECTIVES: To determine implementation rates of a broad range of interventions while accounting for valid reasons for non-use, predictors of implementation and feasibility of generating new indices to describe quality of care. METHODS: Based on a review of current practice guidelines and clinical trials related to five common conditions, implementation rates of 17 interventions were assessed retrospectively. Subjects were a complete sample of 150 adults with target medical conditions discharged from general medical units at an urban community hospital. RESULTS: The Ideal Intervention Index (3I), which described the proportion of ideal intervention opportunities that were implemented, was 0.74 (95% CI 0.70 to 0.78). The Justified Non-Use Index (JNUI), which described the proportion of all the interventions not implemented that were justified by a valid reason for non-use, was 0.49 (95% CI 0.41 to 0.55). Smoking cessation therapy in high-risk patients had the lowest indices (3I 0.30, 95% CI 0.00 to 0.60; JNUI 0.00), and aspirin for secondary stroke prevention had the highest (3I 1.0; JNUI 1.0). CONCLUSIONS: Overall, proven interventions are underused among the patients studied, and the reasons for non-use are frequently not readily discernible. There is potential for improvement, but research is required to further investigate reasons for non-use. It is feasible to measure implementation rates of proven interventions as an indicator of quality of care using the indices developed. PMID- 18678726 TI - Accuracy in the recording of pressure ulcers and prevention after implementing an electronic health record in hospital care. AB - OBJECTIVE: To compare the accuracy in recording of pressure-ulcer prevalence and prevention before and after implementing an electronic health record (EHR) with templates for pressure-ulcer assessment. METHODS: All inpatients at the departments of surgery, medicine and geriatrics were inspected for the presence of pressure ulcers, according to the European Pressure Ulcer Advisory Panel methodology, during 1 day in 2002 (n = 357) and repeated in 2006 (n = 343). The corresponding patient records were audited retrospectively for the presence of documentation on pressure ulcers. RESULTS: In 2002, the prevalence of pressure ulcers obtained by auditing paper-based patient records (n = 413) was 14.3%, compared with 33.3% in physical inspection (n = 357). The largest difference was seen in the geriatric department, where records revealed 22.9% pressure ulcers and skin inspection 59.3%. Four years later, after the implementation of the EHR, there were 20.7% recorded pressure ulcers and 30.0% found by physical examination of patients. The accuracy of the prevalence data had improved most in the geriatric department, where the EHR showed 48.1% and physical examination 43.2% pressure ulcers. Corresponding figures in the surgical department were 22.2% and 14.1%, and in the medical department 29.9% and 10.2%, respectively. The patients received pressure-reducing equipment to a higher degree (51.6%) than documented in the patient record (7.9%) in 2006. CONCLUSIONS: The accuracy in pressure-ulcer recording improved in the EHR compared with the paper-based health record. However, there were still deficiencies, which mean that patient records did not serve as a valid source of information on pressure-ulcer prevalence and prevention. PMID- 18678725 TI - The Team Climate Inventory: application in hospital teams and methodological considerations. AB - OBJECTIVE: To test the validity, reliability and discriminating capacity of an instrument to assess team climate, the Team Climate Inventory (TCI), in a sample of Dutch hospital teams. The TCI is based on a four-factor theory of team climate for innovation. DESIGN: Validation study. SETTING: Hospital teams in The Netherlands. PARTICIPANTS: 424 healthcare professionals; 355 nurses working in 22 nursing teams and 69 nurses and doctors working in 14 quality-improvement teams. MAIN OUTCOME MEASURES: Exploratory and confirmatory factor analyses, Pearson's product moment correlations, internal homogeneity of the TCI scales based on Cronbach alpha, and the TCI capability to discriminate between two types of healthcare teams, namely nursing teams and quality-improvement teams. RESULTS: The validity test revealed the TCI's five-factor structure and moderate data fit. The Cronbach alphas of the five scales showed acceptable reliabilities. The TCI discriminated between nursing teams and quality-improvement teams. The mean scores of quality-improvement teams were all significantly higher than those of the nursing teams. CONCLUSION: Patient care teams are essential for high-quality patient care, and team climate is an important characteristic of successful teams. This study shows that the TCI is a valid, reliable and discriminating self report measure of team climate in hospital teams. The TCI can be used as a quality-improvement tool or in quality-of-care research. PMID- 18678727 TI - Medication errors reported by US family physicians and their office staff. AB - BACKGROUND: Most medication error studies come from inpatient settings. There is limited information about medication errors in primary care settings. OBJECTIVE: To describe medication errors reported by family physicians and their office staff and to estimate their preventability using currently available electronic prescribing and monitoring tools. Design, setting, participants and study instrument: In two error reporting studies conducted by the American Academy of Family Physicians (AAFP) National Research Network (NRN), 1265 medical errors were voluntarily reported by >440 primary care clinicians and staff from 52 physician offices. The 194 error reports related to medications were abstracted and analysed using a medication error coding tool-Medication Error Types, Reasons, and Informatics Preventability (METRIP). MAIN OUTCOME MEASURES: Type, severity and preventability of medication errors and associated adverse drug events (ADEs). RESULTS: 126 (70%) of the medication errors were prescribing errors, 17 (10%) were medication administration errors, 17 (10%) documentation errors, 13 (7%) dispensing errors and 5 (3%) were monitoring errors. ADEs resulted from 16% of reported medication errors. The severity of harm from reported errors were: prevented and did not reach patients, (72, 41%), reached patients but did not require monitoring (63, 35%), reached patients and required monitoring (15, 8%), reached patients and required intervention (23, 13%) and reached patients and resulted in hospitalisation (5, 3%). No deaths were reported. Of the errors that were prevented from reaching patients, 29 (40%) were prevented by pharmacists, 14 (19%) by physicians, 12 (17%) by patients and 5 (7%) by nurses. 102 (57%) of the reported errors might have been prevented with enhanced electronic prescribing and monitoring tools. CONCLUSIONS: Most medication errors reported from US family physician offices were related to prescribing errors and more than half of the errors reached patients. The errors were prevented by pharmacists, patients and physicians. More than half of the errors could be prevented by electronic tools. PMID- 18678728 TI - Developing quality indicators for general practice care for vulnerable elders; transfer from US to The Netherlands. AB - BACKGROUND: Measurement of the quality of healthcare is a first step for quality improvement. To measure quality of healthcare, a set of quality indicators is needed. We describe the adaptation of a set of systematically developed US quality indicators for healthcare for vulnerable elders in The Netherlands. We also compare the US and the Dutch set to see if quality indicators can be transferred between countries, as has been done in two studies in the UK, with mixed results. METHOD: 108 US quality indicators on GP care for vulnerable elders, covering eight conditions, were assessed by a panel of nine clinical experts in The Netherlands. A modified version of the RAND/UCLA appropriateness method was used. The panel members received US literature reviews, extended with more recent and Dutch literature, summarising the evidence for each quality indicator. RESULTS: 72 indicators (67% of US set) were (nearly) identical in the Dutch and US sets. For some conditions, this percentage was much lower. For undernutrition, only half of the US indicators were included in the Dutch set. For depression, many indicators were discarded or changed in a significant way, with the result that only five of the original 17 indicators (29%) are the same in the Dutch and the US set. CONCLUSIONS: Quality indicators can be transferred between countries, but with caution, because in two of the three studies on transferring indicators between the US and Europe, 33-44% of the indicators were discarded. For some conditions in the current study, this percentage is much higher. For undernutrition, there is hardly any evidence, and differences between the indicator sets can be attributed to differences in expert opinion between the countries. For depression, it seems that different evidence is considered important in the US and in The Netherlands, of which the Dutch body of knowledge is not known in the US. PMID- 18678729 TI - Guideline implementation in allied health professions: a systematic review of the literature. AB - BACKGROUND: Clinical guidelines aim to improve the safety and quality of patient care by providing clinicians with graded recommendations based on evidence of best practice. OBJECTIVE: To evaluate the effects of the introduction of clinical guidelines for allied health professionals, and to estimate the effectiveness of the guideline dissemination and implementation strategies used. METHODS: A comprehensive search of six electronic databases to June 2006 and the Cochrane Effective Practice and Organisation of Care database was conducted. Randomised controlled trials, controlled clinical trials, controlled before and after studies and interrupted time series studies were included if the intervention was aimed at implementing guidelines in the allied health professions. Articles were screened for eligibility and their methodological quality was assessed; data were extracted independently by two reviewers. RESULTS: Fourteen studies met the inclusion criteria. The methodological quality varied greatly, with the proportion of quality criteria met ranging from 0 to 6 out of seven. In most studies, the effects reported for patient and process outcomes were small and in favour of the intervention group. Of the 14 included studies, 10 focused on educational interventions. Six of the 14 studies used a single intervention strategy and seven used a multi-faceted implementation strategy. One study compared both single and multi-faceted strategies. Multi-faceted interventions were no more effective than single intervention strategies and effects of the same strategy varied across trials. CONCLUSIONS: There is no evidence to support a set guideline implementation strategy for allied health professionals. When implementing clinical guidelines it is important to first identify specific barriers to change using theoretical frameworks of behaviour change and then develop strategies that deal with these barriers. When measuring the effectiveness of these strategies, professionals should consider the use of both patient and process outcomes and choose outcomes that reflect their aims. PMID- 18678730 TI - Use of statistical process control charts in stroke medicine to determine if clinical evidence and changes in service delivery were associated with improvements in the quality of care. AB - BACKGROUND AND OBJECTIVE: Monitoring the effect of service changes on quality of care is essential. By using statistical process control (SPC) charts, this study aimed to explore the relationship between changes in the structure of stroke services and the process of care. METHODS: Prospectively acquired data on the process of acute stroke care from three hospitals admitting 2962 patients (July 2001 to June 2004) were charted retrospectively on SPC charts for individual values (I charts) to determine whether or not "special cause variation" followed known changes in stroke service structure and publication of the Medical Research Council (MRC) Heart Protection Study. Unexpected signals of special cause variation were identified and reasons for observed patterns were sought by discussion with clinical teams. RESULTS: Improved brain imaging provision was followed by a reduction in time to imaging and earlier prescription of aspirin for ischaemic stroke. The MRC Heart Protection Study was followed by increased statin prescription. However, increasing beds allocated to stroke had no influence on the proportion of patients receiving stroke unit care. Some unexpected signals of special cause variation could be plausibly explained (eg, breakdown of brain scanner), but others could not. Anecdotal evidence from healthcare professionals suggests that charts may be acceptable in clinical practice. CONCLUSION: SPC charts have the potential to provide valuable insights into the impact of changes in structure of services and of clinical evidence on the process of stroke care. In the present study, the charts were generally well received by healthcare professionals. PMID- 18678731 TI - "Every error counts": a web-based incident reporting and learning system for general practice. AB - BACKGROUND: Incident reporting systems have been established primarily in the inpatient setting. Their goal is the identification of safety risks in healthcare as a precondition for improvements in the overall quality of care. Knowledge about medical errors in general practice is sparse, as are reporting systems for patient safety in this setting. This article describes the development, structure and initial results of an incident reporting system for general practices in German-speaking countries. METHODS: Jeder Fehler Zaehlt (JFZ; www.jeder-fehler zaehlt.de) is a web-based reporting system that receives incident reports from anonymous German-speaking users. Reports are fed into a database, classified and analysed by a team of experts. Exemplary reports are published on the internet and in journals, and the reporting system provides the opportunity for users to comment on and learn from these incidents. RESULTS: The incident reporting system received 199 reports in the 17 months following its launch, of which 188 were classifiable. Of these, 72.9% were classified as process errors and 26.1% as knowledge/skills errors. The most frequently reported process errors were treatment errors (32.2%), communication errors (12.6%) and investigation errors (8.5%). Of the classified errors, 41.5% were associated with harm to the patient. More than 300 comments were made, mostly by anonymous users of the system. CONCLUSIONS: JFZ is a well-functioning and growing incident reporting system. Future efforts to improve the benefits of incident reporting will concentrate on increasing the utilisation of the system and broadening the spectrum of reported incidents. PMID- 18678732 TI - Ticks produce highly selective chemokine binding proteins with antiinflammatory activity. AB - Bloodsucking parasites such as ticks have evolved a wide variety of immunomodulatory proteins that are secreted in their saliva, allowing them to feed for long periods of time without being detected by the host immune system. One possible strategy used by ticks to evade the host immune response is to produce proteins that selectively bind and neutralize the chemokines that normally recruit cells of the innate immune system that protect the host from parasites. We have identified distinct cDNAs encoding novel chemokine binding proteins (CHPBs), which we have termed Evasins, using an expression cloning approach. These CHBPs have unusually stringent chemokine selectivity, differentiating them from broader spectrum viral CHBPs. Evasin-1 binds to CCL3, CCL4, and CCL18; Evasin-3 binds to CXCL8 and CXCL1; and Evasin-4 binds to CCL5 and CCL11. We report the characterization of Evasin-1 and -3, which are unrelated in primary sequence and tertiary structure, and reveal novel folds. Administration of recombinant Evasin-1 and -3 in animal models of disease demonstrates that they have potent antiinflammatory properties. These novel CHBPs designed by nature are even smaller than the recently described single-domain antibodies (Hollinger, P., and P.J. Hudson. 2005. Nat. Biotechnol. 23:1126-1136), and may be therapeutically useful as novel antiinflammatory agents in the future. PMID- 18678734 TI - Facundo Batista: watching B cells spread and grab antigens. Interview by Hema Bashyam. PMID- 18678733 TI - AID expression levels determine the extent of cMyc oncogenic translocations and the incidence of B cell tumor development. AB - Immunoglobulin (Ig) isotype switching is a recombination event that changes the constant domain of antibody genes and is catalyzed by activation-induced cytidine deaminase (AID). Upon recruitment to Ig genes, AID deaminates cytidines at switch (S) recombination sites, leading to the formation of DNA breaks. In addition to their role in isotype switching, AID-induced lesions promote Igh-cMyc chromosomal translocations and tumor development. However, cMyc translocations are also present in lymphocytes from healthy humans and mice, and thus, it remains unclear whether AID directly contributes to the dynamics of B cell transformation. Using a plasmacytoma mouse model, we show that AID(+/-) mice have reduced AID expression levels and display haploinsufficiency both in the context of isotype switching and plasmacytomagenesis. At the Ig loci, AID(+/-) lymphocytes show impaired intra- and inter-switch recombination, and a substantial decrease in the frequency of S mutations and chromosomal breaks. In AID(+/-) mice, these defects correlate with a marked decrease in the accumulation of B cell clones carrying Igh-cMyc translocations during tumor latency. These results thus provide a causality link between the extent of AID enzymatic activity, the number of emerging Igh-cMyc-translocated cells, and the incidence of B cell transformation. PMID- 18678735 TI - Art therapy with serious juvenile offenders: a phenomenological analysis. AB - Forty-six seriously delinquent, incarcerated boys received individual and group therapy for 32 months. The study examined how art therapy addressed the boys' psychological needs via analysis of the boys' self-selected art productions. In descending order of frequency, the eight most frequent need themes were identity issues; need for security and tranquility; need for freedom, adventure, and fun; need for ideal parental relationships; need for affiliation and affection; erotic and sexual needs; expression of depression, childhood trauma, and other psychological problems; and religious or spiritual needs. The boys' perceptions of what was most helpful about art therapy in descending order were stress relief and relaxation, reduction of boredom, pride and self-confidence, positive recognition, working through frustration, enjoyment and fun, improvement of ability to concentrate, and the way they were treated. Three brief case histories and a description of the art therapy procedures are given. Possible implications for cognitive restructuring are discussed. PMID- 18678736 TI - Training adult protective services workers about domestic violence: training needs and strategies. AB - Many cases of elder abuse involve instances of partner abuse. In this study, attention is given to whether adult protective services supervisors think their workers know enough about specific aspects of domestic violence. Attention is also given to the strategies that the supervisors think would be most useful to teach about domestic violence. According to the supervisors, workers need to know more about intervening with perpetrators, planning for their own safety, and dealing with frustrations. The supervisors rated the importance of understanding theoretical perspectives as low. Results also show that agencies with domestic violence training policies participate in domestic violence training programs more often. PMID- 18678737 TI - Membrane trafficking: intracellular highways and country roads. PMID- 18678738 TI - Nanoscale architecture of endoplasmic reticulum export sites and of Golgi membranes as determined by electron tomography. PMID- 18678739 TI - Advances in fluorescent protein-based imaging for the analysis of plant endomembranes. PMID- 18678740 TI - The endosomal system of plants: charting new and familiar territories. PMID- 18678741 TI - What is moving in the secretory pathway of plants? PMID- 18678742 TI - SNAREs: cogs and coordinators in signaling and development. PMID- 18678743 TI - The regulatory RAB and ARF GTPases for vesicular trafficking. PMID- 18678744 TI - Regulation of membrane trafficking, cytoskeleton dynamics, and cell polarity by ROP/RAC GTPases. PMID- 18678745 TI - Vesicle trafficking during somatic cytokinesis. PMID- 18678746 TI - PIN polar targeting. PMID- 18678747 TI - Plasma membrane receptor complexes. PMID- 18678748 TI - Plant receptors go endosomal: a moving view on signal transduction. PMID- 18678749 TI - Secretory pathways in plant immune responses. PMID- 18678750 TI - A proteomics approach to membrane trafficking. PMID- 18678751 TI - Single-stranded DNA-binding protein Whirly1 in barley leaves is located in plastids and the nucleus of the same cell. PMID- 18678752 TI - Factors affecting the incidence of genotoxicity biomarkers in peripheral blood lymphocytes: impact on design of biomonitoring studies. AB - A review of risk factors affecting background rates of micronuclei and chromosomal aberration (CA) formation in peripheral blood lymphocytes (PBLs) was undertaken with a view to aiding the interpretation of genotoxicity biomonitoring studies. Both endogenous factors and those due to methodological variation were evaluated. Background variation of other indices of genotoxicity in PBLs (specifically 8-hydroxy-deoxyguanosine and comet assays) were also considered as these data likely reflect overlapping causes of DNA damage and may provide some indicators for future research areas. A number of host risk factors, namely age, gender, smoking, vitamin B(12) and folate status, were identified for which there is strong or sufficient evidence that they impact on background levels of genotoxicity biomarkers. Evaluation of these factors should be routinely included in genotoxicity biomonitoring studies. Although data on the influence of smoking is somewhat inconsistent, because of its known association with cancer and DNA damage, it is also classified as a high-risk factor. A number of other factors were identified for which there is weak or insufficient evidence including alcohol consumption, disease conditions and infections, physical exercise, body mass index and genotype. The review shows that the evaluation of biomonitoring studies of genotoxicity is complex and there is a need to improve study designs by setting an a priori hypothesis, collecting good exposure data and stratifying groups appropriately, using appropriate power calculations before initiating biomonitoring studies, and collecting information on appropriate risk factors. There is a need for further collaborative work and the establishment of centres of excellence on genotoxicity biomonitoring. If these measures are achieved, then it would be possible to use the data from biomonitoring studies in risk assessments to derive risk management measures. PMID- 18678753 TI - Clues about the genetic basis of adaptation emerge from comparing the proteomes of two Ostreococcus ecotypes (Chlorophyta, Prasinophyceae). AB - We compared the proteomes of two picoplanktonic Ostreococcus unicellular green algal ecotypes to analyze the genetic basis of their adaptation with their ecological niches. We first investigated the function of the species-specific genes using Gene Ontology databases and similarity searches. Although most species-specific genes had no known function, we identified several species specific functions involved in various cellular processes, which could be critical for environmental adaptations. Additionally, we investigated the rate of evolution of orthologous genes and its distribution across chromosomes. We show that faster evolving genes encode significantly more membrane or excreted proteins, consistent with the notion that selection acts on cell surface modifications that is driven by selection for resistance to viruses and grazers, keystone actors of phytoplankton evolution. The relationship between GC content and chromosome length also suggests that both strains have experienced recombination since their divergence and that lack of recombination on the two outlier chromosomes could explain part of their peculiar genomic features, including higher rates of evolution. PMID- 18678754 TI - Hormone- and light-regulated nucleocytoplasmic transport in plants: current status. AB - The gene regulation mechanisms underlying hormone- and light-induced signal transduction in plants rely not only on post-translational modification and protein degradation, but also on selective inclusion and exclusion of proteins from the nucleus. For example, plant cells treated with light or hormones actively transport many signalling regulatory proteins, transcription factors, and even photoreceptors and hormone receptors into the nucleus, while actively excluding other proteins. The nuclear envelope (NE) is the physical and functional barrier that mediates this selective partitioning, and nuclear transport regulators transduce hormone- or light-initiated signalling pathways across the membrane to mediate nuclear activities. Recent reports revealed that mutating the proteins regulating nuclear transport through the pores, such as nucleoporins, alters the plant's response to a stimulus. In this review, recent works are introduced that have revealed the importance of regulated nucleocytoplasmic partitioning. These important findings deepen our understanding about how co-ordinated plant hormone and light signal transduction pathways facilitate communication between the cytoplasm and the nucleus. The roles of nucleoporin components within the nuclear pore complex (NPC) are also emphasized, as well as nuclear transport cargo, such as Ran/TC4 and its binding proteins (RanBPs), in this process. Recent findings concerning these proteins may provide a possible direction by which to characterize the regulatory potential of hormone or light-triggered nuclear transport. PMID- 18678755 TI - A possible association between early apical resorption of primary teeth and ectodermal characteristics of the permanent dentition. AB - The hypothesis of this study is that children with unexpected early apical resorption of the primary teeth are also predisposed to resorption in the permanent dentition. Accordingly, the aim was to perform a longitudinal study focussing on the permanent teeth in children with unexpected early apical resorption in the primary dentition. Panoramic radiographs of 12 children (7 boys and 5 girls) aged 6 years 4 months to 8 years 9 months with unexpected early apical resorption of primary teeth were identified from a dental archive of 588 patients. After written request, follow-up radiographs were obtained (2-15 year interval between early and follow-up radiographs). The radiographs were examined in order to verify the abnormal resorption pattern of the primary teeth and dental deviations in the permanent teeth, known to predispose for root resorption (i.e. invaginations, narrow crowns, abrupt root deflections, slender roots, short roots, taurodontia, agenesis, deviant pattern of eruption). Primary dentition: Two phenotypically different resorption groups were identified: group I, eight patients (resorption of the roots only), and group II, four patients (resorption of root and crown). Permanent dentition: In all 12 children, dental deviations in the permanent dentition were observed. Additionally, idiopathic external apical resorption of the permanent teeth was seen in three children, two of whom had received orthodontic treatment. PMID- 18678756 TI - Stability of hard tissue profile after mandibular setback in sagittal split osteotomies: a longitudinal and long-term follow-up study. AB - The aim of the study was to conduct a long-term follow-up on the stability of the hard tissues after bilateral sagittal split osteotomy (BSSO) with rigid internal fixation (RIF)to set back the mandible and to compare it with that of mandibular advancement performed by the same team of surgeons and with the same examination protocol. Seventeen consecutive patients (6 females and 11 males) could be re examined 12.7 years (T5) after surgery. The previous examinations were before surgery (T1), 5 days (T2), and 6.6 (T3) and 14.4 (T4) months after surgery. Lateral cephalograms were traced by hand, digitized, and evaluated with the Dentofacial Planner software program. The x-axis for the system of co-ordinates ran through sella (point zero) and the line nasion-sella-line minus 7 degrees. The program determined the x- and y-values of each variable and the usual angles and distances. The effects of treatment were determined with Wilcoxon matched pairs, signed ranks test, with Bonferroni adjustment, and the relationship between variables with Spearman rank correlation coefficient. Relapse at point B was 0.94 mm or 15 per cent and at pogonion 1.46 mm or 21 per cent of the initial setback at T5. Relapse was mainly short-term (T4-T2), 13 per cent for point B and 17 per cent for pogonion. Gender correlated significantly with relapse (T5-T2) at point B (P = 0.002) and pogonion (P = 0.021), i.e. females in contrast to males showed further distalization of the mandible instead of relapse. No correlations were seen for age or the amount of surgical setback. The long-term results in mandibular setback patients were more stable when compared with the mandibular advancement patients examined previously. The initial soft tissue profile, the initial growth direction, and the remodelling processes of the hard tissues must be considered as reasons for long-term relapse. Growth direction positively influenced the long-term results in females: further distalization of the mandible occurred. PMID- 18678757 TI - A method for defining targets in contemporary incisor inclination correction. AB - Different craniofacial properties require individual targets in incisor inclination. These requirements are mostly scheduled on the basis of cephalometric diagnosis, but, however, performed using straightwire appliances, which refer to third-order angles and not to cephalometric data. The objective of this study was to analyze the relationship between incisor third-order angles, incisor inclination, and skeletal craniofacial findings in untreated ideal occlusion subjects with natural dentoalveolar compensation of skeletal variation, in order to link the field of cephalometric assessment of incisor inclination with that of contemporary orthodontic incisor inclination correction. This study utilized lateral cephalograms and corresponding dental casts of 69 untreated Caucasians (21 males and 48 females between 12 and 35 years of age) with neutral (Angle Class I) molar and canine relationships and an incisor relationship that was sagittaly and vertically considered as ideal by three orthodontists (i.e. well supported by the antagonistic teeth and without the need for either deep or open bite correction). Upper (U1) and lower (L1) axial incisor inclinations were assessed with reference to the cephalometric lines NA and NL, and NB and ML, respectively. Sagittal and vertical skeletal relationships were classified using SNA (SNB) and NSL-ML (NSL-NL) angles. Third-order angles (U1TA and L1TA) were derived from direct dental cast measurements using an incisor inclination recording appliance. The relationships between cephalometric and third-order measurements evaluated by calculating Pearson product-moment correlation coefficients (a = 0.05) showed strong correlations between cephalometric axial inclination data (U1NA/deg, L1NB/deg, U1NA/mm, L1NB/mm, U1NL, and L1ML) and sagittal-skeletal data, but no significant relationship between skeletal-vertical findings and incisor inclination. The mean U1TA was 4.9 (standard deviation [SD] 5.85) and the mean L1TA -3.0 (SD 6.9) degrees. Regression analyses were used for axial inclination (ANB angle designated as the independent variable) and for third-order data (U1NA, L1NB, U1NL, and L1ML designated as independent variables). Based on the correlations found in this study, a novel method for defining targets in upper and lower incisor third-order correction according to natural standards is presented. As a consequence, third-order movements can be adapted to cephalometric diagnosis with enhanced accuracy. PMID- 18678758 TI - Factors affecting the duration of orthodontic treatment: a systematic review. AB - The aim of this study was to systematically investigate the literature for articles referring exclusively to the duration of orthodontic therapy and to explore the various factors that could affect this. A Medline search from 1990 to the first week of March 2005 was conducted, the Cochrane Database for Systematic Reviews was utilized, five orthodontic journals were hand searched, the abstracts of related articles were reviewed to search for any relevant studies that might have been missed, and the reference lists of the retrieved articles were hand assessed. Eligibility was determined by reading the reports identified by the search. The end result of this search provided 41 articles. Although there is a need for more conclusive research, the present review revealed several conclusions concerning the duration of orthodontic treatment: (1) there are indications that extraction treatment lasts longer than the non-extraction therapy; (2) age does not seem to play a role provided the patients are in the permanent dentition; (3) when Class II division 1 malocclusions are considered, there is evidence that the earlier the orthodontic treatment begins the longer its duration; (4) there is conflicting information regarding treatment duration within public health systems; (5) combined orthodontic-surgical treatment duration is variable and appears to be operator sensitive; (6) various factors, such as the technique employed, the skill and number of operators involved, the compliance of the patients, and the severity of the initial malocclusion, all seem to play a role; and (7) impacted maxillary canines appear to prolong treatment. PMID- 18678759 TI - The effects of a newly designed twin-slot bracket on severely malpositioned teeth -a typodont experimental study. AB - The aims of this study were to design a twin-slot bracket featuring two horizontal slots and to examine its efficiency in tooth displacement. Based on the structure of a traditional edgewise bracket, an additional slot was added to a twin-slot bracket and the prototype products were fabricated for the typodont experiments. The orthodontic correction of malpositioned canines was conducted on a typodont to examine the efficiency of the twin-slot bracket in tooth displacement compared with a single-slot edgewise bracket. Three modalities of tooth movement requiring a heavy force moment, namely, axial rotation, mesiodistal tipping, and bodily translation, were conducted. The canine positions before and after simulation were measured and the changes identified. Statistical analysis was undertaken using a t-test to determine the significance of the differences in canine repositioning between the two bracket types. The results showed that in the twin-slot bracket group, the treatment changes in the canine position by derotation and uprighting were 40 +/- 3 and 25 +/- 2 degrees, respectively, compared with 20 +/- 5 and 10 +/- 2 degrees in the edgewise group (P < 0.01). When retracted into an extraction space with an initial 10 degrees of mesial tip, the mesiodistal angulation of the canines in the twin-slot bracket group remained unchanged while in the edgewise group the canines became distally tipped by 5 +/- 2 degrees (P < 0.01). The twin-slot bracket significantly increased the bracket width without reducing the interbracket span and therefore can generate increased force moments within the bracket, leading to an improved manipulation in tooth repositioning. PMID- 18678760 TI - Influence of different tooth types on the bond strength of two orthodontic adhesive systems. AB - The aim of this investigation was to evaluate the effects of different tooth types on the shear bond strength (SBS) of two orthodontic resin adhesive systems in vitro. Two hundred extracted sound human teeth were used in the study. Ten teeth of each tooth type were the mounted in acrylic resin leaving the buccal surface of the crowns parallel to the base of the moulds. In each experimental group, the adhesives (Transbond XT and Light Bond) were applied to the etched enamel surfaces. The orthodontic composite resins were then applied to the surface in cylindrical-shaped plastic matrices. For SBS testing, a force transducer (Ultradent) was applied at a crosshead speed of 1 mm/minute at the interface between the tooth and composite until failure occurred. Data were analysed using two-way analysis of variance (ANOVA), Kruskal-Wallis one-way ANOVA, a Bonferroni adjusted Mann-Whitney U-test, and an independent t-test. Generally, it was found that tooth type had a significant effect on SBS (P < 0.05) with Light Bond showing a higher SBS than Transbond XT (P < 0.05). The highest bond strengths were observed for the upper central incisor and lower molars with Light Bond (P < 0.05) and the lowest mean bond strengths for the upper molars and lower canine with Transbond XT (P <0.05). The results demonstrated that enamel SBS was significantly altered by both tooth type and adhesive system. Thus, the findings of this study confirm that enamel bond strength is not uniform for all teeth. These results may also explain the variability in the enamel-bonding efficacy of adhesives. PMID- 18678761 TI - The effects of cervical headgear with an expanded inner bow in the permanent dentition. AB - In this study, the effects of cervical headgear (CHG) use on the transverse dimension of the maxillary dental arch were evaluated in patients in the permanent dentition. Thirteen girls and 12 boys (mean age: 13.41 +/- 0.52 years) with a bilateral full cusp Class II molar relationship comprised the study group. Fifteen girls and 10 boys with a Class I normal occlusion comprised the controls. In the treatment group, CHG with an expanded inner bow was used for a mean period of 11.2 +/- 5.6 months. The headgear was used for molar distalization and the force magnitude was 196.1 cN. After CHG treatment, the patients underwent non extraction fixed orthodontic treatment for 14.1 +/- 2.5 months. During this period, the control group received regular dental check-ups. Dental casts obtained at the beginning (T1) and end (T2) of headgear use and at the end of orthodontic treatment (T3) and posteroanterior cephalograms taken at T1 and T2 were evaluated. A Student's t-test was used for intergroup comparison at T1, T2, and T3 and a Mann-Whitney U-test with a Bonferroni correction for comparison of treatment/observation changes. At T2, intercanine (0.96 +/- 0.56 mm), interpremolar (1.6 +/- 0.55 mm for the first premolar, 1.74 +/- 0.65 mm for the second premolar), and intermolar (2.31 +/- 0.75 mm) widths increased, while the distance between the intersection of the zygomatic process and the maxillary alveolar process on the right (JR) and left (JL) did not change. Fixed orthodontic treatment did not have any effect on any of the measurements. With the intentional expansion of the inner bow of CHG, the amount of maxillary dental arch expansion achieved in the permanent dentition was statistically significant (P < 0.017). PMID- 18678763 TI - Social support and symptom distress in adolescents/young adults with cancer. AB - The purpose of this secondary analysis was to describe the relationships of friend, family, and health care provider social support to pain, fatigue, depressed mood, and insomnia in adolescents and young adults with cancer using the Adolescent Resilience Model. Specific aims of the study were to describe the relationships of 3 separate sources of perceived support-friends, family, and health care providers to the symptom-related distress of pain, fatigue, depressed mood, and insomnia. PMID- 18678765 TI - Conceptual knowledge is underpinned by the temporal pole bilaterally: convergent evidence from rTMS. AB - Conceptual knowledge provides the basis on which we bring meaning to our world. Studies of semantic dementia patients and some functional neuroimaging studies indicate that the anterior temporal lobes, bilaterally, are the core neural substrate for the formation of semantic representations. This hypothesis remains controversial, however, as traditional neurological models of comprehension do not posit a role for these regions. To adjudicate on this debate, we conducted 2 novel experiments that used off-line, low-frequency, repetitive transcranial magnetic stimulation to disrupt neural processing temporarily in the left or right temporal poles (TPs). The time required to make semantic decisions was slowed considerably, yet specifically, by this procedure. The results confirm that both TPs form a critical substrate within the neural network that supports conceptual knowledge. PMID- 18678764 TI - Neural correlates of speeded as compared with delayed responses in a stop signal task: an indirect analog of risk taking and association with an anxiety trait. AB - The stop signal task (SST) is widely used to explore neural processes involved in cognitive control. By randomly intermixing stop and go trials and imposing on participants to respond quickly to the go but not the stop signal, the SST also introduces an indirect element of risk, which participants may avert by slowing down or ignore by responding "as usual," during go trials. This "risk-taking" component of the SST has to our knowledge never been investigated. The current study took advantage of variability of go trial reaction time (RT) and compared the post-go go trials that showed a decrease in RT (risk-taking decision) and those post-go go trials that showed an increase in RT ("risk-aversive" decision) in 33 healthy individuals who underwent functional magnetic resonance imaging during the SST. This contrast revealed robust activation in bilateral visual cortices as well as left inferior parietal and posterior cingulate cortices, amygdala, and middle frontal gyrus (P < 0.05, family-wise error [FWE] corrected). Furthermore, we observed that the magnitude of amygdala activity is positively correlated with trait anxiety of the participants. These results thus delineated, for the first time, a neural analog of risk taking during stop signal performance, highlighting a novel aspect and broadening the utility of this behavioral paradigm. PMID- 18678766 TI - Lipid changes in breast cancer patients on exemestane treatment: final results of the TEAM Greek substudy. AB - BACKGROUND: The Greek substudy of the Tamoxifen and Exemestane Adjuvant Multicenter International trial compared the effect of exemestane on the lipid profile of postmenopausal, breast cancer patients to that of tamoxifen in the adjuvant setting. PATIENTS AND METHODS: Lipidemic profile changes were studied in 142 postmenopausal patients randomized to receive either adjuvant exemestane (n=77) or tamoxifen (n=65). Total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL) and serum triglyceride (TRG) levels were measured at baseline and then every 3 months for the first 12 months of treatment and at 18 and 24 months. RESULTS: A trend for a reduction in TC was found in both treatment arms; however, TC and LDL levels were consistently and significantly decreased in tamoxifen arm only. The mean HDL level was higher for the tamoxifen arm compared with the exemestane arm across time. No significant trend was detected throughout the study period on TRG levels on either arm. CONCLUSIONS: Unlike tamoxifen's beneficial effect on TC and LDL levels, exemestane appears to have a neutral effect on lipidemic profile of postmenopausal, breast cancer patients. These data offer additional information with regard to the safety and tolerability of exemestane treatment in the adjuvant setting. PMID- 18678767 TI - A systematic review of the scales used for the measurement of cancer-related fatigue (CRF). AB - BACKGROUND: Fatigue in cancer is very common and can be experienced at all stages of disease and in survivors. There is no accepted definition of cancer-related fatigue (CRF) and no agreement on how it should be measured. A number of scales have been developed to quantify the phenomenon of CRF. These vary in the quality of psychometric properties, ease of administration, dimensions of CRF covered and extent of use in studies of cancer patients. This review seeks to identify the available tools for measuring CRF and to make recommendations for ongoing research into CRF. METHODS: A systematic review methodology was used to identify scales that have been validated to measure CRF. The inclusion criteria required the scale to have been validated for use in cancer patients and/or widely used in this population. Scales also had to meet a minimum quality score for inclusion. RESULTS: The reviewers identified 14 scales that met the inclusion criteria. The most commonly used scales and best validated were the Functional Assessment of Cancer Therapy Fatigue (FACT F), the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ C30) (fatigue subscale) and the Fatigue Questionnaire (FQ). CONCLUSIONS: Unidimensional scales are the easiest to administer and have been most widely used. The authors recommend the use of the EORTC QLQ C30 fatigue subscale or the FACT F. The FQ gives a multidimensional assessment and has also been widely used. A substantial minority of the scales identified have not been used extensively or sufficiently validated in cancer patients and cannot be recommended for routine use without further validation. PMID- 18678768 TI - Physical activity and incidence of atrial fibrillation in older adults: the cardiovascular health study. AB - BACKGROUND: Vigorous exertion and endurance training have been reported to increase atrial fibrillation (AF). Associations of habitual light or moderate activity with AF incidence have not been evaluated. METHODS AND RESULTS: We prospectively investigated associations of leisure-time activity, exercise intensity, and walking habits, assessed at baseline and updated during follow-up visits, with incident AF, diagnosed by annual 12-lead ECGs and hospital discharge records, from 1989 to 2001 among 5446 adults > or =65 years of age in the Cardiovascular Health Study. During 47 280 person-years of follow-up, 1061 new AF cases occurred (incidence 22.4/1000 person-years). In multivariable-adjusted analyses, leisure-time activity was associated with lower AF incidence in a graded manner, with 25% (hazard ratio [HR] 0.75, 95% confidence interval [CI] 0.61 to 0.90), 22% (HR 0.78, 95% CI 0.65 to 0.95), and 36% (HR 0.64, 95% CI 0.52 to 0.79) lower risk in quintiles 3, 4, and 5 versus quintile 1 (P for trend <0.001). Exercise intensity had a U-shaped relationship with AF (quadratic P=0.02): Versus no exercise, AF incidence was lower with moderate-intensity exercise (HR 0.72, 95% CI 0.58 to 0.89) but not with high-intensity exercise (HR 0.87, 95% CI 0.64 to 1.19). Walking distance and pace were each associated with lower AF risk in a graded manner (P for trend <0.001); when we assessed the combined effects of distance and pace, individuals in quartiles 2, 3, and 4 had 25% (HR 0.75, 95% CI 0.56 to 0.99), 32% (HR 0.68, 95% CI 0.50 to 0.92), and 44% (HR 0.56, 95% CI 0.38 to 0.82) lower AF incidence than individuals in quartile 1. Findings appeared unrelated to confounding by comorbidity or indication. After evaluation of cut points of moderate leisure-time activity (approximately 600 kcal/week), walking distance (12 blocks per week), and pace (2 mph), 26% of all new AF cases (95% CI 7% to 43%) appeared attributable to absence of these activities. CONCLUSIONS: Light to moderate physical activities, particularly leisure-time activity and walking, are associated with significantly lower AF incidence in older adults. PMID- 18678769 TI - Contemporary analysis of descending thoracic and thoracoabdominal aneurysm repair: a comparison of endovascular and open techniques. AB - BACKGROUND: Endovascular repair of thoracic aneurysm has demonstrated low risks of mortality and spinal cord ischemia (SCI), but few large series have been published on endovascular thoracoabdominal aneurysm repair, and reports suffer from a lack of accurate comparison with similar open surgical procedures. METHODS AND RESULTS: A consecutive cohort of patients with thoracic and thoracoabdominal aneurysms treated electively with endovascular repair (ER) or surgical repair (SR) techniques between 2001 and 2006 were analyzed. The association between repair technique and SCI was evaluated with univariable analysis. Adjustments for potential confounders and for the propensity to receive ER or SR were also performed in multivariable analysis. A total of 724 patients (352 ER, 372 SR) underwent repair. The mean age was 67 years, and 65% were male. ER patients were on average 9 years older (P<0.001), had more comorbid conditions, and more frequently had prior distal repair (P<0.001) or underwent a type I or IV repair. SR patients more commonly had chronic dissection or required type II or type III repairs (P<0.001). Mortality at 30 days (5.7% ER versus 8.3% SR, P=0.2) and 12 months (15.6% ER versus 15.9% SR, P=0.9) was similar. A borderline difference in SCI was found between repair techniques: 4.3% of ER and 7.5% of SR patients (P=0.08) had SCI. In patients with ER, prior distal aortic operation was associated with the development of SCI in univariable analysis (odds ratio 4.1, 95% confidence interval 1.4 to 11.7). Multivariable analysis showed that the type of required repair (type I, II, III, or IV) was the primary factor associated with the development of SCI in ER and SR patients. CONCLUSIONS: No significant difference in the incidence of mortality or SCI was found between ER and SR techniques. The strongest factor associated with SCI remains the extent of the disease. Further studies are indicated to compare ER with patients considered eligible for SR. PMID- 18678770 TI - Mitral leaflet adaptation to ventricular remodeling: occurrence and adequacy in patients with functional mitral regurgitation. AB - BACKGROUND: Functional mitral regurgitation (MR) is caused by systolic traction on the mitral leaflets related to ventricular distortion. Little is known about whether chronic tethering causes the mitral leaflet area to adapt to the geometric needs imposed by tethering, in part because of inability to reconstruct leaflet area in vivo. Our aim was to explore whether adaptive increases in leaflet area occur in patients with functional MR compared with normal subjects and to test the hypothesis that leaflet area influences MR severity. METHODS AND RESULTS: A new method for 3-dimensional echocardiographic measurement of mitral leaflet area was developed and validated in vivo against 15 sheep heart valves, later excised. This method was then applied in 80 consecutive patients from 3 groups: patients with normal hearts by echocardiography (n=20), patients with functional MR caused by isolated inferior wall-motion abnormality or dilated cardiomyopathy (n=29), and patients with inferior wall-motion abnormality or dilated cardiomyopathy but no MR (n=31). Leaflet area was increased by 35+/-20% in patients with LV dysfunction compared with normal subjects. The ratio of leaflet to annular area was 1.95+/-0.40 and was not different among groups, which indicates a surplus leaflet area that adapts to left-heart changes. In contrast, the ratio of total leaflet area to the area required to close the orifice in midsystole was decreased in patients with functional MR compared with those with normal hearts (1.29+/-0.15 versus 1.78+/-0.39, P=0.001) and compared with patients with inferior wall-motion abnormality or dilated cardiomyopathy but no MR (1.81+/-0.38, P=0.001). After adjustment for measures of LV remodeling and tethering, a leaflet-to-closure area ratio <1.7 was associated with significant MR (odds ratio 23.2, 95% confidence interval 2.0 to 49.1, P=0.02). CONCLUSIONS: Mitral leaflet area increases in response to chronic tethering in patients with inferior wall-motion abnormality and dilated cardiomyopathy, but the development of significant MR is associated with insufficient leaflet area relative to that demanded by tethering geometry. The varying adequacy of leaflet adaptation may explain in part the heterogeneity of this disease among patients. The results suggest the need to understand the mechanisms that underlie leaflet adaptation and whether leaflet area can potentially be modified as part of the therapeutic approach. PMID- 18678771 TI - Smoothelin-B deficiency results in reduced arterial contractility, hypertension, and cardiac hypertrophy in mice. AB - BACKGROUND: Smoothelins are actin-binding proteins that are abundantly expressed in healthy visceral (smoothelin-A) and vascular (smoothelin-B) smooth muscle. Their expression is strongly associated with the contractile phenotype of smooth muscle cells. Analysis of mice lacking both smoothelins (Smtn-A/B(-/-) mice) previously revealed a critical role for smoothelin-A in intestinal smooth muscle contraction. Here, we report on the generation and cardiovascular phenotype of mice lacking only smoothelin-B (Smtn-B(-/-)). METHODS AND RESULTS: Myograph studies revealed that the contractile capacity of the saphenous and femoral arteries was strongly reduced in Smtn-B(-/-) mice, regardless of the contractile agonist used to trigger contraction. Arteries from Smtn-A/B(-/-) compound mutant mice exhibited a similar contractile deficit. Smtn-B(-/-) arteries had a normal architecture and expressed normal levels of other smooth muscle cell-specific genes, including smooth muscle myosin heavy chain, alpha-smooth muscle actin, and smooth muscle-calponin. Decreased contractility of Smtn-B(-/-) arteries was paradoxically accompanied by increased mean arterial pressure (20 mm Hg) and concomitant cardiac hypertrophy despite normal parasympathetic and sympathetic tone in Smtn-B(-/-) mice. Magnetic resonance imaging experiments revealed that cardiac function was not changed, whereas distension of the proximal aorta during the cardiac cycle was increased in Smtn-B(-/-) mice. However, isobaric pulse wave velocity and pulse pressure measurements indicated normal aortic distensibility. CONCLUSIONS: Collectively, our results identify smoothelins as key determinants of arterial smooth muscle contractility and cardiovascular performance. Studies on mutations in the Smtn gene or alterations in smoothelin levels in connection to hypertension in humans are warranted. PMID- 18678772 TI - Cardiac magnetic resonance with T2-weighted imaging improves detection of patients with acute coronary syndrome in the emergency department. AB - BACKGROUND: Cardiac magnetic resonance (CMR) imaging permits early triage of patients presenting to the emergency department with acute chest pain but has been limited by the inability to differentiate new from old myocardial infarction. Our objective was to evaluate a CMR protocol that includes T2 weighted imaging and assessment of left ventricular wall thickness in detecting patients with acute coronary syndrome in the emergency department. METHODS AND RESULTS: In this prospective cohort observational study, we enrolled patients presenting to the emergency department with acute chest pain, negative cardiac biomarkers, and no ECG changes indicative of acute ischemia. The CMR protocol consisted of T2-weighted imaging, first-pass perfusion, cine function, delayed enhancement magnetic resonance imaging, and assessment of left ventricular wall thickness. The clinical outcome (acute coronary syndrome) was defined by review of clinical charts by a consensus panel that used American Heart Association/American College of Cardiology guidelines. Among 62 patients, 13 developed acute coronary syndrome during the index hospitalization. The mean CMR time was 32+/-8 minutes. The new CMR protocol (with the addition of T2-weighted and left ventricular wall thickness) increased the specificity, positive predictive value, and overall accuracy from 84% to 96%, 55% to 85%, and 84% to 93%, respectively, compared with the conventional CMR protocol (cine, perfusion, and delayed-enhancement magnetic resonance imaging). Moreover, in a logistic regression analysis that contained information on clinical risk assessment (c statistic=0.695) and traditional cardiac risk factors (c-statistic=0.771), the new CMR protocol significantly improved the c-statistic to 0.958 (P<0.0001). CONCLUSIONS: The present study indicates that a new CMR protocol improves the detection of patients with acute coronary syndrome in the emergency department and adds significant value over clinical assessment and traditional cardiac risk factors. PMID- 18678773 TI - Electroanatomic mapping of the left ventricle in a porcine model of chronic myocardial infarction with magnetic resonance-based catheter tracking. AB - BACKGROUND: X-ray fluoroscopy constitutes the fundamental imaging modality for catheter visualization during interventional electrophysiology procedures. The minimal tissue discriminative capability of fluoroscopy is mitigated in part by the use of electroanatomic mapping systems and enhanced by the integration of preacquired 3-dimensional imaging of the heart with computed tomographic or magnetic resonance (MR) imaging. A more ideal paradigm might be to use intraprocedural MR imaging to directly image and guide catheter mapping procedures. METHODS AND RESULTS: An MR imaging-based electroanatomic mapping system was designed to assess the feasibility of navigating catheters to the left ventricle in vivo using MR tracking of microcoils incorporated into the catheters, measuring intracardiac ventricular electrograms, and integrating this information with 3-dimensional MR angiography and myocardial delayed enhancement images to allow ventricular substrate mapping. In all animals (4 normal, and 10 chronically infarcted swine), after transseptal puncture under fluoroscopic guidance, catheters were successfully navigated to the left ventricle with MR tracking (13 to 15 frames per second) by both transseptal and retrograde aortic approaches. Electrogram artifacts related to the MR imaging gradient pulses were successfully removed with analog and digital signal processing. In all animals, it was possible to map the entire left ventricle and to project electrogram voltage amplitude maps to identify the scarred myocardium. CONCLUSIONS: It is possible to use MR tracking to navigate catheters to the left ventricle, to measure electrogram activity, and to render accurate 3-dimensional voltage maps in a porcine model of chronic myocardial infarction, completely in the MR imaging environment. Myocardial delayed enhancement guidance provided dense sampling of the proximity of the infarct and accurate localization of complex infarcts. PMID- 18678774 TI - Improvement in left ventricular remodeling by the endothelial nitric oxide synthase enhancer AVE9488 after experimental myocardial infarction. AB - BACKGROUND: Reduced endothelial nitric oxide (NO) bioavailability contributes to the progression of heart failure. In this study, we investigated whether the transcription enhancer of endothelial NO synthase (eNOS) AVE9488 improves cardiac remodeling and heart failure after experimental myocardial infarction (MI). METHODS AND RESULTS: Starting 7 days after coronary artery ligation, rats with MI were treated with placebo or AVE9488 (25 ppm) as a dietary supplement for 9 weeks. AVE9488 therapy versus placebo substantially improved left ventricular (LV) function, reduced LV filling pressure, and prevented the rightward shift of the pressure-volume curve. AVE9488 also attenuated the extent of pulmonary edema, reduced LV fibrosis and myocyte cross-sectional area, and prevented the increases in LV gene expression of atrial natriuretic factor, brain natriuretic peptide, and endothelin-1. eNOS protein levels and calcium-dependent NOS activity were decreased in the surviving LV myocardium from placebo MI rats and normalized by AVE9488. The beneficial effects of AVE9488 on LV dysfunction and remodeling after MI were abrogated in eNOS-deficient mice. Aortic eNOS protein expression and endothelium-dependent NO-mediated vasorelaxation were significantly enhanced by AVE9488 treatment after infarction, whereas increased vascular superoxide anion formation was reduced. Moreover, AVE9488 prevented the marked depression of circulating endothelial progenitor cell levels in rats with heart failure after MI. CONCLUSIONS: Long-term treatment with the eNOS enhancer AVE9488 improved LV remodeling and contractile dysfunction after MI. Molecular alterations, circulating endothelial progenitor cell levels, and endothelial vasomotor dysfunction were improved by AVE9488. Pharmacological interventions designed to increase eNOS-derived NO constitute a promising therapeutic approach for the amelioration of postinfarction ventricular remodeling and heart failure. PMID- 18678775 TI - American heart association newly elected fellows, spring 2008. PMID- 18678776 TI - Images in cardiovascular medicine. Catastrophic thrombus formation during optical coherence tomography. PMID- 18678777 TI - Images in cardiovascular medicine. Bilateral vertical veins from a common confluence in supracardiac total anomalous pulmonary venous connection. PMID- 18678778 TI - Letter by anderson et al regarding article, "abnormal conduction and morphology in the atrioventricular node of mice with atrioventricular canal-targeted deletion of Alk3/Bmpr1a receptor". PMID- 18678779 TI - Letter by Gourdie and Sedmera regarding article, "abnormal conduction and morphology in the atrioventricular node of mice with atrioventricular canal targeted deletion of Alk3/Bmpr1a receptor". PMID- 18678781 TI - What do you need to know before performing a percutaneous coronary intervention? PMID- 18678782 TI - Corticosteroids for recurrent pericarditis: on the road to evidence-based medicine. PMID- 18678783 TI - Evidence mandating earlier and more aggressive treatment of hypercholesterolemia. PMID- 18678784 TI - Periprocedural stroke and cardiac catheterization. PMID- 18678785 TI - Images in cardiovascular medicine. Rupture of infected splenic artery aneurysm secondary to infective endocarditis. PMID- 18678786 TI - Ambulatory blood pressure monitoring in children and adolescents: recommendations for standard assessment: a scientific statement from the American Heart Association Atherosclerosis, Hypertension, and Obesity in Youth Committee of the council on cardiovascular disease in the young and the council for high blood pressure research. PMID- 18678787 TI - Roles of phosphatidylinositol 3-kinase-Akt and NADPH oxidase in adenosine 5' triphosphate-sensitive K+ channel function impaired by high glucose in the human artery. AB - The present study was designed to examine roles of the phosphatidylinositol 3 kinase-Akt pathway and reduced nicotinamide-adenine dinucleotide phosphate oxidases in the reduced ATP-sensitive K(+) channel function via superoxide produced by high glucose in the human artery. We evaluated the activity of the phosphatidylinositol 3-kinase-Akt pathway, as well as reduced nicotinamide adenine dinucleotide phosphate oxidases, the intracellular levels of superoxide and ATP-sensitive K(+) channel function in the human omental artery without endothelium. Levels of the p85-alpha subunit and reduced nicotinamide-adenine dinucleotide phosphate oxidase subunits, including p47phox, p22phox, and Rac-1, increased in the membrane fraction from arteries treated with D-glucose (20 mmol/L) accompanied by increased intracellular superoxide production. High glucose simultaneously augmented Akt phosphorylation at Ser 473, as well as Thr 308 in the human vascular smooth muscle cells. A phosphatidylinositol 3-kinase inhibitor LY294002, as well as tiron and apocynin, restored vasorelaxation and hyperpolarization in response to an ATP-sensitive K(+) channel opener levcromakalim. Therefore, it can be concluded that the activation of the phosphatidylinositol 3-kinase-Akt pathway, in combination with the translocation of p47phox, p22phox, and Rac-1, contributes to the superoxide production induced by high glucose, resulting in the impairment of ATP-sensitive K(+) channel function in the human visceral artery. PMID- 18678788 TI - Role of endothelin-1 in clinical hypertension: 20 years on. PMID- 18678789 TI - Transtelephonic home blood pressure to assess the monoamine oxidase-B inhibitor rasagiline in Parkinson disease. AB - Monoamine oxidase inhibitors are associated with dietary tyramine interactions that can induce hypertensive crises. Rasagiline mesylate is a novel irreversible selective monoamine oxidase type B inhibitor for Parkinson disease that may have a low risk of interaction with dietary tyramine because of its selectivity. To study interactions of rasagiline with diets unrestricted in tyramine-containing foods, we incorporated transtelephonic, self-monitoring of the blood pressure (BP) into a randomized, placebo-controlled trial of rasagiline 0.5 and 1.0 mg daily in 414 levodopa-treated Parkinson patients with motor fluctuations. The proportion of patients with a systolic BP increase of >30 mm Hg was the primary BP end point. In 13 968 self-measured readings at baseline, the proportion of systolic BP values that increased by >30 mm Hg after a meal ranged from 9.5% to 12.9% in the 3 treatment groups. In 25 733 BPs obtained postrandomization, the proportion of values with a >30-mm Hg systolic postprandial increase was 15% in the placebo group, 15% in the rasagiline 0.5-mg group, and 11% in the rasagiline 1-mg group after 3 weeks of double-blind therapy and 13%, 14%, and 12%, respectively, after 26 weeks of treatment (P value was not significant for all of the comparisons among treatment groups). A postprandial increase in systolic BP to >180 mm Hg at any time after randomization was seen in 3.3%, 2.6%, and 2.9% of the placebo, 0.5-mg, and 1.0-mg rasagiline groups, respectively. These data demonstrate that rasagiline did not induce postprandial hypertension in patients with Parkinson disease who were on an unrestricted diet. PMID- 18678790 TI - Olmesartan prevents cardiovascular injury and hepatic steatosis in obesity and diabetes, accompanied by apoptosis signal regulating kinase-1 inhibition. AB - Dietary obesity is associated with type 2 diabetes and cardiovascular diseases, although the underlying mechanism is unknown. This study was undertaken to elucidate the role of angiotensin II and apoptosis signal regulating kinase-1 (ASK1) in obesity/diabetes-associated cardiovascular complications and hepatic steatosis. Mice fed a high-fat diet were treated with olmesartan, an angiotensin II type 1 receptor blocker, to elucidate the role of angiotensin II in diabetic mice. Treatment of mice fed a high-fat diet with olmesartan markedly suppressed cardiac inflammation and fibrosis, as well as vascular endothelial dysfunction and remodeling, induced by obesity/diabetes. Moreover, olmesartan suppressed the disruption of the vascular endothelial NO synthase dimer in diabetic mice. Olmesartan also significantly prevented hepatic steatosis and fibrosis in diabetic mice. These beneficial effects of olmesartan on diabetic mice were associated with the attenuation of ASK1 activation in these mice. ASK1-deficient mice and wild-type mice were compared, regarding the effects of a high-fat diet. A comparison between ASK1-deficient and wild-type mice showed that ASK1 deficiency attenuated cardiac inflammation and fibrosis, as well as vascular endothelial dysfunction and remodeling induced by obesity/diabetes. The amelioration of vascular endothelial impairment by ASK1 deficiency was attributed to the prevention of endothelial NO synthase dimer disruption. ASK1 deficiency also significantly lessened hepatic steatosis in diabetic mice. In conclusion, our work provided the evidence that ASK1 is significantly activated in diet induced diabetic mice and contributes to cardiovascular diseases and hepatic steatosis in diabetic mice. Moreover, the beneficial effects of angiotensin II inhibition on dietary diabetic mice seem to be mediated by the inhibition of ASK1 activation. PMID- 18678791 TI - Edgar Allan Poe: The raven. PMID- 18678792 TI - Selecting among second-step antidepressant medication monotherapies: predictive value of clinical, demographic, or first-step treatment features. AB - CONTEXT: Little is known about selecting among second-step medications for major depressive disorder after intolerance or lack of remission with an initial selective serotonin reuptake inhibitor. OBJECTIVE: To determine whether sociodemographic, clinical, or first-step treatment features predict remission with or intolerance overall or differentially to any 1 of 3 second-step medications after an unsatisfactory outcome with citalopram hydrobromide. DESIGN: An equipoise stratified randomized study. Participants were recruited from July 17, 2001, through April 20, 2004. SETTING: Public or private sector primary care (n = 18) and psychiatric care (n = 23) settings across the United States. PARTICIPANTS: Representative outpatients aged 18 to 75 years with nonpsychotic major depressive disorder (N = 727). INTERVENTIONS: Sustained-release bupropion hydrochloride was started at 150 mg/d and incrementally increased to 400 mg/d. Sertraline hydrochloride was started at 50 mg/d and incrementally increased to 200 mg/d. Extended-release venlafaxine hydrochloride was started at 37.5 mg/d and incrementally increased to 375 mg/d. MAIN OUTCOME MEASURES: The 16-item Quick Inventory of Depressive Symptomatology, Self-Rated and the Frequency, Intensity, and Burden of Side Effects Rating. RESULTS: Remission was more likely among participants who were white, employed, cohabiting or married, or privately insured or who had prior intolerance to citalopram or at least a response to citalopram, and no prior suicide attempts. Remission was less likely among participants with concurrent generalized anxiety, obsessive-compulsive, panic, or posttraumatic stress disorders; social phobia; anxious or melancholic features; or more severe depression. Intolerance was less likely for Hispanic participants, but more likely for participants with previous suicide attempts or intolerance to citalopram. Participants with concurrent substance use were less likely to remit (odds ratio, 0.37) and more likely not to tolerate extended-release venlafaxine. Intolerance to citalopram was associated with intolerance to sertraline (P = .04). CONCLUSIONS: Clinical, demographic, and treatment history were of little value in recommending 1 medication vs another as a second-step treatment for major depressive disorder. Participants most likely to remit in the second step had less Axis I psychiatric disorder comorbidity, less social disadvantage, and at least a response to citalopram in the first step. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00021528. PMID- 18678794 TI - Testing causality in the association between regular exercise and symptoms of anxiety and depression. AB - CONTEXT: In the population at large, regular exercise is associated with reduced anxious and depressive symptoms. Results of experimental studies in clinical populations suggest a causal effect of exercise on anxiety and depression, but it is unclear whether such a causal effect also drives the population association. We cannot exclude the major contribution of a third underlying factor influencing exercise behavior and symptoms of anxiety and depression. OBJECTIVE: To test causal effects of exercise on anxious and depressive symptoms in a population based sample. DESIGN: Population-based longitudinal study (1991-2002) in a genetically informative sample of twin families. SETTING: Causal effects of exercise were tested by bivariate genetic modeling of the association between exercise and symptoms of anxiety and depression, correlation of intrapair differences in these traits among genetically identical twins, and longitudinal modeling of changes in exercise behavior and anxious and depressive symptoms. PARTICIPANTS: A total of 5952 twins from the Netherlands Twin Register, 1357 additional siblings, and 1249 parents. All participants were aged 18 to 50 years. MAIN OUTCOME MEASUREMENTS: Survey data about leisure-time exercise (metabolic equivalent task hours per week based on type, frequency, and duration of exercise) and 4 scales of anxious and depressive symptoms (depression, anxiety, somatic anxiety, and neuroticism, plus a composite score). RESULTS: Cross sectional and longitudinal associations were small and were best explained by common genetic factors with opposite effects on exercise behavior and symptoms of anxiety and depression. In genetically identical twin pairs, the twin who exercised more did not display fewer anxious and depressive symptoms than the co twin who exercised less. Longitudinal analyses showed that increases in exercise participation did not predict decreases in anxious and depressive symptoms. CONCLUSION: Regular exercise is associated with reduced anxious and depressive symptoms in the population at large, but the association is not because of causal effects of exercise. PMID- 18678793 TI - Association of a polymorphism near CREB1 with differential aversion processing in the insula of healthy participants. AB - CONTEXT: Previous functional neuroimaging studies have identified a network of brain regions that process aversive stimuli, including anger. A polymorphism near the cyclic adenosine monophosphate response element binding protein gene (CREB1) has recently been associated with greater self-reported effort at anger control as well as risk for antidepressant treatment-emergent suicidality in men with major depressive disorder, but its functional effects have not been studied. OBJECTIVE: To determine whether this genetic variant is associated with altered brain processing of and behavioral avoidance responses to angry facial expressions. DESIGN AND PARTICIPANTS: A total of 28 white participants (mean age, 29.2 years; 13 women) were screened using the Structured Clinical Interview for DSM-IV to exclude any lifetime Axis I psychiatric disorder and were genotyped for rs4675690, a single-nucleotide polymorphism near CREB1. MAIN OUTCOME MEASURES: Blood oxygenation level-dependent signal by functional magnetic resonance imaging in the amygdala, insula, anterior cingulate, and orbitofrontal cortex during passive viewing of photographs of faces with emotional expressions. To measure approach and avoidance responses to anger, an off-line key-press task that traded effort for viewing time assessed valuation of angry faces compared with other expressions. RESULTS: The CREB1-linked single-nucleotide polymorphism was associated with significant differential activation in an extended neural network responding to angry and other facial expressions. The CREB1-associated insular activation was coincident with activation associated with behavioral avoidance of angry faces. CONCLUSIONS: A polymorphism near CREB1 is associated with responsiveness to angry faces in a brain network implicated in processing aversion. Coincident activation in the left insula is further associated with behavioral avoidance of these stimuli. PMID- 18678795 TI - Apolipoprotein E epsilon4 allele genotype and the effect of depressive symptoms on the risk of dementia in men: the Honolulu-Asia Aging Study. AB - CONTEXT: The apolipoprotein E epsilon4 (APOE epsilon4) allele is a genetic risk factor for Alzheimer disease. Recently, depression has also become recognized as a risk factor for dementia. However, the possible effect of the APOE genotype on the association between depression and dementia is unexamined. OBJECTIVE: To examine the independent and combined effects of depression and APOE epsilon4 on the risk of dementia and its subtypes. DESIGN: The Honolulu-Asia Aging Study, a population-based prospective cohort study of Japanese American men. SETTINGS AND PARTICIPANTS: Depressive symptoms and presence of the APOE epsilon4 allele were assessed between March 1991 and October 1993 in 1932 cognitively healthy men aged 71 to 90 years. Incident cases of dementia were diagnosed during approximately 6 years of follow-up based on neurologic assessment at 2 repeated examinations (April 1994-April 1996 and October 1997-February 1999). MAIN OUTCOME MEASURES: Overall dementia, Alzheimer disease, and vascular dementia. RESULTS: The interaction of depression and APOE epsilon4 was statistically significant in the analytical models. Compared with men with neither APOE epsilon4 nor depression, the risk of dementia in nondepressed men with APOE epsilon4 was not significant (hazard ratio, 1.1; 95% confidence interval [CI], 0.6-1.8); however, depressed men without APOE epsilon4 had a 1.6-fold greater risk (95% CI, 0.8-3.0), whereas depressed men with APOE epsilon4 had a 7.1-fold greater risk (95% CI, 3.0-16.7) of dementia. For subtypes, we found similar increased risks of Alzheimer disease. CONCLUSIONS: The APOE epsilon4 status modifies the association between depressive symptoms and dementia in elderly men. Because individuals with depressive symptoms and the APOE epsilon4 allele had a markedly increased risk of dementia, one might be especially watchful for early signs of dementia in the older person with depression who is also positive for the APOE epsilon4 allele. Because this cohort includes only men, further investigation in women is required. PMID- 18678796 TI - Capsulotomy for obsessive-compulsive disorder: long-term follow-up of 25 patients. AB - CONTEXT: Capsulotomy is sometimes used as a treatment of last resort in severe and treatment-refractory cases of obsessive-compulsive disorder (OCD). OBJECTIVE: To evaluate the long-term efficacy and safety of capsulotomy in OCD. DESIGN: Noncontrolled, long-term follow-up trial (mean of 10.9 years after surgery). SETTING: University hospital referral center. PATIENTS: Twenty-five consecutive patients with OCD who underwent capsulotomy from 1988 to 2000. INTERVENTION: Unilateral or bilateral capsulotomy. Lesions were created by means of radiofrequency heating (thermocapsulotomy) or gamma radiation (radiosurgery, gammacapsulotomy). MAIN OUTCOME MEASURE: Yale-Brown Obsessive-Compulsive Rating Scale (Y-BOCS) score. RESULTS: The mean Y-BOCS score was 34 preoperatively and 18 at long-term follow-up (P < .001). Response (defined as > or = 35% reduction at long-term follow-up compared with baseline) was seen in 12 patients at long-term follow-up. Nine patients were in remission (Y-BOCS score, < 16) at long-term follow-up. Only 3 patients were in remission without adverse effects at long-term follow-up. Response rates did not differ significantly between surgical methods. A mean weight gain of 6 kg was reported in the first postoperative year. Ten patients were considered to have significant problems with executive functioning, apathy, or disinhibition. Six of these 10 patients had received high doses of radiation or had undergone multiple surgical procedures. Results of our magnetic resonance imaging analysis in 11 patients suggest that the OCD symptom reduction may be increased by reducing the lateral extension of the lesions, and a reduction in the medial and posterior extension may limit the risk of adverse effects (ie, smaller lesions may produce better results). CONCLUSIONS: Capsulotomy is effective in reducing OCD symptoms. There is a substantial risk of adverse effects, and the risk may vary between surgical methods. Our findings suggest that smaller lesions are safer and that high radiation doses and multiple procedures should be avoided. PMID- 18678797 TI - Rapid eye movement sleep in relation to overweight in children and adolescents. AB - CONTEXT: Short sleep duration is associated with obesity, but few studies have examined the relationship between obesity and specific physiological stages of sleep. OBJECTIVE: To examine specific sleep stages, including rapid eye movement (REM) sleep and stages 1 through 4 of non-REM sleep, in relation to overweight in children and adolescents. DESIGN, SETTING, AND PARTICIPANTS: A total of 335 children and adolescents (55.2% male; aged 7-17 years) underwent 3 consecutive nights of standard polysomnography and weight and height assessments as part of a study on the development of internalizing disorders (depression and anxiety). MAIN OUTCOME MEASURES: Body mass index (calculated as weight in kilograms divided by height in meters squared) z score and weight status (normal, at risk for overweight, overweight) according to the body mass index percentile for age and sex. RESULTS: The body mass index z score was significantly related to total sleep time (beta = -0.174), sleep efficiency (beta = -0.027), and REM density (beta = -0.256). Compared with normal-weight children, overweight children slept about 22 minutes less and had lower sleep efficiency, shorter REM sleep, lower REM activity and density, and longer latency to the first REM period. After adjustment for demographics, pubertal status, and psychiatric diagnosis, 1 hour less of total sleep was associated with approximately 2-fold increased odds of overweight (odds ratio = 1.85), 1 hour less of REM sleep was associated with about 3-fold increased odds (odds ratio = 2.91), and REM density and activity below the median increased the odds of overweight by 2-fold (odds ratio = 2.18) and 3-fold (odds ratio = 3.32), respectively. CONCLUSIONS: Our results confirm previous epidemiological observations that short sleep time is associated with overweight in children and adolescents. A core aspect of the association between short sleep duration and overweight may be attributed to reduced REM sleep. Further studies are needed to investigate possible mechanisms underpinning the association between diminished REM sleep and endocrine and metabolic changes that may contribute to obesity. PMID- 18678798 TI - CRH haplotype as a factor influencing cerebrospinal fluid levels of corticotropin releasing hormone, hypothalamic-pituitary-adrenal axis activity, temperament, and alcohol consumption in rhesus macaques. AB - CONTEXT: Both highly stress-reactive and novelty-seeking individuals are susceptible to alcohol use disorders. Variation in stress reactivity, exploration, and response to novelty have been attributed to differences in corticotropin-releasing hormone (CRH) system function. As such, CRH gene variation may influence risk for alcohol use and dependence. OBJECTIVE: To determine whether CRH variation influences relevant intermediate phenotypes, behavior, and alcohol consumption in rhesus macaques. DESIGN: We sequenced the rhesus macaque CRH locus (rhCRH) and performed cladistic clustering of haplotypes. In silico analysis, gel shift, and in vitro reporter assays were performed to identify functional variants. Cerebrospinal fluid (CSF) and blood samples were obtained, and levels of CRH and corticotropin (ACTH) were measured by radioimmunoassay. Behavioral data were collected from macaques during infancy. Among adolescent/adult animals, we recorded responses to an unfamiliar conspecific and measured levels of ethanol consumption. SETTING: National Institutes of Health Animal Center. PARTICIPANTS: Rhesus macaques. MAIN OUTCOME MEASURES: Animals were genotyped for a single-nucleotide polymorphism disrupting a glucocorticoid response element, rhCRH -2232 C>G, and the effects of this allele on CSF levels of CRH, plasma levels of ACTH, behavior, and ethanol consumption were assessed by analysis of variance. RESULTS: We show that -2232C>G alters DNA x protein interactions and confers decreased sensitivity of the CRH promoter to glucocorticoids in vitro. Consistent with the known effects of glucocorticoids on CRH expression in the brain, carriers of the G allele had lower CSF levels of CRH but higher levels of ACTH. Infants carrying the G allele were more exploratory and bold, and among adolescent and adult male macaques, the G allele was associated with exploratory/bold responding to an unfamiliar male. Adults with the C/G genotype also exhibited increased alcohol consumption in the social group, a model for high-risk alcohol-seeking behavior. CONCLUSION: Haplotypes that differ in terms of corticosteroid sensitivity have been identified in humans. Our data may suggest that functionally similar CRH variants could influence risk for externalizing disorders in human subjects. PMID- 18678799 TI - Absence of preferential looking to the eyes of approaching adults predicts level of social disability in 2-year-old toddlers with autism spectrum disorder. AB - CONTEXT: Within the first week of life, typical human newborns give preferential attention to the eyes of others. Similar findings in other species suggest that attention to the eyes is a highly conserved phylogenetic mechanism of social development. For children with autism, however, diminished and aberrant eye contact is a lifelong hallmark of disability. OBJECTIVE: To quantify preferential attention to the eyes of others at what is presently the earliest point of diagnosis in autism. DESIGN: We presented the children with 10 videos. Each video showed an actress looking directly into the camera, playing the role of caregiver, and engaging the viewer (playing pat-a-cake, peek-a-boo, etc). Children's visual fixation patterns were measured by eye tracking. PARTICIPANTS: Fifteen 2-year-old children with autism were compared with 36 typically developing children and with 15 developmentally delayed but nonautistic children. MAIN OUTCOME MEASURE: Preferential attention was measured as percentage of visual fixation time to 4 regions of interest: eyes, mouth, body, and object. Level of social disability was assessed by the Autism Diagnostic Observation Schedule. RESULTS: Looking at the eyes of others was significantly decreased in 2-year-old children with autism (P < .001), while looking at mouths was increased (P < .01) in comparison with both control groups. The 2 control groups were not distinguishable on the basis of fixation patterns. In addition, fixation on eyes by the children with autism correlated with their level of social disability; less fixation on eyes predicted greater social disability (r = -0.669, P < .01). CONCLUSIONS: Looking at the eyes of others is important in early social development and in social adaptation throughout one's life span. Our results indicate that in 2-year-old children with autism, this behavior is already derailed, suggesting critical consequences for development but also offering a potential biomarker for quantifying syndrome manifestation at this early age. PMID- 18678800 TI - Estrogen in severe mental illness: a potential new treatment approach. AB - CONTEXT: Accumulating evidence suggests that estrogens may have therapeutic effects in severe mental illnesses, including schizophrenia, via neuromodulatory and neuroprotective activity. OBJECTIVE: To compare the efficacy of adjunctive transdermal estradiol with that of adjunctive placebo in the treatment of acute psychotic symptoms. DESIGN: Randomized, double-blind study. SETTING: Patients were recruited from inpatient acute hospital wards and outpatient clinics of 2 metropolitan Melbourne general hospitals. PARTICIPANTS: One hundred two women of childbearing age with schizophrenia. All participants were in an acute or chronic phase of their illness; 73 participants were outpatients and the rest were inpatients. Intervention Patients were randomized to receive 100 microg of transdermal estradiol (n = 56) or transdermal placebo (n = 46) for 28 days. MAIN OUTCOME MEASURES: Psychopathological symptoms were assessed weekly with the Positive and Negative Syndrome Scale. RESULTS: The addition of 100 microg of transdermal estradiol significantly reduced positive (P < .05) and general psychopathological (P < .05) symptoms during the 28-day trial period compared with women receiving antipsychotic medication alone. CONCLUSION: Estradiol appears to be a useful treatment for women with schizophrenia and may provide a new adjunctive therapeutic option for severe mental illness. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00206570. PMID- 18678801 TI - National trends in psychotherapy by office-based psychiatrists. AB - CONTEXT: In addition to prescribing medications, providing psychotherapy has long been a defining characteristic of the practice of clinical psychiatry. However, there are indications that the role of psychiatrists in providing psychotherapy may have diminished in recent years. OBJECTIVE: To examine recent national trends in the provision of psychotherapy by office-based psychiatrists. DESIGN: Data from the 1996 through 2005 cross-sectional National Ambulatory Medical Care Survey were analyzed to examine trends in psychotherapy provision within nationally representative samples of visits to office-based psychiatrists. Multivariate analyses examined the time trend, adjusting for patient, visit, and setting characteristics. Practice-level analyses examined time trends in the percentage of psychiatrists who provided psychotherapy to all, some, or none of their patients during a typical week. SETTING: Office-based psychiatry practices in the United States. PARTICIPANTS: Patients with psychiatric diagnoses visiting outpatient psychiatrists. MAIN OUTCOME MEASURE: Provision of psychotherapy in visits longer than 30 minutes. RESULTS: Psychotherapy was provided in 5597 of 14,108 visits (34.0% [weighted]) sampled during a 10-year period. The percentage of visits involving psychotherapy declined from 44.4% in 1996-1997 to 28.9% in 2004-2005 (P < .001). This decline coincided with changes in reimbursement, increases in managed care, and growth in the prescription of medications. At the practice level, the decrease in providing psychotherapy corresponded with a decline in the number of psychiatrists who provided psychotherapy to all of their patients from 19.1% in 1996-1997 to 10.8% in 2004-2005 (P = .001). Psychiatrists who provided psychotherapy to all of their patients relied more extensively on self-pay patients, had fewer managed-care visits, and prescribed medications in fewer of their visits compared with psychiatrists who provided psychotherapy less often. CONCLUSIONS: There has been a recent significant decline in the provision of psychotherapy by psychiatrists in the United States. This trend is attributable to a decrease in the number of psychiatrists specializing in psychotherapy and a corresponding increase in those specializing in pharmacotherapy--changes that were likely motivated by financial incentives and growth in psychopharmacological treatments in recent years. PMID- 18678802 TI - Effect of pediatric physical therapy on deformational plagiocephaly in children with positional preference: a randomized controlled trial. AB - OBJECTIVE: To study the effect of pediatric physical therapy on positional preference and deformational plagiocephaly. DESIGN: Randomized controlled trial. SETTING: Bernhoven Hospital, Veghel, the Netherlands. PARTICIPANTS: Of 380 infants referred to the examiners at age 7 weeks, 68 (17.9%) met criteria for positional preference, and 65 (17.1%) were enrolled and followed up at ages 6 and 12 months. INTERVENTION: Infants with positional preference were randomly assigned to receive either physical therapy (n = 33) or usual care (n = 32). MAIN OUTCOME MEASURES: The primary outcome was severe deformational plagiocephaly assessed by plagiocephalometry. The secondary outcomes were positional preference, motor development, and cervical passive range of motion. RESULTS: Both groups were comparable at baseline. In the intervention group, the risk for severe deformational plagiocephaly was reduced by 46% at age 6 months (relative risk, 0.54; 95% confidence interval, 0.30-0.98) and 57% at age 12 months (0.43; 0.22-0.85). The numbers of infants with positional preference needed to treat were 3.85 and 3.13 at ages 6 and 12 months, respectively. No infant demonstrated positional preference at follow-up. Motor development was not significantly different between the intervention and usual care groups. Cervical passive range of motion was within the normal range at baseline and at follow-up. When infants were aged 6 months, parents in the intervention group demonstrated significantly more symmetry and less left orientation in nursing, positioning, and handling. CONCLUSION: A 4-month standardized pediatric physical therapy program to treat positional preference significantly reduced the prevalence of severe deformational plagiocephaly compared with usual care. CLINICAL TRIAL REGISTRATION: isrctn.org Identifier: ISRCTN84132771. PMID- 18678803 TI - Nonsurgical treatment of deformational plagiocephaly: a systematic review. AB - OBJECTIVE: To evaluate and summarize the evidence comparing nonsurgical therapies in the treatment of infants with deformational plagiocephaly. DATA SOURCES: Scientific articles and abstracts published in English between January 1978 and August 2007 were searched from 5 online literature databases, along with a manual search of conference proceedings. STUDY SELECTION: Studies were selected and appraised for methodological quality by 2 reviewers independently using a Critical Appraisal Skills Programme form (cohort criteria). INTERVENTIONS: Molding helmet therapy vs head repositioning therapy. MAIN OUTCOME MEASURE: Success rate of the treatment. RESULTS: A total of 3793 references were retrieved. There were no randomized controlled trials. Only 7 cohort studies met the inclusion criteria. Five of the 7 studies presented evidence that molding therapy is more effective than repositioning, even with the biases favoring the repositioning groups. In the molding groups, the asymmetry was more severe and the infants were older. The infants who failed to respond to repositioning therapy were also switched to molding therapy. The treatment outcomes from the other 2 studies were difficult to assess because of flaws in their study design. Finally, the relative improvement of using molding therapy was calculated from one study. It was about 1.3 times greater than with repositioning therapy. CONCLUSION: The studies showed considerable evidence that molding therapy may reduce skull asymmetry more effectively than repositioning therapy. However, definitive conclusions on the relative effectiveness of these treatments were tempered by potential biases in these studies. Further research is warranted. PMID- 18678804 TI - Prolonged sedation and/or analgesia and 5-year neurodevelopment outcome in very preterm infants: results from the EPIPAGE cohort. AB - OBJECTIVE: To describe the long-term outcome of very preterm infants receiving prolonged sedation and/or analgesia and examine the relationship between prolonged sedation and/or analgesia and this long-term outcome. DESIGN: A prospective population-based study (Etude EPIdemiologique sur les Petits Ages GEstationnels [EPIPAGE]). To reduce bias, the propensity score method was used. SETTING: Nine regions of France. PARTICIPANTS: The study population included very preterm infants of fewer than 33 weeks' gestational age, born in 1997, who received mechanical ventilation and/or surgery. Main Exposure Prolonged exposure to sedative and/or analgesic drugs in the neonatal period, defined as exposure of more than 7 days to sedative and/or opioid drugs. MAIN OUTCOME MEASURE: Presence of moderate or severe disability at 5 years of age. RESULTS: The analysis concerns 1572 premature infants who received mechanical ventilation for whom information about exposure to prolonged sedation and/or analgesia in the neonatal period was available. A total of 115 were exposed and 1457 were not exposed. There was no significant difference between the number of patients lost to follow up from the group of very preterm infants who were exposed to prolonged sedation and/or analgesia and the group who were not. Exposed very preterm infants had severe or moderate disability at 5 years (41/97; 42%) more often than those who were not exposed (324/1248; 26%). After adjustment for gestational age and propensity score, this association was no longer statistically significant (adjusted relative risk, 1.0; 95% confidence interval, 0.8-1.2). CONCLUSION: Prolonged sedation and/or analgesia is not associated with a poor 5-year neurological outcome after adjustment for the propensity score. PMID- 18678805 TI - Propensity scores. PMID- 18678806 TI - Cardiovascular follow-up at school age after perinatal glucocorticoid exposure in prematurely born children: perinatal glucocorticoid therapy and cardiovascular follow-up. AB - OBJECTIVE: To study whether antenatal or neonatal glucocorticoid therapy to reduce the incidence and severity of chronic lung disease in preterm infants is associated with long-term adverse cardiac effects and hypertension. DESIGN: Retrospective matched-cohort study. SETTING: Outpatient clinic of a tertiary care hospital. PARTICIPANTS: One hundred ninety-three children aged 7 to 10 years who had been born prematurely between December 2, 1993, and September 15, 1997. Main Exposure Neonatal treatment with dexamethasone disodium phosphate(n = 48) or the clinically equally effective glucocorticoid hydrocortisone (n = 51), or only antenatal treatment with betamethasone disodium phosphate and betamethasone acetate (n = 51). These 3 groups were compared with a reference group of prematurely born children who had not been exposed to perinatal glucocorticoid therapy (n = 43). MAIN OUTCOME MEASURES: General hemodynamic data (heart rate and blood pressure), cardiovascular function as assessed at echocardiography, intima media thickness of the carotid arteries, and cardiac biochemical features as early markers of expansion and volume overload of the cardiac left ventricle (B type natriuretic peptide and N-terminal pro-B-type natriuretic peptide). RESULTS: No significant group differences were found for heart rate, blood pressure, biochemical features, intima-media thickness, or systolic or diastolic left ventricular function. CONCLUSIONS: Although no differences were found in blood pressure and cardiovascular function at school age in children antenatally or neonatally treated with glucocorticoids, further cardiovascular follow-up may be advisable because cardiovascular dysfunction may become apparent only later in life. PMID- 18678808 TI - High levels of physical inactivity and sedentary behaviors among US immigrant children and adolescents. AB - OBJECTIVE: To examine the prevalence and correlates of physical inactivity and sedentary behavior among immigrant and US-born children. DESIGN: Cross-sectional analysis using data from the 2003 National Survey of Children's Health, a telephone survey conducted between January 29, 2003, and July 1, 2004. SETTING: United States. PARTICIPANTS: Multivariate logistic and least squares regression models were used to analyze immigrant differentials among 68 288 children aged 6 through 17 years. Main Exposure Ethnic-immigrant status. MAIN OUTCOME MEASURES: Prevalence and odds of regular physical activity, inactivity, television watching, and lack of sports participation. RESULTS: Physical inactivity and sedentary behaviors varied widely among children in various ethnic-immigrant groups. For example, 22.5% of immigrant Hispanic children were physically inactive compared with 9.5% of US-born white children with US-born parents. Approximately 67% of immigrant Hispanic children did not participate in sports compared with 30.2% of native Asian children. Overall, immigrant children were significantly more likely to be physically inactive and less likely to participate in sports than native children; they were, however, less likely to watch television 3 or more hours per day than native children, although the nativity gap narrowed with increasing acculturation levels. Compared with native white children, the adjusted odds of physical inactivity and lack of sports participation were both 2 times higher for immigrant Hispanic children with foreign-born parents, and the odds of television watching were 1.5 and 2.3 times higher for native Hispanic and black children, respectively. CONCLUSIONS: Immigrant children in each ethnic minority group generally had higher physical inactivity and lower sports participation levels than native children. To reduce disparities, health education programs need to promote physical activity among children in immigrant families. PMID- 18678809 TI - Insulin resistance status: predicting weight response in overweight children. AB - OBJECTIVE: To assess the relationship between insulin resistance and body mass index (BMI) z score associated with dietary modification that focuses on limiting sweetened beverage consumption in overweight children. DESIGN: A retrospective cohort study conducted between July 1, 2004, and April 28, 2006. SETTING: Community-based primary care practices treating children. PATIENTS: Forty-five children (aged < or =18 years) at or above the 95th percentile of BMI for age and sex. Intervention Children and parents were instructed by a dietitian regarding caloric reduction and modification of sweetened beverage intake. MAIN OUTCOME MEASURES: Insulin resistance at baseline was calculated from fasting insulin and glucose levels (ie, homeostatic model assessment [HOMA]) and change in BMI z score from baseline to 12-week follow-up. RESULTS: Change in BMI z score in response to a decrease in sweetened beverages correlated (Pearson product moment correlation coefficient = 0.42; P < .01) with baseline insulin resistance. Those with a decrease in or an unchanged BMI z score had significantly greater insulin resistance than those whose BMI z score increased (mean [SD] HOMA, 6.2 [4.2] vs 2.6 [2.0], P < .01). Linear regression confirmed that HOMA was a significant predictor of change in BMI z score when controlling for age, race, and sex. CONCLUSIONS: Among overweight children in primary care practices, a significant relationship was found between insulin resistance and the change in BMI z score associated with a dietitian-mediated intervention that includes a focus on decreasing sweetened beverage consumption. Estimating insulin resistance may inform dietary recommendations for overweight children. PMID- 18678810 TI - Occult depressive symptoms in adolescent emergency department patients. AB - OBJECTIVES: To estimate the frequency of occult depressive symptoms in adolescent emergency department (ED) patients (aged 13-17 years) and to determine patient characteristics associated with depressive symptoms. DESIGN: A cross-sectional study of adolescent ED patients. SETTING: The pediatric ED of Hennepin County Medical Center in Minneapolis, Minnesota, and the EDs of the Children's Hospitals and Clinics of Minnesota. PATIENTS: Medically stable adolescent ED patients with nonpsychiatric concerns. INTERVENTIONS: Patients completed the Beck Depression Inventory II and answered questions about their attitudes, activities, and lifestyle choices. Guardians were asked about family demographics, living situations, and other patient characteristics. MAIN OUTCOME MEASURES: The frequency of moderate and severe depressive symptoms as measured by the Beck Depression Inventory II. Group results were analyzed with descriptive statistics; patient characteristics associated with depressive symptoms were determined by multivariate analysis. RESULTS: A total of 967 patients were enrolled. According to the Beck Depression Inventory II, 20% (197 patients) had moderate to severe depressive symptoms. Of these, 58% recognized their depressive symptoms and 50% were recognized by their guardians as having depressive symptoms. When compared with nondepressed patients, adolescents with depressive symptoms more often were female, were not involved in organized social activities, knew someone who intentionally hurt himself or herself or died a violent death, were currently involved in a sexual relationship, or used street drugs. Race, family income, family stability, and witnessing violence were not associated with a positive depression screen result. CONCLUSIONS: Depressive symptoms occur frequently among adolescents and are often unrecognized. Efforts to increase awareness of depression among ED physicians, adolescents, and parents of adolescents may be beneficial. PMID- 18678807 TI - Community supports after surviving extremely low-birth-weight, extremely preterm birth: special outpatient services in early childhood. AB - OBJECTIVE: To determine special outpatient services (SOS) use, need, associated factors, and neurodevelopmental and functional outcomes among extremely preterm infants at 18 to 22 months' corrected age. DESIGN: Retrospective analysis. SETTING: National Institute of Child Health and Human Development (NICHD) Neonatal Research Network. PARTICIPANTS: Infants younger than 28 weeks' gestational age who had been born weighing less than 1000 g at an NICHD Neonatal Research Network center from January 1, 1997, to December 31, 2000, and who were receiving follow-up at 18 to 22 months' corrected age. INTERVENTIONS: Questionnaires were administered at the 18- to 22-month follow-up visit regarding SOS use since hospital discharge and the current need for SOS (social work, visiting nurse, medical specialty, early intervention, speech and language services, occupational therapy and physical therapy, and neurodevelopmental and behavioral services). MAIN OUTCOME MEASURES: The use of and need for SOS were analyzed by gestational age. Logistic regression analysis identified factors independently associated with the use of more than 5 services and with the need for any services. RESULTS: Of 2315 infants, 54.7% used more than 3 SOS by 18 to 22 months, and 19.1% used 6 to 7 SOS. The need for any SOS was reported by approximately 37%. The following variables that were commonly associated with adverse neurodevelopmental outcomes were also associated with the use of more than 5 SOS: sepsis, birth weight, postnatal corticosteroid use, bronchopulmonary dysplasia, and cystic periventricular leukomalacia or grade 3 or 4 intraventricular hemorrhage. Male sex was associated with the need for any SOS. Although high SOS use was more likely among children with adverse neurodevelopmental outcomes, a reported need for SOS was common even among those with mild developmental impairment (39.7%) and mild cerebral palsy (42.2%). CONCLUSIONS: High SOS use is common, has identifiable neonatal risk factors, and is associated with neurodevelopmental impairment. Extremely preterm survivors have substantial need for community supports regardless of their impairment level. Efforts to improve comprehensive delivery of family-centered community based services are urgently needed. PMID- 18678811 TI - Childhood respiratory disease and the risk of anxiety disorder and major depression in adulthood. AB - OBJECTIVE: To examine the association between early childhood respiratory disease and the risk of anxiety and depression in adulthood. DESIGN: Cohort study. SETTING: Providence cohort of the National Collaborative Perinatal Project. PARTICIPANTS: Offspring of 1062 mothers selected for follow-up from birth through adulthood. MAIN EXPOSURE: Childhood respiratory disease. MAIN OUTCOME MEASURE: Odds of developing anxiety and depression by age 34 years. RESULTS: Respiratory disease in childhood was associated with an elevated risk of receiving treatment for anxiety disorders but not mood disorders. Specifically, higher respiratory rate at age 4 months was associated with significantly increased odds of receiving treatment for anxiety by age 34 years (odds ratio, 2.2; 95% confidence interval, 1.1-4.4; P < .05). Respiratory disease at age 1 year was associated with significantly increased odds of receiving treatment for anxiety (odds ratio, 2.8; 95% confidence interval, 1.2-6.5; P = .04). In addition, having respiratory disease at age 1 year only was associated with increased odds of receiving treatment for anxiety (odds ratio, 3.1; 95% confidence interval, 1.1-8.7; P < .05), whereas having both respiratory disease at age 1 year and suspect (parent reported but not physician diagnosed) respiratory disease at age 7 years was associated with the greatest odds of receiving treatment for anxiety (odds ratio, 19.8; 95% confidence interval, 2.8-141.9; P < .05). CONCLUSIONS: These results are consistent with and extend previous findings and provide prospective evidence of a link between respiratory disease in early childhood and increased risk of anxiety disorders by age 34 years. These findings may have implications for prevention or early intervention with groups at high risk for anxiety disorders. PMID- 18678812 TI - Positive parenting and early puberty in girls: protective effects against aggressive behavior. AB - OBJECTIVE: To determine whether positive parenting practices are associated with less aggressive and delinquent behavior in early-maturing girls. DESIGN: Cross sectional survey. SETTING: Interviews with a community sample of children and their caregivers were conducted in their homes or in a research setting. PARTICIPANTS: An ethnically diverse cohort of 330 fifth-grade girls (mean age, 11.25 years) from 3 metropolitan areas. MAIN EXPOSURE: Early onset of menarche, parental nurturance, knowledge of the child's activities, and communication. MAIN OUTCOME MEASURES: Physical, relational, and nonphysical aggression and delinquent behavior. RESULTS: A total of 25% of girls could be reliably classified as early maturers. Early maturation was associated with delinquency (b = 0.53) but not aggression. Low levels of maternal nurturance were associated with delinquency and relational aggression (both b = -0.04). Early maturation was associated with higher relational aggression only at low levels of nurturance (b = 0.94), communication (b = 1.36), and knowledge (b = 1.06) (P < .05 for each interaction). Also, early maturation only predicted physical aggression when combined with low maternal nurturance (b = 0.93). CONCLUSIONS: Early puberty is a risk factor for delinquency, and early puberty combined with low parental nurturance, communication, or parental knowledge of the child's activities presents a risk for aggressive behavior in early adolescent girls. Early-maturing girls may benefit from increased parental nurturance, communication, and knowledge. PMID- 18678813 TI - Picture of the month. Congenital Dislocation of the Knee. PMID- 18678815 TI - p-Phenylenediamine in black henna tattoos: a practice in need of policy in children. PMID- 18678816 TI - Effect of long-acting OROS methylphenidate on routine driving in young adults with attention-deficit/hyperactivity disorder. PMID- 18678817 TI - Discussing illness at well-child care visits. PMID- 18678818 TI - Teaching NeuroImage: corticospinal tract. PMID- 18678819 TI - Teaching video NeuroImage: the "Fonzarelli" sign: focal thumb dystonia as an early manifestation of Parkinson disease. PMID- 18678821 TI - The muddle of mycophenolate mofetil in myasthenia. PMID- 18678822 TI - The meaning of interictal spikes in temporal lobe epilepsy: should we count them? PMID- 18678823 TI - Dehydroepiandrosterone for myotonic dystrophy type 1. AB - BACKGROUND: Myotonic dystrophy type 1 may be associated with low circulating dehydroepiandrosterone (DHEA) levels. This study was aimed at investigating the efficacy and safety of DHEA in myotonic dystrophy type 1 patients. METHODS: This was a prospective, multicenter, randomized, double-blind, placebo-controlled trial conducted from February 2005 to January 2006 at 10 university-affiliated neuromuscular disease centers in France. Seventy-five ambulatory adults with myotonic dystrophy type 1 received an oral replacement dose (100 mg/d) or a pharmacologic dose (400 mg/d) of DHEA, or placebo. The primary endpoint was the relative change in the manual muscle testing (MMT) score from baseline to week 12. Secondary outcome measures included changes from baseline to week 12 in quantitative muscle testing and timed functional testing, respiratory and cardiac function, and quality of life. This study was registered with ClinicalTrials.gov identifier NCT00167609. RESULTS: The median (1st, 3rd quartile) relative changes in MMT score from baseline to week 12 after randomization were 3.1 (-0.9, 6.7), 1.9 (-2.7, 3.5), and 2.2 (0, 7.9), in the DHEA 100 mg, DHEA 400 mg, and placebo groups, respectively. There were no differences between placebo and combined DHEA groups (p = 0.34), placebo and DHEA 100 mg (p = 0.86), or placebo and DHEA 400 mg (p = 0.15). There were also no evidence for a difference between groups for the changes from baseline to week 12 in any secondary outcome. CONCLUSIONS: There is no evidence that a 12-week treatment with replacement or pharmacologic doses of dehydroepiandrosterone improves muscle strength in ambulatory myotonic dystrophy type 1 patients. PMID- 18678824 TI - Network atrophy in temporal lobe epilepsy: a voxel-based morphometry study. AB - BACKGROUND: Recent evidence suggests that temporal lobe epilepsy affects a neuronal network rather than a single circumscribed structure within the temporal lobe. Thus, in mesial temporal lobe epilepsy (mTLE) with hippocampal sclerosis gray matter abnormalities have been found beyond the hippocampus in extrahippocampal regions by means of voxel-based morphometry (VBM). On the contrary, in cryptogenic or MRI-negative TLE (cTLE) no consistent gray matter abnormalities in VBM studies have been reported. METHODS: We used optimized VBM with modulation to detect gray matter abnormalities compared to healthy controls in patients with mTLE and cTLE. Twenty-two patients with mTLE (right/left TLE 13:9), 17 patients with cTLE (right/left TLE 7:10), and 12 healthy controls were enrolled in the study. RESULTS: In mTLE we found decreased gray matter volume (GMV) beyond the hippocampus in the ipsilateral thalamus. GMV decrease was more widespread in patients with left-sided seizure focus including the left parahippocampal and superior temporal gyrus, frontal regions, cerebellum, and the right cingulum. In cTLE, decreased GMV was observed in the frontal and orbitofrontal cortex, the cerebellum, neocortical temporal regions, and in the right parahippocampal cortex. Again, patients with left-sided seizure focus had a more widespread and extensive GMV decrease including regions such as the right and left cingulum. CONCLUSION: We found evidence for distinct neuronal network damage in mesial temporal lobe epilepsy (mTLE) and cryptogenic TLE (cTLE) which is more widespread in patients with left-sided seizure focus. Atrophy of the cingulum was a common feature in left- but not in right-sided mTLE and cTLE. PMID- 18678825 TI - Acute ophthalmoplegia (without ataxia) associated with anti-GQ1b antibody. AB - BACKGROUND: Anti-GQ1b antibody has been found in Miller Fisher syndrome (MFS), Guillain-Barre syndrome (GBS) with ophthalmoplegia, Bickerstaff brainstem encephalitis (BBE), and acute ophthalmoplegia without ataxia (AO). The aim of this study was to determine the clinical features of AO associated with anti-GQ1b antibody. METHODS: We retrospectively collected 34 patients with anti-GQ1b antibody syndrome. Of these patients, 31 patients had ophthalmoplegia. The patients with ophthalmoplegia were classified into MFS (n = 13), AO (n = 11), GBS with ophthalmoplegia (n = 6), and BBE (n = 1). We analyzed clinical features and patterns of external and internal ophthalmoplegia of AO, and neuro-ophthalmologic findings were compared with those of other anti-GQ1b syndromes with ophthalmoplegia. RESULTS: AO was observed in 11 (32.4%) of the 34 patients with anti-GQ1b antibody. External ophthalmoparesis was present in all the patients and included mixed horizontal-vertical (n = 7), pure horizontal (n = 3), and pure vertical gaze palsy (n = 1). Binocular involvement was common, but unilateral ophthalmoparesis was also observed in 27.3%. Other findings included ptosis (n = 5, 45.5%) and internal ophthalmoplegia (n = 6, 54.5%). Other anti-GQ1b antibody syndromes had prominent neurologic signs including ataxia, weakness, and facial palsy in addition to ophthalmoplegia. The patterns of neuro-ophthalmologic findings did not differ between AO and other anti-GQ1b antibody syndromes with ophthalmoplegia. CONCLUSIONS: Acute ophthalmoplegia (AO) commonly occurs in anti GQ1b antibody syndrome and manifests as various combinations of external and internal ophthalmoplegia. Internal ophthalmoplegia is fairly common and unilateral involvement may occur in AO. PMID- 18678826 TI - Effect of fish oil on cognitive performance in older subjects: a randomized, controlled trial. AB - BACKGROUND: High intake of n-3 polyunsaturated fatty acids may protect against age-related cognitive decline. However, results from epidemiologic studies are inconclusive, and results from randomized trials in elderly subjects without dementia are lacking. OBJECTIVE: To investigate the effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) supplementation on cognitive performance. METHODS: Double-blind, placebo-controlled trial involving 302 cognitively healthy (Mini-Mental State Examination score > 21) individuals aged 65 years or older. Participants were randomly assigned to 1,800 mg/d EPA-DHA, 400 mg/d EPA-DHA, or placebo capsules for 26 weeks. Cognitive performance was assessed using an extensive neuropsychological test battery that included the cognitive domains of attention, sensorimotor speed, memory, and executive function. RESULTS: The mean age of the participants was 70 years, and 55% were male. Plasma concentrations of EPA-DHA increased by 238% in the high-dose and 51% in the low-dose fish oil group compared with placebo, reflecting excellent compliance. Baseline scores on the cognitive tests were comparable in the three groups. Overall, there were no significant differential changes in any of the cognitive domains for either low-dose or high-dose fish oil supplementation compared with placebo. CONCLUSIONS: In this randomized, double-blind, placebo controlled trial, we observed no overall effect of 26 weeks of eicosapentaenoic acid and docosahexaenoic acid supplementation on cognitive performance. PMID- 18678828 TI - Effects of poverty on cognitive function: a hidden neurologic epidemic. AB - Mental retardation is one of the most prevalent neurologic disorders globally. Surveys in high-income countries show 3 to 5 per 1,000 with severe intellectual disability, i.e., IQ below 55. Estimates from developing countries, however, have found prevalence rates from 5 to as much as 22 per 1,000. Protein-energy malnutrition, dietary micronutrient deficiencies, environmental toxins, and lack of early sensory stimulation or the ability to profit from it may contribute to neurodevelopmental disabilities. Tropical diseases such as parasitosis with resultant anemia, malaria, and other infections are major contributory causes. Reduction of poverty and its effects would reduce the present and future burden of mental retardation and cognitive dysfunction, especially in developing countries. PMID- 18678829 TI - Invited article: Residency Review Committee for neurology: structure and function. PMID- 18678827 TI - Fish consumption and risk of subclinical brain abnormalities on MRI in older adults. AB - OBJECTIVE: To investigate the association between fish consumption and subclinical brain abnormalities. METHODS: In the population-based Cardiovascular Health Study, 3,660 participants age > or =65 underwent an MRI scan in 1992-1994. Five years later, 2,313 were scanned. Neuroradiologists assessed MRI scans in a standardized and blinded manner. Food frequency questionnaires were used to assess dietary intakes. Participants with known cerebrovascular disease were excluded from the analyses. RESULTS: After adjustment for multiple risk factors, the risk of having one or more prevalent subclinical infarcts was lower among those consuming tuna/other fish > or =3 times/week, compared to <1/month (relative risk 0.74, 95% CI = 0.54-1.01, p = 0.06, p trend = 0.03). Tuna/other fish consumption was also associated with trends toward lower incidence of subclinical infarcts. Additionally, tuna/other fish intake was associated with better white matter grade, but not with sulcal and ventricular grades, markers of brain atrophy. No significant associations were found between fried fish consumption and any subclinical brain abnormalities. CONCLUSIONS: Among older adults, modest consumption of tuna/other fish, but not fried fish, was associated with lower prevalence of subclinical infarcts and white matter abnormalities on MRI examinations. Our results add to prior evidence that suggest that dietary intake of fish with higher eicosapentaenoic acid and docosahexaenoic acid content, and not fried fish intake, may have clinically important health benefits. PMID- 18678830 TI - Four new cases of therapy-related acute promyelocytic leukemia after mitoxantrone. PMID- 18678831 TI - Break in binocular fusion during head turning in MS patients with INO. PMID- 18678832 TI - Hypoglossal-vertebral entrapment syndrome. PMID- 18678833 TI - Mixed lineage kinase inhibitor CEP-1347 fails to delay disability in early Parkinson disease. PMID- 18678834 TI - Is it time to use observational data to estimate treatment effectiveness in multiple sclerosis? PMID- 18678835 TI - Chemotherapy in addition to supportive care improves survival in advanced non small-cell lung cancer: a systematic review and meta-analysis of individual patient data from 16 randomized controlled trials. AB - PURPOSE: Since our individual patient data (IPD) meta-analysis (MA) of supportive care and chemotherapy for non-small-cell lung cancer (NSCLC), published in 1995, many trials have been completed. An updated, IPD MA has been carried out to assess newer regimens and determine conclusively the effect of chemotherapy. METHODS: Systematic searches for randomized controlled trials (RCTs) were undertaken, followed by central collection, checking, and reanalysis of updated IPD. Results from RCTs were combined to calculate individual and pooled hazard ratios (HRs). RESULTS: Data were obtained from 2,714 patients from 16 RCTs. There were 1,293 deaths among 1,399 patients assigned supportive care and chemotherapy and 1,240 among 1,315 assigned supportive care alone. Results showed a significant benefit of chemotherapy (HR, 0.77; 95% CI, 0.71 to 0.83; P 6.9 mmol/L [>125 mg/dL] for type 2 diabetes). These cut-points do not adequately reflect disease biology, may inappropriately treat patients on either side of the cut-point as 2 homogenous risk groups, fail to incorporate other risk factors, and are invariable to patient preference. This article discusses risk prediction as an alternative to diagnosis: Patient risk factors (blood pressure, age) are combined into a single statistical model (risk for a cardiovascular event within 10 years) and the results are used in shared decision making about possible treatments. The authors compare and contrast the diagnostic and risk prediction approaches and attempt to identify the types of medical problem to which each is best suited. PMID- 18678848 TI - The deadliest catch: fishing for HIV in new waters. PMID- 18678849 TI - Improving care and outcomes of uninsured persons with chronic disease... now. PMID- 18678850 TI - Plan-do-study-act cycle rejuvenates a marriage. PMID- 18678851 TI - A world no less sublime. PMID- 18678852 TI - Is there enough evidence to support use of N-acetylcysteine in contrast-induced nephropathy? PMID- 18678853 TI - Is there enough evidence to support use of N-acetylcysteine in contrast-induced nephropathy? PMID- 18678854 TI - Is there enough evidence to support use of N-acetylcysteine in contrast-induced nephropathy? PMID- 18678855 TI - Is there enough evidence to support use of N-acetylcysteine in contrast-induced nephropathy? PMID- 18678856 TI - Fever increases the risk for cardiac arrest in the Brugada syndrome. PMID- 18678857 TI - A case of apical ballooning cardiomyopathy associated with duloxetine. PMID- 18678858 TI - The special issue on prevention of violence ignores the primordial violence. PMID- 18678859 TI - Fail or flourish? Cognitive appraisal moderates the effect of solo status on performance. AB - When everyone in a group shares a common social identity except one individual, the one who is different from the majority has solo status. Solo status increases one's visibility and performance pressure, which may result in stress. Stress has divergent effects on performance, and individuals' response to stressful situations is predicted by their cognitive appraisal (challenge or threat) of the situation. Two experiments test the hypothesis that cognitive appraisal moderates the effect of solo status on performance. Experiment 1 finds that at relatively high appraisal levels (resources exceed demands), solo status improves men's and women's performance; at relatively low appraisal levels, solo status hurts performance. Experiment 2 replicates this effect for solo status based on minimal group assignment. Results suggest that for individuals who feel challenged and not threatened by their work, it may help to be a solo. PMID- 18678860 TI - Culture and aesthetic preference: comparing the attention to context of East Asians and Americans. AB - Prior research indicates that East Asians are more sensitive to contextual information than Westerners. This article explored aesthetics to examine whether cultural variations were observable in art and photography. Study 1 analyzed traditional artistic styles using archival data in representative museums. Study 2 investigated how contemporary East Asians and Westerners draw landscape pictures and take portrait photographs. Study 3 further investigated aesthetic preferences for portrait photographs. The results suggest that (a) traditional East Asian art has predominantly context-inclusive styles, whereas Western art has predominantly object-focused styles, and (b) contemporary members of East Asian and Western cultures maintain these culturally shaped aesthetic orientations. The findings can be explained by the relation among attention, cultural resources, and aesthetic preference. PMID- 18678861 TI - Role of xanthine oxidase activation and reduced glutathione depletion in rhinovirus induction of inflammation in respiratory epithelial cells. AB - Rhinoviruses are the major cause of the common cold and acute exacerbations of asthma and chronic obstructive pulmonary disease. We previously reported rapid rhinovirus induction of intracellular superoxide anion, resulting in NF-kappaB activation and pro-inflammatory molecule production. The mechanisms of rhinovirus superoxide induction are poorly understood. Here we found that the proteolytic activation of the xanthine dehydrogenase/xanthine oxidase (XD/XO) system was required because pretreatment with serine protease inhibitors abolished rhinovirus-induced superoxide generation in primary bronchial and A549 respiratory epithelial cells. These findings were confirmed by Western blotting analysis and by silencing experiments. Rhinovirus infection induced intracellular depletion of reduced glutathione (GSH) that was abolished by pretreatment with either XO inhibitor oxypurinol or serine protease inhibitors. Increasing intracellular GSH with exogenous H2S or GSH prevented both rhinovirus-mediated intracellular GSH depletion and rhinovirus-induced superoxide production. We propose that rhinovirus infection proteolytically activates XO initiating a pro inflammatory vicious circle driven by virus-induced depletion of intracellular reducing power. Inhibition of these pathways has therapeutic potential. PMID- 18678862 TI - Post-transcriptional control of CCAAT/enhancer-binding protein beta (C/EBPbeta) expression: formation of a nuclear HuR-C/EBPbeta mRNA complex determines the amount of message reaching the cytosol. AB - In 3T3-L1 cells, HuR is constitutively expressed and prior to induction of differentiation localized predominantly to the nucleus. Within minutes of induction of differentiation, nuclear HuR binds to its target ligand mRNAs, and the complexes appear to move to the cytosol. One ligand mRNA is the CCAAT/enhancer-binding protein beta (C/EBPbeta) message. To examine the function and importance of the HuR-C/EBPbeta interaction, retroviral expression constructs were created in which the HuR binding site was altered by deletion (betadel) or deletion and substitution (betad/s). Expression of these constructs in murine embryonic fibroblasts resulted in significant adipose conversion relative to those cells expressing wild type C/EBPbeta. C/EBPbeta protein content was increased markedly in both betadel and betad/s, which correlated with the acquisition of the adipocyte phenotype. Analysis of the betad/s cell line demonstrated a robust expression of C/EBPalpha coincident with peroxisome proliferator-activated receptor gamma expression. Total C/EBPbeta mRNA accumulation indicated no difference between cells harboring either the wild type C/EBPbeta cDNA or betad/s construct. However, cytosolic C/EBPbeta mRNA in the cells expressing the betad/s construct was maintained at levels between 2- and 7 fold greater than in the cells expressing the wild type construct. Alteration in mRNA half-life was not responsible for the increased accumulation. Mechanistically, these data suggest that HuR binding results in nuclear retention of the C/EBPbeta mRNA and is consistent with HuR control, at least in part, of mRNA processing. PMID- 18678863 TI - Identification of Mg2+ -dependent neutral sphingomyelinase 1 as a mediator of heat stress-induced ceramide generation and apoptosis. AB - Neutral sphingomyelinases (SMases) are involved in the induction of ceramide mediated proapoptotic signaling under heat stress conditions. Although ceramide is an important mediator of apoptosis, the neutral SMase that is activated under heat stress has not been identified. In this study, we cloned an Mg(2+)-dependent neutral SMase from a zebrafish embryonic cell cDNA library using an Escherichia coli expression-cloning vector. Screening of the clones using an SMase activity assay with C(6)-7-nitro-2-1,3-benzoxadiazol-4-yl-sphingomyelin as the substrate resulted in the isolation of one neutral SMase cDNA clone. This cDNA encoded a polypeptide of 420 amino acids (putative molecular weight: 46,900) containing two predicted transmembrane domains in its C-terminal region. The cloned neutral SMase 1 acted as a mediator of stress-induced apoptosis. Bacterially expressed recombinant neutral SMase 1 hydrolyzed [choline-methyl-(14)C]sphingomyelin optimally at pH 7.5 in the presence of an Mg(2+) ion. In zebrafish embryonic cells, the endogenous SMase enzyme was localized in the microsomal fraction. In FLAG-tagged SMase-overexpressing cells, neutral SMase 1 colocalized with a Golgi marker in a cytochemical analysis. Inactivation of the enzyme by an antisense phosphorothioate oligonucleotide repressed the induction of ceramide generation, caspase-3 activation, and apoptotic cell death by heat stress. Thus, neutral SMase 1 participates in an inducible ceramide-mediating, proapoptotic signaling pathway that operates in heat-induced apoptosis in zebrafish embryonic cells. PMID- 18678864 TI - Step-size analyses of the mitochondrial Hsp70 import motor reveal the Brownian ratchet in operation. AB - Newly synthesized mitochondrial precursor proteins have to become unfolded by the mitochondrial Hsp70 (mtHsp70) import motor to cross the mitochondrial membranes. To assess the mechanism of unfolding of precursor proteins by mtHsp70, we designed a system to measure step sizes of the mtHsp70 import motor, which are distances at which the motor system moves along polypeptide chains during a single turnover of ATP. We made a series of fusion proteins consisting of a mitochondrial presequence containing the first mtHsp70 binding site, a spacer sequence containing an Hsp70 avoidance segment followed by the second mtHsp70 binding site, and different folded mature domains. Analyses of the dependence of the import rates of those fusion proteins on the lengths of Hsp70 avoidance segments allowed us to estimate the step sizes, which differ for different mature domains and different lengths of the spacers. These results suggest that the mtHsp70 import motor functions at least as a molecular Brownian ratchet to unfold mitochondrial precursor proteins. PMID- 18678865 TI - Activity determinants and functional specialization of Arabidopsis PEN1 syntaxin in innate immunity. AB - In eukaryotes, proteins of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) family are believed to have a general role for the fusion of intracellular transport vesicles with acceptor membranes. Arabidopsis thaliana PEN1 syntaxin resides in the plasma membrane and was previously shown to act together with its partner SNAREs, the adaptor protein SNAP33, and endomembrane-anchored VAMP721/722 in the execution of secretory immune responses against powdery mildew fungi. We conducted a structure-function analysis of PEN1 and show that N-terminal phospho-mimicking and non phosphorylatable variants neither affected binary nor ternary SNARE complex formation with cognate partners in vitro. However, expression of these syntaxin variants at native protein levels in a pen1 mutant background suggests that phosphorylation is required for full resistance activity in planta. All tested site-directed substitutions of SNARE domain or "linker region" residues reduced PEN1 defense activity. Two of the variants failed to form ternary complexes with the partner SNAREs in vitro, possibly explaining their diminished in planta activity. However, impaired pathogen defense in plants expressing a linker region variant is likely because of PEN1 destabilization. Although Arabidopsis PEN1 and SYP122 syntaxins share overlapping functions in plant growth and development, PEN1 activity in disease resistance is apparently the result of a complete functional specialization. Our findings are consistent with the hypothesis that PEN1 acts in plant defense through the formation of ternary SNARE complexes and point to the existence of unknown regulatory factors. Our data indirectly support structural inferences that the four-helical coiled coil bundle in ternary SNARE complexes is formed in a sequential order from the N- to C-terminal direction. PMID- 18678866 TI - Ergot alkaloid biosynthesis in Aspergillus fumigatus. Overproduction and biochemical characterization of a 4-dimethylallyltryptophan N-methyltransferase. AB - The putative gene fgaMT was identified in the biosynthetic gene cluster of fumigaclavines in Aspergillus fumigatus. The coding region of fgaMT was amplified by PCR from a cDNA library, cloned into pQE60, and overexpressed in Escherichia coli. FgaMT comprises 339 amino acids with a molecular mass of about 38.1 kDa. The soluble dimeric His(6)-FgaMT was purified to near homogeneity and characterized biochemically. FgaMT was found to catalyze the N-methylation of 4 dimethylallyltryptophan in the presence of S-adenosylmethionine, resulting in the formation of 4-dimethylallyl-l-abrine, which was identified by NMR and mass spectrometry analysis. Therefore, FgaMT represents the second pathway-specific enzyme in the biosynthesis of ergot alkaloids. The enzyme did not require metal ions for its enzymatic reaction and showed a relatively high specificity toward the prenyl moiety at position C-4 of the indole ring. 4-Dimethylallyltryptophan derivatives with modification at the indole ring were also accepted by FgaMT as substrates. K(m) values for 4-dimethylallyltryptophan and S-adenosylmethionine were determined at 0.12 and 2.4 mm, respectively. The turnover number was 2.0 s( 1). PMID- 18678867 TI - Life span extension and neuronal cell protection by Drosophila nicotinamidase. AB - The life span of model organisms can be modulated by environmental conditions that influence cellular metabolism, oxidation, or DNA integrity. The yeast nicotinamidase gene pnc1 was identified as a key transcriptional target and mediator of calorie restriction and stress-induced life span extension. PNC1 is thought to exert its effect on yeast life span by modulating cellular nicotinamide and NAD levels, resulting in increased activity of Sir2 family class III histone deacetylases. In Caenorhabditis elegans, knockdown of a pnc1 homolog was shown recently to shorten the worm life span, whereas its overexpression increased survival under conditions of oxidative stress. The function and regulation of nicotinamidases in higher organisms has not been determined. Here, we report the identification and biochemical characterization of the Drosophila nicotinamidase, D-NAAM, and demonstrate that its overexpression significantly increases median and maximal fly life span. The life span extension was reversed in Sir2 mutant flies, suggesting Sir2 dependence. Testing for physiological effectors of D-NAAM in Drosophila S2 cells, we identified oxidative stress as a primary regulator, both at the transcription level and protein activity. In contrast to the yeast model, stress factors such as high osmolarity and heat shock, calorie restriction, or inhibitors of TOR and phosphatidylinositol 3 kinase pathways do not appear to regulate D-NAAM in S2 cells. Interestingly, the expression of D-NAAM in human neuronal cells conferred protection from oxidative stress-induced cell death in a sirtuin-dependent manner. Together, our findings establish a life span extending the ability of nicotinamidase in flies and offer a role for nicotinamide-modulating genes in oxidative stress regulated pathways influencing longevity and neuronal cell survival. PMID- 18678868 TI - Pollen proteins bind to the C-terminal domain of Nicotiana alata pistil arabinogalactan proteins. AB - Pollen tube growth is influenced by interaction between pollen proteins and the pistil extracellular matrix. The transmitting tract-specific glycoprotein (NaTTS) and 120-kDa glycoprotein (120K) are two pistil arabinogalactan proteins (AGPs) that share a conserved C-terminal domain (CTD) and directly influence pollen tubes in Nicotiana alata. 120K and other extracellular matrix proteins are taken up and transported to vacuoles of growing pollen tubes. We hypothesize that signaling and trafficking processes inside pollen tubes are important for controlling pollen tube growth. We performed a yeast two-hybrid screen of pollen cDNAs using sequences from 120K and NaTTS as baits. We found that an S-RNase binding protein (SBP1), a C2 domain-containing protein (NaPCCP), and a putative cysteine protease bound to the AGP baits. SBP1 from Petunia hybrida and Solanum chacoense is a putative E3 ubiquitin ligase that binds to S-RNase and other proteins. C2 domain-containing proteins bind lipids and can regulate myriad cellular processes. Cysteine proteases are often associated with the degradation of vacuolar proteins. Expression analysis revealed that transcripts for these proteins are expressed in mature pollen. NaPCCP and NaSBP1 were characterized further because of their potential roles in signaling and trafficking. In vitro pull-down assays verified binding between maltose-binding protein (MBP) fusions, MBP::NaPCCP or MBP::NaSBP1 and glutathione S-transferase (GST), GST::AGP CTD fusions. NaSBP1 binds to the AGP CTDs through its helical and RING domains. NaPCCP binds through its C-terminal region. Binding between NaPCCP and NaSBP1 and the pistil AGPs may contribute to signaling and trafficking inside pollen tubes growing in planta. PMID- 18678869 TI - Proteinase-activated receptor 2 stimulates Na,K-ATPase and sodium reabsorption in native kidney epithelium. AB - Proteinase-activated receptors 2 (PAR2) are expressed in kidney, but their function is mostly unknown. Since PAR2 control ion transport in several epithelia, we searched for an effect on sodium transport in the cortical thick ascending limb of Henle's loop, a nephron segment that avidly reabsorbs NaCl, and for its signaling. Activation of PAR2, by either trypsin or a specific agonist peptide, increased the maximal activity of Na,K-ATPase, its apparent affinity for sodium, the sodium permeability of the paracellular pathway, and the lumen positive transepithelial voltage, featuring increased NaCl reabsorption. PAR2 activation induced calcium signaling and phosphorylation of ERK1,2. PAR2-induced stimulation of Na,K-ATPase Vmax was fully prevented by inhibition of phospholipase C, of changes in intracellular concentration of calcium, of classical protein kinases C, and of ERK1,2 phosphorylation. PAR2-induced increase in paracellular sodium permeability was mediated by the same signaling cascade. In contrast, increase in the apparent affinity of Na,K-ATPase for sodium, although dependent on phospholipase C, was independent of calcium signaling, was insensitive to inhibitors of classical protein kinases C and of ERK1,2 phosphorylation, but was fully prevented by the nonspecific protein kinase inhibitor staurosporine, as was the increase in transepithelial voltage. In conclusion, PAR2 increases sodium reabsorption in rat thick ascending limb of Henle's loop along both the transcellular and the paracellular pathway. PAR2 effects are mediated in part by a phospholipase C/protein kinase C/ERK1,2 cascade, which increases Na,K-ATPase maximal activity and the paracellular sodium permeability, and by a different phospholipase C-dependent, staurosporine sensitive cascade that controls the sodium affinity of Na,K-ATPase. PMID- 18678870 TI - Inactivation of UCP1 and the glycerol phosphate cycle synergistically increases energy expenditure to resist diet-induced obesity. AB - Our current paradigm for obesity assumes that reduced thermogenic capacity increases susceptibility to obesity, whereas enhanced thermogenic capacity protects against obesity. Here we report that elimination of two major thermogenic pathways encoded by the mitochondrial uncoupling protein (Ucp1) and mitochondrial glycerol-3-phosphate dehydrogenase (Gdm) result in mice with increased resistance to diet-induced obesity when housed at 28 degrees C, provided prior adaptation occurred at 20 degrees C. Obesity resistant Gdm(-/ ).Ucp1(-/-) mice maintained at 28 degrees C have increased energy expenditure, in part through conversion of white to brown adipocytes in inguinal fat. Increased oxygen consumption in inguinal fat cell suspensions and the up-regulation of genes of mitochondrial function and fat metabolism indicated increased thermogenic activity, despite the absence of UCP1, whereas liver and skeletal muscle showed no changes in gene expression. Additionally, comparisons of energy expenditure in UCP1-deficient and wild type mice fed an obesogenic diet indicates that UCP1-based brown fat-based thermogenesis plays no role in so-called diet induced thermogenesis. Accordingly, a new paradigm for obesity emerges in which the inactivation of major thermogenic pathways force the induction of alternative pathways that increase metabolic inefficiency. PMID- 18678871 TI - ADP competes with FAD binding in putrescine oxidase. AB - Putrescine oxidase from Rhodococcus erythropolis NCIMB 11540 (PuO(Rh)) is a soluble homodimeric flavoprotein of 100 kDa, which catalyzes the oxidative deamination of putrescine and some other aliphatic amines. The initial characterization of PuO(Rh) uncovered an intriguing feature: the enzyme appeared to contain only one noncovalently bound FAD cofactor per dimer. Here we show that this low FAD/protein ratio is the result of tight binding of ADP, thereby competing with FAD binding. MS analysis revealed that the enzyme is isolated as a mixture of dimers containing two molecules of FAD, two molecules ADP, or one FAD and one ADP molecule. In addition, based on a structural model of PuO(Rh) that was built using the crystal structure of human monoamine oxidase B (MAO-B), we constructed an active mutant enzyme, PuO(Rh) A394C, that contains covalently bound FAD. These findings show that the covalent FAD-protein linkage can be formed autocatalytically and hint to a new-found rationale for covalent flavinylation: covalent flavinylation may have evolved to prevent binding of ADP or related cellular compounds, which would prohibit formation of flavinylated and functional enzyme. PMID- 18678872 TI - Analysis of a membrane interacting region of herpes simplex virus type 1 glycoprotein H. AB - Glycoprotein H (gH) of herpes simplex virus type I (HSV-1) is involved in the complex mechanism of membrane fusion of the viral envelope with the host cell. Membrane interacting regions and potential fusion peptides have been identified in HSV-1 gH as well as glycoprotein B (gB). Because of the complex fusion mechanism of HSV-1, which requires four viral glycoproteins, and because there are only structural data for gB and glycoprotein D, many questions regarding the mechanism by which HSV-1 fuses its envelope with the host cell membrane remain unresolved. Previous studies have shown that peptides derived from certain regions of gH have the potential to interact with membranes, and based on these findings we have generated a set of peptides containing mutations in one of these domains, gH-(626-644), to investigate further the functional role of this region. Using a combination of biochemical, spectroscopic, and nuclear magnetic resonance techniques, we showed that the alpha-helical nature of this stretch of amino acids in gH is important for membrane interaction and that the aromatic residues, tryptophan and tyrosine, are critical for induction of fusion. PMID- 18678873 TI - Role of SUV3 helicase in maintaining mitochondrial homeostasis in human cells. AB - In yeast mitochondria, RNA degradation takes place through the coordinated activities of ySuv3 helicase and yDss1 exoribonuclease (mtEXO), whereas in bacteria, RNA is degraded via RNaseE, RhlB, PNPase, and enolase. Yeast lacking the Suv3 component of the mtEXO form petits and undergo a toxic accumulation of omega intron RNAs. Mammalian mitochondria resemble their prokaryotic origins by harboring a polyadenylation-dependent RNA degradation mechanism, but whether SUV3 participates in regulating RNA turnover in mammalian mitochondria is unclear. We found that lack of hSUV3 in mammalian cells subsequently yielded an accumulation of shortened polyadenylated mtRNA species and impaired mitochondrial protein synthesis. This suggests that SUV3 may serve in part as a component of an RNA degradosome, resembling its yeast ancestor. Reduction in the expression levels of oxidative phosphorylation components correlated with an increase in reactive oxygen species generation, whereas membrane potential and ATP production were decreased. These cumulative defects led to pleiotropic effects in mitochondria such as decreased mtDNA copy number and a shift in mitochondrial morphology from tubular to granular, which eventually manifests in cellular senescence or cell death. Thus, our results suggest that SUV3 is essential for maintaining proper mitochondrial function, likely through a conserved role in mitochondrial RNA regulation. PMID- 18678874 TI - Suppression of lipopolysaccharide-stimulated tumor necrosis factor-alpha production by adiponectin is mediated by transcriptional and post-transcriptional mechanisms. AB - Adiponectin is an adipokine with potent anti-inflammatory properties. Treatment of macrophages with adiponectin results in a suppression of lipopolysaccharide (LPS)-stimulated cytokine production. Here we investigated the transcriptional and post-transcriptional mechanisms by which adiponectin suppresses LPS stimulated tumor necrosis factor (TNF)-alpha production. Treatment of RAW 264.7 macrophages with LPS increased TNF-alpha promoter-driven luciferase activity (TNF alpha promoter/Luc activity) by 20-fold over basal. After culture with 1 mug/ml globular adiponectin (gAcrp) for 18 h, TNF-alpha promoter/Luc activity was increased even in the absence of LPS; further challenge with LPS only increased TNF-alpha promoter/Luc activity by 1.4-fold. Treatment with gAcrp decreased LPS stimulated ERK1/2 phosphorylation and IkappaB degradation and suppressed the ability of LPS to increase the DNA binding activity of Egr-1 and p65. gAcrp also suppressed LPS-mediated stabilization of TNF-alpha mRNA. In controls cells, the half-life of TNF-alpha mRNA was increased from approximately 30 min at base line to approximately 80 min in response to LPS. After treatment with gAcrp for 18 h, LPS failed to increase TNF-alpha mRNA stability. This gAcrp-mediated loss of stimulus-induced stabilization of TNF-alpha mRNA required the presence of the TNF alpha 3'-untranslated region and was associated with an increase in expression and RNA binding activity of tristetraprolin, an mRNA-binding protein that destabilizes TNF-alpha mRNA. In summary, these data characterize the complex transcriptional and post-transcriptional effects of gAcrp on LPS-stimulated TNF alpha expression in macrophages. gAcrp treatment profoundly suppressed the ability of LPS to increase TNF-alpha transcription and reduced the stimulus induced stabilization of TNF-alpha mRNA in response to LPS. PMID- 18678875 TI - A novel protein kinase A-independent, beta-arrestin-1-dependent signaling pathway for p38 mitogen-activated protein kinase activation by beta2-adrenergic receptors. AB - A growing body of evidence has demonstrated that p38 mitogen-activated protein kinase (MAPK) has a crucial role in various physiological and pathological processes mediated by beta(2)-adrenergic receptors (beta(2)-ARs). However, the detailed mechanism of beta(2)-ARs-induced p38 MAPK activation has not yet been fully defined. The present study demonstrates a novel kinetic model of p38 MAPK activation induced by beta(2)-ARs in human embryonic kidney 293A cells. The beta(2)-AR agonist isoproterenol induced a time-dependent biphasic phosphorylation of p38 MAPK: the early phase peaked at 10 min, and was followed by a delayed phase that appeared at 90 min and was sustained for 6 h. Interestingly, inhibition of the cAMP/protein kinase A (PKA) pathway failed to affect the early phosphorylation but abolished the delayed activation. By contrast, silencing of beta-arrestin-1 expression by small interfering RNA inhibited the early phase activation of p38 MAPK. Furthermore, the NADPH oxidase complex is a downstream target of beta-arrestin-1, as evidenced by the fact that isoproterenol-induced Rac1 activation was also suppressed by beta-arrestin-1 knockdown. In addition, early phase activation of p38 MAPK was prevented by inactivation of Rac1 and NADPH oxidase by pharmacological inhibitors, overexpression of a dominant negative mutant of Rac1, and p47(phox) knockdown by RNA interference. Of note, we demonstrated that only early activation of p38 MAPK is involved in isoproterenol-induced F-actin rearrangement. Collectively, these data suggest that the classic cAMP/PKA pathway is responsible for the delayed activation, whereas a beta-arrestin-1/Rac1/NADPH oxidase-dependent signaling is a heretofore unrecognized mechanism for beta(2)-AR-mediated early activation of p38 MAPK. PMID- 18678876 TI - Modulation of the regulatory activity of bacterial two-component systems by SlyA. AB - Activation of the transcriptional regulator SlyA by the PhoP/PhoQ two-component system controls intracellular expression of numerous factors influencing Salmonella virulence. By dissecting the SlyA regulon using stable isotope labeling with amino acids in cell culture analysis, we found that SlyA enhances overall transcription of PhoP-activated loci. This amplification of cellular responses to Mg2+ occurs when SlyA binds to the phoPQ promoter thereby activating phoP autoregulation via a positive feedback mechanism. SlyA footprints a DNA region located one helical turn upstream of the PhoP box, which overlaps the H-NS binding motif required for signal-dependent phoP repression in high Mg2+ conditions. Therefore, binding of SlyA likely antagonizes H-NS and facilitates the interaction of PhoP to its own promoter, subsequently activating the phoPQ operon. Establishment of this regulatory circuit allows SlyA to exert its effect on the PhoP/PhoQ system specifically in Salmonella, which may confer an additional transcriptional regulation. Thus, our results provide a molecular mechanism that determines SlyA-dependent activation of PhoP-regulated genes in modulating Salmonella virulence. Evidence from this study also suggests a function of SlyA as a mediator in signal transduction from the PhoP/PhoQ system to other bacterial two-component systems in Salmonella. PMID- 18678877 TI - Time-resolved mechanism of extracellular gate opening and substrate binding in a glutamate transporter. AB - Glutamate transporters, also referred to as excitatory amino acid transporters (EAATs), are membrane proteins that regulate glutamatergic signal transmission by clearing excess glutamate after its release at synapses. A structure-based understanding of their molecular mechanisms of function has been elusive until the recent determination of the x-ray structure of an archaeal transporter, Glt(Ph). Glt(Ph) exists as a trimer, with each subunit containing a core region that mediates substrate translocation. In the present study a series of molecular dynamics simulations have been conducted and analyzed in light of new experimental data on substrate binding properties of EAATs. The simulations provide for the first time a full atomic description of the time-resolved events that drive the recognition and binding of substrate. The core region of each subunit exhibits an intrinsic tendency to open the helical hairpin HP2 loop, the extracellular gate, within tens of nanoseconds exposing conserved polar residues that serve as attractors for substrate binding. The NMDGT motif on the partially unwound part of the transmembrane helix TM7 and the residues Asp-390 and Asp-394 on TM8 are also distinguished by their important role in substrate binding and close interaction with mediating water molecules and/or sodium ions. The simulations reveal a Na+ binding site comprised in part of Leu-303 on TM7 and Asp 405 on TM8 and support a role for sodium ions in stabilizing substrate-bound conformers. The functional importance of Leu-303 or its counterpart Leu-391 in human EAAT1 (hEAAT1) is confirmed by site-directed mutagenesis and Na+ dependence assays conducted with hEAAT1 mutants L391C and L391A. PMID- 18678878 TI - Neuroprotection of GluR5-containing kainate receptor activation against ischemic brain injury through decreasing tyrosine phosphorylation of N-methyl-D-aspartate receptors mediated by Src kinase. AB - Previous studies indicate that cerebral ischemia breaks the dynamic balance between excitatory and inhibitory inputs. The neural excitotoxicity induced by ionotropic glutamate receptors gain the upper hand during ischemia-reperfusion. In this paper, we investigate whether GluR5 (glutamate receptor 5)-containing kainate receptor activation could lead to a neuroprotective effect against ischemic brain injury and the related mechanism. The results showed that (RS)-2 amino-3-(3-hydroxy-5-tert-butylisoxazol-4-yl) propanoic acid (ATPA), a selective GluR5 agonist, could suppress Src tyrosine phosphorylation and interactions among N-methyl-D-aspartate (NMDA) receptor subunit 2A (NR2A), postsynaptic density protein 95 (PSD-95), and Src and then decrease NMDA receptor activation through attenuating tyrosine phosphorylation of NR2A and NR2B. More importantly, ATPA had a neuroprotective effect against ischemia-reperfusion-induced neuronal cell death in vivo. However, four separate drugs were found to abolish the effects of ATPA. These were selective GluR5 antagonist NS3763; GluR5 antisense oligodeoxynucleotides; CdCl(2), a broad spectrum blocker of voltage-gated calcium channels; and bicuculline, an antagonist of gamma-aminobutyric acid A (GABA(A)) receptor. GABA(A) receptor agonist muscimol could attenuate Src activation and interactions among NR2A, PSD-95 and Src, resulting the suppression of NMDA receptor tyrosine phosphorylation. Moreover, patch clamp recording proved that the activated GABA(A) receptor could inhibit NMDA receptor-mediated whole-cell currents. Taken together, the results suggest that during ischemia-reperfusion, activated GluR5 may facilitate Ca(2+)-dependent GABA release from interneurons. The released GABA can activate postsynaptic GABA(A) receptors, which then attenuates NMDA receptor tyrosine phosphorylation through inhibiting Src activation and disassembling the signaling module NR2A-PSD-95-Src. The final result of this process is that the pyramidal neurons are rescued from hyperexcitability. PMID- 18678879 TI - The apolipoprotein CIII enhancer regulates both extensive histone modification and intergenic transcription of human apolipoprotein AI/CIII/AIV genes but not apolipoprotein AV. AB - The apolipoprotein (apo) AI/CIII/AIV/AV cluster genes are expressed at different levels in the liver and intestine. The apoCIII enhancer, a common regulatory element, regulates the tissue-specific expression of apoAI, apoCIII, and apoAIV but not apoAV. To study this regulation at the chromatin level, the histone modifications and intergenic transcription in the human apoAI/CIII/AIV/AV cluster were investigated in HepG2 and Caco-2 cells and in the livers of transgenic mice carrying the human gene cluster constructs with or without the apoCIII enhancer. We found that both the promoters and the intergenic regions of the apoAI/CIII/AIV genes were hyperacetylated and formed an open subdomain that did not include the apoAV gene. Hepatic and intestinal intergenic transcripts were identified to transcribe bidirectionally with strand preferences along the cluster. The deletion of the apoCIII enhancer influenced both histone modification and intergenic transcription in the apoAI/CIII/AIV gene region. These results demonstrate that the apoCIII enhancer contributes to the maintenance of an active chromatin subdomain of the apoAI/CIII/AIV genes, but not apoAV. PMID- 18678880 TI - Evaluation of peak exposures in the dutch flour processing industry: implications for intervention strategies. AB - OBJECTIVES: To effectively decrease occupational exposure to flour dust and related allergens, detailed information on exposure determinants and effectiveness of control measures is essential. In this paper, we use personal real-time exposure measurements to get more insight into the relationship between specific work characteristics, including the use of control measures, and (peak) exposure to flour dust. The study has three objectives: (i) identify tasks and activities related to peak exposure, (ii) identify control measures and other important exposure determinants and (iii) assess the potential impact of these control measures on the (peak) exposure to flour dust. METHODS: A data set containing 82 real-time exposure measurements in combination with information from detailed observations was used to study the association between peak exposures and different tasks, activities and other determinants such as control measures. Descriptive statistics of peak exposure on job level were generated as well as information on contribution of task-specific peak exposures to time weighted average (TWA) exposure levels. Finally, we evaluated the efficacy of a variety of control measures on task exposure by comparing exposure levels of groups of workers with and without controls. RESULTS: In workers included in this study, >75% of TWA exposure is directly associated with peak exposures during a limited set of well-defined tasks/activities. The impact of a single task on population TWA exposure is generally limited (<40%). Worker behavior seems an important determinant in effective exposure control for many tasks. CONCLUSIONS: Data from real-time measurements provide important detailed information with respect to exposure determinants and control measures, not obtainable from conventional measurement studies focusing at TWA exposure. This information is essential to perform prospective impact assessments of intervention strategies on the populations' exposure distribution. PMID- 18678881 TI - Respiratory exposure to components of water-miscible metalworking fluids. AB - Water-miscible metalworking fluids (MWFs) are capable of causing respiratory symptoms and diseases. Recently, much emphasis has been put on developing new methods for assessing respiratory exposure to MWF emulsions. The air concentrations of ingredients and contaminants of MWF and inhalable dust were measured in 10 metal workshops in southern Finland. Oil mist was determined by infra red spectroscopy analysis after tetrachloroethylene extraction from the filter. Aldehydes were collected on Sep-Pak chemosorbents and analysed by liquid chromatography. Volatile organic compounds (VOCs) were collected on Tenax adsorbents and analysed by gas chromatography with mass spectrometric detection after thermal desorption. Endotoxins were collected on glass fibre filter and analysed by enzyme-based spectrophotometry, and viable microbes were collected on polycarbonate filter and cultured. Inhalable dust was collected on cellulose acetate filter and quantified gravimetrically. Associations between the different exposures were calculated with Spearman's correlations. The mean concentration of oil mist was 0.14 (range <0.010-0.60) mg m(-3). The mean total concentration of aldehydes was 0.095 (0.026-0.38) mg m(-3), with formaldehyde as the main aldehyde. The average total concentration of VOC was 1.9 (0.34-4.5) mg m(-3) consisting mainly of high-boiling aliphatic hydrocarbons. Several potential sensitizing chemicals such as terpenes were found in small quantities. The concentration of microbial contaminants was low. All the measured air concentrations were below the Finnish occupational exposure limits. The exposure in machine shops was quantitatively dominated by volatile compounds. Additional measurements of MWF components such as aldehydes, alkanolamines and VOCs are needed to get more information on the chemical composition of workshops' air. New air cleaning methods should be introduced, as oil mist separators are insufficient to clean the air of small molecular impurities. PMID- 18678882 TI - Increased 5-lipoxygenase immunoreactivity in the hippocampus of patients with Alzheimer's disease. AB - The proinflammatory enzyme 5-lipoxygenase (5-LOX) is upregulated in Alzheimer's disease (AD), but its localization and association with the hallmark lesions of the disease, beta-amyloid (Abeta) plaques and neurofibrillary tangles (NFTs), is unknown. This study examined the distribution and cellular localization of 5-LOX in the medial temporal lobe from AD and control subjects. The spatial relationship between 5-LOX immunoreactive structures and AD lesions was also examined. We report that, in AD subjects, 5-LOX immunoreactivity is elevated relative to controls, and its localization is dependent on the antibody-targeted portion of the 5-LOX amino acid sequence. Carboxy terminus-directed antibodies detected 5-LOX in glial cells and neurons, but less frequently in neurons with dystrophic (NFT) morphology. In contrast, immunoreactivity observed using 5-LOX amino terminus-directed antibodies was virtually absent in neurons and abundant in NFTs, neuritic plaques, and glia. Double-labeling studies showed a close association of 5-LOX-immunoreactive processes and glial cells with Abeta immunoreactive plaques and vasculature and also detected 5-LOX in tau immunoreactive and amyloid containing NFTs. Different immunolabeling patterns with antibodies against carboxy vs amino terminus of 5-LOX may be caused by post translational modifications of 5-LOX protein in Abeta plaques and NFTs. The relationship between elevated intracellular 5-LOX and hallmark AD pathological lesions provides further evidence that neuroinflammatory pathways contribute to the pathogenesis of AD. PMID- 18678883 TI - The heterozygous disproportionate micromelia (dmm) mouse: morphological changes in fetal cartilage precede postnatal dwarfism and compared with lethal homozygotes can explain the mild phenotype. AB - The disproportionate micromelia (Dmm) mouse has a mutation in the C-propeptide coding region of the Col2a1 gene that causes lethal dwarfism when homozygous (Dmm/Dmm) but causes only mild dwarfism observable approximately 1-week postpartum when heterozygous (Dmm/+). The purpose of this study was 2-fold: first, to analyze and quantify morphological changes that precede the expression of mild dwarfism in Dmm/+ animals, and second, to compare morphological alterations between Dmm/+ and Dmm/Dmm fetal cartilage that may correlate with the marked skeletal differences between mild and lethal dwarfism. Light and electron transmission microscopy were used to visualize structure of chondrocytes and extracellular matrix (ECM) of fetal rib cartilage. Both Dmm/+ and Dmm/Dmm fetal rib cartilage had significantly larger chondrocytes, greater cell density, and less ECM per unit area than +/+ littermates. Quantitative RT-PCR showed a decrease in aggrecan mRNA in Dmm/+ vs +/+ cartilage. Furthermore, the cytoplasm of chondrocytes in Dmm/+ and Dmm/Dmm cartilage was occupied by significantly more distended rough endoplasmic reticulum (RER) compared with wild-type chondrocytes. Fibril diameters and packing densities of +/+ and Dmm/+ cartilage were similar, but Dmm/Dmm cartilage showed thinner, sparsely distributed fibrils. These findings support the prevailing hypothesis that a C-propeptide mutation could interrupt the normal assembly and secretion of Type II procollagen trimers, resulting in a buildup of proalpha1(II) chains in the RER and a reduced rate of matrix synthesis. Thus, intracellular entrapment of proalpha1(II) seems to be primarily responsible for the dominant-negative effect of the Dmm mutation in the expression of dwarfism. PMID- 18678884 TI - The effect of passive movement on denervated soleus highlights a differential nerve control on SERCA and MyHC isoforms. AB - The sarco-endoplasmic reticulum Ca2+ ATP-ase (SERCA) and myosin heavy chain (MyHC) levels were measured in hindlimb-denervated and selectively denervated rat soleus muscles. Selective denervation allowed passive movement of the soleus, whereas hindlimb denervation rendered it to passivity. To minimize chronic effects, we followed the changes only for 2 weeks. Selective denervation resulted in less muscle atrophy, a faster slow-to-fast transition of MyHC isoforms, and less coordinated expressions of the slow vs fast isoforms of MyHC and SERCA. Generally, expression of the slow-twitch type SERCA2a was found to be less dependent, whereas the slow-twitch type MyHC1 was the most dependent on innervation. Our study shows that passive movement is able to ameliorate denervation-induced atrophy of the soleus and that it also accentuates the dyscoordination in the expression of the corresponding slow and fast isoforms of MyHC and SERCA. PMID- 18678885 TI - Emergence and robustness of a community discussion network on mercury contamination and health in the Brazilian Amazon. AB - Information exchanges, debates, and negotiations through community social networks are essential to ensure the sustainability of the development process initiated in participatory research. The authors analyze the structural properties and robustness of a discussion network about mercury issues in a community in the Brazilian Amazon involved in a participatory research aimed at reducing exposure to the pollutant. Most of the villagers are connected in a large network and are separated from other individuals by few intermediaries. The structure of the discussion network displays resilience to the random elimination of villagers but shows vulnerability to the removal of one villager who has been a long-term collaborator of the project. Although the network exhibits a structure likely to favor an efficient flow of information, results show that specific actions should be taken to stimulate the emergence of a pool of opinion leaders and increase the redundancy of discussion channels. PMID- 18678886 TI - Factors affecting the diffusion of solar water disinfection: a field study in Bolivia. AB - This study examines a broad array of theory-based factors derived from diffusion research that affect the current and intended use of solar water disinfection (SODIS), a simple, low-cost technology for treating drinking water at the household level. The perceived attributes of an innovation, the nature of the social system in which it is diffused, the extent of change agents' promotional efforts in diffusing it, and the nature of the communication channels used were operationalized by 16 variables. The aim of the study is to determine the influence of each factor and its predictive power. Eight areas in Bolivia were visited, and 644 families were interviewed on the basis of a structured questionnaire. Simultaneous multiple regression analysis showed that 9 of the 16 factors derived from diffusion research contributed significantly to predicting the current use of SODIS. The implications of the findings for customizing future SODIS diffusion activities are outlined. PMID- 18678887 TI - Demyelination arrest and remyelination induced by glatiramer acetate treatment of experimental autoimmune encephalomyelitis. AB - The interplay between demyelination and remyelination is critical in the progress of multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). In the present study, we explored the capacity of glatiramer acetate (GA, Copaxone) to affect the demyelination process and/or lead to remyelination in mice inflicted by chronic EAE, using both scanning electron microscopy and immunohistological methods. Spinal cords of untreated EAE mice revealed substantial demyelination accompanied by tissue destruction and axonal loss. In contrast, in spinal cords of GA-treated mice, in which treatment started concomitantly with disease induction (prevention), no pathology was observed. Moreover, when treatment was initiated after the appearance of clinical symptoms (suppression) or even in the chronic disease phase (delayed suppression) when substantial demyelination was already manifested, it resulted in a significant decrease in the pathological damage. Detection of oligodendrocyte progenitor cells (OPCs) expressing the NG2 or O4 markers via colocalization with the proliferation marker BrdU indicated their elevated levels in spinal cords of GA treated mice. The mode of action of GA in this system is attributed to increased proliferation, differentiation, and survival of OPCs along the oligodendroglial maturation cascade and their recruitment into injury sites, thus enhancing repair processes in situ. PMID- 18678888 TI - An immunotoxin with greatly reduced immunogenicity by identification and removal of B cell epitopes. AB - Recombinant immunotoxins are hybrid proteins composed of an Fv that binds to a tumor antigen fused to a bacterial or plant toxin. Immunotoxin BL22 targets CD22 positive malignancies and is composed of an anti-CD22 Fv fused to a 38-kDa fragment of Pseudomonas exotoxin A (PE38). BL22 has produced many complete remissions in drug-resistant Hairy cell leukemia, where many treatment cycles can be given, because neutralizing antibodies do not form. In marked contrast, only minor responses have been observed in trials with immunotoxins targeting solid tumors, because only a single treatment cycle can be given before antibodies develop. To allow more treatment cycles and increase efficacy, we have produced a less immunogenic immunotoxin by identifying and eliminating most of the B cell epitopes on PE38. This was accomplished by mutation of specific large hydrophilic amino acids (Arg, Gln, Glu, Lys) to Ala, Ser, or Gly. The new immunotoxin (HA22 8X) is significantly less immunogenic in three strains of mice, yet retains full cytotoxic and anti-tumor activities. Elimination of B-cell epitopes is a promising approach to the production of less immunogenic proteins for therapeutic purposes. PMID- 18678889 TI - Y-chromosomal evidence of a pastoralist migration through Tanzania to southern Africa. AB - Although geneticists have extensively debated the mode by which agriculture diffused from the Near East to Europe, they have not directly examined similar agropastoral diffusions in Africa. It is unclear, for example, whether early instances of sheep, cows, pottery, and other traits of the pastoralist package were transmitted to southern Africa by demic or cultural diffusion. Here, we report a newly discovered Y-chromosome-specific polymorphism that defines haplogroup E3b1f-M293. This polymorphism reveals the monophyletic relationship of the majority of haplotypes of a previously paraphyletic clade, E3b1-M35*, that is widespread in Africa and southern Europe. To elucidate the history of the E3b1f haplogroup, we analyzed this haplogroup in 13 populations from southern and eastern Africa. The geographic distribution of the E3b1f haplogroup, in association with the microsatellite diversity estimates for populations, is consistent with an expansion through Tanzania to southern-central Africa. The data suggest this dispersal was independent of the migration of Bantu-speaking peoples along a similar route. Instead, the phylogeography and microsatellite diversity of the E3b1f lineage correlate with the arrival of the pastoralist economy in southern Africa. Our Y-chromosomal evidence supports a demic diffusion model of pastoralism from eastern to southern Africa approximately 2,000 years ago. PMID- 18678890 TI - MDC1 regulates intra-S-phase checkpoint by targeting NBS1 to DNA double-strand breaks. AB - The product of the Nijmegen breakage syndrome gene (NBS1) plays crucial roles in DNA damage response through its association with many proteins, including MRE11 and RAD50. However, it remains to be determined exactly how NBS1 accumulates at or near DNA double-strand breaks. Here we report that MDC1 directly binds to NBS1 and targets NBS1 to the sites of DNA damage. The MDC1-NBS1 interaction occurs through a specific region (residues 200-420) of MDC1, which contains multiple consensus casein kinase 2 (CK2) phosphorylation sites. In addition, this interaction requires both the forkhead-associated (FHA) and tandem BRCA1 C terminal (BRCT) domains of NBS1. Disruption of the MDC1-NBS1 interaction results in failure of NBS1 accumulation at DNA double-strand breaks and impairment of intra-S checkpoint activation. These studies provide important mechanistic insights as to how MDC1 regulates NBS1 and the intra-S-phase checkpoint in response to DNA damage. PMID- 18678891 TI - Influence of nonlinear electrostatics on transfer energies between liquid phases: charge burial is far less expensive than Born model. AB - The widely used Born model describes the electrostatic response of continuous media using static dielectric constants. However, when applied to a liquid environment, a comparison of Born model predictions with experimental values (e.g., transfer free energies and pK(a) shifts) found that agreement is only achieved by using physically unrealistic dielectric constants for proteins, lipids, etc., and/or equally unrealistic atomic radii. This leads to questions concerning the physical origins for this failure of the Born model. We partially resolve this question by applying the Langevin-Debye (LD) model of a continuous distribution of point, polarizable dipoles, a model that contains an added dependence of the electrostatic response on the solvent's optical dielectric constant and both gas- and liquid-phase dipole moments, features absent in the Born model to which the LD model reduces for weak fields. The LD model is applied to simple representations of three biologically relevant systems: (i) globular proteins, (ii) lipid bilayers, and (iii) membrane proteins. The linear Born treatment greatly overestimates both the self-energy and the transfer free energy from water to hydrophobic environments (e.g., a protein interior). By using the experimental dielectric constant, the energy cost of charge burial in either globular or membrane proteins of the Born model is reduced by almost 50% with the nonlinear theory as is the pK(a) shift, and the shifts agree well with experimental trends. PMID- 18678892 TI - Cost-effective priorities for global mammal conservation. AB - Global biodiversity priority setting underpins the strategic allocation of conservation funds. In identifying the first comprehensive set of global priority areas for mammals, Ceballos et al. [Ceballos G, Ehrlich PR, Soberon J, Salazar I, Fay JP (2005) Science 309:603-607] found much potential for conflict between conservation and agricultural human activity. This is not surprising because, like other global priority-setting approaches, they set priorities without socioeconomic objectives. Here we present a priority-setting framework that seeks to minimize the conflicts and opportunity costs of meeting conservation goals. We use it to derive a new set of priority areas for investment in mammal conservation based on (i) agricultural opportunity cost and biodiversity importance, (ii) current levels of international funding, and (iii) degree of threat. Our approach achieves the same biodiversity outcomes as Ceballos et al.'s while reducing the opportunity costs and conflicts with agricultural human activity by up to 50%. We uncover shortfalls in the allocation of conservation funds in many threatened priority areas, highlighting a global conservation challenge. PMID- 18678893 TI - Hydraulic integration and shrub growth form linked across continental aridity gradients. AB - Both engineered hydraulic systems and plant hydraulic systems are protected against failure by resistance, reparability, and redundancy. A basic rule of reliability engineering is that the level of independent redundancy should increase with increasing risk of fatal system failure. Here we show that hydraulic systems of plants function as predicted by this engineering rule. Hydraulic systems of shrubs sampled along two transcontinental aridity gradients changed with increasing aridity from highly integrated to independently redundant modular designs. Shrubs in humid environments tend to be hydraulically integrated, with single, round basal stems, whereas dryland shrubs typically have modular hydraulic systems and multiple, segmented basal stems. Modularity is achieved anatomically at the vessel-network scale or developmentally at the whole plant scale through asymmetric secondary growth, which results in a semiclonal or clonal shrub growth form that appears to be ubiquitous in global deserts. PMID- 18678894 TI - Capillarity-like growth of protein folding nuclei. AB - A full structural description of transition state ensembles in protein folding includes the specificity of the ordered residues composing the folding nucleus as well as spatial density. To our knowledge, the spatial properties of the folding nucleus and interface of specific nuclei have yet to receive significant attention. We analyze folding routes predicted by a variational model in terms of a generalized formalism of the capillarity scaling theory that assumes the volume of the folded core of the nucleus grows with chain length as V(f) approximately N(3nu). For 27 two-state proteins studied, the scaling exponent nu ranges from 0.2 to 0.45 with an average of 0.33. This average value corresponds to packing of rigid objects, although generally the effective monomer size in the folded core is larger than the corresponding volume per particle in the native-state ensemble. That is, on average, the folded core of the nucleus is found to be relatively diffuse. We also study the growth of the folding nucleus and interface along the folding route in terms of the density or packing fraction. The evolution of the folded core and interface regions can be classified into three patterns of growth depending on how the growth of the folded core is balanced by changes in density of the interface. Finally, we quantify the diffuse versus polarized structure of the critical nucleus through direct calculation of the packing fraction of the folded core and interface regions. Our results support the general picture of describing protein folding as the capillarity-like growth of folding nuclei. PMID- 18678895 TI - Induction of angiogenesis in tissue-engineered scaffolds designed for bone repair: a combined gene therapy-cell transplantation approach. AB - One of the fundamental principles underlying tissue engineering approaches is that newly formed tissue must maintain sufficient vascularization to support its growth. Efforts to induce vascular growth into tissue-engineered scaffolds have recently been dedicated to developing novel strategies to deliver specific biological factors that direct the recruitment of endothelial cell (EC) progenitors and their differentiation. The challenge, however, lies in orchestration of the cells, appropriate biological factors, and optimal factor doses. This study reports an approach as a step forward to resolving this dilemma by combining an ex vivo gene transfer strategy and EC transplantation. The utility of this approach was evaluated by using 3D poly(lactide-co-glycolide) (PLAGA) sintered microsphere scaffolds for bone tissue engineering applications. Our goal was achieved by isolation and transfection of adipose-derived stromal cells (ADSCs) with adenovirus encoding the cDNA of VEGF. We demonstrated that the combination of VEGF releasing ADSCs and ECs results in marked vascular growth within PLAGA scaffolds. We thereby delineate the potential of ADSCs to promote vascular growth into biomaterials. PMID- 18678896 TI - A triallelic system of S5 is a major regulator of the reproductive barrier and compatibility of indica-japonica hybrids in rice. AB - Hybrid sterility is a major form of postzygotic reproductive isolation. Although reproductive isolation has been a key issue in evolutionary biology for many decades in a wide range of organisms, only very recently a few genes for reproductive isolation were identified. The Asian cultivated rice (Oryza sativa L.) is divided into two subspecies, indica and japonica. Hybrids between indica and japonica varieties are usually highly sterile. A special group of rice germplasm, referred to as wide-compatibility varieties, is able to produce highly fertile hybrids when crossed to both indica and japonica. In this study, we cloned S5, a major locus for indica-japonica hybrid sterility and wide compatibility, using a map-based cloning approach. We show that S5 encodes an aspartic protease conditioning embryo-sac fertility. The indica (S5-i) and japonica (S5-j) alleles differ by two nucleotides. The wide compatibility gene (S5-n) has a large deletion in the N terminus of the predicted S5 protein, causing subcellular mislocalization of the protein, and thus is presumably nonfunctional. This triallelic system has a profound implication in the evolution and artificial breeding of cultivated rice. Genetic differentiation between indica and japonica would have been enforced because of the reproductive barrier caused by S5-i and S5-j, and species coherence would have been maintained by gene flow enabled by the wide compatibility gene. PMID- 18678897 TI - GABAA transmission is a critical step in the process of triggering homeostatic increases in quantal amplitude. AB - When activity levels are altered over days, a network of cells is capable of recognizing this perturbation and triggering several distinct compensatory changes that should help to recover and maintain the original activity levels homeostatically. One feature commonly observed after activity blockade has been a compensatory increase in excitatory quantal amplitude. The sensing machinery that detects altered activity levels is a central focus of the field currently, but thus far it has been elusive. The vast majority of studies that reduce network activity also reduce neurotransmission. We address the possibility that reduced neurotransmission can trigger increases in quantal amplitude. In this work, we blocked glutamatergic or GABA(A) transmission in ovo for 2 days while maintaining relatively normal network activity. We found that reducing GABA(A) transmission triggered compensatory increases in both GABA and AMPA quantal amplitude in embryonic spinal motoneurons. Glutamatergic blockade had no effect on quantal amplitude. Therefore, GABA binding to the GABA(A) receptor appears to be a critical step in the sensing machinery for homeostatic synaptic plasticity. The findings suggest that homeostatic increases in quantal amplitude may normally be triggered by reduced levels of activity, which are sensed in the developing spinal cord by GABA, via the GABA(A) receptor. Therefore, GABA appears to be serving as a proxy for activity levels. PMID- 18678898 TI - CXCR4 antagonism increases T cell trafficking in the central nervous system and improves survival from West Nile virus encephalitis. AB - The migration of lymphocytes into the CNS during viral encephalitis is hindered by the blood-brain barrier (BBB) such that most infiltrating cells remain localized to perivascular spaces. This sequestration of leukocytes away from the parenchyma is believed to protect the CNS from immunopathologic injury. Infections of the CNS with highly cytopathic neurotropic viruses, such as West Nile virus (WNV), however, require the parenchymal penetration of T lymphocytes for virus clearance and survival, suggesting that perivascular localization might hinder antiviral immune responses during WNV encephalitis. Using human and murine brain specimens from individuals with WNV encephalitis, we evaluated the expression of CXCL12 and its receptor, CXCR4, at the BBB and tested the hypothesis that inhibition of CXCR4 would promote T lymphocyte entry into the CNS parenchyma and increase viral clearance. Antagonism of CXCR4 significantly improved survival from lethal infection through enhanced intraparenchymal migration of WNV-specific CD8(+) T cells within the brain, leading to reduced viral loads and, surprisingly, decreased immunopathology at this site. The benefits of enhanced CD8(+) T cell infiltration suggest that pharmacologic targeting of CXCR4 may have therapeutic utility for the treatment of acute viral infections of the CNS. PMID- 18678899 TI - The Fezf2-Ctip2 genetic pathway regulates the fate choice of subcortical projection neurons in the developing cerebral cortex. AB - Pyramidal neurons in the deep layers of the cerebral cortex can be classified into two major classes: callosal projection neurons and long-range subcortical neurons. We and others have shown that a gene expressed specifically by subcortical projection neurons, Fezf2, is required for the formation of axonal projections to the spinal cord, tectum, and pons. Here, we report that Fezf2 regulates a decision between subcortical vs. callosal projection neuron fates. Fezf2(-/-) neurons adopt the fate of callosal projection neurons as assessed by their axonal projections, electrophysiological properties, and acquisition of Satb2 expression. Ctip2 is a major downstream effector of Fezf2 in regulating the extension of axons toward subcortical targets and can rescue the axonal phenotype of Fezf2 mutants. When ectopically expressed, either Fezf2 or Ctip2 can alter the axonal targeting of corticocortical projection neurons and cause them to project to subcortical targets, although Fezf2 can promote a subcortical projection neuron fate in the absence of Ctip2 expression. PMID- 18678900 TI - Dynamic energy landscape view of coupled binding and protein conformational change: induced-fit versus population-shift mechanisms. AB - Allostery, the coupling between ligand binding and protein conformational change, is the heart of biological network and it has often been explained by two representative models, the induced-fit and the population-shift models. Here, we clarified for what systems one model fits better than the other by performing molecular simulations of coupled binding and conformational change. Based on the dynamic energy landscape view, we developed an implicit ligand-binding model combined with the double-basin Hamiltonian that describes conformational change. From model simulations performed for a broad range of parameters, we uncovered that each of the two models has its own range of applicability, stronger and longer-ranged interaction between ligand and protein favors the induced-fit model, and weaker and shorter-ranged interaction leads to the population-shift model. We further postulate that the protein binding to small ligand tends to proceed via the population-shift model, whereas the protein docking to macromolecules such as DNA tends to fit the induced-fit model. PMID- 18678901 TI - A high-throughput screening strategy identifies cardiotonic steroids as alternative splicing modulators. AB - Alternative splicing has emerged as a promising therapeutic target in a number of human disorders. However, the discovery of compounds that target the splicing reaction has been hindered by the lack of suitable high-throughput screening assays. Conversely, the effects of known drugs on the splicing reaction are mostly unclear and not routinely assessed. We have developed a two-color fluorescent reporter for cellular assays of exon inclusion that can accommodate nearly any cassette exon and minimizes interfering effects from changes in transcription and translation. We used microtubule-associated protein tau (MAPT) exon 10, whose missplicing causes frontotemporal dementia, to test the reporter in screening libraries of known bioactive compounds. These screens yielded several compounds that alter the splicing of the exon, both in the reporter and in the endogenous MAPT mRNA. One compound, digoxin, has long been used in the treatment of heart failure, but was not known to modulate splicing. The positive compounds target different signal transduction pathways, and microarray analysis shows that each compound affects the splicing of a different set of exons in addition to MAPT exon 10. Our results identify currently prescribed cardiotonic steroids as modulators of alternative splicing and demonstrate the feasibility of screening for drugs that alter exon inclusion. PMID- 18678902 TI - Apoptosis leads to a degradation of vital components of active nuclear transport and a dissociation of the nuclear lamina. AB - Apoptosis, a physiologically critical process, is characterized by a destruction of the cell after sequential degradation of key cellular components. Here, we set out to explore the fate of the physiologically indispensable nuclear envelope (NE) in this process. The NE mediates the critical nucleocytoplasmic transport through nuclear pore complexes (NPCs). In addition, the NE is involved in gene expression and contributes significantly to the overall structure and mechanical stability of the cell nucleus through the nuclear lamina, which underlies the entire nucleoplasmic face of the NE and thereby interconnects the NPCs, the NE, and the genomic material. Using the nano-imaging and mechanical probing approach atomic force microscopy (AFM) and biochemical methods, we unveiled the fate of the NE during apoptosis. The doomed NE sustains a degradation of both the mediators of the critical selective nucleocytoplasmic transport, namely NPC cytoplasmic filaments and basket, and the nuclear lamina. These observations are paralleled by marked softening and destabilization of the NE and the detection of vesicle-like nuclear fragments. We conclude that destruction of the cell nucleus during apoptosis proceeds in a strategic fashion. Degradation of NPC cytoplasmic filaments and basket shuts down the critical selective nucleocytoplasmic cross talk. Degradation of the nuclear lamina disrupts the pivotal connection between the NE and the chromatin, breaks up the overall nuclear architecture, and softens the NE, thereby enabling the formation of nuclear fragments at later stages of apoptosis. PMID- 18678903 TI - Mid-Miocene cooling and the extinction of tundra in continental Antarctica. AB - A major obstacle in understanding the evolution of Cenozoic climate has been the lack of well dated terrestrial evidence from high-latitude, glaciated regions. Here, we report the discovery of exceptionally well preserved fossils of lacustrine and terrestrial organisms from the McMurdo Dry Valleys sector of the Transantarctic Mountains for which we have established a precise radiometric chronology. The fossils, which include diatoms, palynomorphs, mosses, ostracodes, and insects, represent the last vestige of a tundra community that inhabited the mountains before stepped cooling that first brought a full polar climate to Antarctica. Paleoecological analyses, (40)Ar/(39)Ar analyses of associated ash fall, and climate inferences from glaciological modeling together suggest that mean summer temperatures in the region cooled by at least 8 degrees C between 14.07 +/- 0.05 Ma and 13.85 +/- 0.03 Ma. These results provide novel constraints for the timing and amplitude of middle-Miocene cooling in Antarctica and reveal the ecological legacy of this global climate transition. PMID- 18678904 TI - Vibrational vs. translational energy in promoting a prototype metal-hydrocarbon insertion reaction. AB - The reaction Y + CH(4) --> HYCH(3) --> YCH(2) + H(2) is initiated by C-H insertion involving a 20 +/- 3 kcal/mol potential energy barrier. The reaction is studied in crossed molecular beams under two different conditions with nearly the same total energy. One experiment is carried out at a collision energy of 15.1 kcal/mol with one quantum of CH(4) antisymmetric (nu(3)) stretching vibrational excitation (8.63 kcal/mol), the other at a collision energy of 23.8 kcal/mol. The reaction cross-section for C-H stretch excited methane (sigma(s)) is found to be at least a factor of 2.2 times larger than for ground-state methane (sigma(g)) at the same total energy. PMID- 18678905 TI - Spectroscopic identification and stability of the intermediate in the OH + HONO2 reaction. AB - The reaction of nitric acid with the hydroxyl radical influences the residence time of HONO(2) in the lower atmosphere. Prior studies [Brown SS, Burkholder JB, Talukdar RK, Ravishankara AR (2001) J Phys Chem A 105:1605-1614] have revealed unusual kinetic behavior for this reaction, including a negative temperature dependence, a complex pressure dependence, and an overall reaction rate strongly affected by isotopic substitution. This behavior suggested that the reaction occurs through an intermediate, theoretically predicted to be a hydrogen-bonded OH-HONO(2) complex in a six-membered ring-like configuration. In this study, the intermediate is generated directly by the association of photolytically generated OH radicals with HONO(2) and stabilized in a pulsed supersonic expansion. Infrared action spectroscopy is used to identify the intermediate by the OH radical stretch (nu(1)) and OH stretch of nitric acid (nu(2)) in the OH-HONO(2) complex. Two vibrational features are attributed to OH-HONO(2): a rotationally structured nu(1) band at 3516.8 cm(-1) and an extensively broadened nu(2) feature at 3260 cm(-1), both shifted from their respective monomers. These same transitions are identified for OD-DONO(2). Assignments of the features are based on their vibrational frequencies, analysis of rotational band structure, and comparison with complementary high level ab initio calculations. In addition, the OH (v = 0) product state distributions resulting from nu(1) and nu(2) excitation are used to determine the binding energy of OH-HONO(2), D(0) 2 years in a monkeypox virus challenge model. If variola virus were used as a biological weapon, however, the ability of a vaccine to quickly induce immunity would be essential. Here, we demonstrate more rapid immune responses after a single vaccination with MVA compared to the licensed Dryvax vaccine. To determine the kinetics of protection of the two vaccines, macaques were challenged intravenously with monkeypox virus at 4, 6, 10, and 30 days after immunization. At 6 or more days after vaccination with MVA or Dryvax, the monkeys were clinically protected (except for 1 of 16 animals vaccinated with MVA), although viral loads and number of skin lesions were generally higher in the MVA vaccinated group. With only 4 days between immunization and intravenous challenge, however, MVA still protected whereas Dryvax failed. Protection correlated with the more rapid immune response to MVA compared to Dryvax, which may be related to the higher dose of MVA that can be tolerated safely. PMID- 18678912 TI - Four enzymes define the incorporation of coenzyme A in thienamycin biosynthesis. AB - The enzymatic activities of three proteins encoded by the thienamycin gene cluster of Streptomyces cattleya (ThnR, ThnH, and ThnT) have been shown to incrementally cleave CoA to afford the active side-chain component of the beta lactam antibiotic thienamycin. These results supersede proposals based on earlier radiochemical incorporation experiments. For 20 years it has been thought that cysteine was directly incorporated into the antibiotic. Specific, stepwise truncation of CoA to 4-phosphopantetheine, pantetheine, and finally cysteamine was observed with ThnR, ThnH, and ThnT, respectively, in a series of coupled enzymatic assays. Pantetheinylated carbapenams were synthesized to address possible thienamycin biosynthetic intermediates and were shown to be effective substrates for the pantetheine-cleaving enzyme ThnT. Finally, a fourth gene, thnF, was shown to encode a protein capable of N-acetylating a model compound containing cysteamine in the presence of acetyl-CoA, consistent with the production of the S. cattleya cometabolite, N-acetylthienamycin. Taken together, these four enzymes are proposed to siphon CoA from primary metabolism to create the side chains for the predominant S. cattleya carbapenems, thienamycin and N acetylthienamycin, in a process likely to be general for the broader class of these antibiotics. PMID- 18678913 TI - Pharmacologic doses of ascorbate act as a prooxidant and decrease growth of aggressive tumor xenografts in mice. AB - Ascorbic acid is an essential nutrient commonly regarded as an antioxidant. In this study, we showed that ascorbate at pharmacologic concentrations was a prooxidant, generating hydrogen-peroxide-dependent cytotoxicity toward a variety of cancer cells in vitro without adversely affecting normal cells. To test this action in vivo, normal oral tight control was bypassed by parenteral ascorbate administration. Real-time microdialysis sampling in mice bearing glioblastoma xenografts showed that a single pharmacologic dose of ascorbate produced sustained ascorbate radical and hydrogen peroxide formation selectively within interstitial fluids of tumors but not in blood. Moreover, a regimen of daily pharmacologic ascorbate treatment significantly decreased growth rates of ovarian (P < 0.005), pancreatic (P < 0.05), and glioblastoma (P < 0.001) tumors established in mice. Similar pharmacologic concentrations were readily achieved in humans given ascorbate intravenously. These data suggest that ascorbate as a prodrug may have benefits in cancers with poor prognosis and limited therapeutic options. PMID- 18678914 TI - Pitx2 regulates gonad morphogenesis. AB - Organ shape and size, and, ultimately, organ function, relate in part to the cell and tissue spatial arrangement that takes place during embryonic development. Despite great advances in the genetic regulatory networks responsible for tissue and organ development, it is not yet clearly understood how specific gene functions are linked to the specific morphogenetic processes underlying the internal organ asymmetries found in vertebrate animals. During female chick embryogenesis, and in contrast to males where both testes develop symmetrically, asymmetrical gonad morphogenesis results in only one functional ovary. The disposition of paired organs along the left-right body axis has been shown to be regulated by the activity of the homeobox containing gene pitx2. We have found that pitx2 regulates cell adhesion, affinity, and cell recognition events in the developing gonad primordium epithelia. This in turn not only allows for proper somatic development of the gonad cortex but also permits the proliferation and differentiation of primordial germ cells. We illustrate how Pitx2 activity directs asymmetrical gonad morphogenesis by controlling mitotic spindle orientation of the developing gonad cortex and how, by modulating cyclinD1 expression during asymmetric ovarian development, Pitx2 appears to control gonad organ size. All together our observations indicate that the effects elicited by Pitx2 during the development of the female chick ovary are critical for cell topology, growth, fate, and ultimately organ morphogenesis and function. PMID- 18678915 TI - Novelties of the flowering plant pollen tube underlie diversification of a key life history stage. AB - The origin and rapid diversification of flowering plants has puzzled evolutionary biologists, dating back to Charles Darwin. Since that time a number of key life history and morphological traits have been proposed as developmental correlates of the extraordinary diversity and ecological success of angiosperms. Here, I identify several innovations that were fundamental to the evolutionary lability of angiosperm reproduction, and hence to their diversification. In gymnosperms pollen reception must be near the egg largely because sperm swim or are transported by pollen tubes that grow at very slow rates (< approximately 20 microm/h). In contrast, pollen tube growth rates of taxa in ancient angiosperm lineages (Amborella, Nuphar, and Austrobaileya) range from approximately 80 to 600 microm/h. Comparative analyses point to accelerated pollen tube growth rate as a critical innovation that preceded the origin of the true closed carpel, long styles, multiseeded ovaries, and, in monocots and eudicots, much faster pollen tube growth rates. Ancient angiosperm pollen tubes all have callosic walls and callose plugs (in contrast, no gymnosperms have these features). The early association of the callose-walled growth pattern with accelerated pollen tube growth rate underlies a striking repeated pattern of faster and longer-distance pollen tube growth often within solid pathways in phylogenetically derived angiosperms. Pollen tube innovations are a key component of the spectacular diversification of carpel (flower and fruit) form and reproductive cycles in flowering plants. PMID- 18678916 TI - Deciphering the genetic determinants for aerobic nicotinic acid degradation: the nic cluster from Pseudomonas putida KT2440. AB - The aerobic catabolism of nicotinic acid (NA) is considered a model system for degradation of N-heterocyclic aromatic compounds, some of which are major environmental pollutants; however, the complete set of genes as well as the structural-functional relationships of most of the enzymes involved in this process are still unknown. We have characterized a gene cluster (nic genes) from Pseudomonas putida KT2440 responsible for the aerobic NA degradation in this bacterium and when expressed in heterologous hosts. The biochemistry of the NA degradation through the formation of 2,5-dihydroxypyridine and maleamic acid has been revisited, and some gene products become the prototype of new types of enzymes with unprecedented molecular architectures. Thus, the initial hydroxylation of NA is catalyzed by a two-component hydroxylase (NicAB) that constitutes the first member of the xanthine dehydrogenase family whose electron transport chain to molecular oxygen includes a cytochrome c domain. The Fe(2+) dependent dioxygenase (NicX) converts 2,5-dihydroxypyridine into N-formylmaleamic acid, and it becomes the founding member of a new family of extradiol ring cleavage dioxygenases. Further conversion of N-formylmaleamic acid to formic and maleamic acid is catalyzed by the NicD protein, the only deformylase described so far whose catalytic triad is similar to that of some members of the alpha/beta hydrolase fold superfamily. This work allows exploration of the existence of orthologous gene clusters in saprophytic bacteria and some pathogens, where they might stimulate studies on their role in virulence, and it provides a framework to develop new biotechnological processes for detoxification/biotransformation of N-heterocyclic aromatic compounds. PMID- 18678917 TI - Deletion of the mouse RegIIIbeta (Reg2) gene disrupts ciliary neurotrophic factor signaling and delays myelination of mouse cranial motor neurons. AB - A large number of cytokines and growth factors support the development and subsequent maintenance of postnatal motor neurons. RegIIIbeta, also known as Reg2 in rat and HIP/PAP1 in humans, is a member of a family of growth factors found in many areas of the body and previously shown to play an important role in both the development and regeneration of subsets of motor neurons. It has been suggested that RegIIIbeta expressed by motor neurons is both an obligatory intermediate in the downstream signaling of the leukemia inhibitory factor/ciliary neurotrophic factor (CNTF) family of cytokines, maintaining the integrity of motor neurons during development, as well as a powerful influence on Schwann cell growth during regeneration of the peripheral nerve. Here we report that in mice with a deletion of the RegIIIbeta gene, motor neuron survival was unaffected up to 28 weeks after birth. However, there was no CNTF-mediated rescue of neonatal facial motor neurons after axotomy in KO animals when compared with wild-type. In mice, RegIIIbeta positive motor neurons are concentrated in cranial motor nuclei that are involved in the patterning of swallowing and suckling. We found that suckling was impaired in RegIIIbeta KO mice and correlated this with a significant delay in myelination of the hypoglossal nerve. In summary, we propose that RegIIIbeta has an important role to play in the developmental fine-tuning of neonatal motor behaviors mediating the response to peripherally derived cytokines and growth factors and regulating the myelination of motor axons. PMID- 18678918 TI - Ion permeation through a Cl--selective channel designed from a CLC Cl-/H+ exchanger. AB - The CLC family of Cl(-)-transporting proteins includes both Cl(-) channels and Cl(-)/H(+) exchange transporters. CLC-ec1, a structurally known bacterial homolog of the transporter subclass, exchanges two Cl(-) ions per proton with strict, obligatory stoichiometry. Point mutations at two residues, Glu(148) and Tyr(445), are known to impair H(+) movement while preserving Cl(-) transport. In the x-ray crystal structure of CLC-ec1, these residues form putative "gates" flanking an ion-binding region. In mutants with both of the gate-forming side chains reduced in size, H(+) transport is abolished, and unitary Cl(-) transport rates are greatly increased, well above values expected for transporter mechanisms. Cl(-) transport rates increase as side-chain volume at these positions is decreased. The crystal structure of a doubly ungated mutant shows a narrow conduit traversing the entire protein transmembrane width. These characteristics suggest that Cl(-) flux through uncoupled, ungated CLC-ec1 occurs via a channel-like electrodiffusion mechanism rather than an alternating-exposure conformational cycle that has been rendered proton-independent by the gate mutations. PMID- 18678920 TI - Knowledge and attitudes of criminal justice professionals in relation to Fetal Alcohol Spectrum Disorder. AB - Results of a provincial survey of Judges and Crown Prosecutors to determine specifically, their attitudes, knowledge, behaviors and training needs related to Fetal Alcohol Spectrum Disorders. In general, the survey results suggest that while aware of some aspects of FASD, Judges and Prosecutors both desire and need more education and training to support them in their work with individuals with FASD who come into conflict with the law. The findings also suggest that access to accurate and timely assessment and diagnoses of FASD would be beneficial. Survey findings point to the need for specific action to improve the ability of Judges and Prosecutors to recognize and to work with people affected by FASD in the Criminal Justice System. The results further indicate the need for changes and improvements in several areas regarding legal policy issues, research, and professional education and practice. PMID- 18678919 TI - Allosteric modulation of the muscarinic M4 receptor as an approach to treating schizophrenia. AB - Current antipsychotics provide symptomatic relief for patients suffering from schizophrenia and related psychoses; however, their effectiveness is variable and many patients discontinue treatment due to side effects. Although the etiology of schizophrenia is still unclear, a leading hypothesis implicates an imbalanced dopaminergic system. Muscarinic acetylcholine (ACh) receptors regulate dopamine levels in key areas of the brain involved in psychosis, with the M(4) subtype emerging as a key regulator of dopaminergic hyperactivity. Unfortunately, no selective small molecule tools exist to provide pharmacological validation of this hypothesis. Here, we describe the discovery of a small molecule modulator, LY2033298, that is highly selective for human M(4) receptors by virtue of targeting an allosteric site on this receptor. Pharmacological assays confirmed the selectivity of LY2033298 for the M(4) receptor and revealed the highest degree of positive allosteric enhancement of ACh potency thus far identified. Radioligand binding assays also show this compound to directly potentiate agonist binding while having minimal effects on antagonist binding. Mutational analysis identified a key amino acid (D(432)) in the third extracellular loop of the human M(4) receptor to be critical for selectivity and agonist potentiation by LY2033298. Importantly, LY2033298 was active in animal models predictive of clinical antipsychotic drug efficacy indicating its potential use as a first-in class, selective, allosteric muscarinic antipsychotic agent. PMID- 18678921 TI - Cross-sectional silica exposure measurements at two Zambian copper mines of Nkana and Mufulira. AB - We measured the quartz content of 20 bulk settled dust and 200 respirable dust samples in a cross-sectional dust exposure assessment that is part of an epidemiological study to ascertain the risk of nonmalignant respiratory diseases among Zambian copper miners. Dust samples were collected from the copper mines of Mopani Copper Mine plc (Mufulira and Nkana Mines). Analytical measurements employed NIOSH Method 0600 for gravimetric analysis of respirable dust and NIOSH Method 7500 for quartz analysis in bulk and respirable dust samples. The measured quartz content of respirable dust showed that 59% and 26% of Mufulira and Nkana Mine samples, respectively, were above the calculated U.S. Occupational Safety and Health Administration permissible exposure limit. The mean intensities of respirable dust exposure at Mufulira and Nkana were 0.992 mg/m3 (range 0-7.674) and 0.868 mg/m3 (range 0-6.944), respectively while the mean intensities of respirable quartz at Mufulira and Nkana were 0.143 mg/m3 (range 0-1.302) and 0.060 mg/m3 (range 0-0.317), respectively. These results indicate weak dust monitoring at these mines which may increase the risk of nonmalignant disease in many miners. Since Zambian mining regulations do not have crystalline silica exposure limits, these results accord with the recommendation that Zambian mining houses and the government establish crystalline silica analysis laboratory capacity and adopt dust mass concentration occupational exposure limits for more protective dust monitoring of workers. PMID- 18678922 TI - Heavy metals in water of the San Pedro River in Chihuahua, Mexico and its potential health risk. AB - The objective of this study was to determine the seasonal and downstream water quality variations of the San Pedro River in Chihuahua, Mexico. Water samples were collected monthly from October 2005 to August 2006 in triplicate, totaling 165 water samples. The five sampling locations were: below the Francisco I. Madero dam (LP); between Rosales and Delicias (RD); Meoqui (M); El Torreon (ET), and Julimes (LJ). The levels of As, Be, Ca, Cd, Co, Cu, Cr, Fe, Li, Mg, Mn, Mo, Ni, Pb, Sb, Se, Sr, Ti, Ta, V and Zn were measured using an Inductively Coupled Plasma- Optical Emission Spectrometry (ICP-OES) Perkin Elmer 2100. In addition, temperature, pH, electrical conductivity and total and fecal coliformes were determined. The statistical analysis considered a factorial treatment design; where factor A was the location point and factor B was sampling date. In addition, a multivariate technique looking for principal components was performed. The results indicated that some samples exceeded Mexican standards for As, Be, Ca, Cd, Co, Cr, Fe, Mn, Ni, Pb, Sb, Se, Sr and Zn. The As level must be considered for a red flag to the communities along the Rio San Pedro because both the monthly average level (0.10 mg L-1) and location (0.10 mg L-1) exceeded the Mexican and International norms. The multivariate analysis showed a predominant aggregation at the LP location, meaning that there was a predominance of As, Sr, Fe and Li. At the rest of the locations the elements did not present a tendency for aggregation. Statistics applied to sampling month showed that December, January, March and April were aggregated in a negative quadrant of component 1 indicating a predominance of V, Ni, Be, Fe and As. Overall, the results confirmed that this stretch of the San Pedro River is contaminated with heavy metals and other contaminants that might affect human health as well as the health of the ecosystem. PMID- 18678923 TI - Responses of antioxidant enzymes in catfish exposed to liquid crystals from E waste. AB - Liquid crystals (LCs) are typically elongated organic molecules with a non uniform distribution of electrical charges leading to a dipole. LCs are widely used in displays of computers and other electronic devices. The rapid obsolescence rate of electronics results in large amounts of liquid crystal displays (LCDs) entering the environment. Data on health effects of LCs on living creatures are currently limited to some acute toxicity tests by a few major LC manufacturers. These tests concluded that the vast majority of LCs are not acutely toxic. Since the amount of LCs in electronic devices is very small, the health effects of LCs at low concentrations or doses become important. Catfish were used as the test animals in this study. Four major enzymes of the fish's antioxidant defense system catalase (CAT), superoxide dismutase (SOD), selenium dependent glutathione peroxidase (Se-GPx), and glutathione-S-transferase (GST) were chosen as biomarkers to examine effects of LCs, which were taken from obsolete laptop personal computers made in the early 1990s. The catfish were fed with food containing different contents of LCs for 40 days. Activities of the four chosen enzymes in fish livers were assayed. The results showed that there were significant inductions of CAT, SOD, and Se-GPx activities in response to the LC doses. The plots of the enzyme activities versus LC doses suggested an occurrence of oxidative stress when the dose reached about 20 microg LC/g fish.d. It was concluded that LCs can cause pollutant-induced stress to catfish at low doses. CAT, SOD and Se-GPx are effective biomarkers to give early warning on potential health effects of LCs on some aquatic lives including catfish. PMID- 18678924 TI - Evaluation of the effect of ecologic on root knot nematode, Meloidogyne incognita, and tomato plant, Lycopersicon esculenum. AB - Nonchemical methods and strategies for nematode management including cultural methods and engineered measures have been recommended as an alternative to methyl bromide (a major soil fumigant), due to its role in the depletion of the ozone layer. Hence, an international agreement has recently been reached calling for its reduced consumption and complete phasing out. This present research evaluates the potential of Ecologic, a biological, marine shell meal chitin material, as a soil amendment management agent for root knot nematode, Meloidogyne incognita, control, and its effect on the growth of Floradel tomato plant, Lycopersicon esculentum. To accomplish this goal, studies were conducted during which, experimental pots were set up in greenhouse environments using sterilized soil inoculated with 5,000 root-knot eggs per 1500 g soil. There were 4 treatments and 5 replications. Treatments were: No chitin; 50 g chitin; 100 g chitin; and 200 g chitin. A two-week wait period following Ecologic amendment preceded Floradel tomato planting to allow breakdown of the chitin material into the soil. Fresh and dry weights of shoot and root materials were taken as growth end-points. A statistically significant difference (p < or = 0.05) was obtained with regard to the growth rate of L. esculentum at 100 g chitin treatment compared to the control with no chitin. Mean fresh weights of Floradel tomato were 78.0 +/- 22.3 g, 81.0 +/- 20.3 g, 109.0 +/- 25.4 g and 102.0 +/- 33.3 g at 0, 50, 100 and 200 g chitin, respectively. The analysis of root knot nematode concentrations indicated a substantial effect on reproduction rate associated with chitin amendment. Study results showed a significant decrease in both root knot nematode eggs and juveniles (J2) at 100g and 200 g Ecologic chitin levels, however, an increase in nematode concentrations was recorded at the 50 g Ecologic chitin level (p < or = 0.05). The mean amounts of J2 population, as expressed per 1500 cm3 soil, were 49,933 +/- 38,819, 86,050 +/- 25248, 103 +/- 133 and 103 +/- 133 for 0, 50, 100 and 200 g chitin, respectively. Similarly, the mean numbers of root knot nematode eggs (per 1500 cm3 of soil) were 40,759 +/- 36,712, 66,048 +/- 39,730, 9,904 +/- 16,591 and 9,257 +/- 17,204. Root gall rating was also significantly lower (p < or = 0.05) at the 100 g and 200 g chitin levels compared to the control. Percent gall ratings were 3.3 +/- 1.0%, 3.2 +/- 1.0%, 1.0 +/- 0.5%, and 1.0% +/- 0.6% for amendment levels of 0, 50, 100, and 200 g chitin, respectively. PMID- 18678925 TI - Preliminary molecular dynamic simulations of the estrogen receptor alpha ligand binding domain from antagonist to apo. AB - Estrogen receptors (ER) are known as nuclear receptors. They exist in the cytoplasm of human cells and serves as a DNA binding transcription factor that regulates gene expression. However the estrogen receptor also has additional functions independent of DNA binding. The human estrogen receptor comes in two forms, alpha and beta. This work focuses on the alpha form of the estrogen receptor. The ERalpha is found in breast cancer cells, ovarian stroma cells, endometrium, and the hypothalamus. It has been suggested that exposure to DDE, a metabolite of DDT, and other pesticides causes conformational changes in the estrogen receptor. Before examining these factors, this work examines the protein unfolding from the antagonist form found in the 3ERT PDB crystal structure. The 3ERT PDB crystal structure has the estrogen receptor bound to the cancer drug 4 hydroxytamoxifen. The 4-hydroxytamoxifen ligand was extracted before the simulation, resulting in new conformational freedom due to absence of van der Waals contacts between the ligand and the receptor. The conformational changes that result expose the binding clef of the co peptide beside Helix 12 of the receptor forming an apo conformation. Two key conformations in the loops at either end of the H12 are produced resulting in the antagonist to apo conformation transformation. The results were produced over a 42ns Molecular Dynamics simulation using the AMBER FF99SB force field. PMID- 18678926 TI - Compartmentalization of aquaporins in the human intestine. AB - Improper localization of water channel proteins called aquaporins (AQP) induce mucosal injury which is implicated in Crohn's disease and ulcerative colitis. The amino acid sequences of AQP3 and AQP10 are 79% similar and belong to the mammalian aquaglyceroporin subfamily. AQP10 is localized on the apical compartment of the intestinal epithelium called the glycocalyx while AQP3 is selectively targeted to the basolateral membrane. Despite the high sequence similarity and evolutionary relatedness, the molecular mechanism involved in the polarity, selective targeting and function of AQP3 and AQP10 in the intestine is largely unknown. Our hypothesis is that the differential polarity and selective targeting of AQP3 and AQP10 in the intestinal epithelial cells is influenced by amino acid signal motifs. We performed sequence and structural alignments to determine differences in signals for localization and posttranslational glycosylation. The basolateral sorting motif "YRLL" is present in AQP3 but absent in AQP10; while Nglycosylation signals are present in AQP10 but absent in AQP3. Furthermore, the C-terminal region of AQP3 is longer compared to AQP10. The sequence and structural differences between AQP3 and AQP10 provide insights into the differential compartmentalization and function of these two aquaporins commonly expressed in human intestines. PMID- 18678927 TI - Juvenile Hormone Analogues, methoprene and fenoxycarb dose-dependently enhance certain enzyme activities in the silkworm Bombyx mori (L). AB - Use of Juvenile Hormone Analogues (JHA) in sericulture practices has been shown to boost good cocoon yield; their effect has been determined to be dose dependent. We studied the impact of low doses of JHA compounds such as methoprene and fenoxycarb on selected key enzymatic activities of the silkworm Bombyx mori. Methoprene and fenoxycarb at doses of 1.0 microg and 3.0 fg/larvae/48 hours showed enhancement of the 5th instar B. mori larval muscle and silkgland protease, aspartate aminotransaminase (AAT) and alanine aminotransaminase (ALAT), adenosine triphosphate synthase (ATPase) and cytochrome-c-oxidase (CCO) activity levels, indicating an upsurge in the overall oxidative metabolism of the B.mori larval tissues. PMID- 18678928 TI - Density heterogeneities and the influence of multiple Coulomb and nuclear scatterings on the Bragg peak distal edge of proton therapy beams. AB - Density heterogeneities in the path of proton beams are known to cause degradation of the Bragg peak and, thus, widening of its distal fall-off. Inadequate accounting for this effect may lead to unwanted dose delivered to normal tissue distal to the target volume. In low-density regions, such as the thorax, this may lead to large volumes of healthy tissue receiving unnecessary dose. Although it is known that multiple Coulomb scattering within the density heterogeneities is the main cause of Bragg peak degradation, no systematic attempt has been made to quantify the contribution of multiple Coulomb scattering and nuclear scattering. Through a systematic study using a 220 MeV proton beam, we show that nuclear scattering contributes to about 5% of the distal fall-off width and is only slightly dependent on heterogeneity complexity. Furthermore, we also show that the energy spectra of the proton fluence downstream of various heterogeneity volumes are well correlated with the Bragg peak distal fall-off widths. Based on this correlation, a novel method for predicting distal fall-offs is suggested. This method is tested for three clinical setups of a voxelized model of a human head based on computer tomography data. Results are within 3% of the distal fall-off values obtained using Monte Carlo simulations. PMID- 18678929 TI - PENLINAC: extending the capabilities of the Monte Carlo code PENELOPE for the simulation of therapeutic beams. AB - In this work we present PENLINAC, a code package developed to facilitate the use of the Monte Carlo code PENELOPE for the simulation of therapeutic beams, including high-energy electrons, photons and 60Co beams. The code simplifies the creation of the treatment machine geometry, allowing the modeling of their components from elementary geometric bodies and their further conversion to the quadric functions-based structure handled by PENELOPE. The code is implemented in various subroutines that allow the user to handle several models of radiation sources and phase spaces. The phase spaces are not part of the geometry and can store many variables of the particle in a relatively small data space. The set of subroutines does not alter the PENELOPE algorithms; thus, the main program implemented by the user can maintain its kind-of-particle-independent structure. A support program can handle and analyze the phase spaces to generate, among others, last interaction maps and probability distributions that can be used as sources in simulation. Results from simulations of a Clinac linear accelerator head are presented in order to demonstrate the package capabilities. Dose distributions calculated in a water phantom for a variety of beams of this accelerator showed good agreement with measurements. PMID- 18678930 TI - Accuracy of quantitative reconstructions in SPECT/CT imaging. AB - The goal of this study was to determine the quantitative accuracy of our OSEM APDI reconstruction method based on SPECT/CT imaging for Tc-99m, In-111, I-123, and I-131 isotopes. Phantom studies were performed on a SPECT/low-dose multislice CT system (Infinia-Hawkeye-4 slice, GE Healthcare) using clinical acquisition protocols. Two radioactive sources were centrally and peripherally placed inside an anthropometric Thorax phantom filled with non-radioactive water. Corrections for attenuation, scatter, collimator blurring and collimator septal penetration were applied and their contribution to the overall accuracy of the reconstruction was evaluated. Reconstruction with the most comprehensive set of corrections resulted in activity estimation with error levels of 3-5% for all the isotopes. PMID- 18678931 TI - Cloning, expression and purification of cytochrome c(6) from the brown alga Hizikia fusiformis and complete X-ray diffraction analysis of the structure. AB - The primary sequence of cytochrome c(6) from the brown alga Hizikia fusiformis has been determined by cDNA cloning and the crystal structure has been solved at 1.6 A resolution. The crystal belonged to the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = b = 84.58, c = 232.91 A and six molecules per asymmetric unit. The genome code, amino-acid sequence and crystal structure of H. fusiformis cytochrome c(6) were most similar to those of red algal cytochrome c(6). These results support the hypothesis that brown algae acquired their chloroplasts via secondary endosymbiosis involving a red algal endosymbiont and a eukaryote host. PMID- 18678932 TI - Malonate-bound structure of the glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) and characterization of the native Fe2+ metal-ion preference. AB - The structure of a malonate-bound form of the glycerophosphodiesterase from Enterobacter aerogenes, GpdQ, has been refined at a resolution of 2.2 A to a final R factor of 17.1%. The structure was originally solved to 2.9 A resolution using SAD phases from Zn2+ metal ions introduced into the active site of the apoenzyme [Jackson et al. (2007), J. Mol. Biol. 367, 1047-1062]. However, the 2.9 A resolution was insufficient to discern significant details of the architecture of the binuclear metal centre that constitutes the active site. Furthermore, kinetic analysis revealed that the enzyme lost a significant amount of activity in the presence of Zn2+, suggesting that it is unlikely to be a catalytically relevant metal ion. In this communication, a higher resolution structure of GpdQ is presented in which malonate is visibly coordinated in the active site and analysis of the native metal-ion preference is presented using atomic absorption spectroscopy and anomalous scattering. Catalytic implications of the structure and its Fe2+ metal-ion preference are discussed. PMID- 18678933 TI - Structure of the Brachydanio rerio Polo-like kinase 1 (Plk1) catalytic domain in complex with an extended inhibitor targeting the adaptive pocket of the enzyme. AB - Polo-like kinase 1 (Plk1) is a member of the Polo-like kinase family of serine/threonine kinases involved in the regulation of cell-cycle progression and cytokinesis and is an attractive target for the development of anticancer therapeutics. The catalytic domain of this enzyme shares significant primary amino-acid homology and structural similarity with another mitotic kinase, Aurora A. While screening an Aurora A library of ATP-competitive compounds, a urea containing inhibitor with low affinity for mouse Aurora A but with submicromolar potency for human and zebrafish Plk1 (hPlk1 and zPlk1, respectively) was identified. A crystal structure of the zebrafish Plk1 kinase domain-inhibitor complex reveals that the small molecule occupies the purine pocket and extends past the catalytic lysine into the adaptive region of the active site. Analysis of the structures of this protein-inhibitor complex and of similar small molecules cocrystallized with other kinases facilitates understanding of the specificity of the inhibitor for Plk1 and documents for the first time that Plk1 can accommodate extended ATP-competitive compounds that project toward the adaptive pocket and help the enzyme order its activation segment. PMID- 18678934 TI - Expression, purification and preliminary X-ray diffraction studies of the transcriptional factor PyrR from Bacillus halodurans. AB - The PyrR transcriptional regulator is widely distributed in bacteria. This RNA binding protein is involved in the control of genes involved in pyrimidine biosynthesis, in which uridyl and guanyl nucleotides function as effectors. Here, the crystallization and preliminary X-ray diffraction analysis of two crystal forms of Bacillus halodurans PyrR are reported. One of the forms belongs to the monoclinic space group P2(1) with unit-cell parameters a = 59.7, b = 87.4, c = 72.1 A, beta = 104.4 degrees , while the other form belongs to the orthorhombic space group P22(1)2(1) with unit-cell parameters a = 72.7, b = 95.9, c = 177.1 A. Preliminary X-ray diffraction data analysis and molecular-replacement solution revealed the presence of four and six monomers per asymmetric unit; a crystallographic tetramer is formed in both forms. PMID- 18678935 TI - Crystallization and preliminary X-ray analysis of L-methionine gamma-lyase 1 from Entamoeba histolytica. AB - L-Methionine gamma-lyase (MGL) is a pyridoxal phosphate-dependent enzyme that is involved in the degradation of sulfur-containing amino acids. MGL is an attractive drug target against amoebiasis because the mammalian host of its causative agent Entamoeba histolytica lacks MGL. For the development of anti amoebic agents based on the structure of MGL, one of two MGL isoenzymes (EhMGL1) was crystallized in the monoclinic space group P2(1), with unit-cell parameters a = 99.12, b = 85.38, c = 115.37 A, beta = 101.82 degrees . The crystals diffract to beyond 2.0 A resolution. The presence of a tetramer in the asymmetric unit (4 x 42.4 kDa) gives a Matthews coefficient of 2.8 A(3) Da(-1) and a solvent content of 56%. The structure was solved by the molecular-replacement method and structure refinement is now in progress. PMID- 18678936 TI - Crystallization and preliminary X-ray analysis of the inducible lysine decarboxylase from Escherichia coli. AB - The decameric inducible lysine decarboxylase (LdcI) from Escherichia coli has been crystallized in space groups C2 and C222(1); the Ta6Br12(2+) cluster was used to derivatize the C2 crystals. The method of single isomorphous replacement with anomalous scattering (SIRAS) as implemented in SHELXD was used to solve the Ta6Br12(2+)-derivatized structure to 5 A resolution. Many of the Ta6Br12(2+) binding sites had twofold and fivefold noncrystallographic symmetry. Taking advantage of this feature, phase modification was performed in DM. The electron density map of LdcI displays many features in agreement with the low-resolution negative-stain electron-density map [Snider et al. (2006), J. Biol. Chem. 281, 1532-1546]. PMID- 18678937 TI - Crystallization of Spatzle, a cystine-knot protein involved in embryonic development and innate immunity in Drosophila melanogaster. AB - The Spatzle protein is involved in both the definition of the dorsal-ventral axis during embryonic development and in the adult innate immune response. The disulfide-linked dimeric cystine-knot protein has been expressed as a proprotein in inclusion bodies in Escherichia coli and refolded in vitro by rapid dilution. Initial orthorhombic crystals that diffracted to 7 A resolution were obtained after three months by the sitting-drop vapour-diffusion method. Optimization of the crystallization conditions resulted in orthorhombic crystals (space group P2(1)2(1)2(1), with unit-cell parameters a = 53.0, b = 59.2, c = 62.5 A) that diffracted to 2.8 A resolution in-house. The small volume of the asymmetric unit indicated that it was not possible for the crystals to contain the complete pro Spatzle dimer. Mass spectrometry, N-terminal sequencing and Western-blot analysis revealed that the crystals contained the C-terminal disulfide-linked cystine-knot dimer. Comparison of various crystallization experiments indicated that degradation of the N-terminal prodomain was dependent on the buffer conditions. PMID- 18678938 TI - Crystallization and preliminary X-ray diffraction analysis of the ternary human GM-CSF receptor complex. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a haemopoietic growth factor that acts though a ternary receptor signalling complex containing specific alpha (GMRalpha) and common beta (betac) receptor subunits. Human GM-CSF is encoded by the gene csf2, while the genes for GMRalpha and betac are csf2ra and csf2rb, respectively. Crystals of the ternary ectodomain complex comprising GM-CSF and the soluble extracellular regions of both the GMRalpha subunit and either betac or its glutamine-substitution mutant N346Q were obtained using the hanging-drop vapour-diffusion method. The best diffracting crystals of the ternary complex were obtained using the N346Q mutation of the betac subunit. These crystals grew using polyethylene glycol 3350 with a high concentration of proline, belonged to space group P6(3)22 and diffracted to 3.3 A resolution. PMID- 18678939 TI - Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-beta-D-xylanase 10B in complex with xylohexaose. AB - The cellulosome of Clostridium thermocellum is a highly organized multi-enzyme complex of cellulases and hemicellulases involved in the hydrolysis of plant cell wall polysaccharides. The bifunctional multi-modular xylanase Xyn10B is one of the hemicellulase components of the C. thermocellum cellulosome. The enzyme contains an internal glycoside hydrolase family 10 catalytic domain (GH10) and a C-terminal family 1 carbohydrate esterase domain (CE1). The N-terminal moiety of Xyn10B (residues 32-551), comprising a carbohydrate-binding module (CBM22-1) and the GH10 E337A mutant, was crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3(2)21 and contain a dimer in the asymmetric unit. The crystals diffracted to beyond 2.0 A resolution. PMID- 18678940 TI - Purification, crystallization and preliminary X-ray analysis of the membrane bound [NiFe] hydrogenase from Allochromatium vinosum. AB - The membrane-bound [NiFe] hydrogenase is a unique metalloprotein that is able to catalyze the reversible oxidation of hydrogen to protons and electrons during a complex reaction cycle. The [NiFe] hydrogenase was isolated from the photosynthetic purple sulfur bacterium Allochromatium vinosum and its crystallization and preliminary X-ray analysis are reported. It was crystallized by the hanging-drop vapour-diffusion method using sodium citrate and imidazole as crystallization agents. The crystals belong to space group P2(1)2(1)2, with unit cell parameters a = 205.00, b = 217.42, c = 120.44 A. X-ray diffraction data have been collected to 2.5 A resolution. PMID- 18678941 TI - Expression, purification, crystallization and preliminary X-ray analysis of an NADP-dependent glyceraldehyde-3-phosphate dehydrogenase from Helicobacter pylori. AB - The classical glycolytic pathway contains an NAD-dependent glyceraldehyde-3 phosphate dehydrogenase, with NADP-dependent forms reserved for photosynthetic organisms and archaea. Here, the cloning, expression, purification, crystallization and preliminary X-ray analysis of an NADP-dependent glyceraldehyde-3-phosphate dehydrogenase from Helicobacter pylori is reported; crystals of the protein were grown both in the presence and the absence of NADP. PMID- 18678942 TI - Expression, purification, crystallization and preliminary X-ray analysis of an NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from Helicobacter pylori. AB - Helicobacter pylori is a dangerous human pathogen that resides in the upper gastrointestinal tract. Little is known about its metabolism and with the onset of antibiotic resistance new treatments are required. In this study, the expression, purification, crystallization and preliminary X-ray diffraction of an NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from H. pylori are reported. PMID- 18678943 TI - Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of chlorite dismutase: a detoxifying enzyme producing molecular oxygen. AB - Chlorite dismutase, a homotetrameric haem-based protein, is one of the key enzymes of (per)chlorate-reducing bacteria. It is highly active (>2 kU mg(-1)) in reducing the toxic compound chlorite to the innocuous chloride anion and molecular oxygen. Chlorite itself is produced as the intermediate product of (per)chlorate reduction. The chlorite dismutase gene in Azospira oryzae strain GR 1 employing degenerate primers has been identified and the active enzyme was subsequently overexpressed in Escherichia coli. Chlorite dismutase was purified, proven to be active and crystallized using sitting drops with PEG 2000 MME, KSCN and ammonium sulfate as precipitants. The crystals belonged to space group P2(1)2(1)2 and were most likely to contain six subunits in the asymmetric unit. The refined unit-cell parameters were a = 164.46, b = 169.34, c = 60.79 A. The crystals diffracted X-rays to 2.1 A resolution on a synchrotron-radiation source and a three-wavelength MAD data set has been collected. Determination of the chlorite dismutase structure will provide insights into the active site of the enzyme, for which no structures are currently available. PMID- 18678944 TI - Purification, identification and preliminary crystallographic studies of a 2S albumin seed protein from Lens culinaris. AB - Lens culinaris (lentil) is a widely consumed high-protein-content leguminous crop. A 2S albumin protein (26.5 kDa) has been identified using NH(2)-terminal sequencing from a 90% ammonium sulfate saturation fraction of total L. culinaris seed protein extract. The NH(2)-terminal sequence shows very high homology to PA2, an allergy-related protein from Pisum sativum. The 2S albumin protein was purified using a combination of size-exclusion and ion-exchange chromatography. Crystals of the 2S seed albumin obtained using the hanging-drop vapour-diffusion method diffracted to 2.5 A resolution and were indexed in space group P4(1) (or P4(3)), with unit-cell parameters a = b = 78.6, c = 135.2 A. PMID- 18678945 TI - Crystallization and preliminary X-ray diffraction studies of maleylacetate reductase from Rhizobium sp. strain MTP-10005. AB - Maleylacetate reductase (EC 1.3.1.32), which catalyzes the reduction of maleylacetate to 3-oxoadipate, plays an important role in the aerobic microbial catabolism of resorcinol. The enzyme has been crystallized at 293 K by the sitting-drop vapour-diffusion method supplemented with a microseeding technique, using ammonium sulfate as the precipitating agent. The crystal belonged to the monoclinic space group C2, with unit-cell parameters a = 56.85, b = 121.13, c = 94.09 A, beta = 101.48 degrees , and contained one dimeric molecule in the asymmetric unit. It diffracted to 1.79 A resolution. PMID- 18678946 TI - Expression, purification and crystallization of Helicobacter pylori L asparaginase. AB - The L-asparaginases from Escherichia coli and Erwinia chrysanthemi are effective drugs that have been used in the treatment of acute childhood lymphoblastic leukaemia for over 30 years. However, despite their therapeutic potential, they can cause serious side effects as a consequence of their intrinsic glutaminase activity, which leads to L-glutamine depletion in the blood. Consequently, new asparaginases with low glutaminase activity, fewer side effects and high activity towards L-asparagine are highly desirable as better alternatives in cancer therapy. L-Asparaginase from Helicobacter pylori was overexpressed in E. coli and purified for structural studies. The enzyme was crystallized at pH 7.0 in the presence of 16-19%(w/v) PEG 4000 and 0.1 M magnesium formate. Data were collected to 1.6 A resolution at 100 K from a single crystal at a synchrotron-radiation source. The crystals belong to space group I222, with unit-cell parameters a = 63.6, b = 94.9, c = 100.2 A and one molecule of L-asparaginase in the asymmetric unit. Elucidation of the crystal structure will provide insight into the active site of the enzyme and a better understanding of the structure-activity relationship in L-asparaginases. PMID- 18678947 TI - Crystallization and preliminary crystallographic analysis of tRNA (m(7)G46) methyltransferase from Escherichia coli. AB - Transfer RNA (tRNA) (m(7)G46) methyltransferase (TrmB) belongs to the Rossmann fold methyltransferase (RFM) family and uses S-adenosyl-L-methionine (SAM) as the methyl-group donor to catalyze the formation of N(7)-methylguanosine (m(7)G) at position 46 in the variable loop of tRNAs. After attempts to crystallize full length Escherichia coli TrmB (EcTrmB) failed, a truncated protein lacking the first 32 residues of the N-terminus but with an additional His(6) tag at the C terminus was crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 3350 (PEG 3350) as precipitant at 283 K. An X-ray diffraction data set was collected using a single flash-cooled crystal that belonged to space group P2(1). PMID- 18678948 TI - Cloning, expression, purification and preliminary crystallographic analysis of the RNase HI domain of the Mycobacterium tuberculosis protein Rv2228c as a maltose-binding protein fusion. AB - The predicted ribonuclease (RNase) HI domain of the open reading frame Rv2228c from Mycobacterium tuberculosis has been cloned as a hexahistidine fusion and a maltose-binding protein (MBP) fusion. Expression was only observed for the MBP fusion protein, which was purified using amylose affinity chromatography and gel filtration. The RNase HI domain could be cleaved from the MBP-fusion protein by factor Xa digestion, but was very unstable. In contrast, the fusion protein was stable, could be obtained in high yield and gave crystals which diffracted to 2.25 A resolution. The crystals belong to space group P2(1) and have unit-cell parameters a = 73.63, b = 101.38, c = 76.09 A, beta = 109.0 degrees. Two fusion protein molecules of 57,417 Da were present in each asymmetric unit. PMID- 18678949 TI - Expression, crystallization and preliminary X-ray crystallographic analysis of XometC, a cystathionine gamma-lyase-like protein from Xanthomonas oryzae pv. oryzae. AB - Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight of rice (Oryza sativa L.), one of the most devastating diseases of rice in most rice-growing countries. XometC, a cystathionine gamma-lyase (CGL) like protein that is an antibacterial drug-target protein against Xoo, was cloned, expressed, purified and crystallized. CGL catalyzes the second step in the reverse-transsulfuration pathway, which is essential for the metabolic interconversion of the sulfur containing amino acids cysteine and methionine. Crystals of two different shapes, plate-shaped and pyramid-shaped, diffracted to 2.9 and 3.2 A resolution and belonged to the primitive orthogonal space group P2(1)2(1)2(1) and the tetragonal space group P4(1) (or P4(3)), with unit-cell parameters a = 73.0, b = 144.9, c = 152.3 A and a = b = 78.2, c = 300.7 A, respectively. For the P2(1)2(1)2(1) crystals, three or four monomers exist in the asymmetric unit with a corresponding V(M) of 3.02 or 2.26 A(3) Da(-1) and a solvent content of 59.3 or 45.7%. For the P4(1) (or P4(3)) crystals, four or five monomers exist in the asymmetric unit with a corresponding V(M) of 2.59 or 2.09 A(3) Da(-1) and a solvent content of 52.5 or 40.6%. PMID- 18678950 TI - Crystallization and preliminary X-ray analysis of a 1,3-1,4-beta-glucanase from Paecilomyces thermophila. AB - In this study, the crystallization and preliminary X-ray analysis of a thermostable 1,3-1,4-beta-glucanase produced by Paecilomyces thermophila is described. The purified 1,3-1,4-beta-glucanase was crystallized using the hanging drop vapour-diffusion method. The crystal belongs to the hexagonal space group P6(3)22, with unit-cell parameters a = b = 154.54, c = 87.62 A. X-ray diffraction data were collected to a resolution of 2.54 A and gave a data set with an overall R(merge) of 7.3% and a completeness of 94.6%. The Matthews coefficient (V(M)) and the solvent content are 2.38 A(3) Da(-1) and 48%, respectively. PMID- 18678951 TI - Crystallization and preliminary crystallographic analysis of recombinant immunoglobulin G-binding protein from Streptococcus suis. AB - Streptococcus suis, an important zoonotic pathogen, expresses immunoglobulin G binding protein, which is thought to be helpful to the organism in eluding the host defence system. Recombinant IgG-binding protein expressed in Escherichia coli has been crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 38.98, b = 43.94, c = 78.17 A and one molecule in the asymmetric unit. Diffraction data were collected to 2.60 A resolution. PMID- 18678952 TI - Crystallization and preliminary crystallographic analysis of mannosyl-3 phosphoglycerate synthase from Rubrobacter xylanophilus. AB - Rubrobacter xylanophilus is the only Gram-positive bacterium known to synthesize the compatible solute mannosylglycerate (MG), which is commonly found in hyperthermophilic archaea and some thermophilic bacteria. Unlike the salt dependent pattern of accumulation observed in (hyper)thermophiles, in R. xylanophilus MG accumulates constitutively. The synthesis of MG in R. xylanophilus was tracked from GDP-mannose and 3-phosphoglycerate, but the genome sequence of the organism failed to reveal any of the genes known to be involved in this pathway. The native enzyme was purified and its N-terminal sequence was used to identify the corresponding gene (mpgS) in the genome of R. xylanophilus. The gene encodes a highly divergent mannosyl-3-phosphoglycerate synthase (MpgS) without relevant sequence homology to known mannosylphosphoglycerate synthases. In order to understand the specificity and enzymatic mechanism of this novel enzyme, it was expressed in Escherichia coli, purified and crystallized. The crystals thus obtained belonged to the hexagonal space group P6(5)22 and contained two protein molecules per asymmetric unit. The structure was solved by SIRAS using a mercury derivative. PMID- 18678953 TI - High crystallizability under air-exclusion conditions of the full-length LysR type transcriptional regulator TsaR from Comamonas testosteroni T-2 and data-set analysis for a MIRAS structure-solution approach. AB - The full-length LysR-type transcriptional regulator TsaR from Comamonas testosteroni T-2 was heterologously overexpressed in Escherichia coli, purified and stabilized under conditions that favoured its rapid crystallization using the microbatch-under-oil technique. The purified protein was highly crystallizable and two different crystal forms were readily obtained. However, only monoclinic crystals gave diffraction beyond 2 A and there was a slight variation in unit cell parameters between crystals. The only other LysR-type regulator for which a full-length crystal form is available is CbnR, but no solution could be obtained when this was used as a model in molecular replacement. Mercury and xenon derivatives were therefore produced in order to phase the structure using a MIRAS approach. PMID- 18678956 TI - Strength deficits identified with concentric action of the hip extensors and eccentric action of the hamstrings predispose to hamstring injury in elite sprinters. AB - STUDY DESIGN: Prospective cohort study. OBJECTIVES: In this prospective cohort study of elite sprinters, muscle strength of the hip extensors, as well as of the knee extensors and flexors, was measured to determine a possible relationship between strength deficits and subsequent hamstring injury within 12 months of testing. The method used for testing muscle strength simulated the specific muscle action during late swing and early contact phases when sprinting. BACKGROUND: There have been no prospective studies in elite sprinters that examine the concentric and eccentric isokinetic strength of the hip extensors and the quadriceps and hamstring muscles in a manner that reflects their actions in late swing or early contact phases of sprinting. Consequently, the causal relationship between hip and thigh muscle strength and hamstring injury in elite sprinters may not be fully understood. METHODS AND MEASURES: Isokinetic testing was performed on 30 male elite sprinters to assess hip extensors, quadriceps, and hamstring muscle strength. The occurrence of hamstring injury among the subjects was determined during the year following the muscle strength measurements. The strength of the hip extensors, quadriceps, and hamstring muscles, as well as the hamstrings-quadriceps and hip extensors- quadriceps ratios were compared. RESULTS: Hamstring injury occurred in 6 subjects during the 1-year period. Isokinetic testing at a speed of 60 degrees /s revealed weakness of the injured limb with eccentric action of the hamstring muscles and during concentric action of the hip extensors. When performing a side-to-side comparison for the injured sprinters, the hamstring injury always occurred on the weaker side. Differences in the hamstrings-quadriceps and hip extensors-quadriceps strength ratios were also evident between uninjured and injured limbs, and this was attributable to deficits in hamstring strength. CONCLUSION: Hamstring injury in elite sprinters was associated with weakness during eccentric action of the hamstrings and weakness during concentric action of the hip extensors, but only when tested at the slower speed of 60 degrees /s. PMID- 18678957 TI - Proximal and distal influences on hip and knee kinematics in runners with patellofemoral pain during a prolonged run. AB - STUDY DESIGN: Cross-sectional experimental laboratory study. OBJECTIVES: To investigate the relationships between hip strength and hip kinematics, and between arch structure and knee kinematics during prolonged treadmill running in runners with and without patellofemoral pain syndrome (PFPS). BACKGROUND: Hip weakness can lead to excessive femoral motions that adversely affect patellofemoral joint mechanics. Similarly, foot mechanics, which are influenced by foot structure, are also known to influence patellofemoral joint mechanics. Thus, proximal and distal factors should be considered when studying individuals with PFPS. METHODS AND MEASURES: Twenty recreational runners with PFPS (5 male, 15 female) and 20 matched uninjured runners participated in the study. Hip abduction and hip external rotation isometric strength measurements were collected before and after a prolonged run, while the arch height index was recorded on all runners before the run. Lower extremity kinematic data were collected at the beginning and end of the run. Two-way repeated-measures analyses of variance (ANOVAs) were used for analysis. RESULTS: Both groups displayed decreases in hip abductor and external rotator strengths at the end of the run. The PFPS group displayed significantly lower hip abduction strength [(kg x cm)/body mass] compared to controls (PFPS group: begin 15.3, end 13.5; uninjured group: begin 17.3, end 15.4). At the end of the run, the level of association between hip abduction strength and the peak hip adduction angle for the PFPS group was statistically significant, indicating a strong relationship (r = 0.74). No other associations with hip strength were observed in either group. Arch height did not differ between groups and no significant association was observed between arch height and peak knee adduction angle during running. CONCLUSIONS: Runners with PFPS displayed weaker hip abductor muscles that were associated with an increase in hip adduction during running. This relationship became more pronounced at the end of the run. LEVEL OF EVIDENCE: Therapy, level 5. PMID- 18678958 TI - The role of pulse duration and stimulation duration in maximizing the normalized torque during neuromuscular electrical stimulation. AB - STUDY DESIGN: Controlled laboratory study OBJECTIVES: To determine the effects of pulse duration and stimulation duration on the evoked torque after controlling for the activated area by using magnetic resonance imaging (MRI). BACKGROUND: Neuromuscular electrical stimulation (NMES) is commonly used in the clinic without considering the physiological implications of its parameters. METHODS AND MEASURES: Seven able-bodied, college students (mean +/- SD age, 28 +/- 4 years) participated in this study. Two NMES protocols were applied to the knee extensor muscle group in a random order. Protocol A applied 100-Hz, 450-microsecond pulses for 5 minutes in a 3-seconds-on 3-seconds-off duty cycle. Protocol B applied 60 Hz, 250-microsecond pulses for 5 minutes in a 10-seconds-on 20-seconds-off duty cycle. The amplitude of the current was similar in both protocols. Torque, torque time integral, and normalized torque for the knee extensors were measured for both protocols. MRI scans were taken prior to, and immediately after, each protocol to measure the cross-sectional area of the stimulated muscle. RESULTS: The skeletal muscle cross-sectional areas activated after both protocols were similar. The longer pulse duration in protocol A elicited 22% greater torque output than that of protocol B (P<.05). After considering the activated area in both protocols, the normalized torque with protocol A was 38% greater than that with protocol B (P<.05). Torque time integral was 21% greater with protocol A (P = .029). Protocol B failed to maintain torque at the start and the end of the 10 second activation. CONCLUSIONS: Longer pulse duration, but not stimulation duration, resulted in a greater evoked and normalized torque compared to the shorter pulse duration, even after controlling for the activated muscular cross sectional areas with both protocols. LEVEL OF EVIDENCE: Therapy, level 5. PMID- 18678959 TI - Decreased quadriceps activation measured hours prior to a noncontact anterior cruciate ligament tear. AB - STUDY DESIGN: Case report. BACKGROUND: Decreased quadriceps activation has been shown to be present following anterior cruciate ligament (ACL) injury, but its presence prior to ACL injury is unknown. The purpose of this case report was to describe the level of quadriceps activation measured hours before a noncontact ACL injury in an individual who previously demonstrated known biomechanical risk factors for ACL injury. CASE DESCRIPTION: A 23-year-old female (height, 176.9 cm; mass, 72.4 kg), sustained a left noncontact ACL injury while landing from a jump stop during a recreational basketball game. This case was unique because data regarding landing biomechanics and quadriceps force and activation were gathered in 2 separate, unrelated studies prior to injury. OUTCOMES: Peak external knee abduction moment (-65.3 Nm) during a drop jump landing 8 months prior to injury indicated elevated risk for ACL injury. Involved quadriceps central activation ratios (CAR) were obtained 1 week (CAR, 0.81) and 4 hours (CAR, 0.77) prior to injury. Strength and CAR (0.76) measurements changed very little within 36 hours of injury and both strength, and activation (CAR, 0.90) improved following surgical reconstruction and formal rehabilitation. DISCUSSION: An individual with known biomechanical risk factors for ACL injury may compound risk for noncontact ACL injury if decreased quadriceps activation is also present. LEVEL OF EVIDENCE: Prognosis, level 4. PMID- 18678960 TI - A primer on selected aspects of evidence-based practice relating to questions of treatment. Part 1: asking questions, finding evidence, and determining validity. AB - The process of evidence-based practice (EBP) guides clinicians in the integration of individual clinical expertise, patient values and expectations, and the best available evidence. Becoming proficient with this process takes time and consistent practice, but should ultimately lead to improved patient outcomes. The EBP process entails 5 steps: (1) formulating an appropriate question, (2) performing an efficient literature search, (3) critically appraising the best available evidence, (4) applying the best evidence to clinical practice, and (5) assessing outcomes of care. This first commentary in a 2-part series will review principles relating to steps 1, 2, and 3 of this 5-step model. The purpose of this commentary is to provide a perspective to assist clinicians in formulating foreground questions, searching for the best available evidence, and determining validity of results in studies of interventions for orthopaedic and sports physical therapy. PMID- 18678961 TI - A primer on selected aspects of evidence-based practice to questions of treatment. Part 2: interpreting results, application to clinical practice, and self-evaluation. AB - The process of evidence-based practice (EBP) guides clinicians in the integration of individual clinical expertise, patient values and expectations, and the best available evidence. Becoming proficient with this process takes time and consistent practice, but should ultimately lead to improved patient outcomes. The EBP process entails 5 steps: (1) formulating an appropriate question, (2) performing an efficient literature search, (3) critically appraising the best available evidence, (4) applying the best evidence to clinical practice, and (5) assessing outcomes of care. This second commentary in a 2-part series will review principles relating to steps 3 through 5 of this 5-step model. The purpose of this commentary is to provide a perspective to assist clinicians in interpreting results, applying the evidence to patient care, and evaluating proficiency with EBP skills in studies of interventions for orthopaedic and sports physical therapy. PMID- 18678962 TI - Upper cervical ligament testing in a patient with os odontoideum presenting with headaches. AB - STUDY DESIGN: Resident's case problem. BACKGROUND: The role of premanipulative testing of the cervical spine is an area of controversy, and there are very few data to inform and guide practitioners on the use of ligamentous stability tests when assessing the upper cervical spine. DIAGNOSIS: A 23-year-old female was referred to physical therapy by a neurologist for the management of intractable headaches of possible musculoskeletal origin. Her Neck Disability Index score was 54% and she rated her headache pain from 3/10 to 9/10 on a Numerical Pain Rating Scale. She reported a 2-year history of intermittent lower extremity paresthesias without a known mechanism or current symptoms. She was treated in physical therapy for 11 visits with improvements in cervical range of motion, strength, and intensity of her headaches, but noted no change in the frequency of headaches. She was subsequently referred to the primary author for a second opinion and potential manual therapy interventions. Initial neurological screening examination for upper and lower motor neuron lesions was unremarkable. Assessment of the transverse ligament, using the anterior shear test in supine, brought on paresthesias in both feet and her toes. The paresthesias continued after the cessation of the test. The Sharp-Purser test performed in sitting, immediately after the transverse ligament test, abolished the paresthesias. She was then referred back to her primary care physician for further evaluation. Subsequent radiographs and magnetic resonance imaging revealed that the patient had a C2-C3 Klippel-Feil congenital fusion and os odontoideum. The patient was examined by a neurosurgeon who concluded that she was not a surgical candidate. Her neurological symptoms completely resolved, but she continued to have headaches. DISCUSSION: Os odontoideum is a clinically important condition, given that the mobile dens may render the transverse ligament incompetent, leading to atlantoaxial instability. Both the role and sequencing of upper cervical ligamentous testing is controversial. The results of this case report suggest that physical therapists should be cognizant of this condition and consider screening the upper cervical ligaments prior to manual or mechanical interventions to this region. LEVEL OF EVIDENCE: Differential diagnosis, level 4. PMID- 18678963 TI - [Evaluation of new complete denture treatment by dentists and patients]. AB - PURPOSE: With an increase in the aged population in Japan, it is important that clinicians evaluate the outcome of complete denture treatment based on evaluation by dentists and patients to provide high-quality treatment. The purpose of this study was to evaluate the evaluation and to clarify the relationship between them by dentists and patients. METHODS: Forty patients wearing complete dentures were selected, after obtaining informed consent. Scores of complete denture, supporting tissue, chewing function, satisfaction and QOL were the factors used for evaluation. These were evaluated at four stages in complete denture treatment: at the initial visit, before insertion of new dentures (just before), first visit after insertion of new dentures (1st visit) and 1 month after insertion. We compared scores obtained just before and at the 1st visit using t test, and analyzed changes in scores with correlation coefficient. In addition, the relationship between dentists and patients was analyzed with kappa statistic. RESULTS: Evaluation of the new denture was influenced by the evaluation of the old denture. In case of high evaluation of old dentures, evaluation of new dentures tended to decrease. Conversely, in case of low evaluation of the old dentures, evaluation of new dentures tended to increase (p < 0.01) . There was no significant correlation between the evaluation by the dentists and the patients. CONCLUSIONS: Changes after wearing new dentures and the relationships were clarified. The disagreement of the evaluation by dentists and patients suggests that both evaluations should be done comprehensively. PMID- 18678964 TI - [Studies on the standardization of parameters for jaw movement analysis--jaw movement analysis at the incisal point]. AB - PURPOSE: To establish standardized evaluating methods for jaw movements analysis independently of the reference coordinate system. In this paper, we focused on the analysis of incisor point paths of border movements. Then we investigated parameters that are independent of the reference coordinate system and can be utilized among different systems. METHODS: Recorded data of jaw border movements from 20 healthy male adults were employed as basic samples. First, the effects of changes of the reference coordinate systems on the analysis were investigated. Second, the line between incisor points at the intercuspal position and the maximal mandibular opening position was used as the reference line. Then the area index of incisor paths was calculated with the length of the perpendicular line dropped from the incisor point to the reference line and the displacement of the foot of the perpendicular. RESULTS: Changes of the spatial relation between a reference plane of the coordinate system and border movements pathways had an effect on a measured area of projected pathways, about 2% - 5% differences of projected area on the frontal plane with 10 degrees rotation around a horizontal axis. The area index of incisor paths showed close correlation with the actual area of a 3-dimensional plane bounded by the incisor path [sagittal border movement: r = 0.53 (p = 0.016), left border opening pathway: r = 0.97 (p < 0.001), right border opening pathway: r = 0.98 (p < 0.001), Spearman rank correlation]. CONCLUSION: The results suggest that new parameters could measure jaw border movements quantitatively independent of the reference coordinate system. PMID- 18678965 TI - [Studies on the standardization of parameters for jaw movement analysis--6 degree of-freedom jaw movements analysis]. AB - PURPOSE: To establish standardized evaluating methods for jaw movements analysis. In this paper, we investigated evaluating parameters for 6 degree-of-freedom jaw movements data. METHODS: Recorded data of jaw border movements from 20 male adults were employed as basic samples. The main parameters were as follows: 1. The displacement of an intercondylar midpoint: the length of a straight line between 2 positions of this point, the intercuspal position and other jaw position. 2. The angle of intercondylar axes: the angle between 2 position of the intercondylar axis, the intercuspal position and other jaw position. 3. The angle of incisal-condylar planes: the angle between 2 position of the plane, the intercuspal position and other jaw position (this plane was defined with the incisal point and condylar points of both sides 4. The mandibular motion range index: quantitative values calculated with 2 of 3 parameters described above. RESULTS: The mandibular motion range index showed a close correlation with respective projected areas of the incisal paths, with the projected area of sagittal border movements on the sagittal plane r = 0.82 (p < 0.001), with the projected area of lateral border movements on the frontal plane: left lateral border movements r = 0.92 (p < 0.001), right lateral border movements r = 0.84 (p < 0.001), Spearman rank correlation. CONCLUSION: New parameters were based on 6 degree-of-freedom jaw movements data and relative relationship between the intercuspal position and other jaw position. They were independent of reference coordinate systems and could measure jaw movement quantitatively. PMID- 18678966 TI - [Bonding of resin cement to bleached dental enamel]. AB - PURPOSE: The effects of bleaching times, types of etching agent and storage period of bleached bovine tooth on the shear bond strength of resin cement to the enamel were examined. MATERIALS AND METHODS: Bovine teeth were repeatedly bleached 0, 1, 3, and 5 times then stored in 37 degrees C water for 1 week. The effect of bleaching number of the bovine tooth on the bond strength of resin cement to the enamel was investigated using 40% phosphoric acid (EG) etching technique. Next, the effects of types of etching agent and of storage period of bleached bovine tooth with three times in 37 degrees C water on the bond strength were studied using 10% citric acid-3% ferric chloride (10-3) or 10% citric acid (10-0) solution. RESULTS: The bleaching of bovine tooth allowed for a dramatic decrease in the bond strength from 18.3 MPa to 9.8 MPa (1 time), and 3.9 MPa (3 times), even though the bovine enamel was etched by EG. However, when 10-3 or 10 0 solution was applied to the three times bleached enamel, bond strengths were 13.9 and 10.0 MPa, respectively. Furthermore, prolonging of the storage period of the three times bleached bovine tooth in water to 2 months resulted in a increase in the bond strength from 3.9 to 10.1 MPa, even if bovine enamel was etched by EG, and close to that obtained from the 10-3 etching. CONCLUSION: To obtain the expected bond strength to bleached enamel, it is better to wait for 2 months for a restoration and use the 10-3 etching. PMID- 18678967 TI - [Establishment of simultaneous measurement method of 8 salivary components using urinary test paper and clinical evaluation of oral environment]. AB - PURPOSE: Clinical findings were compared with glucose, protein, albumin, bilirubin, creatinine, pH, occult blood, ketone body, nitrite, and white blood cells contained in whole saliva to investigate the components that most markedly reflect the periodontal condition. MATERIAL AND METHOD: The subjects were staff of the Prosthodontics Department, Showa University, and patients who visited for dental treatments (57 subjects in total). At the first time, saliva samples were gargled with 1.5 ml of distilled water for 15 seconds and collected by spitting out into a paper cup. At the second time, saliva samples were collected by the same method. At the third time, saliva samples after chewing paraffin gum for 60 seconds were collected by spitting out into a paper cup. Thus whole saliva collecting that was divided on three times. After sampling, 8 mul of the saliva sample was dripped in reagent sticks for the 10 items of urinary test paper and the reflectance was measured using a specific reflectometer. In the periodontal tissue evaluation, the degree of alveolar bone resorption, probing value, and tooth mobility and the presence or absence of lesions in the root furcation were examined and classified into 4 ranks. The mean values in each periodontal disease rank and correlation between the periodontal disease ranks and the components were statistically analyzed. RESULTS: Bilirubin and ketone body were not measurable. The components density of the 8 items was increased as the periodontal disease rank increased. Regarding the correlation between the periodontal disease ranks and the components, high correlations were noted for protein, albumin, creatinine, pH, and white blood cells. CONCLUSION: The simultaneous measurement method of 8 salivary components using test paper may be very useful for the diagnosis of periodontal disease of abutment teeth. PMID- 18678968 TI - [Studies evaluating measurement accuracy of CMS-JAW, a jaw motion tracking device with six degrees of freedom using an ultrasonic recording system]. AB - PURPOSE: For diagnoses of temporomandibular dysfunction, reports indicate the importance of analyzing jaw movements using a jaw motion tracking device with six degrees of freedom. However, such devices are not widely applied in clinical practice, due to difficulty of use. To determine whether CMS-JAW (a newly developed six degrees of freedom jaw motion tracking device that features easy analysis of jaw movements) offers the measurement accuracy required for clinical use, we evaluated the device by measuring condyle translation and rotation during jaw movement. METHODS: In this study, we evaluated measurement accuracy by comparing measurements of condyle movements obtained from the Win Jaw System, a comparable jaw motion tracking device with six degrees of freedom based on an ultrasonic recording system; the Gnathohexagraph II, a device based on a different measurement principle; and the CMS-JAW. In the experiment, we confirmed the condyle translations and rotations obtained by each device by measuring distances and angles, respectively, with a micro slider for three-dimensional measurements (to an accuracy of 0.01 mm and 0.02 mm) and a digital protractor (to an accuracy of 0.1 degrees). RESULTS: No significant distortions were observed in the space replicated based on the range of condyle translations obtained with CMS JAW. Deviations from theoretical values were greatest along the z-axis, with a maximum error of 0.20 mm and root mean square (RMS) error of 0.10 mm. The maximum error for angles of condyle rotation was 0.50 degrees; RMS errors ranged from 0.05 to 0.30 degrees. CONCLUSION: Our analysis of condyle movement demonstrates that the CMS-JAW provides acceptable measurement accuracy for clinical applications. PMID- 18678969 TI - Clinical evaluation of semi-adjustable articulators: reproducibility of sagittal condylar path inclination assessed by a jaw-tracking system with six degrees of freedom. AB - PURPOSE: To validate whether the data for individual patients are correctly reproduced in sagittal condylar path inclination of the articulator, the data obtained by the conventional anterior check bite method and the data obtained by a jaw-tracking system with six degrees of freedom were compared. METHODS: In 5 subjects (4 males and 1 female) with healthy tooth alignments, 5 anterior check bites were obtained from each subject. Sagittal condylar path inclination was measured using seven types of semi-adjustable articulator. Next, the anterior gliding movement was measured 5 times in all 5 subjects using a jaw-tracking system with six degrees of freedom (MMJI- E, Shofu Inc. Kyoto, Japan. The sagittal condylar path inclination data obtained with semi-adjustable articulators were compared to those obtained by the jaw-tracking system using the two-way analysis of variance and Fisher's PLSD method (alpha = 0.05). RESULTS: In measurements of the same subjects for five types of articulators with the check bite method, significant differences were observed by two-way analysis of variance, and differences in measurements among articulators were found (p < 0.01). Regarding the sagittal condylar path inclinations measured with Hanau-184 Wide-View and Dentatus ARL articulators, significant differences were observed on both right and left sides compared with the jaw-tracking system. With the Denar Centri-Check System and Denar Mark II, significant differences were observed on one side (p < 0.05). CONCLUSION: Denar Cadiax Compact, Hanau Condyle Repositioner, and Hanau H2O are recommended for more accurate measurements of the sagittal condylar path inclination. PMID- 18678970 TI - Co-treatment with basic fibroblast growth factor and 17beta-estradiol in the presence of dexamethasone accelerates bone formation by rat bone marrow stromal cell culture. AB - PURPOSE: Bone marrow stromal cells (BMSCs) are a promising cell source in applications for tissue engineering and regenerative medicine. Optimization and control of the growth and differentiation of cultivated cells can be achieved by the administration of growth factors and hormones in vitro. This study provided experimental information on the enhancement of the osteogenic potential of rat BMSCs in vitro and in vivo. METHODS: Mineralized nodule formation of rat BMSCs in culture for 3 weeks with dexamethasone (Dex)-treated media supplemented with both basic fibroblast growth factor (bFGF) and 17beta -estradiol (E2) was examined by histology. In porous beta-tricalcium phosphate (beta - TCP), proliferation, migration, and differentiation of BMSCs were examined by histology and transmission electron microscopy. After culturing, the composites were subcutaneously implanted into syngeneic rats. The tissues with implants were harvested after 4 weeks and evaluated microscopically by using histological stain. RESULTS: Dex-treated media supplemented with both bFGF and E2 was the most effective in mineralized nodule formation of BMSCs in vitro. Light and electron microscopy revealed the presence of many cells with developed rough endoplasmic reticulum. Bone formation in the BMSC/beta -TCP composites in cultures in vitro for 3 weeks was observed histologically at 4 weeks after implantation. When BMSC/beta -TCP composites were cultured in Dex-treated media supplemented with both bFGF and E2, the amount of bone formation at implants was substantially greater than that of composites cultured in Dex-treated media supplemented with bFGF. CONCLUSION: The combined use of bFGF and E2 could effectively improve the bone-forming ability of BMSCs. PMID- 18678971 TI - Association between stress and temporomandibular disorder. AB - PURPOSE: To evaluate the causes of temporomandibular disorder (TMD) by examining the relationships between 3 major TMD symptoms, parafunction, and stress, a questionnaire survey was performed during dental examinations in corporations. METHODS: The survey was performed using 6 questions on the following topics: 1. Trismus; 2. Joint noise; 3. Pain; 4. Clenching in the daytime; 5. Nocturnal bruxism; and 6. Stress. There were 3,225 subjects, 2,809 males and 416 females and the mean age of the subjects was 40.12 years. The relationships between questions 1 to 5 and question 6 were examined by the chi-square test. RESULTS: There were significant correlations (question 1, p = 0.001; questions 2-5, p < 0.001). CONCLUSION: Clearly, there was an influence of psychological factors, such as stress, on TMD, and such factors were considered to play important roles in its etiology, progression, and treatment. The results of this study suggest that well-controlled studies of TMD are necessary. PMID- 18678972 TI - Relationship of pressure pain thresholds between the masticatory muscles and styloid process. Part 2. Healthy subjects and patients with temporomandibular disorders. AB - PURPOSE: We previously reported correlations between the pressure pain threshold (PPT) at the styloid process, which represents individual pain sensitivity, and PPTs of the masticatory muscles in healthy subjects, and proposed the 95% confidence interval (95%CI) of the PPT at the styloid process as a reference range. In this study, we evaluated its usefulness as a reference range. METHODS: Serial changes in the PPT relative to the 95%CI were studied in 7 healthy subjects. Next, the determination of abnormal sites in the PPT relative to the 95%CI was studied in 7 patients with temporomandibular disorders (TMD). RESULTS: In the healthy subjects, the PPT remained within the 95%CI at all sites. Since the PPT deviated from the 95%CI at some sites in TMD patients, abnormal sites could be determined. CONCLUSION: The 95%CI of the PPT at the styloid process is useful as a means for the evaluation of muscle symptoms of TMD patients. PMID- 18678973 TI - [Application of the palatal ramp in a reconstructed mandibulectomy patient]. AB - PATIENT: A 70-year-old patient with reconstructed mandibulectomy presented to our clinic with a complaint of denture instability. Due to left mandibular gingival cancer, she had received segmental resection of the left mandible and reconstruction with the fibula. An intraoral examination revealed few intermaxillary occlusal contacts due to the deviation of the mandibular dental arch. A mandibular denture and maxillary prosthesis with palatal ramp were fabricated to provide sufficient occlusal contacts and bilateral balanced occlusion. DISCUSSION: Regardless of reconstruction, a careful case history, combined with meticulous clinical evaluation was demanded in this mandibulectomy patient. The ramp on maxillary prosthesis could provide not only proper occlusal scheme but also stability of mandibular denture. CONCLUSION: To improve masticatory function in a reconstructed mandibulectomy patient with deviated mandibular movement and disagreement of occlusion, maxillary prosthesis with palatal ramp was effective. PMID- 18678974 TI - [Clinical report of Branemark implant treatment for lower anterior teeth defect with severe resorption of alveolar bone]. AB - PATIENT: A 20-year-old female patient with lower anterior teeth defect was referred from the Department of Oral and Maxillofacial Surgery and Orthodontics, Tokushima University Dental Hospital. The patient had severe resorption of alveolar bone around lower anterior edentulous and was treated with Branemark implant system. DISCUSSION: The implant prosthesis made fine outcome and was satisfactory for the functional and esthetic requirement of the patient. The superstructure, which was constructed with a combination of inner and outer crowns, resulted in over counter design. Maintenance of the implant and dental hygiene was continued with the cooperation of the Department of Preventive Dentistry. CONCLUSION: The dental implant prosthesis could satisfy the functional and esthetic expectations of this patient sufficiently. PMID- 18678975 TI - [Case report of occlusal treatment with full mouth reconstruction]. AB - PATIENT: A 70-year-old woman presented with a complaint of masticatory pain and disturbance. She had collapse of occlusal support and vertical dimension decrease. This case used provisional restoration for increasing vertical dimension and reconstruction of anterior guidance. After confirming occlusal stability, she was treated with fixed prosthesis and removable partial dentures. DISCUSSION: This patient was followed for 6 and a half years with no prosthetic complications, because the reconstruction of bilateral occlusal support and anterior guidance were performed. CONCLUSION: It is important to reconstruct the occlusal support and anterior guidance in cases of prosthetic treatment of occlusal collapse. PMID- 18678976 TI - [Case of prosthodontic treatment for loss of occlusal support in molar regions defect with severe resorption of alveolar bone]. AB - PATIENT: The patient's chief complaint was difficulty of mastication in molar regions. The diagnosis was masticatory dysfunction caused by loss of occlusal contacts. Initially provisional restorations were fabricated, then disharmonious contours were corrected during the following treatments including endodontic and periodontic treatments and tooth extraction. Final restorations were made with fixed partial dentures. DISCUSSION: Preservation of the left incisor of mandible enabled recontouring the provisional restoration and consequently, determining a definitive occlusal scheme for the final restoration. CONCLUSION: For a patient with collapse of posterior occlusal contacts, hopeless teeth which should be extracted were used as temporary abutment for the provisional restoration. This treatment plan minimized trouble for the patient during treatment, resulting in successful final occlusal reconstruction. PMID- 18678977 TI - [Application of sublingual bar to removable partial denture of a patient complaining of major discomfort to lingual plate]. AB - PATIENT: A 62-year-old man complained of discomfort during insertion of lower removable partial denture (RPD). Existing denture was highly objectionable to tongue and intolerable to wear due to volume and contour of its lingual aspect. The treatment plan included fabrication of an interim denture and altering its shape until it did not lead to discomfort, followed by fabrication of a final denture. Consequently, RPD using sublingual bar was found to be the most comfortable to wear and was fabricated as final denture. The patient was ready to insert the final denture and restoration of masticatory function was accomplished. DISCUSSION: In cases with unfavorable anatomic conditions such as a lack of mouth-floor depth, lingual plate should be applied as first choice, and when it is not well tolerated by the patient, sublingual bar should be applied. CONCLUSION: Sublingual bar is suggested to be one of the least objectionable major connectors. PMID- 18678978 TI - [A clinical report of application of implant-supported fixed partial denture with a removable gingival veneer to a patient who has severe resorption of maxillary anterior ridge]. AB - PATIENT: A 39-year-old female patient was admitted with mobility of maxillary anterior fixed partial denture. She was diagnosed as severe chronic periodontitis of abutment teeth of the denture, left alveolus cleft, and anterior/posterior crossbites. The crossbites were improved by prosthetic approach using an implant supported fixed partial denture with a removable gingival veneer at the anterior and crowns at the posterior. The implant-supported fixed partial denture and most teeth have been kept in a stable condition. DISCUSSION: Sufficient lip support and proper phonetic function were recovered by implant-supported fixed partial denture with removable gingival veneer. The long-term stability of implants may be due to easiness of toothbrushing and connection of implants rendered by the fixed prosthesis. CONCLUSION: This case suggests that implant-supported fixed partial denture with removable gingival veneer could be useful for severe ridge resorption. PMID- 18678979 TI - [Case report: long-term follow-up of a transitional denture]. AB - PATIENT: A 70-year-old woman visited our clinic with a chief complaint of chewing difficulty due to serious periodontitis and an improper RPD. A poor prognosis of the maxillary anterior residual teeth was predicted. However, the patient' s consent for tooth extraction could not be obtained. Consequently, the RPD was designed for the future additional tooth. DISCUSSION: The maxillary RPD has been functioning for 9 years through repeated additional teeth and repairs. Designing of the metal plate for the predicted prognosis allowed for additional teeth and repairs after losing of the residual teeth. CONCLUSION: Designing of the denture for the predicted prognosis allowed long-term continuous use of the denture. PMID- 18678980 TI - [Case of functional and esthetic rehabilitation by increased occlusal vertical dimension with prosthetic treatment]. AB - PATIENT: The patient was a 72-year-old woman who presented with chief complaints of masticatory and esthetic impairment due to detachment of a bridge in the anterior tooth region. These symptoms were attributed to displacement of the mandibular position caused by an untreated defect in the mandibular molar region and extrusion of the mandibular anterior tooth region associated with reductions in the occlusal vertical dimension of the molar region. The occlusal vertical dimension was restored using a denture with an occlusal splint as treatment denture, and functional and esthetic recovery was observed. DISCUSSION: Establishment of stable occlusal support in the molar region is thought to be utmost importance in stabilizing prosthesis of the anterior tooth region. CONCLUSION: Although a unilateral defect was observed in the mandibular molar region, use of a denture with an occlusal splint enabled bilateral elevation in addition to creation of denture space in the maxilla. The occlusal splint region was then replaced with prosthesis to obtain a stable occlusal relationship in the molar region and simultaneously to facilitate functional and esthetic recovery of the anterior tooth region. PMID- 18678981 TI - Modulation of circadian rhythm of DNA synthesis in tumor cells by inhibiting platelet-derived growth factor signaling. AB - Circadian synchronization of cell proliferation is observed not only in normal healthy tissues but also in malignant solid tumors. However, the proliferation rhythm of tumor cells is often different from that of normal cells. We reported here that the peculiar rhythm of tumor cell proliferation was modulated by inhibition of platelet-derived growth factor (PDGF) signaling. DNA synthesis in tumor cells implanted in mice showed a 24-h oscillation apparently differing from that of normal bone marrow cells. Continuous administration of AG1295 (10 microg/h, s.c.), a PDGF receptor tyrosine kinase inhibitor, substantially suppressed DNA synthesis in the implanted tumor cells but not in the healthy bone marrow cells. During the administration of this drug, the rhythm of DNA synthesis in the tumor cells was synchronized with that in bone marrow cells. The present results suggest that the circadian rhythm of DNA synthesis in tumor cells is modulated by PDGF receptor signaling, which is activated following tumor progression. Because the rhythmic patterns of clock gene expression in tumor cells did not differ significantly from those in other healthy tissues, the enhanced signal transduction of PDGF receptor may cause an alteration in the rhythmicity of tumor cell proliferation without changing in the intracellular molecular clockwork. PMID- 18678982 TI - Effects of injectable propofol emulsion on singlet oxygen released from activated human neutrophils and that chemically generated. AB - Effects of an injectable emulsion of propofol and its emulsifier on singlet oxygen (1O2) were examined. 1O2 released from activated human neutrophils was detected by chemiluminescence, and chemically generated 1O2 was detected by electron paramagnetic resonance (EPR). Both the propofol emulsion and the emulsifier suppressed 1O2 release from neutrophils. However, the emulsifier did not quench chemically generated 1O2, while the propofol emulsion quenched it. These results indicated that the emulsifier did not scavenge 1O2 released from neutrophils but inhibited 1O2 generation. The suppressive effects of propofol emulsion on 1O2 release from neutrophils consist of 1O2 scavenging and inhibition of 1O2 generation. PMID- 18678983 TI - Cell-culture-dependent change of Ca2+ response of rat aortic myocytes to sphingosine-1-phosphate. AB - We characterized the effects of sphingosine-1-phosphate (S1P) on rat aortic myocytes with or without culture. Application of S1P induced a small Ca2+ response in 40% freshly dispersed aortic myocytes, whereas S1P caused a larger Ca2+ response in 90% myocytes cultured for 72 h. Concentration-response relationships of S1P in cultured myocytes were significantly different from that in non-cultured myocytes. Analysis of the expression of S1P-receptor mRNA transcripts revealed that S1P-receptor type 3 (S1P3) was significantly increased when myocytes were cultured for 24 h. Neither the removal of serum from culture medium nor pretreatment with pharmacological agents, such as ERK, Rho, and PI3 kinase inhibitors, affected the progression of the S1P-induced Ca2+ response during culture. The sustained component of the Ca2+ response to S1P was sensitive to the removal of external Ca2+ and was effectively inhibited by inorganic Ca2+ channel blockers such as Gd3+, Cd2+, and Ni2+. However, application of S1P did not induce any contraction in organ-cultured as well as the intact aorta muscle strip. Aortic myocytes freshly dispersed from the organ-cultured muscle were also ineffective against S1P. Taken together, cell-culture changes the S1P3-mediated Ca2+ response to S1P in rat aortic myocytes. PMID- 18678984 TI - Functional role of Na+/H+ exchanger in Ca2+ influx mediated via human endothelin type A receptor stably expressed in Chinese hamster ovary cells. AB - This study examines the functional role of Na+/H+ exchanger (NHE) in Ca2+ influx mediated by human endothelin type A receptor (ET(A)R) expressed in Chinese hamster ovary (CHO) cells. Endothelin-1 (ET-1) increased extracellular acidification rate (ECAR), which was abolished by 5-(N-ethyl-N isopropyl)amiloride (EIPA), an NHE inhibitor. EIPA and KB-R7943, a Na+/Ca2+ exchanger (NCX) inhibitor, inhibited ET-1-induced sustained increases in intracellular Ca2+ concentration ([Ca2+]i), and EIPA had no effect on [Ca2+]i after KB-R7943 treatment. ET-1-elicited sustained [Ca2+]i increase was suppressed by reducing extracellular Na+ concentration. These results suggest that possible coupling of NHE with NCX via Na+ transport is involved in ET(A)R-mediated sustained [Ca2+]i increase. PMID- 18678985 TI - Epileptogenic activity induced by teicoplanin and effects of some antiepileptics in mice. AB - The present study was undertaken to clarify the epileptogenic activity induced by intracerebroventricular injection (i.c.v.) of methicillin-resistant Staphylococcus aureus (MRSA) antibiotics in mice. Teicoplanin (200 microg, i.c.v.) caused dose-related behavioral seizures such as head twitch and forelimb clonus. At the same time, the drug caused electroencephalographic (EEG) seizures characterized by spike-and-wave complex and a continuous spike with high amplitude. At a high dose (500 microg, i.c.v.), the drug caused a severe clonic convulsion followed by continuous spike and spike-and-wave complex on EEG. On the other hand, vancomycin caused no or almost no epileptogenic activity in both behavior and on EEG. Diazepam and sodium valproate dose-dependently antagonized epileptic seizures in behavior and on EEG induced by teicoplanin (500 microg, i.c.v.). In contrast, carbamazepine and ethosuximide caused no significant changes in both behavioral and EEG seizures induced by teicoplanin. From these findings, it can be concluded that teicoplanin may cause potent epileptogenic activity different from vancomycin when used clinically at extremely high doses. In addition, it may be that teicoplanin-induced seizure is closely related with the gamma-amino butyric acid (GABA)-ergic mechanism. PMID- 18678986 TI - Effects of the beta2-agonist clenbuterol on beta1- and beta2-adrenoceptor mRNA expressions of rat skeletal and left ventricle muscles. AB - The beta2-agonist clenbuterol [4-amino-alpha(t-butyl-amino)methyl-3,5 dichlorobenzyl alcohol] is used as a non-steroidal anabolic drug for sports doping. The effects of clenbuterol on the transcriptional process and mRNA stability of beta-adrenoceptor (beta-AR) in skeletal and cardiac muscles are still unknown. Therefore, we investigated the effects of clenbuterol on beta1- and beta2-AR mRNA expressions of fast-twitch fiber-rich extensor digitorum longus (EDL), slow-twitch fiber-rich soleus (SOL), and left ventricle (LV) muscles by real-time RT-PCR. Adult male Sprague Dawley rats were divided into the clenbuterol-administered group and control group. The administration (dose = 1.0 mg/kg body weight/day, s.c.) of clenbuterol was maintained for 10 days. The administration of clenbuterol significantly increased the weight, RNA concentration, and total RNA content of EDL muscle. No effects of clenbuterol on those of SOL and LV muscles, however, were observed. The administration of clenbuterol significantly decreased beta1-AR mRNA expression of LV muscle. Furthermore, the administration of clenbuterol significantly decreased beta2-AR mRNA expression of EDL and LV muscles. No effect of clenbuterol on beta2-AR mRNA expression of SOL muscle, however, was observed. These results suggest that the effects of clenbuterol on beta1- and beta2-AR mRNA expressions and muscle hypertrophy depend on muscle fiber types. PMID- 18678987 TI - Regulation of purinergic signaling by prostaglandin E2 in murine macrophages. AB - Extracellular nucleotides are primary signals for tissue injury, acting together with various chemical mediators such as prostanoids at the inflammatory site. We investigated whether prostaglandin E2 (PGE2) affects purinergic signaling in murine J774 macrophages. J774 cells expressed four different purinoceptor mRNAs: the ionotropic P2X4 and P2X7 receptors and G-protein-coupled P2Y2 and P2Y6 receptors. Functional responses mediated by these purinoceptor subtypes were confirmed by measurement of intracellular Ca2+ concentration ([Ca2+]i) in fura-2 loaded cells. Thus, low concentrations (10 microM) of ATP (P2Y2 agonist) and UDP (P2Y6 agonist) evoked Ca2+ transient in a phospholipase C (PLC)-dependent manner, whereas the P2X7 agonist benzoylbenzoyl-ATP (BzATP, 500 microM) caused a sustained rise in [Ca2+]i. Furthermore, ivermectin, an activator of the P2X4 receptor channel, enhanced the ATP-induced [Ca2+]i elevation. PGE2 inhibited ATP- and UDP-induced [Ca2+]i elevation, without affecting the BzATP-induced sustained [Ca2+]i elevation. Stimulation of J774 cells by UDP or BzATP increased the production of macrophage inflammatory peptide-alpha (MIP-alpha). PGE2 abolished the UDP-induced MIP-alpha production, but not the BzATP-induced one. These results demonstrate that purinergic signalings in macrophages were regulated by PGE2 in a subtype-specific manner. The different inhibitory effects on distinct purinoceptor functions may be related to the anti-inflammatory property of PGE2. PMID- 18678988 TI - Selective up-regulation of P2X4-receptor gene expression by interferon-gamma in vascular endothelial cells. AB - Extracellular nucleotides are involved in the development of vascular inflammation. However, little is known about whether effects of nucleotides are modulated under inflammatory states. We investigated effects of interferon-gamma (INF-gamma) on ATP-induced responses in vascular endothelial cells. Treatment of human umbilical vein endothelial cells (HUVECs) with IFN-gamma for 24 h resulted in an enhancement of the ATP-induced increase in intracellular Ca2+ concentration ([Ca2+]i) without affecting the UTP-induced one. The increased Ca2+ response to ATP in IFN-gamma-treated cells was dependent on the extracellular Ca2+, and was not inhibited by the phospholipase C inhibitor U73122. RT-PCR and Western blotting revealed that HUVECs dominantly expressed P2X4 receptor. IFN-gamma increased P2X4-receptor mRNA and protein, accompanied by an increase in ATP triggered membrane current. IFN-gamma did not affect P2X4-receptor mRNA stability, but increased P2X4-receptor gene transcription in a cycloheximide insensitive manner. IFN-gamma stimulated phosphorylation of signal transducer and activator of transcription-1 (STAT1). Epigallocatechin gallate (EGCG), an inhibitor of STAT1-mediated signaling, and AG490, a Janus kinase (JAK) inhibitor, impaired P2X4-receptor mRNA up-regulation by IFN-gamma. These results indicate that INF-gamma selectively increases P2X4-receptor gene expression, leading to an up-regulation of purinergic signaling in vascular endothelial cells. PMID- 18678989 TI - Concentrations of isoflavones and their metabolites in the blood of pregnant and non-pregnant heifers fed soy bean. AB - The present study compared the changes in isoflavones (daidzein and genistein) and their metabolite (equol and para-ethyl-phenol) concentrations in the blood plasma of cyclic and pregnant heifers after feeding with soy bean. Twelve healthy heifers were divided into three groups: cyclic heifers (days 8-12 of the estrous cycle; control group; n=4), an early pregnancy group (2 months pregnant; n=4) and a late pregnancy group (8 months pregnant; n=4). All heifers were fed a single dose of 2.5 kg of soy bean and then blood samples were taken from the jugular vein for 8 h at predetermined intervals. The concentrations of soy bean-derived isoflavones and their active metabolites were measured in the blood plasma on an HPLC system. In the blood plasma of the early- and late-pregnant heifers, we found lower concentrations and time-dependent decreases in daidzein and genistein in comparison to cyclic heifers (P<0.05). Moreover, we noticed significant increases of equol and para-ethyl-phenol in the blood plasma of the early pregnant heifers (P<0.05). In contrast, in the blood plasma of the late-pregnant heifers, we did not find an increase in the isoflavone metabolite concentrations compared with the early-pregnant heifers (P>0.05). In conclusion, physiological status (cyclicity or pregnancy) of the females influenced the concentrations of isoflavone metabolites in the blood plasma of the heifers. The stage of pregnancy affects isoflavone absorption, biotransformation and metabolism differently and results in higher concentrations of active metabolites of isoflavones during early pregnancy in comparison to their lower concentrations during late pregnancy. Therefore, we surmise that cows are more sensitive to active isoflavone metabolite actions during early pregnancy than cyclic heifers and heifers in late pregnancy. PMID- 18678990 TI - [Mechanism of onset and aggravation of acute pancreatitis]. PMID- 18678991 TI - [New staging system for the severity of acute pancreatitis]. PMID- 18678992 TI - [Significance of revised version of prognostic scoring system for severity of acute pancreatitis in Japan]. PMID- 18678993 TI - [Japanese criteria for severity assessment of acute pancreatitis: CT Grade using contrast-enhanced computed tomography]. PMID- 18678994 TI - [Clinical features of esophago-gastric varices caused by pancreatic cancer]. AB - We investigated the endoscopic and clinical features of esophageal-gastric varices in patients with pancreatic cancer. Among 96 patients who underwent upper gastrointestinal endoscopy at the time of diagnosis of pancreatic cancer, 26% were found to have varices caused by pancreatic cancer. The number of complications associated with varices of inoperable cancers was higher than that of operable cancers, probably due to the progression of pancreatic cancer. Management of these varices is important because they could reduce the quality of life and sudden bleeding could lead to interruption of chemotherapy. PMID- 18678995 TI - [Case of esophageal cancer successfully performed in early response evaluation for preoperative chemotherapy by FDG-PET]. AB - We present a case of esophageal cancer with multiple lymph node metastases successfully performed early response evaluation for preoperative chemotherapy by FDG-PET. The decrease of SUV from baseline to 11 days after initiation of low dose FP chemotherapy were 32.8% in the primary lesion, 60.4% in the cervical lymph node and 13.5% in the abdominal lymph node. He underwent extended radical esophagectomy 4 weeks after the end of chemotherapy. The histopathologic response was Grade 1 in the primary lesion, Grade 3 in the cervical lymph node and Grade 0 in the abdominal lymph node. The early response evaluation by FDG-PET in each lesions were consistent with histopathologic response evaluation of after chemotherapy. PMID- 18678996 TI - [Case of gastric cancer with isolated brain metastasis successfully managed by gamma knife radiotherapy and chemotherapy]. AB - We describe a 51-year-old man case of a huge gastric cancer with an initial presentation of isolated brain metastasis. The patient was successfully managed by gamma knife radiotherapy, followed by chemotherapy using 5-fluorouracil and cisplatin. He has continued oral administration of TS-1 on an outpatient basis with disease free survival for more than one year. PMID- 18678997 TI - [Autopsy case of abdominal compartment syndrome in a patient with schizophrenia]. AB - A 61-year-old man who had taken several kinds of psychotropic agents for schizophrenia from eighteen was admitted due to acute abdomen. In spite of any treatment he died after arrival. The autopsy revealed marked dilation of gastrointestinal tracts without necrosis through stomach to rectum and pathological examination disclosed hypoganglionosis of whole gastrointestinal wall. We thought that he died of abdominal compartment syndrome as a result of acute on chronic secondary pseudo-obstruction of gastrointestinal tracts due to acquired hypoganglionosis, megacolon, and aerophagia. PMID- 18678998 TI - [Intestinal tuberculosis of the terminal ileum causing obstructive ileus and tuberculous peritonitis and presenting numerous peritoneal small red nodules: a case report]. AB - A 38-year-old woman suffering from lower abdominal pain was referred to our hospital. Abdominal computed tomography showed marked thickening of the terminal ileum to the cecum, localized collection of ascites, and multiple mesenteric lymphadenopathy. A barium contrast small bowel series showed solitary severe stenosis of the terminal ileum with marked swelling of the ileocecal valve, where colonoscopy could not pass through, suggesting that ileal stenosis was caused by intestinal tuberculosis. She also showed strongly positive tuberculin skin test. Laparoscopy-assisted ileocecal resection was performed for confirmation of diagnosis and removal of the stenotic intestinal lesion. Laparoscopically, numerous small red nodules scattered on the stenotic ileal serosa, peritoneum, and mesenterium. Histopathological examination revealed ileal tuberculosis causing ulcerative stricture, and mesenteric tuberculous lymphadenitis. The small red nodules were formed of hemorrhagic tuberculous nodules. PMID- 18678999 TI - [Case with hepatic artery aneurysm discovered after suffering from infectious endocarditis]. AB - A 53-year-old man suffering from infectious endocarditis developed a rupture of the mitral valve tendinous cord. Consequently a mitral valvoplasty was performed. Forty days later, the patient presented with sudden and severe epigastralgia and hematemesis, and was rushed to our hospital. An arterial phase of an abdominal contrast-enhanced CT showed a mass 3 cm in diameter which was strongly enhanced along the side of the hepatic portal region and therefore it was thought to be an aneurysm. An abdominal angiography revealed an aneurysm of the right hepatic artery. As a result, an embolization with coils was performed. Nine months after treatment, CT examination of the abdomen revealed that the aneurysm had completely disappeared. PMID- 18679000 TI - [Complete necrosis of a giant tumor in liver by transcatheter arterial embolization and percutaneous transhepatic portal embolization before liver resection]. AB - A 67-year-old man admitted for abdominal pain. Abdominal CT showed a liver tumor 14 cm in diameter in the right lobe, and a low density area spreading along the left branch of the Glisson's sheath. Serum protein induced by vitamin K absence or antagonist-II (PIVKA-II) level was found extremely high. Transcatheter arterial embolization (TAE) was performed three times, resulting the lesion reduced to 7cm in diameter. As the last TAE showed little effectiveness, we planed to percutaneous transhepatic portal embolization (PTPE) followed by extended right hepatectomy. After PTPE, PIVKA-II was normalized. Histologically, complete necrosis of the tumor lesions was observed in the resected liver. PMID- 18679001 TI - [Drug-induced liver injury caused by an herbal medicine, bofu-tsu-sho-san]. AB - A 37-year-old woman was admitted to a hospital with jaundice. Within a couple of weeks, her liver function improved with only symptomatic therapy. About 30 to 60 days before admission, she had taken a herbal medicine, bofu-tsu-sho-san. A diagnosis of drug-induced liver injury was made according to the diagnostic scale proposed at the Digestive Disease Week-Japan 2004. A drug-lymphocyte stimulation test for each ingredient of bofu-tsu-sho-san; the results were positive for Cnidii Rhizoma, Angelicae Radix and Menthae Herba. The liver biopsy specimen revealed features of acute hepatitis. Physicians should be aware that bofu-tsu sho-san can cause liver injury, as this drug is commonly used as an over-the counter medicine. PMID- 18679002 TI - Angiotensin-converting enzyme gene in Alzheimer's disease. AB - Dementia has been increasing exponentially in recent years, especially in Asia. This increasing prevalence calls for the necessity of antecedent biomarkers. The angiotensin-converting enzyme (ACE) gene, located on chromosome 17q23, has been regarded as a candidate susceptibility gene for Alzheimer's disease (AD), because ACE could degrade beta-amyloid, the pathological hallmark of AD, thereby inhibiting its aggregation. The level and activity of ACE, in part, may be modulated by the insertion or deletion (indel) polymorphism of ACE gene. The indel polymorphism, consisting of the presence or absence of a 287-bp DNA fragment, has been considered the biomarker of AD, although its validity varies with race. In the Japanese, seemingly different results have been reported. One report shows significant association of insertion homozygote with AD, whereas other shows no association of indel polymorphism with AD. In the Taiwanese, the significant association of deletion homozygote with AD was found. Moreover, clinical studies have shown that using ACE inhibitors could slow the deterioration of cognitive function in AD patients, despite that ACE can degrade beta-amyloid. These heterogeneous results on the association of ACE gene with AD and clinical significance of using ACE inhibitor in AD highlight the necessity of exploring detailed mechanisms from the ACE gene to the development of AD. These detailed mechanisms and findings may serve as the basis for further study. PMID- 18679003 TI - Imatinib mesilate inhibits neointimal hyperplasia via growth inhibition of vascular smooth muscle cells in a rat model of balloon injury. AB - Restenosis is a major problem in percutaneous catheter intervention (PCI) for coronary artery stenosis in patients with acute myocardial infarction. Coronary restenosis arises from intimal hyperplasia, i.e., hyperplasia of the vascular smooth muscle cells (SMCs) caused by endothelial cell (EC) damage due to PCI. Drug eluting stent (DES), a novel stent coated with a cell-growth inhibitor, such as rapamycin, has been utilized to block SMC proliferation, but DES also blocks EC repair and thus requires the administration of anti-platelets for a long time to prevent thrombus formation after PCI. Moreover, insufficient prevention of platelet aggregation sometimes induces restenosis after PCI. One of the signal transduction inhibitors, imatinib mesilate, blocks tyrosine kinase activity of platelet-derived growth factor receptor (PDGFR), and therefore it may block the development of neointima through growth inhibition of SMCs without the obstructive effect on EC-repair. We therefore studied the effects of imatinib on neointimal hyperplasia in a balloon injury model of rat carotid arteries. Rats were orally administered with imatinib for 14 days after balloon injury, and sacrificed to analyze the neointimal formation. Intimal hyperplasia was inhibited by imatinib in a dose-dependent manner. Therefore imatinib presumably obstructed the growth of SMCs via interception on growth-signaling of PDGFR. The administration of imatinib after coronary stenting or the use of an imatinib eluting stent may further reduce the risk of restenosis in patients. PMID- 18679004 TI - A patient with focal fibrocartilaginous dysplasia in the distal femur and review of the literature. AB - Focal fibrocartilaginous dysplasia (FFCD) is a rare and benign bone lesion that induces bowing deformity of the long bones in young children. Excessive production of fibrocartilage by abnormal differentiation in the metaphysis or by trauma during delivery or after birth is thought to cause growth disturbance. Radiologically, the lesion is characterized by a lucent defect with marginal sclerosis in the medial metaphysis of the long bone. However, there have been few reports about the initial radiological changes of FFCD before bowing started. We report a patient with FFCD in the left distal femur in whom the radiological changes were serially observed during the course of the disorder. A 2-week-old boy first visited our hospital because of left thigh pain. Plain radiographs did not show any abnormal findings at that time. At 10 weeks, a well-defined lucent defect with bony fragment inside was observed in the distal femoral medial cortex. At 1 year, this bony fragment gradually vanished but varus deformity progressed and reached approximately 40 degrees at the age of 2. After removal of the lesion, osteotomy and immobilization was performed with Ilizarov external fixator comprising rings, rods and wires. Complete bone union was achieved 3 months after operation. It is noteworthy that we could observe the initial radiological changes of FFCD before varus deformity occurred. As far as we know, there have been no descriptions of the bony fragment inside a lucent defect of the lesion. Radiological features may vary in the early phase of FFCD. PMID- 18679005 TI - Aquaporin 4 mRNA levels in neuromuscular tissues of wild-type and dystrophin deficient mice. AB - Aquaporin (AQP) 4 is a water-specific channel protein and is abundant in central nervous tissues and skeletal muscles. Recently, the AQP4 molecule has been increasingly highlighted in its pathophysiological role of several neurological diseases, such as stroke, muscular dystrophy and neuromyelitis optica. We therefore measured the levels of AQP4 mRNA and glyceraldehyde-3 phosphate dehydrogenase mRNA (an internal control) in muscle and brain tissues of wild-type mice (C57BL10/ScSn) and age-matched dystrophin-deficient mdx mice (C57BL10/ScSn mdx) by real-time quantitative RT-PCR. The relative AQP4 mRNA level was highest in the spinal cord among the neuromuscular tissues examined in wild-type mice. Among the muscle tissues of wild-type mice, the relative AQP4 mRNA level was higher in extensor digitorum longus (EDL) muscles, and its descending order was EDL, quadriceps femoris, soleus and heart muscles. It is noteworthy that there was no difference in the relative AQP4 mRNA levels in the brain tissues between wild-type mice and age-matched mdx mice. In contrast, the AQP4 mRNA level in the quadriceps femoris muscle was significantly lower in mdx mice than in wild-type mice. The fact that the spinal cord contains the highest AQP4 mRNA may be related to the pathogenesis of neuromyelitis optica, in which AQP4 protein is the target antigen. In addition, the low expression level of AQP4 mRNA in the mdx mouse muscle suggests a functional link between AQP4 and dystrophin in the muscle tissue. We suggest that a similar pathomechanism may underlie the phenotypic consequences of the mdx mouse and Duchenne muscular dystrophy. PMID- 18679006 TI - Intra-articular injection of hyaluronan diminishes loss of chondrocytes in a rat immobilized-knee model. AB - Joint immobilization is a useful and common treatment modality in orthopedics. However, it also causes unfavorable outcome such as articular cartilage degeneration. Intra-articular injection of hyaluronan has been accepted as a treatment of osteoarthritis, but its effects on immobilized joint remain to be clarified. Hyaluronan is a polysaccharide, distributed ubiquitously in various tissues. In this study, we examined the effect of hyaluronan on the articular cartilage in immobilized joints. The unilateral knee joints of adult male rats were immobilized at 150 degrees in flexion with an internal plate and screws for 1, 2, 4, 6, 8, 12, or 16 weeks (n = 84). Hyaluronan or saline (50 microl/each injection) was administered intra-articularly on the day of surgery and once a week. The articular cartilage from the medial midcondylar region of the knee was obtained, and divided into non-contact, contact and transitional areas (between the non-contact and the contact areas). In each area, a degree of degeneration was evaluated by histomorphometric grading, and measurements of thickness and number of chondrocytes. Histological grading scores in the hyaluronan group were smaller at 12 and 16 weeks compared with those in the saline group. The thickness of the articular cartilage increased in the transitional area in both groups. The number of chondrocytes in the contact and transitional areas gradually decreased, but their number in the hyaluronan group was greater at 12 and 16 weeks compared with that in the saline group. Hyaluronan showed chondroprotective effects on the articular cartilage in a rat immobilized-knee model. PMID- 18679007 TI - Beta-phenylethylamine inhibits K+ currents in neocortical neurons of the rat: a possible mechanism of beta-phenylethylamine-induced seizures. AB - beta-Phenylethylamine (beta-PEA), an endogenous amine synthesized in the brain, serves as a neuromodulator and is involved in the pathophysiology of various neurological disorders such as depression, schizophrenia, and attention-deficit hyperactivity disorder. beta-PEA fully exerts the physiological effects within the nanomolar concentration range via the trace amine receptors, but beta-PEA also causes convulsions at much higher concentrations via an as yet unknown mechanism. To investigate the electrophysiological mechanism by which beta-PEA induces convulsions, we examined the effect of beta-PEA on ionic currents passing through the cell membrane of dissociated rat cerebral cortical neurons, using a patch-clamp technique. The external application of beta-PEA suppressed ionic currents which continuously flowed when the membrane potential was held at -25 mV. The suppression was in a concentration-dependent manner and a half-maximal effective concentration was 540 muM. These currents suppressed by beta-PEA consisted of two K(+) currents: a time- and voltage-dependent K(+) current (M current) and a leakage K(+) current. The suppression of the M-current reduces the efficacy of the current in limiting excessive neuronal firing, and the suppression of the leakage K(+) current can cause membrane depolarization and thus promote neuronal excitation. Reducing both of these currents in concert may produce neuronal seizing activity, which could conceivably underlie the convulsions induced by high-dose beta-PEA. PMID- 18679008 TI - Severity of virilization of external genitalia in Japanese patients with salt wasting 21-hydroxylase deficiency. AB - Females with salt-wasting (SW) 21-hydroxylase deficiency (21OHD) may present with mild external genitalia virilization, despite complete or almost complete enzyme inactivation. We therefore analyzed genotype/phenotype correlation in 13 Japanese female patients with SW 21OHD. Criteria for classification into the SW phenotype included history of a salt-losing crisis with documented hyponatremia, hyperkalemia, and markedly elevated plasma renin activity. Urologists and pediatricians determined the Prader genital stage and classified the location of the vaginal entrance into the common urogenital sinus as low, moderate, or high. CYP21A2 gene, coding for 21-hydroxylase, was analyzed with Southern blotting and direct sequencing. Genotypes were categorized into four mutation groups, based on the degree of enzymatic activity (N, complete enzyme inactivation; groups A, < 2%, B, 3-7%, and C > 30%). Basal androgen levels were available from only six out of thirteen patients, so we could not relate androgen levels with the severity of external genitalia virilization. We compared the degree of external genitalia virilization with genotype. The severity of external genitalia virilization varied from Prader stage 1 to 4. One patient who presented with Prader 1 had a genotype consistent with Group B. In addition, discordance between Prader classification and the location of the vaginal entrance was noted; one patient classified as Prader 4 showed low vaginal entrance, while another patient classified as Prader 3 showed high vaginal entrance. The degree of the impairment of 21-hydroxylase activity does not correlate with the severity of virilization of the external genitalia in female patients with the SW type of 21OHD. PMID- 18679009 TI - Prevalence of complex sleep apnea among Japanese patients with sleep apnea syndrome. AB - Sleep apnea syndrome (SAS) is basically divided into two types: obstructive and central SAS. Recently, the concept of complex SAS has been advocated. Complex SAS is defined as SAS that initially manifests as primarily obstructive SAS, but is characterized by the frequent central apneas after the removal of upper airway obstruction. To determine the prevalence and clinical significance of complex SAS among Japanese patients with SAS, 1,312 patients with SAS were enrolled in this study. Diagnosis of central SAS was made based on diagnostic polysomnography, and differentiation of obstructive SAS from complex SAS was made from polysomnographic findings for treatment with continuous positive airway pressure, which resolved upper airway obstruction. As a result, obstructive SAS was found in 1,232 of 1,312 patients with SAS (93.9%) and central SAS was found in 14 patients (1.1%). The overall prevalence of complex SAS was 5.0% (n = 66). The prevalence of complex SAS among 1,218 male and 94 female patients with SAS were 5.3% and 1.1%, respectively. Patients with complex SAS had significantly higher apnea/hypopnea indices than patients with either obstructive or central SAS, but were similar in both mean age and average body mass index to obstructive SAS patients. There were no significant between-group differences in numbers of patients with clinical complications including hypertension, cardiac diseases, or cerebrovascular diseases. In conclusion, the prevalence of complex SAS in Japanese SAS patients is 5.0%, which is lower than previously reported prevalence of complex SAS in the USA and Australia. PMID- 18679010 TI - Gentle exercise of 40 minutes with dietary counseling is effective in treating metabolic syndrome. AB - Obese people may succeed in reducing their weight temporarily, but most of them regain the lost weight within a few years. We, therefore, recommend moderate long term changes in habit rather than a strict temporary program. The purpose of this study was to examine whether physical activity of 40 minutes, mainly walking, was effective in improving or preventing metabolic syndrome when combined with dieting. Participants, all of whom had a waist circumference >or= 85 cm (men) or >or= 90 cm (women), were assigned into one of two groups: the control group (10 men and 11 women, age: 64.2 +/- 3.4 [SD] years) and the intervention group (10 men and 12 women, age: 62.4 +/- 4.2 years). Participants (n = 22) enrolled in a 52-week program; their diets were assessed using a 3-day dietary record and daily steps were measured with a pedometer. No significant change was seen in their dietary records, but the mean number of daily steps increased from 5,806 to 8,000 9,000. In the intervention group, BMI, waist circumference, and systolic and diastolic blood pressures decreased significantly by week 12 (all p < 0.001) and remained low until week 52. In fact, no significant change was seen between weeks 12 and 52. The control group showed no significant changes in all variables. These results indicate that gentle exercise of about 40 minutes in combination with dieting is effective in preventing metabolic syndrome among a sedentary population, although its effect may be limited. PMID- 18679011 TI - Treatment of pregnant women with a betamimetic and verapamil increases the micronuclei frequency in umbilical cord blood lymphocytes. AB - In prevention of preterm labor, betamimetics are used in gynecological practice mostly combined with antiarrhythmic verapamil because of their therapeutic cardiovascular side effects. The aim of this study was to investigate the influence of a betamimetic (ritodrine hydrochloride, fenoterol or hexoprenaline) and verapamil (administered to mothers) on the frequency of micronuclei (MN) in umbilical cord blood lymphocytes of neonates, using cytokinesis-block micronucleus test. The analyzed sample included 23 babies whose mothers received the therapy and 30 control babies whose mothers received no therapy. The average MN frequency was significantly higher in the neonates whose mothers received the therapy (8.13 +/- 2.69 MN/1000 BN cells), in comparison with the baseline frequency in untreated controls (3.30 +/- 2.63 MN/1000 BN cells), with probability p < 0.05. The highest MN frequency was found in neonates whose mothers received fenoterol and verapamil (2.8-fold i.e. 9.10 +/- 3.00 MN/1000 BN cells), while ritodrine hydrochloride and hexoprenaline combined with verapamil induced 2.3-fold and 2.2-fold higher MN values than in controls (7.50 +/- 3.33 MN/1000 BN cells and 7.29 +/- 0.95 MN/1000 BN cells). Multiple linear regression analysis showed that MN frequency was affected only by the maternal therapeutic treatment, while the neonates' sex, maternal age, cigarette smoking, and therapeutic doses did not affect the MN frequency in umbilical lymphocytes of neonates. We conclude that the treatment of pregnant women with a betamimetic and verapamil significantly increases the MN frequency in umbilical cord blood lymphocytes of neonates, regardless to therapeutic doses. PMID- 18679012 TI - Increased serum levels of leptin in retinal vein occlusion. AB - Retinal vein occlusion is an important cause of visual loss. Several ocular and systemic conditions have been reported for retinal vein occlusion. The pathogenesis of thrombus formation in the retinal vein, which results in retinal vein occlusion, is unclear. The aim of this study was to investigate the correlation between increased serum leptin levels and the occurrence of retinal vein occlusion (RVO). The study group consisted of 40 patients with RVO (58.1 +/- 6 years old; 17 males and 23 females): 15 patients with central RVO, 23 with branch RVO, and 2 with hemispheric RVO. The patients who had any ocular or systemic pathology were not included in the study. The control group consisted of 40 healthy individuals of similar gender, age, date and type of health survey, and geographic region. The blood samples of the RVO patients (n = 40) and controls (n = 40) were obtained antecubitally. Leptin levels were measured by an enzyme-linked immunosorbent assay (ELISA) method, and Student's t-test was used to determine differences between the groups. The mean serum leptin levels were 12.5 +/- 1.64 ng/ml in patients with RVO and 8.4 +/- 1.22 ng/ml in the control subjects; namely, the mean serum leptin levels were significantly higher in the patients with RVO (p < 0.001). These results suggest that leptin may be involved in the pathogenesis of venous thrombosis in the retina probably through its effects on homeostasis of the vessel wall. PMID- 18679013 TI - Novel mutations of mitochondrial DNA associated with type 2 diabetes in Chinese Han population. AB - Mitochondrial single nucleotide polymorphisms (mtSNPs) have been reported to associate with type-2 diabetes mellitus (T2DM), but mtSNPs appear to be considerably different among different populations and regions. To determine mtSNPs in Chinese Han patients with T2DM, the entire sequences of the mitochondrial genomes from 72 T2DM Chinese (59 +/- 4 years) and 50 age-matched healthy subjects (controls) in Chongqing region of Western China were directly sequenced and mtSNPs were analyzed. We found that M8, M9, D, G, R and A haplogroups exist in Chinese Han population and the frequency of haplogroup M9 was significantly higher in patients with T2DM than in the controls (p = 0.0006, OR 0.06 [95% CI 0.008-0.476]). MtSNPs T3394C in NADH dehydrogenase subunit 1 (ND1), G4491A in ND2, T16189C and T16519C were found with significantly higher frequency in patients with T2DM than in the controls (T16189C, p = 0.0045; T16519C, p < 0.0001; T3394C, p = 0.0015; G4491A, p = 0.0015). In contrast, the frequency of C5178A in ND2 and A10398G in ND3 was higher in the controls than in patients with T2DM (C5178A, p = 0.014; A10398G, p = 0.0011). Our results indicate that mtSNPs T3394C, G4491A, T16189C and T16519C show susceptible tendency to T2DM and mtSNPs C5178A and A10398G seem to be genetic factors for against T2DM. These mtSNPs determined in our study is useful and could be used for early diagnosis and prevention of T2DM in Chinese Han population. PMID- 18679014 TI - The role of PI3/Akt pathway in the protective effect of insulin against corticosterone cell death induction in hippocampal cell culture. AB - Corticosterone induces neuroanatomical and neurochemical changes in the hippocampus that are associated with cognitive impairments. In the present study corticosterone induced cell death in primary hippocampal neurons cultured in Neurobasal + B27 medium. Insulin prevents neuronal cell death induced in a concentration dependent manner. The neuroprotective effect of insulin was reversed by LY294002, a phosphatidylinositol 3'-kinase (PI3 kinase) inhibitor, whereas the mitogen-activated protein kinase (MAPK) inhibitor PD98059, an upstream blocker of MAPK had no effect. Western blot analyses showed that insulin induced the activation of protein kinase B (Akt). These results suggest that insulin can prevent neuronal cell death induced by corticosterone in hippocampal neurons by modulating the activity of the PI3 kinase/Akt pathway. PMID- 18679015 TI - Left ventricular response to continuous positive airway pressure: role of left ventricular geometry. AB - BACKGROUND: Continuous positive airway pressure (CPAP) may be a useful adjunct in patients with congestive heart failure. OBJECTIVES: To evaluate the relationship between left ventricular geometry and hemodynamic response to CPAP. METHODS: Right heart catheter studies were performed in 26 patients before, during and after application of CPAP (8 cm H(2)O) over 15 min. Response to therapy was defined as an increase in stroke volume using CPAP. RESULTS: Cardiac output decreased from 6.9 +/- 1.9 to 6.2 +/- 1.4 liters/min (p = 0.01) with a slight increase after cessation of CPAP (not significant). There was no significant change in stroke volume (92 +/- 34 vs. 90 +/- 31 ml, p = 0.584) or pulmonary capillary wedge pressure (14.7 +/- 7.0 vs. 14.2 +/- 6.5 mm Hg, p = 0.26). There was a correlation between hemodynamic effects of CPAP therapy and left ventricular end-diastolic volume (r = 0.515, p = 0.01), mass-volume ratio (r = 0.41, p = 0.04) and pulmonary capillary wedge pressure (r = 0.654, p = 0.001) at baseline. Half the patients (n = 13) were categorized as responders with an average increase in stroke volume of 11.5 +/- 2.1%. Responders showed significantly higher left ventricular end-diastolic volume, pulmonary capillary wedge pressure and lower mass-volume ratio. CONCLUSION: Patients with high pulmonary capillary wedge pressure, elevated end-diastolic volumes and a low left ventricular mass-volume ratio might profit from CPAP therapy. PMID- 18679016 TI - A 62-year-old woman with bilateral pleural effusions and pulmonary infiltrates caused by extramedullary hematopoiesis. AB - A 62-year-old female presented with a 1-month history of irritating cough and increasing dyspnea. A chronic idiopathic myelofibrosis had been diagnosed 5 years ago. CT of the chest and abdomen showed bilateral pleural effusions with a thickened pleura, nodular infiltrations in both lungs, enlarged intraabdominal lymph nodes and splenomegaly. Pleuroscopy (medical thoracoscopy) on the left side revealed dense tumorous nodules mainly on the posterior chest wall pleura, but also on the diaphragm and the lung. Biopsies taken from the chest wall pleura revealed extramedullary hematopoiesis (EMH) with abnormal megakaryocytes as well as myeloid and erythroid precursors. After unsuccessful tetracycline pleurodesis, talcum slurry was instilled via the chest tube without recurrence of the pleural effusion. Furthermore, treatment with hydroxyurea was started, and the disease regressed and then remained stable over the next 24 months. In conclusion, the pleuropulmonary findings were caused by EMH due to chronic idiopathic myelofibrosis. The definite diagnosis was established by pleuroscopy followed by successful pleurodesis with talc slurry, after tetracycline pleurodesis had failed. PMID- 18679018 TI - Acetate metabolism in Escherichia coli strains lacking phosphoenolpyruvate: carbohydrate phosphotransferase system; evidence of carbon recycling strategies and futile cycles. AB - The ptsHIcrr operon was deleted from Escherichia coli wild-type JM101 to generate strain PB11 (PTS(-)). In a mutant derived from PB11 that partially recovered its growth capacity on glucose by an adaptive evolution process (PB12, PTS(-)Glc(+)), part of the phosphoenolpyruvate not used in glucose transport has been utilized for the synthesis of aromatic compounds. In this report, it is shown that on acetate as a carbon source, PB11 displayed a specific growth rate (mu) higher than PB12 (0.21 and 0.13 h(-1), respectively) while JM101 had a mu of 0.28 h(-1). To understand these growth differences on acetate, we compared the expression profiles of central metabolic genes by RT-PCR analysis. Obtained data revealed that some gluconeogenic genes were downregulated in both PTS(-) strains as compared to JM101, while most glycolytic genes were upregulated in PB12 in contrast to PB11 and JM101. Furthermore, inactivation of gluconeogenic genes, like ppsA, sfcA, and maeB,and poxB gene that codes for pyruvate oxidase, has differential impacts in the acetate metabolism of these strains. Results indicate that growth differences on acetate in the PTS(-) derivatives are due to potential carbon recycling strategies, mainly in PB11, and futile carbon cycles, especially in PB12. PMID- 18679019 TI - Evolution of family 18 glycoside hydrolases: diversity, domain structures and phylogenetic relationships. AB - Chitin and its derivates have many industrial and medical uses. There is a demand for chitin-modifying enzymes with new or modified properties and as microorganisms are the primary degraders of chitin in the environment, they provide a source of chitin-modifying enzymes with novel properties. We have analyzed the diversity, domain structure and phylogenetic relationships between family 18 chitinases based on complete genome sequences of bacteria, archaea, viruses, fungi, plants and animals. Our study shows that family 18 chitinases are divided into three main clusters, A, B and C. Clusters A and B both contain family 18 chitinases from bacteria, fungi and plants, suggesting that the differentiation of cluster A and B chitinases preceded the appearance of the eukaryotic lineage. Subgroups within clusters can have specific domain structures, as well as specific amino acid replacements in catalytic sites, which imply functional adaptation. This work provides a comprehensive overview of the evolutionary relationships of family 18 chitinases and provides a context for further investigations on functional aspects of family 18 chitinases in ecology and biotechnology. PMID- 18679017 TI - Estrogen reduces aldosterone, upregulates adrenal angiotensin II AT2 receptors and normalizes adrenomedullary Fra-2 in ovariectomized rats. AB - We studied the effect of ovariectomy and estrogen replacement on expression of adrenal angiotensin II AT1 and AT2 receptors, aldosterone content, catecholamine synthesis, and the transcription factor Fos-related antigen 2 (Fra-2). Ovariectomy increased AT1 receptor expression in the adrenal zona glomerulosa and medulla, and decreased adrenomedullary catecholamine content and Fra-2 expression when compared to intact female rats. In the zona glomerulosa, estrogen replacement normalized AT1 receptor expression, decreased AT1B receptor mRNA, and increased AT2 receptor expression and mRNA. Estrogen treatment decreased adrenal aldosterone content. In the adrenal medulla, the effects of estrogen replacement were: normalized AT1 receptor expression, increased AT2 receptor expression, AT2 receptor mRNA, and tyrosine hydroxylase mRNA, and normalized Fra-2 expression and catecholamine content. We demonstrate that the constitutive adrenal expression of AT1 receptors, catecholamine synthesis and Fra-2 expression are partially under the control of reproductive hormones. Our results suggest that estrogen treatment decreases aldosterone production through AT1 receptor downregulation and AT2 receptor upregulation. AT2 receptor upregulation and modulation of Fra-2 expression may participate in the estrogen-dependent normalization of adrenomedullary catecholamine synthesis in ovariectomized rats. The AT2 receptor upregulation and the decrease in AT1 receptor function and in the production of the fluid-retentive, pro-inflammatory hormone aldosterone partially explain the protective effects of estrogen therapy. PMID- 18679020 TI - Predictors of improvement or decline in instrumental activities of daily living among community-dwelling older Japanese. AB - BACKGROUND: To explore predictors of functional improvement or decline in instrumental activities of daily living (IADL) among community-dwelling older people. METHODS: Of all the residents (>or=65 years, n = 1,673) living in Yoita town, Niigata prefecture, Japan, in 2000, 1,544 subjects who participated in the baseline interview survey (T1) were followed up until 2002 (T2). A wide range of variables obtained in T1 was entered into the model predicting functional improvement or decline in IADL. RESULTS: Among 1,274 individuals who participated in both T1 and T2, 105 subjects showed improvement, 194 decline, and the remaining 975 showed stability in IADL. Multiple logistic regression analyses indicated that good self-rated health and cognitive function (MMSE >or=24) at T1 were good predictors of both improvement and protection against decline in IADL. Independence in basic activities of daily living predicted improvement in IADL. Advanced age and poor walking ability predicted decline in IADL. CONCLUSIONS: Good self-rated health and cognitive function predicted both improvement and protection against decline in IADL. Thus, intervention programs aiming at maintaining both good self-rated health and cognitive function should be able to improve IADL. PMID- 18679021 TI - Parathyroid hormone-related peptide-secreting pancreatic neuroendocrine tumours: case series and literature review. AB - BACKGROUND/AIMS: Parathyroid hormone-related peptide (PTHrP) was discovered in 1987 as the tumour product responsible for humoral hypercalcaemia of malignancy (HHM). Pancreatic neuroendocrine tumours (NETs) are uncommon malignancies with an incidence of 1:100,000 population. PTHrP-secreting NETs are rare but are being recognised as a cause of HHM in NETs. We describe the largest series to date and the management of these rare tumours. METHODS: One male and 4 female patients were seen in our unit from 1998 to 2006. The average duration of follow-up was 86 months from initial diagnosis. All patients had a histologically confirmed diagnosis of pancreatic NET, all of which were low-grade tumours. PTHrP assay was performed in 4 of 5 cases and was elevated in these patients. RESULTS: Initial management involved surgery in 2 of 5 cases, whilst 2 cases underwent chemotherapy and 1 case was commenced on interferon-alpha. We discuss the further therapies that all patients underwent and the complexities that arose in managing disease progression as well as resistant hypercalcaemia. CONCLUSION: PTHrP secreting tumours should be considered in the differential diagnosis of patients with NETs who present with hypercalcaemia and a disproportionately low PTH. PTHrP tumours may well be underestimated as the assay is difficult to perform and may not be requested. In hypercalcaemic patients the standard management acutely includes intravenous fluids, diuretics and intravenous bisphosphonate. In the further management, somatostatin analogue therapy is the first medical option to consider and we discuss the role of other therapies. PMID- 18679022 TI - List of drugs in development for neurodegenerative diseases. Update June 2008. PMID- 18679023 TI - Depression, symptoms and the quality of life in patients on hemodialysis for end stage renal disease. AB - BACKGROUND: End-stage renal disease (ESRD) is a progressive, debilitating, chronic illness requiring nursing and medical interventions. The goal of this study was to explore the level of depression experienced by patients receiving hemodialysis (n = 146), and to compare the symptoms with the quality of life (QoL) between patients that were depressed and those that were not depressed. METHODS: For this descriptive, cross-sectional survey, participants aged 18 and above were recruited from three different regions in the Republic of Korea. The level of depression, symptoms and QoL of the participants were measured by questionnaires from October to December 2006. The data was analyzed with descriptive statistics, the chi(2) and t test using the SPSS WIN 14.0 program. RESULTS: The prevalence of depression (PHQ-9) among the participants was 25.34%. There were more symptoms reported in the depressed group of patients than in those that were not depressed. In addition, the QoL was not as good in the depressed group when compared to patients that were not depressed. CONCLUSION: Therefore, the evaluation for depression may be an important part of the management of patients with ESRD. Further research is needed to understand the causal relationship between depression and health outcomes. PMID- 18679024 TI - Pentoxifylline inhibits transforming growth factor-beta signaling and renal fibrosis in experimental crescentic glomerulonephritis in rats. AB - BACKGROUND/AIMS: Pentoxifylline (PTX) has been shown to inhibit renal inflammation in a rat model of crescentic glomerulonephritis. The present study investigated the role of PTX in renal fibrosis in rats with crescentic glomerulonephritis. METHODS: A rat model of accelerated anti-glomerular basement membrane glomerulonephritis was induced and treated with PTX or vehicle control for 3, 7, 14 and 28 days. The therapeutic effect and mechanism of PTX on renal fibrosis were examined by Northern blot and immunohistochemistry. RESULTS: Diseased rats treated with vehicle control developed a severe crescentic glomerulonephritis with progressive renal fibrosis identified by a marked accumulation of alpha-SMA+ myofibroblasts and collagen matrix. This was associated with tubular epithelial-myofibroblast transition as evident by de novo expression of alpha-SMA and a loss of E-cadherin on damaged tubular epithelial cells. Further studies revealed that severe renal fibrosis was associated with upregulation of renal TGF-beta1 and activation of TGF-beta/Smad signaling, which was blocked by treatment with PTX. CONCLUSIONS: PTX may be an anti-fibrosis agent capable of inhibiting renal fibrosis in a rat model of crescentic glomerulonephritis. Blockade of TGF-beta1 expression and Smad2/3 activation may be a mechanism by which PTX inhibits renal fibrosis. PMID- 18679025 TI - Retinal ganglion cell distribution and spatial resolving power in elasmobranchs. AB - The total number, distribution and peak density of presumed retinal ganglion cells was assessed in 10 species of elasmobranch (nine species of shark and one species of batoid) using counts of Nissl-stained cells in retinal wholemounts. The species sampled include a number of active, predatory benthopelagic and pelagic sharks that are found in a variety of coastal and oceanic habitats and represent elasmobranch groups for which information of this nature is currently lacking. The topographic distribution of cells was heterogeneous in all species. Two benthic species, the shark Chiloscyllium punctatum and the batoid Taeniura lymma, have a dorsal or dorso-central horizontal streak of increased cell density, whereas the majority of the benthopelagic and pelagic sharks examined exhibit a more concentric pattern of increasing cell density, culminating in a central area centralis of higher cell density located close to the optic nerve head. The exception is the shark Alopias superciliosus, which possesses a ventral horizontal streak. Variation in retinal ganglion cell topography appears to be related to the visual demands of different habitats and lifestyles, as well as the positioning of the eyes in the head. The upper limits of spatial resolving power were calculated for all 10 species, using peak ganglion cell densities and estimates of focal length taken from cryo-sectioned eyes in combination with information from the literature. Spatial resolving power ranged from 2.02 to 10.56 cycles deg(-1), which is in accordance with previous studies. Species with a lower spatial resolving power tend to be benthic and/or coastal species that feed on benthic invertebrates and fishes. Active, benthopelagic and pelagic species from more oceanic habitats which feed on larger, more active prey, possess a higher resolving power. Additionally, ganglion cells in a juvenile of C. punctatum, were retrogradely-labeled from the optic nerve with biotinylated dextran amine (BDA). A comparison of the BDA- labeled material and tissue stained for Nissl substance indicates that 76% of the cells in the retinal ganglion cell and inner plexiform layers of the central retina in this species are non-ganglion cells. PMID- 18679026 TI - Effects of autoantibodies against M2 muscarinic acetylcholine receptors on rabbit atria in vivo. AB - BACKGROUND: Evidence has shown that autoantibodies against M2 muscarinic acetylcholine receptors may play a role in the development of atrial fibrillation. The goal of this study was to evaluate the effects of anti-M2 receptor autoantibodies on rabbit atria in vivo. METHODS: Rabbits were immunized monthly with a synthetic peptide corresponding to the M2 receptor. The atrial electrophysiology of the isolated perfused rabbit hearts was studied. Western blots and RT-PCR were performed to determine the expression of the atrial muscarinic receptor and the acetylcholine-activated potassium channel. Atrial tissue was stained with Masson's trichrome stain for fibrosis detection. RESULTS: Autoantibodies were persistently detected in immunized rabbits. M2 rabbits showed a significantly shorter atrial effective refractory period and a longer intra atrial activation time than control rabbits. Electrical stimuli induced a significantly larger number of repetitive atrial responses in M2 rabbits. The protein levels of the M2 receptor and GIRK4 were upregulated in M2 rabbits. The mRNA levels of GIRK1 and GIRK4 were also upregulated. Histological examination revealed significantly increased diffuse fibrotic deposition in M2 rabbit atria compared with control rabbits. CONCLUSION: The M2 receptor autoantibody-positive rabbits showed altered atrial electrophysiology, overexpression of the M2 receptor-I(K,ACh) pathway and atrial fibrosis, which indicates that the autoantibodies against M2 receptors may participate in the induction and perpetuation of atrial fibrillation. PMID- 18679027 TI - Noninvasive coronary angiography: the holy grail for assessment of chest pain in patients presenting to the emergency department? PMID- 18679028 TI - Patient and caregiver perspectives of quality of life in dementia. An investigation of the relationship to behavioural and psychological symptoms in dementia. AB - BACKGROUND/AIMS: Behavioural and psychological symptoms have a high prevalence amongst patients with dementia and can be a significant source of distress to both patients and carers. The present study explored the relationships between quality of life and behavioural and psychological symptoms in dementia (BPSD) from both patient and carer perspectives. Contextual factors surrounding the occurrence of BPSD were explored. METHODS: Forty-six patients and 116 carers completed questionnaire measures of BPSD and quality of life. RESULTS: BPSD were negatively associated with both patient and carer ratings of patient quality of life. The symptoms related to lower quality of life differed between patient and carer ratings: depression and irritability were found to predict lower carer ratings of quality of life, whilst delusions and apathy indicated lower patient ratings. Carers were found to be poor at identifying antecedents and consequences of BPSD. CONCLUSIONS: The presence of BPSD is associated with lower quality of life in dementia. Interventions designed to improve the quality of life for patients should focus on the BPSD specifically associated with the patient's rating of quality of life. Information regarding the role of contextual factors in behaviour management should be made available to carers. PMID- 18679029 TI - Early-onset dementia is associated with higher mortality. AB - BACKGROUND/AIMS: Our objective was to compare the mortality risks of patients with early- and late-onset dementia with non-demented controls of the same age range and to analyse the mortality risks in subtypes of dementia. METHODS: We included 1,203 subjects from our memory clinic. Patients with dementia were subdivided into 2 groups, with early- (<65 years) or late-onset dementia (>or=65 years), and compared with non-demented controls of the same age range. We used Cox proportional hazard models to estimate mortality risks. RESULTS: When compared to non-demented controls of the same age range, the patients with early onset dementia had a strongly elevated mortality risk [hazard ratio (95% confidence interval) = 43.3 (3.1-600.4)], while those with late-onset dementia had a moderately increased mortality risk compared to older controls [hazard ratio (95% confidence interval) = 3.4 (1.8-6.2)]. An additional analysis showed that, adjusted for age, Alzheimer's disease seemed to have the most benign course, with a fourfold increased mortality risk. Dementia with Lewy bodies and vascular dementia (frequently seen at older age) and frontotemporal lobar degeneration and 'other dementias' (often found at younger age) had a six- to eightfold increased mortality risk. CONCLUSION: Dementia is a risk factor for death. Especially in young patients the impact of dementia on mortality is high. PMID- 18679031 TI - Illicit methadone use and abuse in young people accessing treatment for opiate dependence. AB - The current study looked at illicit methadone use in a group of young people attending a Dublin clinic for treatment of opiate dependence. A structured questionnaire was designed and administered to eligible participants (aged 25 years or under on treatment for opiate dependence). Of the total number of participants (n = 81), 73% reported illicit methadone use before treatment entry and the main reason for use was to manage opiate withdrawals. During treatment 55% reported illicit methadone use and failure to get to the clinic was the main reason given. Some participants reported use for hedonic effects (33% prior to treatment and 12% in treatment). Despite strict controls, most participants reported that illicit methadone was readily available at low cost (EUR 23 per 80 mg). Despite legislative and administrative efforts to curtail methadone diversion in Ireland, we found that it is widespread. Although it is generally used to self-medicate withdrawal symptoms in established opiate addicts, the extent of its use raises concerns as a risk for opiate overdose in the community. PMID- 18679030 TI - Cohort changes in cognitive function among Danish centenarians. A comparative study of 2 birth cohorts born in 1895 and 1905. AB - BACKGROUND/AIM: The objective was to examine cohort changes in cognitive function in 2 cohorts of centenarians born 10 years apart. METHODS: The Longitudinal Study of Danish Centenarians comprises all Danes reaching the age of 100 in the period April 1, 1995 through May 31, 1996. A total of 207 out of 276 persons participated (75%). The Danish 1905 Cohort Survey includes all individuals born in 1905. In total, 225 out of 364 persons who reached the age of 100 in the cohort participated in the most recent 2005 follow-up (62%). In both cohorts, cognitive function was assessed using the Mini-Mental State Examination. RESULTS: There were no significant differences in cognitive score between the two centenarian birth cohorts. However, modest tendencies were seen towards better cognitive functioning for the centenarians in the 1905 cohort living at home compared to the home-dwelling ones in the 1895 cohort and worse cognitive performance for the centenarians in the 1905 group living in nursing homes compared to the nursing home dwellers in the 1895 cohort. CONCLUSION: The increasing number of centenarians may not entail larger proportions of cognitively impaired individuals in this extreme age group. PMID- 18679032 TI - Pain therapy in german neurology. Structures and standards of evaluation. AB - In order to analyze aspects of pain patient care in neurology, we conducted a survey among German neurology departments that aimed to determine different structural aspects of neurological pain medicine. A 5-page questionnaire was sent to 391 neurological departments, and a return rate of 59.8% was achieved. Some 70% of university-based neurology departments have established their own outpatient clinic, and some 80% of these departments actively take part in interdisciplinary pain services. University hospitals operate an interdisciplinary pain clinic in 94.7%. Almost all neurological departments admit pain patients, especially for further diagnosis and neurological treatment. These fields are accepted as important neurological tasks. The quality of care is reported to be excellent. Routine questioning for pain of all admitted patients is carried out by 85% of all hospitals, and an extensive pain history is taken by almost 90% of departments. Our survey data confirm that the documentation of medical, psychological and psychosocial pain histories and the process of pain patient care are partly fulfilled, yet need improvement. Routine application of validated instruments and regular inquiry of the presence/course of pain may improve the process of care and--as the basis of outcome--pain management in neurology. PMID- 18679033 TI - Stereopsis and color vision impairment in patients with right extrastriate cerebral lesions. AB - BACKGROUND AND AIM: The extrastriate cortex is a visual processing structure beyond the striate cortex. This region contains cells that differ in selectivity for different features of stimuli such as color, motion and stereopsis. We studied stereopsis and color vision in patients with right extrastriate cerebral lesions. METHODS: Patients with right extrastriate cerebral lesions determined by MRI were recruited among consecutive inpatients. Patients with cognitive impairment (Mini Mental Status Examination score <24) were excluded. An ophthalmologist performed cortical visual testing: visual acuity, stereopsis, color vision and strabismus within 1 week after the insult. RESULTS: Cortical visual testing was performed in 12 patients. Among the patients studied, 11 demonstrated abnormalities: reduced stereoacuity (8/12), color vision abnormalities (7/12). Seven patients showed complex cortical visual abnormalities. However, only 2 patients complained of subjective clinical symptoms. The MRI lesions in patients with disturbance of stereopsis were located in the dorsal occipitoparietal area and the ventral occipitotemporal area. Color vision abnormalities corresponded to lesions in the ventral occipitotemporal area. CONCLUSIONS: Patients with lesions in the right extrastriate cortex tend to have disturbance of stereopsis and color vision abnormalities, even in the absence of visual complaints. PMID- 18679034 TI - Disturbance of predictive response initiation of eye and head movements in cerebellar patients. AB - The predictability of target occurrence has influence upon the latency of motor initiation. Higher frequencies (>0.5 Hz) of periodically paced target presentation lead to decreased latencies of movement initiation. In this study we investigated the initiation of head movements and saccades during predictable and random target presentation in cerebellar patients. During the predictive condition, in particular during higher temporal frequencies and greater amplitudes, patients showed increased latencies compared to healthy subjects. The latencies between the onset of the agonistic and antagonistic muscle pulses were correlated with the severity of cerebellar impairment. We conclude that the cerebellum is involved in predictive response initiation of eye and head movements which is emphasized at higher temporal frequencies. PMID- 18679035 TI - Multifactorial etiology of recurrent miscarriage and its scientific and clinical implications. AB - A considerable proportion of recurrent miscarriage (RM) cases are caused by recurrent chromosomally abnormal conceptions. However, in younger patients and patients with multiple miscarriages, maternal causes seem to dominate. No single biomarker with a high predictive value of maternally caused RM has been identified. Non-genetic biomarkers in RM may not reflect conditions in the pregnant uterus and we rarely know whether they are causes or consequences of miscarriage. Studies of genetic biomarkers are probably the best way to reveal the pathophysiological mechanisms behind RM. Epidemiological and genetic studies suggest that RM due to maternal causes has a multifactorial background. The risk of RM in each patient is probably determined by the interaction of many genetic variants and environmental factors but only few of these have so far been identified. The genetic biomarkers for RM can probably be classified into three groups: (1) variants associated with excessive inflammatory responses and autoimmunity; (2) variants of importance for insulin and androgen sensitivity and turn-over, and (3) variants associated with thrombophilia. Identification of these markers will require whole genome association studies comprising thousands of individuals. Acknowledgement of the multifactorial background for RM has important implications for the management of patients in clinical practice. PMID- 18679036 TI - Renal ACE and ACE2 expression in early diabetic rats. AB - BACKGROUND/AIM: The role of angiotensin-converting enzyme (ACE)-related carboxypeptidase-2 (ACE2) in the regulation of the renin-angiotensin system is not well characterized. This study investigated the changes in the expression of ACE and ACE2 in the kidney in early diabetic rats. METHODS: Streptozotocin induced diabetic rats were examined. The concentrations of angiotensin II in plasma, urine, and renal cortex were measured by radioimmunoassay. The mRNA expression of ACE, ACE2, angiotensin II type 1 receptor (AT1R), and Mas receptor (MasR) in glomeruli and cortex was assessed using real-time PCR. The glomerular and cortical expression of ACE and ACE2 was assessed by immunohistochemistry. RESULTS: For renal cortical tissue, the angiotensin II level was more intensified in the 8-week diabetic rats. Immunohistochemical experiments showed that ACE was increased, but ACE2 was decreased in the glomeruli of 8-week diabetic rats, while both ACE and ACE2 in the tubules were increased. The AT1R mRNA in the glomeruli was decreased, while the MasR mRNA was increased in 2-week diabetic rats. CONCLUSION: The combined effects of increased ACE and decreased ACE2 in glomeruli may be associated with the activation of the renin-angiotensin system in early diabetic rats, which is known to increase proteinuria. PMID- 18679037 TI - Nonventilatory strategies for prevention and treatment of bronchopulmonary dysplasia--what is the evidence? AB - This review briefly summarizes the evidence for a number of mainly drug-related strategies to prevent or treat bronchopulmonary dysplasia (BPD). Oxygen supplementation is frequently used in neonatal units and oxygen toxicity plays an important role in the pathogenesis of BPD. However, current evidence for an optimal oxygen saturation for extremely premature infants is scarce. This gap in knowledge will hopefully be closed by a number of ongoing or prospective trials addressing this issue. The role of inhalational nitric oxide in the prevention of BPD is still unclear despite existing data from a number of large randomized trials. Early administration of caffeine seems to confer a benefit with regard to BPD. Prophylactic or early application of surfactant may also be beneficial. High intramuscular doses of vitamin A slightly reduce the incidence of the disease. There is currently no evidence supporting other nutritional interventions to prevent BPD. Anti-inflammatory drugs, like alpha(1)-proteinase inhibitor, pentoxifylline and azithromycin, and antioxidants, like N-acetylcysteine and superoxide dismutase, have not been proven effective yet. Diuretics can ameliorate lung function, but there is no evidence supporting their long-term use. Ureaplasma urealyticum colonization of airways is associated with an increased risk of BPD. However, there is no proof for an effect of erythromycin on BPD. The potential roles for therapies like bombesin-like peptide-blocking antibodies or Clara cell 10-kDa protein have yet to be defined. PMID- 18679038 TI - Plasma brain-derived neurotrophic factor as a peripheral marker for the action mechanism of antidepressants. AB - Numerous studies have demonstrated that depression is associated with a decreased expression of brain-derived neurotrophic factor (BDNF). BDNF shows antidepressant like effects in animal models. Therefore, we tested the hypothesis that BDNF might be a peripheral marker for the mechanism of action of antidepressant agents in humans. Thirty-two patients meeting the DSM-IV criteria for major depressive disorder and 50 normal control subjects were recruited for this study. Plasma BDNF levels and Hamilton Depression Rating Scales were measured at baseline and 6 weeks after antidepressant administration. At baseline, the mean plasma BDNF level was lower in the depressive patients (698.1 +/- 537.7 pg/ml) than in the control subjects (830.7 +/- 624.8 pg/ml), although the difference was not statistically significant (p = 0.33). The plasma BDNF levels in depressive patients significantly increased from 698.1 +/- 537.7 to 1,028.9 +/- 744.5 after 6 weeks of antidepressant treatment (p = 0.01). Moreover, plasma BDNF levels were significantly increased after 6 weeks of treatment in the responder group, while there was no statistically significant change in the unresponsive group. These results suggest that the therapeutic response after antidepressant administration might be attributable to the increase in BDNF levels. BDNF may play a critical role in the action mechanism of antidepressant drugs. Further studies with a larger number of subjects are needed to verify these findings. PMID- 18679039 TI - Heart rate variability in schizophrenic patients switched from typical antipsychotic agents to amisulpride and olanzapine. 3-month follow-up. AB - Schizophrenia is a severe mental disorder that requires lifelong treatment, and therefore information on the cardiovascular safety and tolerance of antipsychotics is of significant clinical importance. Atypical antipsychotics have been used to treat schizophrenia patients since the 1990s, and more and more patients have been switched to these from typical antipsychotics; however, there is still no accessible evaluation tool for assessing cardiovascular safety. In this study, we used a computer-assisted 5-min measurement of resting heart rate variability (HRV) in schizophrenia patients who were switched to atypical antipsychotic agents (amisulpride and olanzapine) due to severe side effects (tardive dyskinesia). In 15 patients who switched to amisulpride and 18 to olanzapine, HRV was evaluated before the medication was switched, and patients were followed up every month for 3 months after the switch. Frequency-domain analyses of short-term and stationary respiratory rate (RR) intervals were performed to evaluate low-frequency power (LF; 0.04-0.15 Hz), high-frequency power (HF; 0.15-0.40 Hz), the ratio of LF to HF (LF/HF), and LF in normalized units (LF%). Our results showed significant increases in the mean, variance and HF of RR intervals in the amisulpride group, but not in the olanzapine group. These results indicate that amisulpride has a more vagotonic effect, suggesting greater cardiovascular safety as compared with olanzapine when subjects are switched from typical antipsychotic agents. PMID- 18679040 TI - Indirect comparison of once daily insulin detemir and glargine in reducing weight gain and hypoglycaemic episodes when administered in addition to conventional oral anti-diabetic therapy in patients with type-2 diabetes. AB - AIMS: Basal insulin administered to type-2 diabetic patients with poor glycaemic control when managed with oral anti-diabetics (OADs) alone can lead to an increased risk of weight gain and hypoglycaemia. In the absence of head-to-head trials, an indirect comparison of the once-daily insulin detemir with insulin glargine was conducted on the following outcomes: weight gain, hypoglycaemic episodes, and HbA(1c). METHODS: Parallel-group randomised controlled trials of at least 20 weeks duration that compared once-daily evening glargine or detemir with a common comparator, neutral protamine Hagedorn insulin (evening), were selected. Trials focused on insulin-naive, type-2 diabetic patients poorly controlled with OAD. Five open-label trials were identified (n = 2,092 patients; n = 1 detemir and n = 4 glargine trials), with an indirect comparison of glargine (n = 869 patients) and detemir trials (n = 169 patients) carried out using meta-regression to control for covariates. Weight gain was analysed as weighted mean differences (WMD), hypoglycaemic episodes as odds ratios (OR), and HbA(1c) at the end of study as standardised mean differences (SMD). RESULTS: Patients receiving evening detemir gained significantly less weight (unadjusted WMD -1.22 kg, 95% CI -2.15, 0.29 kg; p = 0.010) and significantly fewer of them experienced hypoglycaemic episodes versus evening glargine (unadjusted OR 0.52, 95% CI 0.28, 0.98; p = 0.044). There was no significant difference between treatments for the mean HbA(1c) level at study endpoint (unadjusted SMD 0.09, 95% CI -0.16, 0.33; p = 0.480). CONCLUSIONS: Once-daily use of insulin detemir resulted in significantly less weight gain and fewer hypoglycaemic episodes than glargine, while maintaining clinically appropriate HbA(1c) levels in type-2 diabetic patients currently receiving OAD. PMID- 18679041 TI - Rapid inhibition of ATP-induced currents by corticosterone in rat dorsal root ganglion neurons. AB - The effects of corticosterone (CORT), a natural glucocorticoid hormone, on ATP induced currents in rat dorsal root ganglion (DRG) neurons and the underlying signaling mechanism were studied by using patch-clamp techniques. Three types of currents (fast, slow and mixed) were evoked by ATP in cultured DRG neurons. Pretreatment with CORT (0.01-10 mumol/l) for 30 s could inhibit the fast current and the fast component of the mixed current. In contrast, CORT had no significant effect on the slow current evoked by ATP. The inhibitory effects were concentration dependent, reversible and could be blocked by glucocorticoid receptor antagonist RU38486 (10 micromol/l), but not by GDP-beta-S (0.2 mmol/l), a blocker of G protein activation. Membrane-impermeable bovine serum albumin conjugated corticosterone failed to mimic the effects of CORT. The inhibitory effects of CORT on ATP-induced currents diminished after adding protein kinase A inhibitor H89 (10 micromol/l), but were not influenced by protein kinase C inhibitor chelerythrine chloride (10 micromol/l). These results suggest that glucocorticoid hormones might participate in the control of pain by modulating P2X(3) receptor-mediated events in sensory neurons, and the effect is mediated by glucocorticoid receptors and the downstream activation of protein kinase A. PMID- 18679042 TI - Production and perception of temporal patterns in native and non-native speech. AB - Two experiments examined production and perception of English temporal patterns by native and non-native participants. Experiment 1 indicated that native and non native (L1 = Chinese) talkers differed significantly in their production of one English duration pattern (i.e., vowel lengthening before voiced versus voice-less consonants) but not another (i.e., tense versus lax vowels). Experiment 2 tested native and non-native listener identification of words that differed in voicing of the final consonant by the native and non-native talkers whose productions were substantially different in experiment 1. Results indicated that differences in native and non-native intelligibility may be partially explained by temporal pat-tern differences in vowel duration although other cues such as presence of stop releases and burst duration may also contribute. Additionally, speech intelligibility depends on shared phonetic knowledge between talkers and listeners rather than only on accuracy relative to idealized production norms. PMID- 18679043 TI - Effects of accentual fall on phrase-final vowel duration in Japanese. AB - This study explores the effect of accentual fall on phrase-final vowel duration in read declarative sentences of Standard Japanese. The results show that an intonational phrase-final vowel is significantly shorter when the final phrase has an accentual pitch fall than when it does not. Previous studies have reported a vowel-shortening effect for the final position of Japanese declarative sentences; the new finding reported in this paper is that this shortening effect is enhanced by the pitch fall of an accent in the sentence-final phrase. PMID- 18679044 TI - Effect of speaking rate on the identification of word boundaries. AB - Two experiments were conducted to determine whether listeners' ability to use allophonic variation to identify word boundaries is influenced by speaking rate. Listeners in both experiments were presented two-word sequences (such as great eyes) spoken by naturally fast and naturally slow talkers; in one experiment the sequences were presented in quiet and in the other they were presented in noise. The listeners' task was to identify the intended sequence from among four choices with alternative segmentations (e.g. great eyes, gray ties, great ties, gray eyes). In both experiments performance was worse for the sequences produced by the naturally fast talkers than for those produced by the naturally slow talkers. This finding suggests that the extent to which allophonic variation contributes to the identification of word boundaries may depend on the rate at which the speech was produced. PMID- 18679045 TI - Role of physical chemical properties in drug relay into skin compartments. AB - The ability of a drug to reach the interstitial fluid is an important aspect of drug efficacy - as a possible indicator of skin and cell compartment concentration. This overview addresses the relationship of the physical properties of several antibiotics to their ability to enter the interstitial fluid utilizing a cantharidin blister model. By collecting pharmacokinetic data for 12 antibiotics administered orally and 11 intravenously, we compared the fraction of drug that reaches the interstitial fluid (AUC(blister)/AUC(serum)) to partition coefficients. Following data analysis, we found no correlation (p = 0.98 and 0.09, respectively) between hydrophobicity and the ability to reach the interstitium. Both orally and intravenously administered antibiotics display a strong linear correlation (p < 0.001 and p = 0.006, respectively) in the total concentration found in the serum and interstitial fluid indicating that serum concentration may be an important factor in dictating interstitial fluid concentration. This correlation may prove useful in clinical application as a means of determining interstitial fluid concentration by measuring only serum levels. PMID- 18679046 TI - The contributions of Paul Ehrlich to pharmacology: a tribute on the occasion of the centenary of his Nobel Prize. AB - On the centenary of Paul Ehrlich's Nobel Prize, this German researcher deserves to be remembered as a pioneer in a large number of scientific disciplines. As a result of his enthusiasm and scientific abilities, dedication, and contacts with other scientists of his time, he was able to make countless contributions in fields as diverse as histology, haematology, immunology, oncology, microbiology and pharmacology, among others. Although the Swedish award was meant to recognize the standardization of the manufacture of antidiphtheria serum, it was the discovery of arsphenamine (Salvarsan) for the treatment of syphilis which won him wider international acclaim. From a pharmacological perspective, Ehrlich's outstanding contributions include dissemination of the 'magic bullet' concept for the synthesis of antibacterials, introduction of concepts such as chemoreceptor and chemotherapy, and linking the chemical structure of compounds to their pharmacological activity. These achievements took place within the framework he established for the transition from experimental pharmacology to therapeutic pharmacology. He introduced a modern research system based on the synthesis of multiple chemical structures for pharmacological screening in animal models of disease states. These contributions were undoubtedly decisive in propitiating the wider development of antibiotics decades later. For these reasons, it is fitting to mark the 100th anniversary of the Nobel Prize awarded to this great scientist by commemorating the importance of his contributions to the advance of pharmacology. PMID- 18679047 TI - Immunomodulation by melatonin: its significance for seasonally occurring diseases. AB - Melatonin is not only synthesized by the pineal gland but also in many other organs and tissues of the body, particularly by lymphoid organs such as the bone marrow, thymus and lymphocytes. Melatonin participates in various functions of the body, among which its immunomodulatory role has assumed considerable significance in recent years. Melatonin has been shown to be involved in the regulation of both cellular and humoral immunity. Melatonin not only stimulates the production of natural killer cells, monocytes and leukocytes, but also alters the balance of T helper (Th)-1 and Th-2 cells mainly towards Th-1 responses and increases the production of relevant cytokines such as interleukin (IL)-2, IL-6, IL-12 and interferon-gamma. The regulatory function of melatonin on immune mechanisms is seasonally dependent. This fact may in part account for the cyclic pattern of symptom expression shown by certain infectious diseases, which become more pronounced at particular times of the year. Moreover, melatonin-induced seasonal changes in immune function have also been implicated in the pathogenesis of seasonal affective disorder and rheumatoid arthritis. The clinical significance of the seasonally changing immunomodulatory role of melatonin is discussed in this review. PMID- 18679048 TI - Innate immune markers in mothers and fathers of children newly diagnosed with cancer. AB - OBJECTIVE: The diagnosis of a life-threatening illness in a child is one of the most stressful events imaginable for parents and is associated with increased anxiety and distress. Despite associations between stress and immune function in animal and human models, the immune function in caregivers of children at the time of a child-related potentially traumatic event, like cancer, is not known. METHODS: Nineteen parents (11 mothers, 8 fathers), representing six caregiver pairs, provided blood for natural killer (NK) cell count by flow cytometry and function assays [% NK whole blood, absolute NK whole blood, LU(20) (lytic unit) peripheral blood mononuclear cells, LU(20) NK cells] and completed self-report measures (acute stress) within 2 weeks of learning their child had cancer. The NK cell assay was also completed with a sample of healthy adults, the immune reference group. RESULTS: There were similar levels of NK cell activity between caregivers and the immune reference group. Immune level and psychological outcomes were not associated. LU(20) peripheral blood mononuclear cells and LU(20) NK cells were each correlated at r = 0.83 between mothers and fathers in the same family. CONCLUSIONS: Although based on a small sample, these preliminary results suggest that knowledge about stress responses in parents of children with life-threatening illness may be important and provide novel data regarding the shared impact of stress on immune function within caregiver dyads. PMID- 18679049 TI - The effects of corticosterone and beta-endorphin on adherence, phagocytosis and hydrogen peroxide production of macrophages isolated from Dark Agouti rats exposed to acute stress. AB - BACKGROUND: Given that stressful experiences can change the reaction to a subsequent exposure to stress, we tested the in vitro effects of the stress mediator corticosterone and the opioid peptide beta-endorphin on the function of macrophages isolated from control rats and from rats exposed to electric tail shock stress (ES) or a stress-witnessing procedure (SW) 24 h earlier. METHODS: Peritoneal macrophages isolated from control and stressed rats of the Dark Agouti (DA) strain were treated in vitro with corticosterone or beta-endorphin and tested for adherence, phagocytosis and hydrogen peroxide release. RESULTS: ES diminished adherence and SW decreased phagocytosis. The suppressive effect of corticosterone on phagocytosis was absent in rats exposed to ES and SW, while the suppressive effect of beta-endorphin on adherence was not observed in rats exposed to SW. ES and SW did not affect H(2)O(2) release, neither directly nor indirectly by changing macrophage response to corticosterone and beta-endorphin in this test. CONCLUSIONS: In DA rats early macrophage activation steps, i.e. adherence and phagocytosis, were more sensitive to stress than their effector function, corresponding to H(2)O(2) production. We suggest that neuroendocrine mediators of stress that converge on macrophages might have changed specific macrophage receptors or postreceptor events and alter their response to artificial stressors, represented by corticosterone and beta-endorphin in vitro. PMID- 18679050 TI - Gamma-aminobutyric acid inhibits synergistic interleukin-6 release but not transcriptional activation in astrocytoma cells. AB - OBJECTIVE: A decline in the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) may enhance cytokine release in Alzheimer's disease (AD) resulting in neuroinflammation. We investigated the GABA-mediated suppression of the synergistic release of interleukin (IL)-6 due to interleukin 1-beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha). METHODS: Rat C6 astrocytoma cells were treated with IL-1 beta and TNF-alpha in the absence and presence of GABA. Activation of p38, degradation of I kappaB-alpha and total cellular IL-6 were determined by Western blot analysis. IL-6 release and gene expression were measured by ELISA and RT-PCR, respectively. RESULTS: Although p38 and nuclear factor (NF)-kappaB are essential for the synergistic release of IL-6, GABA did not affect either p38 phosphorylation or I kappaB-alpha degradation. Additionally, GABA suppressed IL-6 release but did not alter cytokine-driven synergistic increases in IL-6 gene expression. Western blot analysis revealed that co-treatments with IL-1 beta and TNF-alpha resulted in an increase in intracellular IL-6 that was prevented by GABA. CONCLUSION: GABA-induced inhibition of IL-6 release appears to coincide with a reduction in cellular IL-6. The GABA-induced suppression of IL-6 release may include inhibition of IL-6 gene translation. PMID- 18679051 TI - Effect of bacterial lipopolysaccharide on the reproductive axis of prepubertal and peripubertal female rats. Ontogenic changes in the immune-neuroendocrine interactions. AB - The immune, endocrine and nervous systems are closely interrelated, which allows the organism to respond to different types of stress such as infection. Chronic infectious and inflammatory conditions are often accompanied by an impaired reproductive function. Leptin, a hormone produced by adipose tissue, exerts a regulatory function on the reproductive axis. It has homology with other proinflammatory cytokines and could be modified by lipopolysaccharide (LPS). Therefore, these studies were designed to investigate the effect of LPS administration on the neuroendocrine mechanisms involved in the regulation of the reproductive axis during sexual maturation. Fifteen- and 30-day-old female rats were injected with a single dose of LPS 250 microg/kg (i.p.) and then nitric oxide synthase (NOS) activity, hypothalamic excitatory/inhibitory amino acids and Gn-RH content, serum LH and leptin concentration were studied. In 15-day-old female rats LPS treatment did not modify hypothalamic inducible (iNOS) and constitutive (cNOS) NOS activity, Gn-RH, glutamate (GLU) and GABA content. Also serum LH and leptin levels were not modified. In 30-day-old rats LPS increased iNOS and cNOS activity (p < 0.001) and hypothalamic Gn-RH content (p < 0.001). At this age hypothalamic GABA content was significantly decreased (p < 0.001) without changes in GLU content, and serum LH (p < 0.001) and leptin (p < 0.0001) decreased significantly. In summary, current studies have demonstrated that LPS administration to 15- and 30-day-old female rats results in a different response of the hypothalamus-pituitary-gonadal axis and of the adipose tissue, demonstrating an ontogenic response of the immune-neuroendocrine system to LPS administration. PMID- 18679052 TI - In vitro macrophage activity: biphasic effect of prolactin and indirect evidence of dopaminergic modulation. AB - OBJECTIVE: Prolactin (PRL), a peptide hormone produced by the pituitary gland, is involved in the interaction between the neuroendocrine and immune system. Since dopamine receptor antagonists increase serum levels of PRL, both PRL and dopamine receptors might be involved in the modulation of macrophage activity, providing means of communication between the nervous and immune systems. This study evaluated the effects of PRL and the dopamine antagonist domperidone (DOMP) on macrophage activity of female rats. METHODS: Oxidative burst and phagocytosis of peritoneal macrophages were evaluated by flow cytometry. Samples of peritoneal liquid from female rats were first incubated with PRL (10 and 100 nM) for different periods. The same procedure was repeated to evaluate the effects of DOMP (10 and 100 nM). RESULTS: In vitro incubation of macrophages with 10 nM DOMP decreased oxidative burst, after 30 min, whereas the PMA-induced burst was decreased by DOMP 10 nM after 2 and 4 h. Treatment with PRL (10 and 100 nM) for 30 min decreased oxidative burst and rate of phagocytosis (10 nM). After 2 h of incubation, 10 nM PRL decreased oxidative burst and phagocytosis intensity, but increased the rate of phagocytosis. On the other hand, after 4 h, PRL 10 and 100 nM increased oxidative burst and the rate of phagocytosis, but decreased intensity of phagocytosis. CONCLUSIONS: These observations suggest that macrophage functions are regulated by an endogenous dopaminergic tone. Our data also suggest that both PRL and dopamine exert their action by acting directly on the peritoneal macrophage. PMID- 18679053 TI - Increased serum levels of inflammatory markers in chronic institutionalized patients with schizophrenia. AB - Activation of the cytokine systems may be involved in the neuropathological changes occurring in the central nervous systems of schizophrenic patients. However, associations between the levels of cytokines and the severity of symptoms have not been completely established. OBJECTIVE: It was the aim of this study to evaluate serum levels of tumor necrosis factor (TNF)-alpha and their soluble receptors (sTNFR) in schizophrenic patients and healthy controls. METHODS: Forty male institutionalized schizophrenic patients (mean age +/- SD, 52.3 +/- 9.9 years) and 20 asymptomatic matched controls were recruited. The severity of symptoms was assessed using the Brief Psychiatric Rating Scale, the Positive and Negative Syndrome Scale and the Abnormal Involuntary Movement Scale. Serum levels of cytokines were measured by ELISAs. RESULTS: Serum levels of sTNFR1 and sTNFR2 were increased in schizophrenic patients when compared with controls (all p < 0.05), but there was no difference in TNF-alpha levels. There was no correlation between the length of disease/hospitalization or the severity of symptoms and the serum levels of these molecules. CONCLUSION: Inflammatory markers are increased in schizophrenia but they do not correlate with symptom severity. PMID- 18679054 TI - Species and sex identification of the Korean goral (Nemorhaedus caudatus) by molecular analysis of non-invasive samples. AB - Korean long-tailed goral (Nemorhaedus caudatus) is one of the most endangered species in South Korea. However, detailed species distribution and sex ratio data on the elusive goral are still lacking due to difficulty of identification of the species and sex in the field. The primary aim of this study was to develop an economical PCR-RFLP method to identify species using invasive or non-invasive samples from five Korean ungulates: goral (N. caudatus), roe deer (Capreolus pygargus), feral goat (Capra hircus), water deer (Hydropotes inermis) and musk deer (Moschus moschiferus). The secondary aim was to find more efficient molecular sexing techniques that may be applied to invasive or non-invasive samples of ungulate species. We successfully utilized PCR-RFLP of partial mitochondrial cytochrome b gene (376 bp) for species identification, and sex specific amplification of ZFX/Y and AMELX/Y genes for sexing. Three species (goral, goat and water deer) showed distinctive band patterns by using three restriction enzymes (XbaI, StuI or SspI). Three different sexing primer sets (LGL331/335 for ZFX/Y gene; SE47/48 or SE47/53 for AMELX/Y gene) produced sex specific band patterns in goral, goat and roe deer. Our results suggest that the molecular analyses of non-invasive samples might provide us with potential tools for the further genetic and ecological study of Korean goral and related species. PMID- 18679055 TI - Overexpression of a rice diacylglycerol kinase gene OsBIDK1 enhances disease resistance in transgenic tobacco. AB - A rice diacylglycerol kinase (DGK) gene, OsBIDK1, which encodes a 499-amino acid protein, was cloned and characterized. OsBIDK1 contains a conserved DGK domain, consisting of a diacylglycerol kinase catalytic subdomain and a diacylglycerol kinase accessory subdomain. Expression of OsBIDK1 in rice seedlings was induced by treatment with benzothiadiazole (BTH), a chemical activator of the plant defense response, and by infection with Magnaporthe grisea, causal agent of blast disease. In BTH-treated rice seedlings, expression of OsBIDK1 was induced earlier and at a higher level than in water-treated control seedlings after inoculation with M. grisea. Transgenic tobacco plants that constitutively express the OsBIDK1 gene were generated and disease resistance assays showed that overexpression of OsBIDK1 in transgenic tobacco plants resulted in enhanced resistance against infection by tobacco mosaic virus and Phytophthora parasitica var. nicotianae. These results suggest that OsBIDK1 may play a role in disease resistance responses. PMID- 18679056 TI - Transcriptional response according to strength of calorie restriction in Saccharomyces cerevisiae. AB - To characterize gene expression that is dependent on the strength of calorie restriction (CR), we obtained transcriptome at different levels of glucose, which is a major energy and carbon source for budding yeast. To faithfully mimic mammalian CR in yeast culture, we reconstituted and grew seeding yeast cells in fresh 2% YPD media before inoculating into 2%, 1%, 0.5% and 0.25% YPD media to reflect different CR strengths. We collected and characterized 160 genes that responded to CR strength based on the rigorous statistical analyses of multiple test corrected ANOVA (adjusted p0.7). Based on the individual gene studies and the GO Term Finder analysis of 160 genes, we found that CR dose-dependently and gradually increased mitochondrial function at the transcriptional level. Therefore, we suggest these 160 genes are markers that respond to CR strength and that might be useful in elucidating CR mechanisms, especially how stronger CR extends life span more. PMID- 18679058 TI - Commentary: sociocultural factors in medicine and gastrointestinal research. AB - (Table is included in full-text article.)The authors examine the work by Gerson et al., with respect to the role of cultural factors in clinical gastrointestinal practice and research. These often underappreciated factors play an important role in the diverse populations that clinicians work with in westernized countries and internationally. This study argues for the importance of addressing culture when examining the perception of symptoms and the roles of physician and patient. Research questions as well as treatment decisions can be compromised if a patient's cultural belief systems are not accommodated. Sociocultural factors affect physician-patient interactions and particularly affect how symptoms are reported and treated. An understanding of specific cultural contributions to symptom experience can enhance the clinician's ability to engage in more effective research and treatment. PMID- 18679059 TI - Leucocytapheresis for inflammatory bowel disease in the era of biologic therapy. AB - (Table is included in full-text article.)The development of biologicals such as infliximab to intercept TNF-alpha validates the current perception that certain cytokines are major factors in the immunopathogenesis of inflammatory bowel disease (IBD), ulcerative colitis and Crohn's disease. Furthermore, major sources of inflammatory cytokines include activated peripheral granulocytes and monocytes (GM), which in patients with IBD are elevated with increased survival time and are found in vast numbers within the inflamed intestinal mucosa. Hence, elevated GM should be appropriate targets of therapy in IBD. Accordingly, in recent years technologies such as the Adacolumn have been developed for selective depletion of elevated GM by extracorporeal adsorption (GMA). Published data show that GMA in patients with steroid-dependent or steroid-refractory IBD is associated with striking efficacy and tapering or discontinuation of steroids, whereas in steroid naive patients GMA spared patients from steroids. Likewise, GMA at appropriate intervals in patients at a high risk of clinical relapse significantly suppressed relapse, thus sparing the patients from the morbidity associated with active IBD. First ulcerative colitis episode, steroid naivety and short disease duration seem to be good predictors of response to GMA and on the basis of our experience, GMA seems to have an excellent safety profile. PMID- 18679060 TI - Quality of Barrett's surveillance in The Netherlands: a standardized review of endoscopy and pathology reports. AB - OBJECTIVE: The quality of Barrett's surveillance relies on an adequate endoscopic inspection, obtaining a sufficient number of biopsy specimens, good communication of the endoscopic findings to the pathologist, and an accurate description of the histological findings by the pathologist. The aim of this study was to assess the quality of Barrett's surveillance in daily practice in The Netherlands. MATERIALS AND METHODS: A structured scoring list was developed to evaluate systematically the quality of endoscopy and pathology reports. From 15 hospitals, endoscopy reports and corresponding pathology reports were selected randomly and evaluated by two observers. In case of disagreement, the observers re-evaluated the reports in a consensus meeting. RESULTS: One hundred and fifty cases were evaluated. The adherence to current standard biopsy protocols (four quadrant biopsies every 2 cm) decreased with increasing Barrett's length: 0-5 cm: 79%; 5-10 cm: 50%; 10-15 cm: 30%. The indication for the endoscopy was mentioned in 28% of the pathology reports, in 4% the presence/absence of oesophagitis was communicated, and in 19% the location and/or aetiology of biopsies was described. The presence/absence of dysplasia was mentioned in 93% of pathology reports. CONCLUSION: Endoscopy reports and pathology reports in current practice do not include all relevant information for an adequate Barrett's surveillance. In short Barrett's oesophagus, the adherence to current standard biopsy protocols is acceptable, but in longer segments (with a higher risk for neoplastic progression) this is clearly insufficient. The communication between endoscopists and pathologist is suboptimal. PMID- 18679061 TI - Inappropriate prescription of proton pump inhibitors on two pulmonary medicine wards. AB - OBJECTIVE: We recently noticed that proton pump inhibitor (PPI) use was high on a pulmonary medicine ward of a university clinic and reasons for this high usage were not clear. Our aim was to determine the indications for PPI use on two pulmonary medicine wards and to assess whether this use was appropriate. METHODS: We assessed prospectively the number of patients on PPIs and the indications for PPI use on two pulmonary medicine wards, one from a university and one from a regional clinic in The Netherlands. RESULTS: On admission, 88 of 300 (29%) patients already used PPIs. The use of PPIs was discontinued in three (1%) patients, whereas PPIs were initiated in 45 (15%) patients, resulting in 130 (43%) patients on PPIs during hospitalization. The most common indication for PPI use was the prevention of medication-associated complications. In 78 of 130 (60%) patients on PPIs, this medication was used for a registered indication, whereas in 52 (40%) patients a registered indication was not present (overuse). In contrast, 19 of 300 (6%) patients were not prescribed PPIs despite the presence of a registered indication for its use (underuse). No differences were found in prescription rate and indications for PPI use between the university and the regional clinic. CONCLUSION: PPI use was very common on two pulmonary medicine wards in the Netherlands. Forty percent of the patients used their PPIs for a nonregistered indication. As use of PPIs is costly and may be associated with side effects, hospital physicians should to be better educated on guidelines for its use. PMID- 18679062 TI - Changes in gene expression of gastric mucosa during therapeutic acid inhibition. AB - BACKGROUND: Long-term therapy with potent acid inhibitors is a common treatment for gastro-esophageal reflux disease. Administration of proton pump inhibitors (PPIs) causes profound and continuous hypochlorhydria by inhibition of the proton pump in gastric parietal cells. Long-term hypergastrinaemia increases mucosal thickness and enterochromaffin-like cell density in oxyntic mucosa. OBJECTIVE: The aim of this study was to see whether this very common clinical intervention induces significant changes in the gastric mucosal gene expression pattern. METHODS: Seven patients suffering from gastro-esophageal reflux disease were included in this study. Endoscopic biopsies were taken from the corpus mucosa before and toward the end of a 3-month treatment with the PPI esomeprazole. RESULTS: Microarray analysis identified 186 differentially expressed genes. A high proportion of genes with changed gene expression levels during PPI treatment are involved in proliferation, apoptosis, and stress response. CONCLUSION: This study identified many genes that were not previously known to be affected by inhibition of gastric acid secretion. Further characterization of the functional roles of genes whose expression is modulated by potent acid inhibition may give new insight into the biological responses to potent acid inhibition, including the mucosal response to the moderately increased gastrin levels encountered in clinical practice. PMID- 18679063 TI - Pneumococcal infection in patients with coeliac disease. AB - OBJECTIVES: Some patients with coeliac disease are hyposplenic. Splenectomy is a risk factor for pneumococcal infection. Our objective was to determine the risk of invasive pneumococcal disease - septicaemia, pneumonia or meningitis - in patients with coeliac disease. METHODS: We analysed routinely collected, linked statistical records of hospital admission to study the risk of pneumococcal infection in patients with coeliac disease, in patients who underwent splenectomy and in a comparison cohort. The main outcome measure was the rate ratio for pneumococcal infection in the coeliac and splenectomized cohorts, compared with the comparison cohort. We undertook the study using linked records in two temporally and geographically distinct populations: the Oxford region (1963-1999) and the whole of England (1998-2003). RESULTS: The rate ratio of pneumococcal infection in patients with coeliac disease was 2.06 (95% confidence interval, 1.27-3.15) in the Oxford population and 1.61 (1.36-1.90) in England as a whole. As a comparison, the rate ratios in splenectomized patients were 3.40 (2.44-4.60) and 3.32 (2.80-3.90) in the Oxford and England populations, respectively. The increased rate ratio in coeliac patients persisted beyond the first year after diagnosis of the coeliac disease. The period covered by the Oxford study was mainly before the widespread availability of pneumococcal vaccination; but the availability of pneumococcal vaccine was widespread during the time of the English study. CONCLUSION: Some patients with coeliac disease have an elevated risk of invasive pneumococcal disease. PMID- 18679064 TI - Comparison of the efficacy of granulocyte and monocyte/macrophage adsorptive apheresis and leukocytapheresis in active ulcerative colitis patients: a prospective randomized study. AB - BACKGROUND: Ulcerative colitis (UC) is a chronic inflammatory bowel disease associated with recurring inflammation of the colorectal mucosa. Recently, cytapheresis has emerged as a new treatment for patients with UC. Removal methods are mainly performed with beads [granulocyte and monocyte/macrophage adsorptive apheresis (GMCAP)] or filters [leukocytapheresis (LCAP)]. Both treatments have been reported to be effective for active UC. There have been few trials, however, comparing the efficacy of GMCAP and LCAP. In this study, we prospectively evaluated the efficacy of LCAP and GMCAP for the treatment of active UC. METHODS: Thirty-nine patients [18 male, 21 female; mean age 38.7 years; duration of disease 6 years; clinical activity index (CAI) >6 points] with moderate-to-severe active UC were randomly assigned to the LCAP (n=21) or GMCAP group (n=17). Adacolumn (cellulose acetate beads; Japan Immunoresearch Laboratories, Takasaki, Japan) for GMCAP and Cellsorba EX (polyethylene phthalate fibers; Asahi Medical Co. Ltd, Tokyo, Japan) for LCAP were used for leukocyte removal. Patients received two sessions of cytapheresis in the first week, followed by four weekly administrations. Steroid doses were tapered if patients achieved clinical improvement. When the CAI score had decreased by 5 points or more, the patient was considered to have improved. RESULTS: Thirteen patients in the GMCAP group and 14 in the LCAP group achieved clinical improvement. No significant difference was found in clinical response and clinical course between LCAP and GMCAP. Hemoglobin levels were significantly decreased immediately after one session of cytapheresis in the LCAP group. No severe adverse effects were observed in any of the patients. No significant differences were observed in any clinical parameters predictive of a response to either LCAP or GMCAP. But in all patients receiving cytapheresis, a high CAI score was a significant risk factor for treatment failure. All of the cytapheresis nonresponders had CAI scores >or=16. CONCLUSION: Both GMCAP and LCAP were effective treatments for active UC. Patients with severe UC and a high CAI score were, however, refractory to treatment. PMID- 18679065 TI - Clinical factors that impair health-related quality of life in ulcerative colitis patients vary with the disease duration. AB - BACKGROUND AND AIM: The health-related quality of life (HRQOL) of patients with ulcerative colitis (UC) can be impaired because of the chronic symptoms. Although UC patients suffer from such symptoms over the long term, there have been few reports on the changes of HRQOL with disease duration. The aim of this study was to clarify these changes. METHODS: The HRQOL of 331 Japanese UC patients was examined using the validated Japanese version of the Inflammatory Bowel Disease Questionnaire (J-IBDQ). HRQOL and factors affecting HRQOL identified using multiple linear regression analysis were stratified by disease duration. RESULTS: Of the 15 clinical factors examined, the clinical activity index score was the strongest determinant (P<0.0001) of all the scores of IBDQ regardless of disease duration. HRQOL did not differ significantly among patients with different disease durations. The factors, however, that affected HRQOL varied according to disease duration. In patients with disease duration of less than 5 years, the clinical activity index score was the predominant factor affecting HRQOL. Being 'on sick leave or hospitalized' was a significant factor impairing HRQOL in patients with disease duration of 5-9 years. Moreover, complications due to corticosteroids significantly impaired all of the IBDQ scores in patients with disease duration of 10 years or more. CONCLUSION: Factors that affected the HRQOL of UC patients varied according to the patients' disease duration. Our findings should assist in the development of a long-term strategy for the treatment of UC patients. PMID- 18679066 TI - Distribution of colon neoplasia in Chinese patients: implications for endoscopic screening strategies. AB - OBJECTIVES: Our aim was to measure the prevalence and distribution of colonic neoplasia in Chinese adults, and to estimate the sensitivity of sigmoidoscopic screening strategies for detecting those with advanced neoplasia. METHODS: Asymptomatic, average-risk Chinese adults aged 50 years or older underwent screening colonoscopy. The prevalence and distribution of colonic neoplasia and advanced neoplasia (defined as an adenoma >or=10 mm or with villous, high-grade dysplastic, or malignant features) were reviewed retrospectively and the outcomes of various sigmoidoscopic screening strategies estimated. RESULTS: Of 1,382 individuals (833 men, 549 women; mean age 58.8 years) included, 243 (18%) had colorectal neoplasia and 72 (5.2%) had advanced neoplasia. Neoplasia prevalence was significantly higher in male and older patients. No significant differences were observed in neoplasia distribution between men and women. Overall, 24 patients had advanced neoplasia in the proximal colon, of whom four had synchronous distal neoplasia. The estimated sensitivity for detecting patients with advanced neoplasia was 72% if we assumed screening sigmoidoscopy was performed, with follow-up colonoscopy for those with distal neoplasia; 165 patients would need to undergo colonoscopy. If, instead, we assumed follow-up colonoscopy was done only for patients with distal advanced neoplasia, the estimated sensitivity would decrease slightly to 71%, but the number of colonoscopies would decrease substantially to 51. CONCLUSION: In average-risk Chinese adults, screening sigmoidoscopy is estimated to detect more than two thirds of patients with advanced neoplasia. In Chinese societies with limited health-care resources, performing colonoscopy only on patients with distal advanced neoplasia is a screening approach that optimizes the return rate on colonoscopic capacity. PMID- 18679067 TI - Paediatric endoscopy performed by adult-service gastroenterologists. AB - BACKGROUND: Children often travel from district hospitals to teaching centres for endoscopic procedures by paediatric gastroenterologists. A 10-year district hospital experience of 'adult-service' gastroenterologists endoscoping children is reported with the aim of quantifying the workload, indications, sedation/anaesthesia practices, findings and safety of paediatric endoscopy performed by adult-service gastroenterologists. METHODS: Data on endoscopic procedures in patients younger than 16 years of age between 1997 and 2006 were obtained from hospital case-notes and computerized endoscopy/histology databases and were analysed. RESULTS: A total of 174 procedures (118 gastroscopies, 41 colonoscopies and 15 flexible sigmoidoscopies) were performed in 162 children. The median (interquartile range) age was 11.5 (5-14) years. Sixty-nine percent of patients were referred by paediatricians and 31% by general practitioners /other adult specialties. Children referred as outpatients waited a total of 50 (23.5 95) days from referral to procedure. Inpatient children waited 3 (1-4) days for their procedure. General anaesthesia was used in 89% (63 of 71) endoscopic procedures in children aged below 11 years and 100% of 47 procedures in children aged below 6 years. In contrast, 96% (99 of 103) procedures in children aged 11 years or older were done in the endoscopy unit with intravenous or no sedation. Organic disease was identified from 90 (52%) procedures. The most common diagnoses were coeliac disease (41), inflammatory bowel disease (26), gastro oesophageal reflux (six) and foreign body removal (seven). No endoscopic complications occurred. CONCLUSION: General gastroenterologists supported by paediatricians can provide endoscopic services for children safely and promptly in their local hospital. This is appropriate for the management of common gastrointestinal problems affecting children. PMID- 18679068 TI - Trends and forecasts of hospital admissions for acute and chronic pancreatitis in the Netherlands. AB - BACKGROUND: The incidence and prevalence of acute and chronic pancreatitis have increased in Western countries. It is likely, the number of hospital admissions has increased correspondingly. AIMS: To analyze the trends in hospital admissions in the Netherlands for acute and chronic pancreatitis from 1992 to 2004 and to forecast the number of admissions up to 2010. METHODS: Analysis of hospital admissions for acute and chronic pancreatitis accumulated in a nationwide database. Curve fitting regression models were used to explore future trends. RESULTS: The number of acute pancreatitis admissions rose in 1992-2004 from 1,785 to 3,120 (74.8% increase). The overall 'annual number' of acute pancreatitis admissions increased from 11.8 to 19.2 per 100,000 person-years. The linear regression model predicted 3,205 [95% confidence intervals (CI), 3,111-3,299] and 3,537 (95% CI, 3,429-3,645) admissions for 2007 and 2010, respectively, a further increase of at least 9.9% in 2010 compared with 2004. In the 12-year time period, chronic pancreatitis admissions showed an increase of 75.4% (from 790 to 1,386). The overall 'annual number' of chronic pancreatitis admissions increased from 5.2 to 8.5 per 100,000 person-years. The cubic regression model predicted 1868 (95% CI, 1,619-2,117) and 3,173 (95% CI, 2,456-3,890) admissions for 2007 and 2010, respectively, an additional increase of 77.2% in 2010 compared with 2004. CONCLUSION: Hospital admissions for acute and chronic pancreatitis have increased substantially from 1992-2004. This trend will most likely continue for the near future and the burden and costs to the Dutch health care system will increase accordingly. PMID- 18679069 TI - Irritable bowel syndrome: an international study of symptoms in eight countries. AB - OBJECTIVES: This report is a preliminary comparative study of irritable bowel syndrome symptoms in eight countries, USA, Mexico, Canada, England, Italy, Israel, India, and China. We also assessed global symptom patterns and correlations and relationships to several psychosocial variables. METHODS: Two hundred and thirty-nine participants completed a bowel symptom scale composed of four symptoms, abdominal pain or discomfort, bloating, diarrhea, and constipation as well as two psychosocial questionnaires, quality of relationship and attribution of symptoms to physical or emotional factors. RESULTS: Pain score in Italy, with the least urban population, was significantly higher than six of the seven other countries whereas it was lowest in India and England. Bloating was highest in Italy and constipation was highest in Mexico, both significantly higher than five other countries. Diarrhea was higher in China than five other countries. All significance values were P<0.05. Globally, diarrhea was less common than constipation, P<0.001 and bloating significantly correlated with constipation as well with pain, P<0.05. Composite analysis of psychosocial variables and symptoms indicated that family conflict correlated directly, P<0.05, whereas family support correlated indirectly, P<0.01, with pain and bloating. Pain, bloating and diarrhea were significantly attributed to physical etiology, P<0.01, whereas only diarrhea was attributed to emotional cause, P<0.05. CONCLUSION: This study suggests that there are significant variations in irritable bowel syndrome symptoms in different geographic locations around the world. Various hypotheses that may explain our data such as cultural beliefs, gut contamination, urban and rural location, dietary practice, and psychosocial factors should be further investigated. PMID- 18679070 TI - The prevalence of occult hepatitis B virus infection in type 2 diabetes mellitus patients. AB - AIM: The prevalence of occult hepatitis B virus (HBV) infection is relatively frequent among patients with immune suppression. The impairment of the immune system is well demonstrated in diabetics. We aimed to investigate the prevalence of occult HBV infection among hepatitis B core antibody (HbcAb)+/- hepatitis B surface antibody (anti-HBs) positive type 2 diabetes mellitus patients. MATERIALS AND METHODS: The study involved 100 HBcAb+/-anti-HBs type 2 diabetes mellitus patients and 100 age and sex matched, HBcAb+/-anti-HBs healthy blood donors. Exclusion criteria were positive serology for HBsAg, hepatitis C virus or HIV, diagnosis of malignancy or earlier organ transplantation history, use of immunosuppressive therapy. All patients were questioned about their past medical history and were tested for serum alanine aminotransferase and HBV DNA level. RESULTS: The diabetic patients did not differ significantly from healthy controls in terms of sex and age. HBV DNA was detected in 11% of the diabetic patients (1 x 10-5 x 10 copies/ml) and in 3% of the controls (4 x 10-1 x 10 copies/ml). The difference between groups was statistically significant (P<0.05). The history of blood transfusion, surgery, and vaccination for HBV and alcohol use were similar in both groups (P>0.05). The serum alanine aminotransferase levels in diabetic patients were close to those of controls (26.2+/-16.4 IU/l vs. 23.9+/-9.7 IU/l; P>0.05). CONCLUSION: These data suggest that the prevalence of occult HBV infection is higher in diabetics compared with healthy controls and this may contribute to the increased prevalence of primary hepatocellular carcinoma in diabetics. PMID- 18679071 TI - Effect of alcohol, cigarette smoking, and diabetes on occurrence of hepatocellular carcinoma in patients with transfusion-acquired hepatitis C virus infection who develop cirrhosis. AB - AIM: Alcohol drinking, cigarette smoking, and diabetes have been claimed as risk factors for hepatocellular carcinoma in case-control studies. The aim of this study was to define the impact of these risk factors on the development of hepatocellular carcinoma in hepatitis C virus-related liver cirrhosis. METHODS: A historical cohort of 138 patients with posttransfusion hepatitis C virus-related cirrhosis was selected by reviewing all files of patients referred to our liver unit. Sixty-three of them (46%) developed hepatocellular carcinoma. RESULTS: At univariate analysis, risk factors for hepatocellular carcinoma were observed in patients aged above 59 years [P=0.004; relative risk (RR): 2.08, 95% confidence interval (CI): 1.19-3.68], male sex (P<0.001; RR: 2.48, 95% CI: 1.59-3.87), habit of alcohol drinking (P=0.001; RR: 1.89, 95% CI: 1.24-2.88), and duration of alcohol consumption of more than 30 years (P=0.02; RR: 2.08, 95% CI: 0.98-4.40). At Cox regression analysis, only male sex was an independent predictive factor (beta=0.86; P=0.002; hazard ratio=2.4, 95% CI: 1.3-4.1). CONCLUSION: Diabetes, smoking, and alcohol drinking were not independently related to the risk of developing hepatocellular carcinoma in hepatitis C virus-related cirrhosis. PMID- 18679072 TI - Peginterferon alpha-2a and ribavirin versus peginterferon alpha-2a monotherapy in early virological responders and peginterferon alpha-2a and ribavirin versus peginterferon alpha-2a, ribavirin and amantadine triple therapy in early virological nonresponders: the SMIEC II trial in naive patients with chronic hepatitis C. AB - OBJECTIVE: The objective of this study was to compare the efficacy of anti hepatitis C virus (anti-HCV) treatment schedules on the basis of an early virological response (EVR), defined as undetectable serum HCV-RNA (<50 IU/ml) after a 12-week induction course of peginterferon alpha-2a (PEG-IFN) 180 mcg/week. METHODS: A total of 210 interferon-naive patients (69% male; median age, 42 years) with histologically proven chronic hepatitis C infection (genotype 1: 62%) received PEG-IFN 180 mcg/week for 12 weeks. Patients with EVR (58%) were randomized to continue PEG-IFN monotherapy (n=64) or to add ribavirin (RBV), 800 mg/day (n=57), for 36 additional weeks. Patients without EVR (42%) were randomized to add RBV (n=42), or RBV plus amantadine, 200 mg/day (n=47), for 36 additional weeks. Sustained virological response (SVR, undetectable HCV-RNA 24 weeks after treatment completion) was compared among treatment groups. RESULTS: Patients with EVR: SVR rate was 60.3% in the PEG-IFN group versus 67.2% in the PEG-IFN+RBV group (NS). In genotypes 2/3, SVR rates were 66.7 versus 73.1% (NS); in genotypes 1/4, SVR rates were 51.6 versus 61.3%, respectively (NS). Patients without EVR: SVR was 16.7% in the PEG-IFN+RBV group versus 31.9% in the triple therapy group (P=0.07). In patients with genotypes 1/4, SVR rates were 9.4 versus 29.7% (P=0.041). CONCLUSION: In genotypes 1/4 patients without EVR, triple therapy results in higher SVR rates than standard dual therapy. This study confirms that addition of amantadine is beneficial in early-recognized 'difficult to-treat' patients. PMID- 18679073 TI - Probiotics in primary sclerosing cholangitis: a randomized placebo-controlled crossover pilot study. AB - BACKGROUND: Beneficial effects of probiotics have been reported in liver disease. Inflammatory bowel disease concurs in 90% of primary sclerosing cholangitis (PSC) patients, suggesting that substances originating from the inflamed gut may damage the biliary tree. OBJECTIVE: To assess potential beneficial effects of probiotics on serum liver tests, pruritus and fatigue in PSC. METHODS: Fourteen patients (13 male/one female, mean age 45 years), with concurrent inflammatory bowel disease were randomized to treatment with probiotics (Ecologic 641, containing four Lactobacillus and two Bifidobacillus strains; Winclove Bio Industries, Amsterdam, The Netherlands) or placebo during 3 months in a double-blind fashion. After a 1 month washout period, crossover was made. RESULTS: No changes in pruritus, fatigue and stool frequency were noted during placebo or probiotics. No significant differences were observed between treatment with probiotics and placebo in bilirubin (at end of probiotic vs. placebo period: -13 vs. -15% change from baseline; P=0.89), alkaline phosphatase (-9 vs. -9%; P=0.99), gamma glutamyl transpeptidase (-11 vs. -5%; P=0.60), aspartate aminotransferase (-16 vs. -15%; P=0.99), alanine aminotransferase (-27 vs. -26%; P=0.97), prothrombin, albumin or bile salts. CONCLUSION: Our data do not support beneficial effects of probiotics on symptoms, liver biochemistry or liver function in PSC. PMID- 18679074 TI - Learning curve and interobserver reproducibility evaluation of liver stiffness measurement by transient elastography. AB - BACKGROUND/AIMS: Fibroscan allows liver stiffness examination (LSE) that is well correlated with fibrosis stages. Our main objective was to evaluate LSE learning curve. METHODS: LSE results of five novice observers with different medical status were compared with those of five expert observers (physicians with >100 examinations) in 250 patients with chronic liver disease. Each novice-expert pair had to blindly examine 50 consecutive patients divided into five consecutive subgroups of 10 patients. RESULTS: In each observer group, novice-expert agreement [intraclass correlation coefficient (Ric)] for LSE results was excellent from the first to the last subgroup. Novice-expert agreement for LSE results varied with liver stiffness level: <9 kPa: Ric=0.49; >or=9 kPa: Ric=0.87. Relative difference (%) between novice and expert LSE results was independently associated with the number of valid LSE measurements, and stabilizes around 20 30% after the fourth valid measurement. In each observer group, novice-expert agreement (Ric) for LSE success rate progressively increased as a function of time. CONCLUSION: LSE requires no learning curve: a novice is able to obtain a reliable result after a single training session, whatever the professional status. However, success rate will progressively increase. An LSE with less than four valid measurements should not be considered as reliable. PMID- 18679075 TI - Exacerbation of oral lichen planus lesions during treatment of chronic hepatitis C with pegylated interferon and ribavirin. AB - It is estimated that 170-200 million people in the world are chronically infected with the hepatitis C virus. This fact indicates that the prevention and treatment of hepatitis C virus infection are a priority in public health care. To date, the best treatment consists of the combination of pegylated interferon (IFN) and ribavirin. IFN is costly and associated with low tolerance and severe side effects. Ribavirin is also problematic because it causes secondary anemia in most patients. It has been described that oral lichen planus (OLP) can appear or be exacerbated during the treatment of chronic hepatitis C. The improvement of the lesions of OLP after the discontinuation of therapy suggests that IFN may induce or worsen these lesions in some patients. This study examines three cases of exacerbation of OLP during the treatment of chronic hepatitis C with pegylated IFN and ribavirin. PMID- 18679076 TI - High intensity focused ultrasound therapy resulted in a complete response in a patient with advanced gastric cancer with liver metastases: a case report. AB - Different therapeutic strategies have been tried when liver metastasis develops after a gastrectomy for gastric cancer, but the disease still has a poor prognosis. We present a 35-year-old woman who achieved complete radiological remission of liver metastases from advanced gastric cancer after a single therapeutic high intensity focused ultrasound session. Our observations suggest that high intensity focused ultrasound combined with chemotherapy may be an additional treatment option for patients with liver metastases from advanced gastric cancer. PMID- 18679078 TI - Alligators and wound healing: the primordial soup. PMID- 18679077 TI - Initially obscure hepatotoxicity attributed to sildenafil. AB - We describe a patient with a hepatotoxic reaction, presenting with general malaise, severe jaundice, and pruritus. This turned out to be caused by the, at first unrevealed, use of sildenafil. The injury seems to be hepatocanalicular, characterized by a hepatocellular liver test pattern, combined with extensive cholestasis on liver biopsy. One should bare in mind that the use of sildenafil may not be readily disclosed by the patient nor his doctor. PMID- 18679083 TI - Are you ready for a Medicare audit? PMID- 18679084 TI - Topical anesthesia for pressure ulcer treatment. PMID- 18679085 TI - Activated, type I collagen (CellerateRx) and its effectiveness in healing recalcitrant diabetic wounds: a case presentation. AB - OBJECTIVE: To review the authors' experience with activated type I collagen in the treatment of recalcitrant wounds in the diabetic population resulting from minor trauma and/or venous stasis disease. With regard to activated collagen, CellerateRx's patented activated collagen fragments are a fraction of the size of the native collagen molecules and particles found in other products, delivering the benefits of collagen to the body immediately. DESIGN: A 2-case presentation wherein patients were treated with CellerateRx (activated, fragmented, and nonintact type I collagen) in a gel and powder form. SUBJECTS: Two middle-aged diabetic male patients with lower extremity wounds refractory to conservative wound care. RESULTS: Complete resolution of recalcitrant wounds in 6 to 7 weeks. CONCLUSIONS: Wound resolution was evident when using the authors' practice protocol, which includes the application of activated collagen. The inherent properties of type I collagen may contribute to a more rapid healing process. PMID- 18679086 TI - A multicenter study involving the use of a human acellular dermal regenerative tissue matrix for the treatment of diabetic lower extremity wounds. AB - This multicenter, retrospective study presents the use of a human acellular dermal regenerative tissue matrix as an alternative treatment for 100 chronic, full-thickness wounds of the lower extremity in 75 diabetic patients. Comorbidities included cardiac disease (86.0%), neuropathy (86.0%), peripheral vascular disease (82.0%), infection (54.0%), obesity (51.0%), and osteomyelitis (37.0%). Wound locations included the foot (86.0%), ankle (8.0%), and lower extremity (6.0%). Mean wound age was 20.4 weeks (1.3-191.4 weeks). University of Texas (UT) wound classifications included 15 (15.0%) 1A, 1 (1.0%) 1B, 1 (1.0%) 1C, 2 (2.0%) 1D, 18 (18.0%) 2A, 8 (8.0%) 2B, 5 (5.0%) 2C, 3 (3.0%) 2D, 3 (3.0%) 3A, 7 (7.0%) 3B, 3 (3.0%) 3C, and 34 (34.0%) 3D. The mean time to matrix incorporation, 100% granulation, and complete healing was 1.5 weeks (0.43-4.4 weeks), 5.1 weeks (0.43-16.7 weeks), and 13.8 weeks (1.7-57.8 weeks), respectively. The overall matrix success rate, as defined by full epithelialization, was 90.0%. One failed wound subsequently healed approximately 7 weeks after matrix reapplication. The healing rate was 91.0%, as 91 of the 100 wounds healed. No statistically significant differences were observed between UT classifications and time to matrix incorporation, 100% granulation, and complete healing. Absence of matrix-related complications and high rates of closure in a wide array of diabetic wounds suggest that this matrix is a viable treatment for complex lower extremity wounds. Lack of any statistically significant differences between UT grades and wound outcome end points lends further support to the universal applicability of this matrix, with successful results in both superficial diabetic wounds and in wounds penetrating to the bone or joint. PMID- 18679087 TI - Perineal dermatitis versus pressure ulcer: distinguishing characteristics. PMID- 18679089 TI - Performing a skin assessment. PMID- 18679090 TI - Financial incentives for lumbar surgery: a critical analysis of physician reimbursement for decompression and fusion procedures. AB - STUDY DESIGN: Retrospective case-control study/economic analysis. OBJECTIVE: To determine the treatment times required for isolated lumbar decompressions and for combined decompression and instrumented fusion procedures to compare the relative reimbursements for each type of operation as a function of time expenditure by the surgeon. SUMMARY OF BACKGROUND DATA: Under current Medicare fee schedules, the payment for a fusion procedure is higher than of an isolated decompression. It has been recently suggested in the lay press that the greater reimbursement for a lumbar arthrodesis may inappropriately influence the manner in which surgeons elect to treat lumbar degenerative conditions, resulting in what they believe to be a substantial number of unnecessary spinal fusions. METHODS: A consecutive series of 50 single-level decompression cases performed by single surgeon were retrospectively analyzed and compared with an equivalent cohort of subjects who underwent single-level decompression and instrumented posterolateral fusion with autogenous iliac crest bone grafting. The operative reports, office charts, and billing records were reviewed to determine the total clinical time invested by the surgeon and the Medicare reimbursement for each surgery. RESULTS: Relative to the corresponding values of the decompression group, combined decompression and fusion procedures were associated with a longer mean surgical time (134.6 min vs. 47.3 min, P<0.0001), a greater number of postoperative visits (1.0 vs. 3.2, P<0.0001), a higher mean total clinical time expenditure (186.6 min vs. 62.2 min, P<0.0001), and a lower mean dollars received per minute of surgeon time ($12.51 vs. $15.51, P<0.001). CONCLUSIONS: These findings challenge the assertion that spine surgeons have an undue financial incentive to recommend a combined decompression and instrumented fusion procedure over an isolated decompression to patients with symptomatic lumbar degeneration, especially when considering the greater time, effort, and risk characteristic of this more complex operation. PMID- 18679091 TI - Lateral mass anchoring screws for cervical laminoplasty: preliminary report of a novel technique. AB - STUDY DESIGN: Preliminary reporting of a group of patients with multiple level cervical spinal stenosis treated with a simple technique using lateral mass anchoring screw and unabsorbable suture line for securing the lamina position after expansive open-door laminoplasty. OBJECTIVE: To develop an improved method for laminoplasty fixation. SUMMARY OF BACKGROUND DATA: Laminoplasty is considered the standard procedure for treating multiple-level cervical spinal stenosis with myelopathy. Keys to successful laminoplasty are expanding and maintaining the spinal canal. There are many techniques for maintaining and securing of the expanded spinal canal such as fascial or joint capsule anchoring suture, spacer interposition, allograft, autograft, or miniplate fixation. However, many reports have indicated that these complicated and/or costly techniques are not superior to other techniques. This study reports a simple, reliable technique using a lateral mass anchoring screw for augmentation of laminoplasty fixation. METHODS: Five patients with multiple level cervical spinal stenosis underwent laminoplasty. A unilateral open door technique was done for the lesion level and the elevated lamina was fixed to lateral mass anchoring screws at each level using unabsorbable suture line. RESULTS: The mean follow-up period was 14.5 months (9 to 34 mo). Postoperatively, the Japanese Orthopedic Association score improved from an average of 8.6 (range: 7 to10 points) to 14.2 points (range: 13 to 15 points). The average recovery rate was 67% (60% to 75%). Follow-up computed tomography scans showed the average improvement in anterioposterior diameter at each level of the cervical canal to be about 4.0 to 7.7 mm. The average open angle at each level was 19.0 to 23.8 degrees. All hinged sides had bony fusion. CONCLUSIONS: Although this is a small series, the preliminary results suggest that this simple lateral mass anchoring screw technique can provide a firm and secure anchor for elevated open lamina in laminoplasty. PMID- 18679092 TI - Motion analysis of bryan cervical disc arthroplasty versus anterior discectomy and fusion: results from a prospective, randomized, multicenter, clinical trial. AB - STUDY DESIGN: Prospective, randomized, multicenter clinical trial. OBJECTIVE: Kinematic analysis of target level and adjacent motion segments after Bryan artificial cervical disc replacement versus anterior cervical fusion. SUMMARY OF BACKGROUND DATA: Disc arthroplasty has been shown to provide short-term clinical results that are comparable or better than those attained with traditional anterior cervical discectomy and fusion. One purported benefit of arthroplasty is the ability to prevent or delay adjacent level operations. METHODS: All patients received either a single-level anterior cervical discectomy and fusion with an anterior cervical plate (Atlantis anterior cervical plate, n=221) or a single level artificial cervical disc replacement (Bryan cervical disc prosthesis, n=242) at C3 to C7. Flexion, extension, and neutral lateral radiographs were obtained preoperatively, and at regular intervals of 24 months. Cervical vertebral bodies were tracked to calculate the functional spinal unit motion parameters, including flexion/extension range of motion and translation. If visible, the functional spinal unit parameters were obtained at the operative level as well as the level above and below. RESULTS: Significantly more motion was retained in the disc replacement group than the plated group at the index level. The disc replacement group retained an average of 7.95 degrees at 24 months. The preoperative motion was 6.43 degrees and there was no evidence of degradation of motion over 24 months. In contrast, the average range of motion in the fusion group was 1.11 degrees at 3-month follow-up and gradually decreased to 0.87 degrees at 24 months. The preoperative motion was 8.39 degrees. The Bryan disc did not migrate. At 24-month follow-up, there was no case of subsidence of the Bryan disc. There was no evidence of bridging bone across any of the Bryan implant disc spaces. CONCLUSIONS: The Bryan disc treatment, on average, maintained flexion/extension range of motion without degradation over 24 months. No ectopic bridging ossification was seen in any of the Bryan discs and no subsidence or displacement of the Bryan disc occurred. PMID- 18679093 TI - Correlation of ProDisc-C failure strength with cervical bone mineral content and endplate strength. AB - STUDY DESIGN: This study was designed to evaluate the effectiveness of preoperative computed tomography (CT) scan in predicting endplate strength. OBJECTIVE: To demonstrate a correlation between the cervical trabecular bone density and the failure strength of the endplate. SUMMARY OF BACKGROUND DATA: Cervical total disc arthroplasty devices have to transmit the force to the endplate for the remainder of the patients' life. One potential complication at this interface is endplate fracture and implant subsidence, which usually occurs early postoperatively and may be related to weakness of the boney endplate. METHODS: Six fresh human cadaver cervical spines were harvested and scanned for the determination of trabecular density using a peripheral quantitative CT (QCT) scanner. The specimens were then disarticulated and the inferior endplate of each vertebral segment was biomechanically tested using a 2-mm indentation probe to determine average endplate strength. The superior endplate of each vertebral body was then fitted with the appropriately sized ProDisc-C endplate and biomechanically tested until failure. Regression analyses were used to compare the interface failure stress of the implant with the bone mineral content and the average endplate stress as measured with the indentation probe. RESULTS: The average bone mineral content of the specimens was 322+/-57 mg/cm. The average endplate strength and stress measured by the indentation test was 176+/-129 N and 56+/-34 N/mm, respectively. The average ProDisc-C/endplate failure load and failure stress were 1875+/-1023 N and 10.2+/-4.1 N/mm, respectively. There was a direct correlation between the ProDisc-C/endplate failure stress and the bone mineral content measured by peripheral QCT (R=0.48, P<0.01). There was also a significant correlation between ProDisc-C/endplate failure stress and the endplate indentation stress. CONCLUSIONS: This study demonstrates the utility of a preoperative QCT scan in predicting the failure stress of the cervical endplate before total disc replacement. This information may potentially decrease early complications of device subsidence or endplate fracture. PMID- 18679094 TI - Effectiveness of repeated radiofrequency neurotomy for cervical facet joint pain. AB - STUDY DESIGN: Retrospective chart review. OBJECTIVES: To determine the success rate and duration of relief of repeat radiofrequency neurotomy (RFN) for cervical facet joint pain. SUMMARY OF BACKGROUND DATA: RFN is an effective but temporary management of cervical facet pain. When pain recurs, RFN is usually repeated, but the outcomes for repeat RFN are not well established. METHODS: Record review of consecutive patients who had an initial successful RFN, recurrence of pain, and then one or more repeat RFN. Responses of repeat RFN were compared with initial RFN for success rates and duration of relief. Success was defined as >/=50% relief of targeted pain and patient sufficiently satisfied with prior RFN to have RFN repeated. RESULTS: There were 14 women and 8 men. Mean age was 47 years (range, 34 to 66 y). Sixty-four RFNs were performed. Mean duration of relief after initial RFN was 12.5 months (range, 3 to 25 mo). Forty-two RFNs were performed after the initial RFN. Forty-one of the 42 RFNs were available to follow-up (98%). Thirty-nine of the 41 RFNs were successful (95%). To date, 11 patients had a series of 2 RFNs performed, 7 had 3, 2 had 4, 1 had 6, and 1 had 7. Twenty-two patients had a second RFN, which was successful in 20 of the 21 available to follow-up (95%) but unsuccessful in 1 (5%). The mean duration of relief in these patients was 12.7 months (range, 3 to 30 mo) and the relief is continuing in 2 patients. Eleven patients had a third RFN, of which 10 (91%) were successful and 1 (9%) was unsuccessful. The mean duration of relief in 8 patients was 9.5 months (range, 3 to 16 mo), and the relief is continuing in the other 2. Four patients had a fourth RFN, which was successful in all 4. The mean duration of relief was 8.75 months (range, 4 to 12 mo). Two patients had a fifth RFN with both having a mean duration of relief of 9 months (11 and 7 mo, respectively). There were 2 patients with a sixth RFN, both were successful, 1 lasted 18 months and in the other relief is continuing. There was 1 patient with a successful seventh RFN, and relief is continuing. The frequency of success and durations of relief remained consistent after each subsequent RFN. CONCLUSIONS: In a carefully selected group of patients with cervical facet joint pain who have been responsive to previous RFN, repeat RFNs are usually successful. PMID- 18679095 TI - The radiation exposure associated with cervical and lumbar spine radiographs. AB - STUDY DESIGN: Cross-sectional study. OBJECTIVE: To calculate the effective radiation doses of routine anteroposterior (AP) and lateral radiographs of the cervical and lumbar spines. SUMMARY OF BACKGROUND DATA: Although plain radiographs are generally used as the initial imaging modality for the evaluation of patients with spinal complaints, the radiation that patients receive during these studies has not been well quantified. The effective radiation dose represents a functional measure of exposure that takes into account the amount of radiation delivered and the radiosensitivity of the exposed organs. Consequently, the effective dose is important to consider from a radiation safety perspective. METHODS: The imaging practices of our radiology department were reviewed and the effective radiation doses for AP and lateral radiographs of the cervical and lumbar spines were calculated using the following variables: emitted radiation dose, source-to-object distance [SOD], film area, and patient tissue dimensions. Values were obtained from both direct measurements and an examination of the established protocols employed at our institution. RESULTS: The effective doses for AP and lateral cervical radiographs were 0.12 and 0.02 mSv, respectively, whereas the corresponding values for AP and lateral lumbar films were much larger (2.20 and 1.50 mSv, respectively). For comparative purposes, a typical chest x ray results in a radiation dose between 0.06 and 0.25 mSv. CONCLUSIONS: In this investigation, cervical spine films gave rise to radiation doses that are similar to those of chest x-rays. However, lumbar spine radiographs generated effective radiation doses that were approximately an order of magnitude greater than these other studies. In both the cervical and lumbar regions, AP views resulted in significantly greater radiation exposure than corresponding lateral images. The effective radiation doses reported here may prove to be valuable for assessing the relative risks and benefits of spine radiographs to establish appropriate guidelines for their use. PMID- 18679096 TI - The awl-staple versus guidewire method for placing vertebral screws in thoracoscopic anterior spinal fusion and instrumentation for adolescent idiopathic scoliosis. AB - STUDY DESIGN: A retrospective chart and radiograph review in a consecutive series of patients at a single institution. OBJECTIVE: To compare 2 methods of placing vertebral body screws in thoracoscopic anterior spinal fusion and instrumentation with respect to radiographic, clinical, and perioperative variables. SUMMARY OF BACKGROUND DATA: Placing vertebral body screws in thoracoscopic anterior spinal fusion and instrumentation for adolescent idiopathic scoliosis can be performed using 2 methods: the awl/staple technique and the guidewire method. There are no current studies that have compared both of these techniques. METHODS: Two groups of patients were analyzed depending on the method of screw placement: The AS group (n=15) used the awl/staple method and the GW group (n=27) used the guidewire method. Surgical times and estimated blood loss were obtained from the medical record. Fluoroscopy times were obtained from the radiology technician. Preoperative posteroanterior standing, bending, and immediate postoperative radiographs were examined to determine the primary curve magnitude, postoperative curve correction, curve flexibility, and the Lenke classification. RESULTS: The AS group had significantly less fluoroscopy times (156.6+/-71.4 vs. 328.4+/-171.6 s) and operative times (416.1+/-65.4 vs. 505.6+/-61.8 min) compared with the GW group (P<0.05). When calculating the duration per fusion level, the AS group was also shorter for fluoroscopy times (27.1+/-13.2 vs. 49.3+/-24.5 s/level) (P<0.05) and surgical times (72.0+/-15.2 vs. 76.9+/-9.8 min/level) (P=0.10). There was a trend toward a lower incidence of screw plow or pullout (0.0% vs. 14.8%) for the AS group (P=0.157). CONCLUSIONS: The awl/staple technique is the preferred technique for placing vertebral body screws when performing thoracoscopic anterior spinal fusion/instrumentation because it has shorter fluoroscopy times, surgical times, and a lower incidence of screw plow/pullout than the traditional guidewire method. PMID- 18679097 TI - Revision lumbar arthrodesis for the treatment of lumbar cage pseudoarthrosis: complications. AB - STUDY DESIGN: A study documenting major complications encountered in revision procedures for lumbar cage pseudoarthrosis. OBJECTIVE: To document the perioperative complications associated with revision surgery for threaded cylindrical cage pseudoarthrosis. SUMMARY OF BACKGROUND DATA: Pseudoarthrosis after cylindrical cage placement manifests as persistent or recurrent pain and disability after surgery. Revision strategies include isolated posterior stabilization and posterior bone grafting, versus circumferential revision where an attempt is made to remove the cages anteriorly, followed by posterior stabilization and fusion. Potential complications associated with these revision procedures have not been adequately documented in the past. METHODS: Forty-seven consecutive patients with the diagnosis of cylindrical cage pseudoarthrosis were surgically treated with either a circumferential revision (AP) or an isolated posterior instrumented fusion (P). All intraoperative and postoperative complications were documented. Radiographic interbody fusion rates and preoperative and postoperative visual analog scale (VAS) scores were documented. RESULTS: Three of the AP patients, all with anterior cage placement at L5-S1, had iliac vein lacerations requiring repair. A fourth patient had a ureteral injury requiring subsequent nephrectomy. Three patients who underwent circumferential revision and 2 patients who had an isolated posterior procedure had postoperative complications, including 2 infections (1 AP and 1 P), 1 radiculopathy (P), and 2 patients with prolonged ileus (both AP). There was a statistically significant decrease in overall VAS scores postoperatively for the 2 groups using the paired t test (P<0.0001). There was no difference in either the preoperative (P=0.22) or 2-year postoperative (P=0.30) VAS scores between the AP and P groups [rank-sum (Mann-Whitney) t tests]. Interbody fusion was achieved in 79% (30 of 38 levels) in the AP group. The interbody fusion rate was 37% (8 of 22) for the P group. CONCLUSIONS: Circumferential revision including cage removal, structural allograft placement, and posterior stabilization is associated with increased perioperative complications. Although an anterior approach showed increased interbody fusion rates, this technique did not lead to more superior clinical outcomes based on VAS scores. It remains to be shown by larger prospective studies if there is a true difference in outcome between these 2 groups that will justify the increased perioperative morbidity associated with attempted cage removal. PMID- 18679098 TI - The effect of parallax on intraoperative positioning of the Charite artificial disc. AB - STUDY DESIGN: Fluoroscopy imaging evaluation for total disc arthroplasty. OBJECTIVES: To describe the effect of fluoroscopic parallax on the relative position of the Charite and identify the most reliable method for evaluating intraoperative disc position. SUMMARY OF BACKGROUND DATA: Results of the Investigational Device Exemption study revealed that clinical outcomes of the Charite artificial disc correlated with accurate placement of the prosthesis. This is the first study to quantitatively evaluate the technique of intraoperative fluoroscopy during positioning of an artificial lumbar disc. METHODS: A Charite artificial disc (DePuy Spine, Raynham, MA) was implanted at the L5-S1 disc space of a radiopaque lumbar spine model in ideal position. A true anteroposterior fluoroscopic image of the L5-S1 disc space was obtained as were additional images as the central ray was moved to the left and right in 1 cm increments and with rotation at 1, 2, and 3 degrees. Images were also evaluated on a poorly placed and minimally displaced disc. Measurements of the distances from the middle tooth of the endplates to the anatomic center (spinous process) and alternative measurements for evaluating disc position were examined from computer-enhanced images. RESULTS: An ideally placed prosthesis appeared more displaced with increasing rotation and distance of the C-arm away from center when the spinous process was used as the midline reference. As little as 3 degrees of rotation of the fluoroscopic ray from the true anteroposterior image, made an ideally placed disc appear in poor position. Conversely, a poorly placed disc seemed to be in ideal position. Calculated displacement using measurements off the vertebral bodies remained more constant. Calculated displacement of an ideally placed prosthesis fell within 2 mm whereas all poorly placed discs had calculated differences greater than 4 mm. CONCLUSIONS: The spinous process is an unreliable anatomic midline marker. In contrast, the borders of the vertebral bodies can more reliably be used to calculate the displacement of the prosthesis from centerline to determine prosthesis position. Fluoroscopic parallax can cause an ideally placed prosthesis to appear more displaced with increasing rotation and distance away from the true anteroposterior image. The spinous process was determined to be an unreliable midline marker. The vertebral borders provided a more dependable anatomic reference point to establish the disc-space midline. PMID- 18679099 TI - Painful disc lesion: can modern biplanar magnetic resonance imaging replace discography? AB - STUDY DESIGN: A new magnetic resonance imaging (MRI) classification of disc degeneration was correlated with discography. It was also studied for intraobserver and interobserver variability in interpreting disc abnormality and for the capability in predicting symptomatic disc. OBJECTIVE: To correlate the results of discography with a new MRI classification for disc degeneration and the presence of the vertebral endplate changes and the high intensity zone (HIZ). SUMMARY OF BACKGROUND DATA: MRI and discography have been compared in numerous studies as methods for detecting disc degeneration. The results were conflicting and the role of MRI in assessing discogenic pain remains ambiguous. METHODS: One hundred thirty-one discs in 55 patients were studied with pain provocation discography and MRI. A new MRI classification was correlated to discography and assessed together with the presence of end plate changes and HIZ, by the sensitivity and specificity in predicting a painful disc. RESULTS: MRI classification had a good intra and interobserver agreement and a significant correlation with discographic findings. The sensitivity and specificity of MRI in predicting painful disc was 94% and 77%. The sensitivities and specificity of HIZs was 27% and 87% and for end plate changes was 32% and 98%, respectively. CONCLUSIONS: Although MRI is an excellent investigation for assessing disc morphology it should be interpreted along with discography findings before planning fusion surgery. The proposed MRI classification is a useful aid in predicting painful degenerative disc. The utility of HIZs and end plate changes is limited due to low sensitivity. PMID- 18679100 TI - The results of beta-tricalcium phosphate coated hydroxyapatite (beta-TCP/HA) grafts for interbody fusion after anterior cervical discectomy. AB - STUDY DESIGN: The efficacy of tricalcium phosphate and hydroxyapatite (beta TCP/HA) grafts was studied after anterior cervical discectomy (ACD). OBJECTIVE: This study presents our observations about the efficacy of beta-TCP/HA grafts after ACD. SUMMARY OF BACKGROUND DATA: Especially in the last 2 decades, fusion materials such as autograft and allograft, as well as different kind of cages were used to maintain fusion after ACD. METHODS: beta-TCP/HA grafts after ACD were used in 17 patients. The cervical and radicular pain was evaluated via visual analog scale (VAS) score preoperatively, at postoperative third week, and after 20 months (range: 18 to 24 mo) after the operation. The radiologic evaluations were done preoperatively, at postoperative first day and at the latest follow-up. The VAS, intervertebral space ratio, height of intervertebral disc space and neural foramen, and cervical and segmental lordosis angles were recorded preoperatively and during the postoperative follow-up period. The presence of fusion was controlled in computed tomography scans taken at the latest follow-up. RESULTS: Both clinical and radiologic evaluations yielded satisfactory results. VAS scores decreased significantly in all patients. The intervertebral space and neural foramen and intervertebral disc heights increased at postoperative day 1 but were found to be decreased at the latest follow-up (P<0.05). On the contrary the cervical and segmental lordosis angles decreased at postoperative day 1 but were found to be increased at the latest follow-up (P<0.05). There was a solid fusion in 16 out of 17 patients (94.11%). CONCLUSIONS: Although there was a loss of the initially obtained neural foraminal and disc height, the application of beta-TCP/HA graft after ACD resulted in a high rate of fusion and patient satisfaction. Additionally, the cervical and segmental lordosis was preserved. We concluded that it is a good alternative to current methods to maintain cervical alignment and fusion after ACD. PMID- 18679101 TI - An in vitro biomechanical investigation: variable positioning of leopard carbon fiber interbody cages. AB - STUDY DESIGN: This study is a biomechanical analysis of intervertebral cage placement, using a biomechanical model that has the appropriate matching geometry of the lumbar spine at the level of L4-L5 based upon prior morphometric studies. OBJECTIVE: The goal of this in vitro biomechanical analysis of interbody cages is to determine the effect of interbody cage position on the mechanics of posterior spinal instrumentation. This biomechanical analysis can potentially be used to guide surgical technique for placement of interbody cage devices. SUMMARY OF BACKGROUND DATA: Lumbar interbody spinal fusion cages are increasingly being used to promote spinal fusion and improve sagittal alignment in patients with degenerative disk disease. The transforaminal approach for placement of these cages has become popular, although the actual position of the cage that will provide optimal mechanical support in the intervertebral space is not known. MATERIALS AND METHODS: Leopard carbon fiber interbody cages (DepuySpine, Raynham, MA) were placed in a spinal fusion model simulating the L4-L5 disk space in one of 3 positions-anterior, middle, or anterolateral. We tested 5 constructs in each of the 3 positions, applying cyclic axial loads of 500 N at a rate of 4 Hz for 100,000 cycles. Strain on the posterior instrumentation and displacement of the cages was measured at periodic intervals. Load to failure of each construct was tested after 100,000 cycles were complete. RESULTS: Statistical analysis of biomechanical indicators show more strain in the anterolateral position as compared with the anterior position (P=0.002) and middle position (P=0.02). No difference was noted between anterior and middle positions (P=1.00). Interval analysis reveals differences in strain at 500 cycles in anterior versus anterolateral (P=0.01) and middle versus anterolateral (P=0.02). At 10,000 cycles, anterolateral strain was significantly higher (P=0.02) than anterior. No significant difference in strain was noted at 50,000 or 100,000 cycles between any of the positions. No significant differences were noted in displacement of the cages between each of the positions. Ultimate load to failure was lower (nonsignificant) in the anterolateral versus anterior position (P=0.06), but no difference was noted between anterior versus middle (P=0.57) or anterolateral versus middle (P=0.69) positions. Linear regression analysis of load-displacement curves shows significance at 500 cycles (P=0.02), approaching significance at 10,000 cycles (P=0.07), and no significant difference at 50,000 (P=0.28) or 100,000 (P=0.28) cycles. CONCLUSIONS: Positioning of interbody cages in an offset position shows higher strain upon posterior instrumentation than a central position, and quicker load to failure than an anteriorly placed cage. Biomechanical studies using shear loading, and testing of adjacent spinal levels, are necessary to further elucidate the biomechanical consequences of variable positioning of interbody cages. PMID- 18679102 TI - Anterior revision of a dislocated ProDisc prosthesis at the L4-5 level. AB - STUDY DESIGN: A case report describing our experience with anterior revision surgery for a dislocated ProDisc artificial disc at the L4-5 level, 2.5 years after the initial surgery. OBJECTIVE: To highlight the difficulties and risks associated with the use of a repeat anterior approach for the revision of a ProDisc that has failed at the L4-5 level. SUMMARY OF BACKGROUND DATA: As the ProDisc has a central vertical keel, more intensive vascular mobilization is required to remove the dislocated implant during surgery than that required in patients who must undergo a revision of the SB-Charite disc. To our knowledge, no prior report addresses whether the anterior removal of a ProDisc at the L4-5 level is possible in patients with severe retroperitoneal fibrosis. METHODS: We describe our surgical experience with the anterior revision of a lumbar ProDisc that failed at the L4-5 level. Revision surgery (anterior lumbar interbody fusion with percutaneous pedicle screw fixation) was performed 2.5 years after the initial surgery because of the anterior dislocation of the artificial disc after the patient had engaged in heavy lifting. The operation was performed with a repeat retroperitoneal approach. The multiple venous injuries that occurred intraoperatively were well controlled with a 5/0 polypropylene (Prolene) suture, but the patient lost a significant amount of blood (3800 mL) during surgery. RESULTS: The patient recovered from surgery without sequelae, and her preoperative symptoms resolved. CONCLUSIONS: A ProDisc revision case at L4-5 level must be attempted with preparation of available venous balloon catheter for emergent bleeding control, cell saver, pulse oxymeter on great toe, and ureteral catheter. In our opinion, usage of adhesion barrier material should be considered in anterior lumbar surgery to make revision exposure easier and safer. PMID- 18679103 TI - Severe hypotension associated with the prone position in a child with scoliosis and pectus excavatum undergoing posterior spinal fusion. AB - STUDY DESIGN: Case report. OBJECTIVE: We report a case of severe hypotension associated with prone positioning in a child with scoliosis and pectus excavatum. SUMMARY OF BACKGROUND DATA: Pectus excavatum is commonly associated with scoliosis and can impact cardiac function. METHODS: Retrospective case analysis. RESULTS: After a modified Ravitch procedure to repair the pectus excavatum, the patient was able to undergo posterior spinal fusion. CONCLUSIONS: Pediatric deformity surgery requires a multidisciplinary approach. PMID- 18679104 TI - Pisa syndrome. AB - STUDY DESIGN: Case report. OBJECTIVE: We report a patient who developed Pisa syndrome (drug-induced pleurothotonus) after treatment with tricyclic antidepressant. SUMMARY OF BACKGROUND DATA: Pisa syndrome is familiar to neuropsychiatrists, but not to orthopedic surgeons and spinal surgery specialists, both of whom have reported few cases of this syndrome. METHODS: A 56 year-old woman had experienced feelings of depression and saw a local doctor, where the tricyclic antidepressant clomipramine hydrochloride (9x25 mg tablets daily) was prescribed. From about 2 months after starting medication, she developed flexion of the trunk to the left. RESULTS: Pisa syndrome was suspected, and then, the daily dose of clomipramine hydrochloride was decreased from 9x to 3x25 mg tablets. Approximately 2 weeks after reducing the dose, abnormal postures gradually improved, and after 1 month, flexion of the trunk resolved. CONCLUSIONS: We reported a case of Pisa syndrome in which pleurothotonus occurred after treatment with a tricyclic antidepressant and improved after dose reduction. When middle-aged or elderly patients who see orthopedic surgeons display abnormal postures of the trunk, physicians need to ask the patient whether they have received a drug that can cause Pisa syndrome, such as an antipsychotic or antiemetic. PMID- 18679105 TI - Editorial announcement: publish ahead of print. PMID- 18679106 TI - The perivascular pool of aquaporin-4 mediates the effect of osmotherapy in postischemic cerebral edema. AB - OBJECTIVE: Osmotherapy with hypertonic saline ameliorates cerebral edema associated with experimental ischemic stroke. We tested the hypothesis that hypertonic saline exerts its antiedema effect by promoting an efflux of water from brain via the perivascular aquaporin-4 pool. We used mice with targeted disruption of the gene encoding alpha-syntrophin (alpha-Syn(-/-)) that lack the perivascular aquaporin-4 pool but retain the endothelial pool of this protein. DESIGN: Prospective laboratory animal study. SETTING: Research laboratory in a university teaching hospital. MEASUREMENTS AND MAIN RESULTS: Halothane anesthetized adult male wildtype C57B/6 and alpha-Syn(-/-) mice were subjected to 90 min of transient middle cerebral artery occlusion and treated with either a continuous intravenous infusion of 0.9% saline or 3% hypertonic saline (1.5 mL/kg/hr) for 48 hr. In the first series of experiments (n = 59), increased brain water content analyzed by wet-to-dry ratios in the ischemic hemisphere of wildtype mice was attenuated after hypertonic saline (79.9% +/- 0.5%; mean +/- SEM) but not after 0.9% saline (82.3% +/- 1.0%) treatment. In contrast in alpha Syn(-/-) mice, hypertonic saline had no effect on the postischemic edema (hypertonic saline: 80.3% +/- 0.7%; 0.9% saline: 80.3% +/- 0.4%). In the second series of experiments (n = 32), treatment with hypertonic saline attenuated postischemic blood-brain barrier disruption at 48 hr in wildtype mice but not in alpha-Syn(-/-) mice; alpha-Syn(-/-) deletion alone had no effect on blood-brain barrier integrity. In the third series of experiments (n = 34), alpha-Syn(-/-) mice treated with either hypertonic saline or 0.9% saline had smaller infarct volume as compared with their wildtype counterparts. CONCLUSIONS: These data demonstrate that 1) osmotherapy with hypertonic saline exerts antiedema effects via the perivascular pool of aquaporin-4, 2) hypertonic saline attenuates blood brain barrier disruption depending on the presence of perivascular aquaporin-4, and 3) deletion of the perivascular pool of aquaporin-4 alleviates tissue damage after stroke, in mice subjected to osmotherapy and in nontreated mice. PMID- 18679107 TI - Malaria and bacterial sepsis: similar mechanisms of endothelial apoptosis and its prevention in vitro. AB - OBJECTIVE AND DESIGN: Apoptotic endothelial damage contributes to multiorgan failure in Plasmodium falciparum malaria and in sepsis. In malaria, endothelial apoptosis is amplified by neutrophils and their secretory products, and reduced by inhibitors of neutrophil-derived substances in vitro. We compared the mechanisms of endothelial apoptosis in malaria and in sepsis, using the human umbilical vein endothelial cell as a model. INTERVENTIONS: Endothelial cells were incubated with patient sera (P. falciparum malaria, Escherichia coli sepsis, Staphylococcus aureus sepsis) or culture supernatants of the respective organisms, with or without neutrophils. Ascorbic acid or ulinastatin was used to neutralize reactive oxygen species or elastase secreted by neutrophils. Transwell sieve inserts or antibodies against leukocyte function antigen 1 or intercellular adhesion molecule 1 was used to study the effect of direct interaction between neutrophils and endothelial cells. The rate of apoptotic endothelial cells was determined by TUNEL and annexin staining. MEASUREMENTS AND MAIN RESULTS: Incubation of endothelial cells with patient sera or culture supernatants (P. falciparum, E. coli, S. aureus) lead to higher apoptosis rates, compared with incubation with control sera or control supernatants. Addition of neutrophils augmented the apoptosis rate further. Addition of ascorbic acid or ulinastatin reduced endothelial apoptosis in the presence of neutrophils. Separation of neutrophils from endothelial cells with Transwell sieve inserts, or addition of anti-leukocyte function antigen-1 antibodies also reduced endothelial cell apoptosis. However, addition of anti-intercellular adhesion molecule-1 antibodies restored high apoptosis rates that had been reduced by Transwell inserts. CONCLUSIONS: These in vitro results show how neutrophils can contribute to endothelial damage in malaria and in sepsis, both by their secretory products and by binding to intercellular adhesion molecule-1 on endothelial cells. The presence of similar pathomechanisms suggests that similar antiapoptotic strategies may offer potential benefit in malaria and in sepsis. PMID- 18679108 TI - Prevention of endotoxin-induced sarcoplasmic reticulum calcium leak improves mitochondrial and myocardial dysfunction. AB - OBJECTIVE: Growing evidence suggests that mitochondria function is impaired in sepsis. Here, we tested the hypothesis that lipopolysaccharide would induce mitochondrial Ca2+ overload and oxygen utilization abnormalities as consequences of sarcoplasmic reticulum Ca2+ handling derangements that are typically observed in sepsis. As lipopolysaccharide-induced sarcoplasmic reticulum dysfunction was mainly characterized by reduced sarcoplasmic reticulum Ca2+ uptake and Ca2+ leak, we tested whether dantrolene, a sarco(endo)plasmic reticulum calcium ATPase leak inhibitor, would prevent mitochondrial and cardiac contractile dysfunction. DESIGN: Randomized controlled trial. SETTING: Experimental laboratory. SUBJECTS: Male Sprague Dawley rats. INTERVENTIONS: Sepsis was induced by injection of endotoxin lipopolysaccharide (10 mg/kg/intravenously). Assessment of contractile function and Ca2+ handling was performed 4 hr after lipopolysaccharide. The relative contribution of the different Ca2+ transporters to relaxation in intact cardiomyocytes was studied during successive electrically evoked twitches and caffeine stimulation. Sarcoplasmic reticulum vesicles and mitochondria from ventricles of rats treated or not with lipopolysaccharide were prepared to evaluate Ca2+ uptake-release and oxygen fluxes, respectively. Effects of dantrolene (10 mg/kg) treatment in rats were evaluated in sarcoplasmic reticulum vesicles, mitochondria, and isolated hearts. MEASUREMENTS AND MAIN RESULTS: Lipopolysaccharide challenge elicited cardiac contractile dysfunction that was accompanied by severe derangements in sarcoplasmic reticulum function, i.e., reduced Ca2+ uptake and increased sarcoplasmic reticulum Ca2+ leak. Functional sarcoplasmic reticulum changes were associated with modification in the status of phospholamban phosphorylation whereas SERCA was unchanged. Rises in mitochondrial Ca2+ content observed in lipopolysaccharide-treated rats coincided with derangements in mitochondrial oxygen efficacy, i.e., reduced respiratory control ratio. Administration of dantrolene in lipopolysaccharide-treated rats prevented mitochondrial Ca2+ overload and mitochondrial oxygen utilization abnormalities. Moreover, dantrolene treatment in lipopolysaccharide rats improved heart mitochondrial redox state and myocardial dysfunction. CONCLUSION: These experiments suggest that sarcoplasmic reticulum Ca2+ handling dysfunction is an early event during endotoxemia that could be responsible for, or contribute to, mitochondrial Ca2+ overload, metabolic failure, and cardiac dysfunction. PMID- 18679109 TI - Wood smoke extract promotes both apoptosis and proliferation in rat alveolar epithelial type II cells: the role of oxidative stress and heme oxygenase-1. AB - OBJECTIVE: Inhalation of toxic smoke causes oxidant lung injury. Alveolar epithelial type II cells are important in the re-epithelialization of alveolar walls after lung injury. We investigated the responses of alveolar epithelial type II cells to insult by wood smoke extract, and we identified the role of reactive oxygen species and heme oxygenase-1 (an oxidative stress protein) in these responses. DESIGN: A randomized, controlled study. SETTING: A research laboratory. SUBJECTS: Cultured rat L2 and primary alveolar epithelial type II cells. INTERVENTIONS AND MAIN RESULTS: Exposure of L2 alveolar epithelial type II cells to smoke extract (60 microg/mL) caused increases in reactive oxygen species, mitogen-activated protein kinases phosphorylation, heme oxygenase-1 expression, apoptosis, proliferation and cell population, all of which were largely reduced by N-acetylcysteine (an antioxidant). Additionally, the smoke extract-induced heme oxygenase-1 induction was significantly attenuated by mitogen-activated protein kinases inhibitors, by small interfering RNA targeting mitogen-activated protein kinases or by N-acetylcysteine. Furthermore, knockdown of heme oxygenase-1 by small interfering RNA prevented heme oxygenase-1 induction whereas increasing smoke extract-induced apoptosis and suppressing smoke extract induced proliferation. Conversely, cobalt protoporphyrin IX (a heme oxygenase-1 inducer) amplified heme oxygenase-1 induction while suppressing smoke extract induced apoptosis and augmenting smoke extract-induced proliferation. Consequently, the smoke extract-induced increase in cell population was changed into a decrease by heme oxygenase-1 small interfering RNA, but was further elevated by cobalt protoporphyrin IX. Smoke extract also caused increases in heme oxygenase-1 expression, apoptosis, proliferation and cell population in primary alveolar epithelial type II cells, and heme oxygenase-1 small interfering RNA similarly augmented smoke extract-induced apoptosis and suppressed smoke extract induced proliferation in these primary cells. CONCLUSIONS: Smoke extract increases intracellular reactive oxygen species, which up-regulates heme oxygenase-1 via the mitogen-activated protein kinase pathways and also promotes both apoptosis and proliferation in rat alveolar epithelial type II cells. Additionally, smoke extract-induced heme oxygenase-1 induction counteracts smoke extract-induced apoptosis, but mediates smoke extract-induced proliferation, resulting in a net increase in cell population. Thus, in response to oxidant smoke insult, alveolar epithelial type II cells have evolved an adaptive mechanism involving heme oxygenase-1 that increases their cell population, presumably to help them perform their function of re-epithelialization following lung injury. PMID- 18679110 TI - Tissue oxygenation during management of cerebral perfusion pressure with phenylephrine or vasopressin. AB - OBJECTIVE: Phenylephrine is often used for management of cerebral perfusion pressure after traumatic brain injury, but can have undesirable actions. Few studies have evaluated alternatives. The hypothesis was that arginine vasopressin was as effective as phenylephrine for maintaining tissue oxygenation during cerebral perfusion pressure management. DESIGN: Prospective randomized, blinded animal study. SETTING: University laboratory. SUBJECTS: Thirty-five anesthetized swine (46 +/- 1 kg). INTERVENTIONS: Blunt trauma to the head and bilateral chests (estimated injury severity score was 25-32) was followed by hypoventilation. Resuscitation was divided into phases to simulate treatment in a typical prehospital, emergency room, and intensive care unit. For 30-45 mins postinjury, 1 L of normal saline was administered. For 45-120 mins, normal saline maintained systolic blood pressure >100 mm Hg plus mannitol for intracranial hypertension. After 120 mins, phenylephrine or arginine vasopressin was titrated to cerebral perfusion pressure >70 mm Hg (randomized and blinded) plus normal saline to maintain filling pressure >12 mm Hg plus glucose to maintain normoglycemia. MEASUREMENTS AND MAIN RESULTS: Mortality rate was 37% (13 of 35) within 2 hrs. Before resuscitation, mean arterial pressure was 61 +/- 5 mm Hg, heart rate was 110 +/- 6 beats/min, PaO2 was 46 +/- 2 mm Hg, and lactate was 5.0 +/- 0.4 mM. Intracranial pressure increased from 8 +/- 1 mm Hg to 20 +/- 1 mm Hg and brain tissue PO2 decreased from 19 +/- 1 mm Hg to 8 +/- 1 mm Hg. Resuscitation corrected most variables, as well as mixed venous, renal, portal, and muscle oxygen saturations, but 90% (20 of 22) required pressor support. After 6 hrs with either pressor, hemodynamics were stable. However, with phenylephrine vs. arginine vasopressin, intracranial pressure averaged >10 mm Hg higher and brain tissue PO2 was 6 mm Hg lower, whereas tissue oxygen saturations were >10% higher in the shoulder and hindlimb muscles (all p < 0.05). CONCLUSIONS: Arginine vasopressin was as effective as phenylephrine for maintaining cerebral perfusion pressure, but intracranial pressure and brain tissue oxygenation were improved at the expense of the periphery. PMID- 18679111 TI - Influence of ventilation strategies on survival in severe controlled hemorrhagic shock. AB - OBJECTIVE: To investigate the effect of different ventilation settings on hemodynamic stability in severe controlled hemorrhagic shock. DESIGN: Prospective, randomized, controlled animal study. SETTING: Research laboratory in a university hospital. SUBJECTS: Approximately 35-45 kg domestic pigs. INTERVENTIONS: Twenty-four domestic pigs were bled 45 mL/kg (estimated 65% of their calculated blood volume) and then ventilated with either 0 cm H2O positive end-expiratory pressure and a respiratory rate of 14 ventilations/min (positive end-expiratory pressure 0 respiratory rate 14), or with 5 cm H2O positive end expiratory pressure, a respiratory rate of 28 ventilations/min, and a tidal volume reduced by half (positive end-expiratory pressure 5 respiratory rate 28), or with 5 cm H2O positive end-expiratory pressure and a respiratory rate of 14 ventilations/min (positive end-expiratory pressure 5 respiratory rate 14). After 1 hr study phase surviving animals, received fluid resuscitation and were monitored for further 1 hr. MEASUREMENTS AND MAIN RESULTS: Pulmonary variables, hemodynamic variables, and short-term survival. There were no significant differences in mean arterial blood pressure and cardiac index after hemorrhage. After 20 mins of different ventilation strategies mean arterial blood pressure was 40 +/- 3 mm Hg in the positive end-expiratory pressure 0 respiratory rate 14 group, vs. 24 +/- 6 mm Hg the positive end-expiratory pressure 5 respiratory rate 28 group (p < 0.05) vs. 19 +/- 3 mm Hg in the positive end-expiratory pressure 5 respiratory rate 14 group (p < 0.01). Cardiac index was 65 +/- 5 mL/min/kg in the positive end-expiratory pressure 0 respiratory rate 14 group vs. 37 +/- 5 mL/min/kg in the positive end-expiratory pressure 5 respiratory rate 28 group(p < 0.01) and 20 +/- 3 mL/min/kg in the positive end-expiratory pressure 5 respiratory rate 14 group (p < 0.01). Mean airway pressure and positive end expiratory pressure correlated strongly with mean arterial blood pressure and cardiac index. None of the positive end-expiratory pressure 0 respiratory rate 14 animals died in the study phase, whereas six of seven positive end-expiratory pressure 5 respiratory rate 28 animals, and all seven positive end-expiratory pressure 5 respiratory rate 14 animals died. CONCLUSIONS: In this porcine model of severe hemorrhagic shock, reduction of positive end-expiratory pressure was the most important ventilation strategy component influencing hemodynamic stability. Reducing mean airway pressure by decreasing tidal volumes and increasing respiratory rates seemed to have less influence on cardiopulmonary function and survival than 0 cm H2O positive end-expiratory pressure. PMID- 18679112 TI - Efficacy of red blood cell transfusion in the critically ill: a systematic review of the literature. AB - BACKGROUND: Red blood cell (RBC) transfusions are common in intensive care unit, trauma, and surgical patients. However, the hematocrit that should be maintained in any particular patient because the risks of further transfusion of RBC outweigh the benefits remains unclear. OBJECTIVE: A systematic review of the literature to determine the association between red blood cell transfusion, and morbidity and mortality in high-risk hospitalized patients. DATA SOURCES: MEDLINE, Embase, Cochrane Register of Controlled Trials, and citation review of relevant primary and review articles. STUDY SELECTION: Cohort studies that assessed the independent effect of RBC transfusion on patient outcomes. From 571 articles screened, 45 met inclusion criteria and were included for data extraction. DATA EXTRACTION: Forty-five studies including 272,596 were identified (the outcomes from one study were reported in four separate publications). The outcome measures were mortality, infections, multiorgan dysfunction syndrome, and acute respiratory distress syndrome. The overall risks vs. benefits of RBC transfusion on patient outcome in each study was classified as (i) risks outweigh benefits, (ii) neutral risk, and (iii) benefits outweigh risks. The odds ratio and 95% confidence interval for each outcome measure was recorded if available. The pooled odds ratios were determined using meta-analytic techniques. DATA SYNTHESIS: Forty-five observational studies with a median of 687 patients/study (range, 63-78,974) were analyzed. In 42 of the 45 studies the risks of RBC transfusion outweighed the benefits; the risk was neutral in two studies with the benefits outweighing the risks in a subgroup of a single study (elderly patients with an acute myocardial infarction and a hematocrit <30%). Seventeen of 18 studies, demonstrated that RBC transfusions were an independent predictor of death; the pooled odds ratio (12 studies) was 1.7 (95% confidence interval, 1.4 1.9). Twenty-two studies examined the association between RBC transfusion and nosocomial infection; in all these studies blood transfusion was an independent risk factor for infection. The pooled odds ratio (nine studies) for developing an infectious complication was 1.8 (95% confidence interval, 1.5-2.2). RBC transfusions similarly increased the risk of developing multi-organ dysfunction syndrome (three studies) and acute respiratory distress syndrome (six studies). The pooled odds ratio for developing acute respiratory distress syndrome was 2.5 (95% confidence interval, 1.6-3.3). CONCLUSIONS: Despite the inherent limitations in the analysis of cohort studies, our analysis suggests that in adult, intensive care unit, trauma, and surgical patients, RBC transfusions are associated with increased morbidity and mortality and therefore, current transfusion practices may require reevaluation. The risks and benefits of RBC transfusion should be assessed in every patient before transfusion. PMID- 18679113 TI - The effect of tracheostomy timing during critical illness on long-term survival. AB - BACKGROUND: Tracheostomy is common in intensive care unit patients, but the appropriate timing is controversial. OBJECTIVE: To determine whether earlier tracheostomy is associated with greater long-term survival. DESIGN: Retrospective cohort analysis. SETTING: Acute care hospitals in Ontario, Canada (n = 114). PATIENTS: All mechanically ventilated intensive care unit patients who received tracheostomy between April 1, 1992 and March 31, 2004, excluding extreme cases (< 2 or > or = 28 days) and children (< 18 yrs). MEASUREMENTS: For crude analyses, tracheostomy timing was classified as early (< or = 10 days) vs. late (> 10 days) with mortality measured at multiple follow-up intervals. Proportional hazards analyses considered tracheostomy as a time-dependent variable to adjust for measurable confounders and possible survivor treatment bias. We used stratification, propensity score, and instrumental variable analyses to adjust for patient differences. RESULTS: A total of 10,927 patients received tracheostomy during the study, of which one-third (n = 3758) received early and two-thirds late (n = 7169). Patients receiving early tracheostomy had lower unadjusted 90-day (34.8% vs. 36.9%; p = 0.032), 1 yr (46.5% vs. 49.8%; p = 0.001), and study mortality (63.9% vs. 67.2%; p < 0.001) than patients receiving late tracheostomy. Multivariable analyses treating tracheostomy as a time dependent variable showed that each additional delay of 1 day was associated with increased mortality (hazard ratio 1.008, 95% confidence interval 1.004-1.012), equivalent to an increase in 90-day mortality from 36.2% to 37.6% per week of delay (relative risk increase 3.9%; number needed to treat, 71 patients to save one life per week delay). LIMITATIONS: This analysis provides guidance regarding timing but not patient selection for tracheostomy. CONCLUSIONS: Physicians performing early tracheostomy should not anticipate a large potential survival benefit. Future research should concentrate on identifying which patients will receive the most benefit. PMID- 18679114 TI - Hypothermia after cardiac arrest does not alter serum inflammatory markers. AB - OBJECTIVE: Hypothermia improves survival and neurologic recovery after cardiac arrest. Cardiac arrest also triggers release of cytokines and inflammatory molecules, and it is unknown whether therapeutic hypothermia alters this inflammatory response. This study tested whether therapeutic hypothermia altered levels of inflammatory markers in serum. DESIGN: Prospective, randomized study. SETTING: University research laboratory. SUBJECTS: Adult, male, Sprague-Dawley rats. INTERVENTIONS: Halothane-anesthetized rats were subjected to 8 mins of asphyxial cardiac arrest and resuscitation. Rat temperature was controlled at 37 degrees C throughout the experiment (normothermia) or reduced to 33 degrees C between 1 and 24 hrs after cardiac arrest (hypothermia). Serum cytokines were measured at baseline, 0.5, 1, 3, 6, 12, and 24 hrs after resuscitation using multiplex analyzer or enzyme-linked immunosorbent assay. MEASUREMENTS AND MAIN RESULTS: Hypothermic rats showed improved neurologic recovery at 12 and 24 hrs. Serum levels of tumor necrosis factor-alpha; macrophage inflammatory protein 1alpha; growth-related oncogene/keratinocyte chemokine; interleukin-2, -9, and 10; monocyte chemotactic protein-1; leptin; and intracellular adhesion molecule-1 increased over time, and the levels of interleukin-18 declined over time. No temporal trends in other molecules were detected. Levels of these molecules did not differ between temperature groups during the hypothermia phase (1-24 hrs). CONCLUSIONS: These data suggest that altering the inflammatory response after cardiac arrest is not necessary for the beneficial effects of hypothermia. These data do not support a specific role of circulating cytokines in the neurologic injury after cardiac arrest. PMID- 18679115 TI - Does granulocyte colony-stimulating factor ameliorate the proinflammatory response in human meningococcal septic shock? AB - OBJECTIVE: To test the hypothesis that granulocyte colony-stimulating factor acts cooperatively with interleukin-10 in down-regulating monocyte function in severe meningococcal septic shock. 1) We quantified the plasma levels of granulocyte colony-stimulating factor, interleukin-10, Neisseria meningitidis lipopolysaccharide and the number of N. meningitidis DNA copies in 28 patients with systemic meningococcal disease. 2) We studied the inhibitory effect of recombinant human granulocyte colony-stimulating factor on normal human monocytes stimulated with purified meningococcal lipopolysaccaride. 3) We monitored the inhibitory effects of endogenously produced granulocyte colony-stimulating factor and interleukin-10 in meningococcal shock plasmas on monocytes. DESIGN: Comparative, experimental study. SETTING: University Hospital and laboratory. SUBJECTS: Twenty-eight patients with systemic meningococcal disease, 13 with persistent shock, 7 died, and 15 without shock. MEASUREMENTS AND MAIN RESULTS: The median levels of granulocyte colony-stimulating factor in shock and nonshock patients were 1.7 x 10(6) and 8.1 x 10(2) pg/mL; interleukin-10, 2.1 x 10(4) and 4 x 10(1) pg/mL; number of N. meningitidis DNA copies, 2.9 x 10(7) and <10(3)/mL; and lipopolysaccharide, 105 and <0.04 endotoxin units/mL, respectively. The plasma levels of granulocyte colony-stimulating factor were reduced by 50% within 4 to 6 hrs after initiation of antibiotic treatment. In model experiments with lipopolysaccharide-stimulated human monocytes, recombinant human granulocyte colony-stimulating factor and interleukin-10 reduced the release of tumor necrosis factor-alpha by mean 30% and 92%, respectively. When plasmas from three shock patients were depleted of native granulocyte colony-stimulating factor or interleukin-10 by immunoprecipitation, no increase in tumor necrosis factor-alpha release occurred after removal of granulocyte colony-stimulating factor, whereas removal of interleukin-10 increased the tumor necrosis factor-alpha release eight fold. CONCLUSIONS: Although granulocyte colony-stimulating factor in plasma increases by five orders of magnitude in patients with meningococcal shock, the anti-inflammatory effect on patients' monocytes is uncertain. PMID- 18679116 TI - Efficacy of the amphibian peptide distinctin in a neutropenic mouse model of staphylococcal sepsis. AB - OBJECTIVE: To investigate the efficacy of distinctin in a neutropenic mouse model of staphylococcal sepsis. DESIGN: Prospective, randomized, and controlled animal study. SETTING: Research laboratory in a University Hospital. SUBJECTS: BALB/c male mice. INTERVENTIONS: Mice were rendered neutropenic by injecting cyclophosphamide (200 mg/kg of body weight/day) on days -4 and -2 preinfection. Infection was induced at time 0 by intraperitoneal injection of 1 x 10(9) colony forming units of the staphylococcal strain. For each model, all animals were randomized to receive intravenous isotonic sodium chloride solution, 1 mg/kg distinctin, and 10 mg/kg imipenem, 10 mg/kg vancomycin, 10 mg/kg teicoplanin or 10 mg/kg linezolid alone, or combined with 1 mg/kg distinctin. MEASUREMENTS AND MAIN RESULTS: Lethality, bacterial growth in blood and peritoneum, spleen, liver, and mesenteric lymph nodes. RESULTS: All combined regimen showed lower lethality rates than singly treated-groups. Distinctin plus vancomycin or teicoplanin exerted the lowest lethality rate. All regimens were significantly superior to controls at reducing blood, spleen, peritoneum, liver and mesenteric lymph node complex bacterial burdens, whereas all combined treated groups were higher effective than singly treated groups. CONCLUSION: Our data indicate that distinctin alone or combined with other antibiotics may be useful in treating severe staphylococcal infections. PMID- 18679117 TI - Hypertonic saline up-regulates A3 adenosine receptor expression of activated neutrophils and increases acute lung injury after sepsis. AB - OBJECTIVE: Hypertonic saline resuscitation reduces tissue damage by inhibiting polymorphonuclear neutrophils. Hypertonic saline triggers polymorphonuclear neutrophils to release adenosine triphosphate that is converted to adenosine, inhibiting polymorphonuclear neutrophils through A2a adenosine receptors. Polymorphonuclear neutrophils also express A3 adenosine receptors that enhance polymorphonuclear neutrophil functions. Here we investigated whether A3 receptors may diminish the efficacy of hypertonic saline in a mouse model of acute lung injury. DESIGN: Randomized animal study and laboratory investigation. SETTING: University research laboratory. INTERVENTIONS: The effect of A3 receptors on the efficacy of hypertonic saline resuscitation was assessed in A3 receptor knockout and wild-type mice. Animals were treated with hypertonic saline (7.5% NaCl, 4 mL/kg) before or after cecal ligation and puncture, and acute lung injury and mortality were determined. The effect of timing of hypertonic saline exposure on A3 receptor expression and degranulation was studied in vitro with isolated human polymorphonuclear neutrophils. MEASUREMENTS AND MAIN RESULTS: Treatment of human polymorphonuclear neutrophils with hypertonic saline before stimulation with formyl methionyl-leucyl-phenylalanine inhibited A3 receptor expression and degranulation, whereas hypertonic saline-treatment after formyl methionyl-leucyl phenylalanine-stimulation augmented A3 receptor expression and degranulation. Acute lung injury in wild-type mice treated with hypertonic saline after cecal ligation and puncture was significantly greater than in wild-type mice pretreated with hypertonic saline. This aggravating effect of delayed hypertonic saline treatment was absent in A3 receptor knockout mice. Similarly, mortality in wild type mice with delayed hypertonic saline-treatment was significantly higher (88%) than in animals treated with hypertonic saline before cecal ligation and puncture (50%). Mortality in A3 receptor knockout mice remained only 50% regardless of timing of hypertonic saline administration. CONCLUSIONS: Polymorphonuclear neutrophil A3 receptors expression determines whether hypertonic saline resuscitation inhibits or aggravates polymorphonuclear neutrophil-induced acute lung injury. These findings suggest that A3 antagonists could improve the efficacy of hypertonic saline resuscitation by reducing side effects in patients whose polymorphonuclear neutrophils are activated before hypertonic saline treatment. PMID- 18679118 TI - Risk factors for short-term mortality from carbon monoxide poisoning treated with hyperbaric oxygen. AB - OBJECTIVE: Carbon monoxide (CO) poisoning is common in the United States, accounting for approximately 2,700 deaths annually. Few publications have described the mortality rate of CO-poisoned patients who survive to reach a hospital and die despite maximal medical care. Further, while risk factors for cognitive sequelae in survivors of CO poisoning have become clearer recently, factors associated with death are less well defined. This study was conducted to 1) determine the short-term mortality risk for patients treated with hyperbaric oxygen for CO poisoning, and 2) determine whether any factors related to the poisoning episode are predictive of mortality. DESIGN/SETTING/PATIENTS: A departmental database and medical records of 1,505 consecutive patients treated with hyperbaric oxygen at a single institution from 1978 to 2005 were reviewed. MEASUREMENTS: Demographic and clinical data were extracted for analysis. Mortality data, including cause of death, were obtained through a search of the National Death Index of the National Center for Health Statistics. MAIN RESULTS: A total of 38 patients experienced short-term mortality from their episode of CO poisoning, yielding a death rate of 2.6% in medically treated patients. Characteristics significantly associated with mortality included fire as a source of CO, loss of consciousness, carboxyhemoglobin level, arterial pH, and presence of endotracheal intubation during hyperbaric treatment. CONCLUSIONS: The mortality rate for medically treated CO-poisoned patients in this series was 2.6%, similar to the limited combined experience previously reported in the literature. Factors most strongly associated with mortality were severe metabolic acidosis and need for endotracheal intubation. PMID- 18679119 TI - Neuroprotection by nitrous oxide: facts and evidence. AB - BACKGROUND AND OBJECTIVE: Preliminary studies have shown that nitrous oxide, like xenon, may possess potentially neuroprotective properties. However, because of its possible neurotoxic and proneurotoxic effects (obtained under particular conditions) and its bad reputation at anesthetic concentrations, no thorough investigations have been performed on the potentially neuroprotective properties of nitrous oxide. The aim of this study was to investigate the possible neuroprotective effects of nitrous oxide at nonanesthetic concentrations on different models of excitotoxic insult and brain ischemia. MEASUREMENTS AND MAIN RESULTS: Here, we show using multiple models of ex vivo and in vivo excitotoxic insults and brain ischemia that nitrous oxide, administered alone at nonanesthetic doses, offers global neuroprotection from reduction of neurotransmitter release induced by ischemia to reduction of subsequent cell injury. In vivo, in rats subjected to transient cerebral ischemia, nitrous oxide at 50 vol% offers full neuroprotection at both the histologic and neurologic outcome levels when administered up to 2 hrs, but not 3 hrs, after ischemia onset. CONCLUSIONS: These data provide experimental evidence that nitrous oxide, which is a cost-efficient and easily available gas, has potentially neuroprotective properties in rodents when given alone at nonanesthetic concentrations. Therefore, because there is a lot at stake for the affected patients and society--in terms of easy access to treatment, profound impact of brain damage, cost of treatment, and subsequent financial cost on society--we believe that further studies should investigate thoroughly the possible potential clinical interest of nitrous oxide for the treatment of ischemic stroke in terms of optimal indications, type of ischemic injury, duration and time points for treatment, and the optimal concentration of gas to be used in clinical circumstances. PMID- 18679121 TI - Extracorporeal membrane oxygenation support can extend the duration of cardiopulmonary resuscitation. AB - OBJECTIVES: To evaluate the use of extracorporeal membrane oxygenation in prolonged cardiopulmonary resuscitation and to estimate how long cardiopulmonary resuscitation can be extended with acceptable results. DESIGN: Review of consecutive adult in-hospital cardiopulmonary resuscitation patients without return of spontaneous circulation in 10 mins and with extracorporeal membrane oxygenation rescue, and analysis of the relationship between outcome and cardiopulmonary resuscitation duration and possible etiologies. The data were collected following the Utstein style guidelines on in-hospital cardiopulmonary resuscitation. Two organ dysfunction scores were incorporated into the analysis for outcome prediction. SETTING: A university-affiliated tertiary referral medical center and extracorporeal membrane oxygenation center. PATIENTS: An observational cohort study in 135 consecutive adult in-hospital cardiopulmonary resuscitation patients without return of spontaneous circulation who received extracorporeal membrane oxygenation during cardiopulmonary resuscitation. MAIN RESULTS: The average cardiopulmonary resuscitation duration was 55.7 +/- 27.0 mins and 56.3% of patients received subsequent interventions to treat underlying etiologies. The successful weaning rate was 58.5% and the survival-to-discharge rate was 34.1%. The majority of survivors (89%) had an acceptable neurologic status on discharge. Risk factors for hospital mortality included longer cardiopulmonary resuscitation duration, etiology of acute coronary syndrome, and a higher organ dysfunction score in the first 24 hrs. Logistic regression analysis revealed the probability of survival was approximately 0.5, 0.3, or 0.1 when the duration of cardiopulmonary resuscitation was 30, 60, or 90 mins, respectively. CONCLUSION: Assisted circulation might extend the presently accepted duration of cardiopulmonary resuscitation in adult in-hospital cardiopulmonary resuscitation patients. PMID- 18679120 TI - Surfactant protein-B polymorphisms and mortality in the acute respiratory distress syndrome. AB - OBJECTIVE: To determine whether polymorphisms of the surfactant protein B gene may be associated with increased mortality in patients with the acute respiratory distress syndrome. DESIGN: A prospective cohort study. SETTING: Four adult intensive care units at a tertiary academic medical center. PATIENTS: Two hundred fourteen white patients who had met criteria for acute respiratory distress syndrome. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients were genotyped for a variable nuclear tandem repeat polymorphism in intron 4 of the surfactant protein B gene and the surfactant protein B gene +1580 polymorphism. For the variable nuclear tandem repeat surfactant protein B gene polymorphism, patients were found to have either a homozygous wild-type genotype or a variant genotype consisting of either a heterozygous insertion or deletion polymorphism. Logistic regression was performed to analyze the relationship of the polymorphisms to mortality in patients with acute respiratory distress syndrome. In multivariate analysis, the presence of variable nuclear tandem repeat surfactant protein B gene polymorphism was associated with a 3.51 greater odds of death at 60 days in patients with acute respiratory distress syndrome as compared to those patients with the wild-type genotype (95% confidence interval 1.39-8.88, p = 0.008). There was no association found between the +1580 variant and outcome (p = 0.15). CONCLUSIONS: In this study, the variable nuclear tandem repeat surfactant protein B gene polymorphism in intron 4 is associated with an increased 60 day mortality in acute respiratory distress syndrome after adjusting for age, severity of illness, and other potential confounders. Additional studies in other populations are needed to confirm this finding. PMID- 18679122 TI - Increased mortality of ventilated patients with endotracheal Pseudomonas aeruginosa without clinical signs of infection. AB - OBJECTIVE: To investigate the frequency and outcomes of ventilated patients with newly acquired large burdens of Pseudomonas aeruginosa and to test the hypothesis that large quantities of bacteria are associated with adverse patient outcomes. DESIGN: A prospective, single-center, observational, cohort study. SETTING: Medical-surgical intensive care units in a tertiary care university hospital. PATIENTS: All adult patients requiring > or = 48 hrs of mechanical ventilation and identified as having newly acquired P. aeruginosa in their lower respiratory tracts between October 2002 and April 2006. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Daily surveillance cultures of endotracheal aspirates were performed on patients intubated > or = 48 hrs; 69 patients with newly acquired P. aeruginosa were enrolled. Daily P. aeruginosa quantification of endotracheal aspirates was performed; clinical signs of infection were noted. Of 45 patients with high P. aeruginosa burdens (> or = 1,000,000 colony-forming units/mL in endotracheal aspirates; > or = 10,000 colony-forming units/mL in bronchoalveolar lavage), 17 (37.8%) patients did not meet clinical criteria for ventilator associated pneumonia and had a statistically significant higher risk of death (adjusted hazard ratio, 37.53; 95% confidence interval, 3.79-371.96; p = 0.002) when compared with the patients who had P. aeruginosa ventilator-associated pneumonia. When excluding the ten patients who had ventilator-associated pneumonia attributed to bacteria other than P. aeruginosa or attributed to multiple bacteria including P. aeruginosa, the risk of death remained statistically significant (adjusted hazard ratio, 23.98; 95% confidence interval: 2.49-230.53; p = 0.006). Furthermore, more patients with high P. aeruginosa burdens secreted the type III secretion facilitator protein, PcrV (p = 0.01). CONCLUSIONS: A group of patients with large burdens of P. aeruginosa who did not meet clinical criteria for ventilator-associated pneumonia had an increased risk of death when compared with patients who had high P. aeruginosa burdens and met ventilator-associated pneumonia criteria. Patients with high P. aeruginosa burden seemed to possess more virulent strains. PMID- 18679123 TI - Bronchoscopy is associated with decreased mesenteric arterial flow. AB - BACKGROUND: We hypothesized that fiberoptic bronchoscopy can contribute to mesenteric ischemia and bacterial translocation. OBJECTIVE: To test this hypothesis we investigated in patients undergoing bronchoscopy mesenteric blood flow and markers in association with ischemia reperfusion injury. PATIENT: Forty seven consecutive patients requiring diagnostic fiberoptic bronchoscopy were studied. MEASUREMENTS: Parameters evaluated were superior mesenteric artery Doppler sonography, oxidative stress mediators, arterial blood gases, blood cultures pre-fiberoptic bronchoscopy, and 1st, 4th, and 24th hr post-fiberoptic bronchoscopy. MAIN RESULTS: After bronchoscopy; PaO2 decreased by 21.8% +/- 1.5% (range 6-40), and remained low at all time points (p = 0.0001, p = 0.0001, p = 0.008). Flow volume decreased by 38.8% +/- 14.9% (range 6-72), and remained low at 1st and 4th hr (p = 0.0001, p = 0.01). Resistive and pulsatile index increased at 1st hr (p = 0.0001, p = 0.001) and 4th hr (p = 0.018, p = 0.045). Myeloperoxidase and malondialdehyde increased at 1st hr (p = 0.0001) and 4th hr (p = 0.037, p = 0.028). Corresponding glutathione and catalase decreased at 1st hr (p = 0.0001), and glutathione remained significant at 4th and 24th hr (p = 0.0001, p = 0.003). Correlation between flow volume and PaO2 (r = .71, p = 0.0001), myeloperoxidase (r = -.39, p = 0.05), glutathione (r = .41, p = 0.03) was significant. Nine of 47 (19.1%) had fever, and 3 of 47 (6.4%) had gram negative bacteremia. CONCLUSION: Fiberoptic bronchoscopy is associated with decreased mesenteric blood flow, which may place the patient at risk for mesenteric ischemia and gastrointestinal bacterial translocation. PMID- 18679124 TI - Serious acute chikungunya virus infection requiring intensive care during the Reunion Island outbreak in 2005-2006. AB - OBJECTIVE: To report the clinical and laboratory findings of adults with serious chikungunya virus acute infection hospitalized in an intensive care unit. DESIGN: Case series study from August 2005 to May 2006. SETTING: Medical intensive care unit, South Reunion Hospital. PATIENTS: We observed 33 episodes of confirmed acute chikungunya virus infection (chikungunya virus-IgM or reverse transcription polymerase chain reaction positive in the serum) admitted to the intensive care unit. INTERVENTIONS: We collected cerebrospinal fluid, serum, and sometimes tissue samples from patients with suspected chikungunya fever in our intensive care unit. These samples underwent viral testing for evidence of acute chikungunya virus infection. MEASUREMENTS AND MAIN RESULTS: Of the 33 patients, 19 (58%) had chikungunya virus specific manifestations, 8 (24%) had associated acute infectious disease and 6 (18%) exacerbations of previous complaints. Among the chikungunya virus specific manifestations, we identified 14 cases of encephalopathy, one case each of myocarditis, hepatitis and Guillain Barre syndrome. Eighty-five percent of patients had a McCabe score = 1 (for nonfatal or no underlying disease). Mortality was 48%. CONCLUSIONS: Chikungunya virus infection may be responsible for very severe clinical presentation, including young patients with unremarkable medical histories. Chikungunya virus infection is strongly suspected to have neurologic, hepatic, and myocardial tropism leading to dramatic complications and high mortality rate. PMID- 18679125 TI - Impaired plasma B-type natriuretic peptide clearance in human septic shock. AB - INTRODUCTION: High B-type natriuretic peptide (BNP) levels are reported in the context of septic shock. We hypothesized that high BNP levels might be related to an alteration in BNP clearance pathway, namely neutral endopeptidase (NEP) 24.11. NEP 24.11 activity was measured in septic shock and in cardiogenic shock patients. We further evaluated whether baseline plasma BNP can predict fluid responsiveness and whether BNP can still be released in plasma despite high initial BNP levels, in response to overloading. MATERIAL AND METHODS: Prospective observational study. Patients in severe sepsis (S) or in septic shock (SS) needing a fluid challenge were included. Stroke volume (SV) and BNP were measured before (SV1, BNP1) and 45 mins after (SV2, BNP2) a standardized fluid challenge. DeltaBNP was defined as the difference between BNP2 and BNP1. NEP 24.11 activity was determined by fluorometry in 12 SS and 4 S patients before fluid challenge and in 5 cardiogenic shock patients. RESULTS: Twenty-three patients (61 +/- 18 years old, Simplified Acute Physiology Score II: 54 +/- 21; 19 SS, 4 S; BNP1: 1371 +/- 1434 pg/mL) were studied. BNP1 concentrations were significantly higher in SS than in S (1643 +/- 1437 vs. 80 +/- 35 pg/mL; p = 0.002). There was no correlation between baseline BNP and fluid responsiveness. Nine of the 11 patients with BNP1 >1000 pg/mL were fluid responders. DeltaBNP was greater in fluid nonresponders than in fluid responders (22 +/- 27% vs. 6 +/- 11%, p = 0.028). Plasma BNP was higher in SS than in cardiogenic shock patients (1367 +/- 1438 vs. 750 +/- 346 respectively; p = 0.027). NEP 24.11 activity was lower in SS than in S patients (0.10 +/- 0.06 nmole/mL/min vs. 0.50 +/- 0.22 nmole/mL/min, p <0.0001) cardiogenic shock patients (0.10 +/- 0.06 nmole/mL/min vs. 0.58 +/- 0.19 nmole/mL/min; p = 0.002). CONCLUSION: High levels of BNP might be related to an alteration in BNP clearance. During sepsis, high BNP levels are not predictive of fluid nonresponsiveness. Nevertheless, in fluid nonresponders, acute ventricular stretching can result in further BNP release. PMID- 18679126 TI - Efficacy of three different valve systems of needle-free closed connectors in avoiding access of microorganisms to endovascular catheters after incorrect handling. AB - OBJECTIVE: Disinfectable needle-free closed connectors were designed to avoid needle-stick injuries and to be easily disinfected before handling. Workloads or lack of knowledge, however, could impede the correct handling of these devices, allowing endoluminal catheter colonization. The aim of our study was to assess the barrier effect of different disinfectable needle-free closed connectors during correct and incorrect handling using an experimental model. DESIGN: We used a model consisting of a blood culture bottle with a peripheral venous catheter inserted under sterile conditions. Three different disinfectable needle free closed connectors with different valve designs (microClave, Bionector, and Smartsite plus) were used to close the catheters. The external surfaces of the disinfectable needle-free closed connectors were contaminated with different concentrations of a Staphylococcus epidermidis culture broth. After contamination, 10 units of each connector and each concentration were assigned to the correct handling group (cleaned with 70% ethylic alcohol before handling) and the same number to the incorrect handling group (handled without disinfection) with a total of 180 bottles. RESULTS: Increases in concentrations of external contamination and incorrect handling of the connectors resulted in an increase in connectors' permeability to the pass of microorganisms to the endoluminal way. MicroClave proved the best barrier in the experimental conditions described. CONCLUSION: The barrier effect of disinfectable needle-free closed connectors is adversely affected by incorrect handling, the quantity of external valve colonization, and the valve design. PMID- 18679127 TI - Growth of intensive care unit resource use and its estimated cost in Medicare. AB - OBJECTIVE: The past 10-15 yrs brought significant changes in the United States healthcare system. Effects on Medicare intensive care unit use and costs are unknown. Intensive care unit costs are estimated using the Russell equation with a ratio of intensive care unit to floor cost per day, or "R value," of 3, which may no longer be valid. We sought to determine contemporary Medicare intensive care unit resource use, costs, and R values; whether these vary by patient and hospital characteristics; and the impact of updated values on estimated intensive care unit costs. DESIGN: Retrospective analysis of Medicare Inpatient Prospective Payment System hospitalizations from 1994 to 2004 using Medicare Provider Analysis and Review files. SETTING: All nonfederal acute care US hospitals paid through the Inpatient Prospective Payment System. SUBJECTS: Inpatient prospective payment system hospitalizations from 1994 to 2004 (n = 121,747, 260). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We examined resource use and costs (adjusted to y2004$), calculating intensive care unit and floor costs directly and using these to generate year-specific R values. By 2004, 33% of Medicare hospitalizations had intensive care unit or coronary care unit care, with more than half of the increase in total hospitalizations because of additional intensive care unit hospitalizations. Adjusted intensive care unit cost per day remained stable ($2,616 vs. $2,575; 1994 vs 2004), yet adjusted floor cost per day rose substantially ($1,027 vs. $1,488) driven by decreased floor length of stay. Annual adjusted Medicare intensive care unit costs increased 36% to $32.3B, largely because of increased utilization. R values decreased progressively from 2.55 to 1.73, were lower for surgical vs. medical admissions and survivors vs. nonsurvivors, but varied little by hospital characteristics. An R value of 3 overestimated Medicare intensive care unit costs by 17.6% ($5.7 billion) in 2004. CONCLUSIONS: Medicare intensive care unit use is rising rapidly and will likely continue to do so. Despite significant healthcare system changes, adjusted daily critical care costs remained stable, yet care outside the intensive care unit became more expensive. To track intensive care unit cost over time, year-specific R values should be used. PMID- 18679128 TI - Negative effects of rofecoxib treatment on cardiac function after ischemia reperfusion injury in APOE3Leiden mice are prevented by combined treatment with thromboxane prostanoid-receptor antagonist S18886 (terutroban). AB - OBJECTIVE: Selective cyclooxygenase-2 inhibition by rofecoxib was associated with increased risk of cardiovascular events. We hypothesized that concomitant treatment with thromboxane prostanoid receptor antagonist S18886 might ameliorate possible negative effects. We evaluated the effects of S18886, rofecoxib, and the interaction of both compounds in a combined treatment on myocardial infarct (MI) size and cardiac function after experimental ischemia/reperfusion injury in hyperlipidemic APOE*3Leiden transgenic mice. DESIGN: Prospective, randomized, control trial. SETTING: Research laboratory. SUBJECTS: Hyperlipidemic APOE*3Leiden transgenic mice. INTERVENTIONS: After four weeks of feeding an atherogenic diet, MI was induced by a 30-min ligation of the left anterior descending coronary artery, followed by reperfusion. Oral compound treatment was initiated 90 mins before MI, and continued daily by gavage for seven days. Four treatment groups (n = 12, each) were studied: solvent (Control), S18886, rofecoxib, and S18886 plus rofecoxib. MEASUREMENTS AND MAIN RESULTS: One week after MI, the mice were anesthetized and cardiac function was quantified by left ventricular (LV) pressure-volume relationships obtained by miniature pressure conductance catheters. The ischemic area was measured by morphometry and expressed as percentage of LV area. No significant differences in infarct size were found between groups. Compared with control, treatment with S18886 did not affect heart function whereas the rofecoxib group had significantly lower cardiac output (4.5 +/- 0.8 vs. 3.2 +/- 1.1 mL/min, p < 0.01), lower ejection fraction (40 +/- 8 vs. 27 +/- 11%, p < 0.005), and increased end-systolic volume (18.6 +/- 5.7 vs. 28.6 +/- 9.0 muL, p < 0.05). The group with combined (S18886+rofecoxib) treatment was not different from control. Statistical analysis showed significant interactive effects between S18886 and rofecoxib indicating that negative effects of rofecoxib on cardiac function were prevented by S18886 treatment. CONCLUSIONS: Rofecoxib treatment reduced global and systolic LV function after ischemia reperfusion injury in APOE*3Leiden mice. These negative effects are prevented by combined treatment with thromboxane prostanoid-receptor antagonist S18886 (terutroban). PMID- 18679131 TI - Fatal course of recurrent primary biliary cirrhosis after liver transplantation. PMID- 18679133 TI - Prophylactic antibiotic usage for Pneumocystis jirovecii pneumonia in patients with systemic lupus erythematosus on cyclophosphamide: a survey of US rheumatologists and the review of literature. AB - BACKGROUND: Use of cyclophosphamide in systemic lupus erythematosus (SLE) is associated with Pneumocystis jirovecii pneumonia (PJP) that has substantial morbidity and mortality. However, the frequency of PJP in these patients is unknown and there are no guidelines for prophylactic antibiotics. OBJECTIVES: The objectives of this study are to evaluate the frequency of PJP and the need for prophylactic antibiotics in these patients. METHODS: We estimated incidence of PJP and use of prophylactic trimethoprim-sulfamethoxazole in these patients by a literature search and an e-mail survey of US rheumatologists. RESULTS: We identified 18 manuscripts dealing with infections in SLE patients treated with cyclophosphamide. In these manuscripts, 121 cases of PJP were identified in 76,156 SLE patients with a frequency of 15.88 per 10,000 patients.Of 264 rheumatologists surveyed, 133 (50.37%) were using prophylactic antibiotics in these patients. One hundred thirty-one (49.63%) respondents did not use prophylactic antibiotics. 5,174 SLE patients received cyclophosphamide in last 5 years with 19.6 +/- 30.6 (mean +/- SD) patients per rheumatologist. 32 cases of PJP were reported. The total cumulative experience of 264 rheumatologists was 4742 years [(17.96 +/- 10.35) (mean +/- SD)] with a PJP rate of 67.48 per 10,000 years of practice. CONCLUSIONS: The frequency of PJP in SLE patients on cyclophosphamide remains low (0.1588%). Therefore, routine use of trimethoprim sulfamethoxazole for PJP prophylaxis in SLE patients on cyclophosphamide does not appear to be substantiated by this study, except in those with elevated risk, ie, with severe leucopenia, lymphopenia, high dose corticosteroids, hypocomplementemia, active renal disease, and higher mean SLEDAI score. There is a need for consensus guidelines addressing prophylactic antibiotics in these patients. PMID- 18679129 TI - Influenza. AB - OBJECTIVE: Influenza is a major concern for intensivists in all communities in the U.S. While there is considerable concern whether or not the country will be ready for a pandemic influenza, even seasonal influenza poses a major challenge to hospitals. The objective of this review is to summarize current knowledge of influenza with emphasis on the issues that intensivist will encounter. SETTING: Intensive care unit in a 450-bed, tertiary care, teaching hospital. METHODS: Source data were obtained from a PubMed search of the medical literature. PubMed "related articles" search strategies were likewise employed frequently. SUMMARY AND CONCLUSIONS: Seasonal influenza causes more than 200,000 hospitalizations and 41,000 deaths in the U.S. every year, and is the seventh leading cause of death in the U.S. Despite this impact there is a shortcoming in knowledge of influenza among many health care workers, and a paucity of clinical data and studies to guide therapy. Intensivists need to recognize the importance of seasonal influenza as a cause of severe morbidity and mortality. This review summarizes current knowledge of the diagnosis, complications, therapy, and infection control measures associated with influenza. PMID- 18679134 TI - Drug allergies may be more frequent in systemic lupus erythematosus than in rheumatoid arthritis. AB - OBJECTIVE: To measure the frequency of drug allergies in patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), and register the causal drug, the type, and severity of the reaction. MATERIAL AND METHODS: Direct interview and chart review in patients with RA and SLE were conducted. We registered demographic data, drug allergies, the causal drug, how causality was assessed, and the type and severity of the allergic reaction. We include as drug allergies only those cases in which the result of exposure and re-exposure was known or in which a physician evaluating the original event established the causality link with the suspected drug. Differences between groups were assessed by chi test. RESULTS: Two hundred ninety-three RA and 58 patients with SLE were included. Fifty-three of the patients with RA (18%) and 20 of the patients with SLE (34.3%, P = 0.049) reported drug allergies. Most of them presented skin rash as their clinical expression of allergy (73%); anaphylaxis was reported in 4 cases (5%). Allergy to sulfa drugs is found more frequently in SLE (P = 0.0079). No differences were found when comparing the frequency of other drug allergies, such as penicillin and metamizole. DISCUSSION: Drug allergies are more frequent in SLE than in RA. Sulfa drugs are still the most frequent cause of drug allergies in SLE. Allergies because of drugs forbidden in the United States but easily available in specific ethnic groups are frequent in patients with SLE and RA. Their specific consumption must be intentionally assessed in cases of suspected drug allergies. PMID- 18679135 TI - Accelerated pulmonary nodulosis and sterile pleural effusion in a patient with psoriatic arthropathy during methotrexate therapy: a case report. AB - Pulmonary nodulosis and sterile pleural exudates are well-known extra-articular manifestations in rheumatoid arthritis patients with a positive rheumatoid factor. In some patients, treatment with methotrexate has been postulated as the trigger of these complications. We report a patient with psoriatic arthropathy, negative RF, negative anticyclic citrulinated peptide antibodies but positive antibodies to cardiolipin who developed massive sterile pleural empyema and multiple cavitary pulmonary nodules during methotrexate treatment. We suggest that awareness of methotrexate-induced lung and pleural complications should be extended to other than rheumatoid arthritis diseases, not necessarily accompanied by rheumatoid factor or anticyclic citrulinated peptide antibodies. PMID- 18679136 TI - Infiltration of histiocytes and multinucleated giant cells in the myocardium of a patient with multicentric reticulohistiocytosis. AB - Multicentric reticulohistiocytosis (MRH) is a rare multisystem disease characterized by skin nodules and a destructive polyarthritis. We describe cardiac disease in a 42-year-old African American female with a history of MRH for 29 years and dilated cardiomyopathy for 5 years. During a congestive heart failure flare, the patient was slow to respond to standard treatment. A left/right heart catheterization and endomyocardial biopsy were performed, and the patient was found to have evidence of myocardial MRH infiltration. PMID- 18679137 TI - Work status among patients with ankylosing spondylitis in Argentina. AB - OBJECTIVE: To evaluate work status and its determinants among ankylosing spondylitis (AS) patients in our country. MATERIALS AND METHODS: We carried out a case-control study, including AS patients older than 16-year-old. Demographic, socioeconomic, and employment data were collected. Individuals from the general population matched by age, sex, and socioeconomic level served as the control group. Functional capacity was evaluated by bath ankylosing spondylitis functional index (BASFI) and health assessment questionnaire (HAQ-S) and disease activity by bath ankylosing spondylitis disease activity index (BASDAI). All patients completed quality of life (ASQol), depression (CES-D), and fatigue (FSS) validated questionnaires. RESULTS: Sixty-four patients with AS (57 men) and 93 controls (83 men) were included. The frequency of retirement due to age was comparable in both groups, however, a significantly larger number of patients with AS were retired due to disability: 6 (9.4%) versus 0 (P = 0.004). Patients with AS had a significantly greater number of comorbidities than controls (78.7% vs. 31.5%, P = 1 x 10(-8)). Sixteen patients (26.2%) were unemployed compared with 4 controls (4.5%) (P = 0.001). Permanence at work was significantly lower in patients (median age 58, SD 54-61 years) versus controls (median age 66, SD 63-68 years) (log Rank P = 0.001). There were less full-time workers (71% vs. 87%, P = 0.03) and higher number of sick leave days (6.56 +/- 13 vs. 2.13 +/- 4.5, P = 0.01) in AS patients than controls. Unemployed patients had significantly worst quality of life, disease activity, and depression scores than employed patients. In the multivariate analysis, the primary variable associated to unemployment was depression [Odds Ratio (OR): 20, 95% confidence interval (CI), 1.69-258, P = 0.001] followed by disease activity (OR: 1.05, 95% CI, 1-1.1, P = 0.04) and age (OR: 1.1, 95% CI, 1.02-1.21, P = 0.01) as secondary variables. CONCLUSIONS: The frequency of work disability among our patients with AS was significantly higher than in age, sex, and socioeconomic matched population. The main variable associated with work disability and unemployment was depression. Whether this factor is contributing to or is a result of disability is not known. PMID- 18679138 TI - Modified and unmodified electroconvulsive therapy: a comparison of attitudes between psychiatrists in Beijing and Hong Kong. AB - OBJECTIVE: Unmodified electroconvulsive therapy (ECT) is still widely practiced in many psychiatric institutions in China. The study reported herein aimed to explore the attitudes of psychiatrists in Beijing toward unmodified and modified ECT and compare them with those of psychiatrists in Hong Kong, where only modified ECT has been used during the past 40 years. METHOD: One hundred five psychiatrists of a university-affiliated psychiatric hospital in Beijing and all psychiatrists (n = 238) currently practicing in Hong Kong were invited to complete a questionnaire exploring their attitudes toward unmodified and modified ECT. RESULTS: The Beijing respondents had significantly more experience with unmodified ECT than their Hong Kong counterparts. Although 56% of the Beijing respondents preferred modified to unmodified treatment, 81% of them regarded unmodified ECT as safe and associated with minimal morbidity and mortality. Patient choice, financial status, and safety considerations were the main factors that dictated the choice of mode of ECT in Beijing. CONCLUSIONS: It seems that unmodified ECT still has a role as an effective psychiatric treatment in developing countries such as China. Any initiative to replace unmodified with modified treatment should take into account economic conditions, the sociocultural context, and the psychiatric traditions of the particular regions of the country. PMID- 18679139 TI - A retrospective study of the factors associated with hypercoagulability in the pediatric patients at a tertiary care children's hospital. AB - OBJECTIVE: The objective of this study was to determine the risk factors of hypercoagulability in children. We explored the interaction of multiple risk factors with the incidence of thrombosis. Our hypothesis was that as the number of risk factors for thrombosis increased the actual incidence of thrombosis would also increase. DESIGN: Retrospective review from 2003 through 2006 based on a search using two electronic medical record databases. SETTING: Pediatric Tertiary Care Children's Hospital. PATIENTS: Two hundred twenty-six patients were identified and analyzed. MEASUREMENTS: Search terms included factor V Leiden polymerase chain reaction, prothrombin gene 20210A mutation, methylene tetrahydrofolate reductase mutation, antithrombin III, and protein C and S levels. Clinical data were compiled for regression analysis. MAIN RESULTS: The presence of one risk factor was not significant. Two risk factors increased the risk of thrombosis (p = 0.005; OR 3.128). Three or more risk factors further increased the risk of thrombosis (p = 0.003; OR 4.861). Older age (>11 yrs) was protective against thrombosis (p = 0.007; OR 0.995), and the presence of a central venous catheter when analyzed against accumulating risk factors showed a higher risk than that found during the regression analysis (p = 0.001; OR 3.638). CONCLUSIONS: The population at our institution is reflective of the previously reported standards for the genetic predispositions toward thrombosis. Although older age is associated with a lower incidence of thrombosis, the presence of a central venous access device is detrimental. Accumulation of factors results in an increased risk of thrombosis. This article suggests that when inserting a central venous access device, consideration of a hypercoagulation workup should occur. Those with any two or more risk factors, genetic or acquired, and the comorbidity of a CVL may warrant consideration for the institution of anticoagulation with an agent like low molecular weight heparin. PMID- 18679140 TI - Cerebrospinal fluid cleaved-tau protein and 9-hydroxyoctadecadienoic acid concentrations in pediatric patients with hydrocephalus. AB - OBJECTIVE: To ascertain if cerebrospinal fluid cleaved-tau protein and 9 hydroxyoctadecadienoic acid, reflecting potential biomarkers of overall neuronal injury and lipid peroxidation, respectively, are elevated in hydrocephalus patients compared with controls, and if cleaved-tau and 9-hydroxyoctadecadienoic acid levels correlate with each other. DESIGN: Prospective clinical observational study. SETTING: Tertiary-care children's hospital. PATIENTS: Children younger than or equal to 18 yrs who underwent ventriculoperitoneal shunt placement or revision surgery for intrinsic hydrocephalus. MEASUREMENTS AND MAIN RESULTS: During the study period 12 patients with intrinsic hydrocephalus required ventriculoperitoneal shunt placement or revision. Cerebrospinal fluid cleaved-tau levels were significantly elevated in patients with hydrocephalus (44.7 +/- 9.6 ng/mL, n = 11) compared with control patients (0.0 +/- 0.0 ng/mL, n = 9; p < 0.0001). Cleaved-tau levels correlated with patient age (r = .609, p = 0.047) and duration of symptoms (r = .755, p = 0.007). No significant difference in cerebrospinal fluid 9-hydroxyoctadecadienoic acid levels between patients with hydrocephalus (24.6 +/- 5.7, n = 8) and control patients (24.9 +/- 9.3 ng/mL, n = 7) was detected (p = 0.25). There was also no statistically significant correlation between 9-hydroxyoctadecadienoic acid levels and duration of symptoms (r = .668, p = 0.07), nor was there a significant correlation between 9 hydroxyoctadecadienoic acid levels and patient age (r = -.011, p > 0.10). There was no significant relationship between 9-hydroxyoctadecadienoic acid levels and signs of elevated intracranial pressure, nor was there a correlation between 9 hydroxyoctadecadienoic acid levels and cleaved-tau levels. CONCLUSION: Children with hydrocephalus who have clinical signs of increased intracranial pressure and require ventriculoperitoneal shunt placement or revision exhibit markedly elevated cerebrospinal fluid cleaved-tau levels, suggesting evidence of axonal damage. However, this axonal injury does not seem to be associated with significant lipid peroxidation, at least as assessed by quantifying cerebrospinal fluid 9-hydroxyoctadecadienoic acid at a single, concurrent time point. The significant relationship between age and cerebrospinal fluid cleaved-tau levels suggest that brain injury associated with hydrocephalus may be more pronounced in older children. PMID- 18679141 TI - Arginine vasopressin to manage hypoxemic infants after stage I palliation of single ventricle lesions. AB - OBJECTIVE: Management of patients with single ventricle physiology following stage I palliation procedures is often challenging, with optimization of the ratio of pulmonary-to-systemic blood flow as an important goal. Persistent hypoxemia may be a manifestation of elevated pulmonary vascular resistance and therefore decreased blood flow to the lungs. In such situations, the use of arginine vasopressin to increase systemic vascular resistance may be an effective strategy to improve pulmonary blood flow and maintain adequate pulmonary-to systemic blood flow ratio. We describe three infants in whom persistent hypoxemia improved after institution of arginine vasopressin. DESIGN: Retrospective chart review. SETTING: Twenty-four bed medical-surgical pediatric intensive care unit at a large tertiary care academic hospital. PATIENTS: Three neonates with single ventricle physiology who received arginine vasopressin in the setting of hypoxemia following stage I palliation. RESULTS: Arginine vasopressin was initiated in all three patients for hypoxemia with a goal to increase systemic vascular resistance and generate a higher driving pressure for pulmonary blood flow. Twelve hours after arginine vasopressin initiation, systemic arterial saturation as determined by pulse oximetry and blood pressure increased, whereas heart rate, inotrope score, and Fio2 decreased in all three patients. Urine output was maintained and arterial lactate decreased during this time. Pulmonary to-systemic flow ratio increased in one patient in whom it could be determined. CONCLUSION: In patients with single ventricle physiology and persistent hypoxemia following stage I palliation, administration of arginine vasopressin could improve oxygenation possibly by increasing systemic vascular resistance and therefore the pulmonary blood flow. PMID- 18679142 TI - Post hoc analysis of calfactant use in immunocompromised children with acute lung injury: Impact and feasibility of further clinical trials. AB - OBJECTIVE: To assess the impact of calfactant (a modified natural bovine lung surfactant) in immunocompromised children with acute lung injury and to determine the number of patients required for a definitive clinical trial of calfactant in this population. DESIGN: Post hoc analysis of data from a previous randomized, control trial. SETTING: Tertiary care pediatric intensive care units. PATIENTS: All children, defined as immunocompromised, enrolled in a multicenter, masked, randomized, control trial of calfactant for acute lung injury conducted between July 2000 and July 2003. INTERVENTIONS: Patients received either an intratracheal instillation of calfactant or an equal volume of air placebo in a protocolized manner. MEASUREMENTS AND MAIN RESULTS: Eleven of 22 (50%) calfactant-treated patients died when compared with 18 of 30 (60%) placebo patients (absolute risk reduction 10.0%, 95% confidence interval [CI] -17.3, 37.3). Among the 23 patients with an initial oxygen index (OI) >/=13 and 0.05) observed for brain natriuretic peptide over time. CONCLUSIONS: Brain natriuretic peptide measured early after admission is increased in children with septic shock, especially in those with cold shock. In addition, the level at 12 hrs correlates with both severity of illness and myocardial dysfunction. Brain natriuretic peptide may be useful in assessing myocardial dysfunction from septic shock, particularly in identifying children with cold shock. Further studies are warranted to determine whether this measurement will be helpful in guiding therapy in pediatric septic shock. PMID- 18679144 TI - Implications of thrombocytopenia and platelet course on pediatric intensive care unit outcomes. AB - INTRODUCTION: Thrombocytopenia has been shown to be an independent predictor of mortality and prolonged hospital length of stay in critically ill adults. Studies are lacking in the pediatric intensive care unit population. We evaluated the relationship between platelet counts at admission, platelet course, and outcomes. HYPOTHESES: 1) Thrombocytopenia at the time of admission to the pediatric intensive care unit is a risk factor for increased mortality and prolonged length of stay. 2) Thrombocytopenia at any point during pediatric intensive care unit stay is associated with increased mortality and length of stay. 3) Falling platelet counts during a pediatric intensive care unit course are associated with greater mortality and longer length of stay. METHOD: Prospective observational study. STUDY POPULATION: All patients admitted to a multidisciplinary tertiary care pediatric intensive care unit in a University Hospital over the course of a year. ANALYSIS OF DATA: Data were analyzed using logistic and linear regression. RESULTS: Thrombocytopenia (platelet count <150 x 10/L) was present in 17.3% of pediatric intensive care unit patients on admission. Mortality was higher in thrombocytopenic patients (17.6% vs. 2.47%, p < 0.001). The median length of stay in the thrombocytopenia and nonthrombocytopenia groups was 4 days vs. 1.6 days, respectively (p < 0.001). The pediatric intensive care unit patients (25.3%) were thrombocytopenic at some point in their stay. They had higher mortality (17.1% vs. 0.9%, odds ratio [OR] 23.8, 95% confidence interval [CI] 5.2-108.6, p < 0.0005) and longer length of stay (median 6.6 days vs. 1.5 days, p < 0.0005) compared with those who were never thrombocytopenic. For every 10% fall in platelet count from the time of admission, the OR for mortality was 1.4 (95% CI 1.1-1.8) and the length of stay was longer (p < 0.0005). Patients with normal platelet counts at admission who later developed thrombocytopenia had increased mortality (OR 18.6, 95% CI 3.2-107.3) and longer length of stay (p < 0.0005) compared with those who did not develop thrombocytopenia. CONCLUSION: Thrombocytopenia and falling platelet counts are associated with increased risk of mortality and length of stay in the pediatric intensive care unit. PMID- 18679145 TI - Myocardial transcriptional profiles in a murine model of sepsis: evidence for the importance of age. AB - BACKGROUND: Age influences outcome of sepsis and septic shock. The mechanism of this age-dependent vulnerability to sepsis remains largely unknown. Because much of the mortality and morbidity associated with sepsis and septic shock is the result of severe derangements in the cardiovascular system, it is possible that the myocardium responds to injury in a developmentally influenced manner. We hypothesized that analysis of cardiac RNA expression profiles may differentiate between the myocardial response to sepsis in young and old mice. METHODS AND RESULTS: Sixteen FVB/N male mice were stratified based on age. Young animals were 6 wks old, correlating to 4 to 6 human years, and aged animals were 20 months old correlating to 70 to 80 human years. Animals underwent either cecal ligation and puncture to produce polymicrobial sepsis or a sham operation. Both ventricles were excised after kill at 24 hrs. There were 53 genes that differed in RNA abundance between the four groups (false discovery rate of 0.005, p < 0.00001). Additionally, four genes were associated with an age-dependent response to sepsis: CYP2B2 (cytochrome P450, family 2, subfamily B, polypeptide 6), VGLL2 (vestigial like 2), and PAH (phenylalanine hydroxylase). The fourth gene is an expressed sequence tag, the function of which is related to the cytochrome P450 family. These genes play roles in phenylalanine, tyrosine, tryptophan, and fatty acid metabolism. CONCLUSIONS: This report describes the transcriptional response of the heart to sepsis. In addition, our findings suggest that these differences are in part age-dependent and serve as hypothesis generation. PMID- 18679146 TI - A comprehensive review of pediatric endotracheal suctioning: Effects, indications, and clinical practice. AB - OBJECTIVE: To provide a comprehensive, evidence-based review of pediatric endotracheal suctioning: effects, indications, and clinical practice. METHODS: PubMed, Cumulative Index of Nursing and Allied Health Literature, and PEDro (Physiotherapy Evidence Database) electronic databases were searched for English language articles, published between 1962 and June 2007. Owing to the paucity of objective pediatric data, all reports dealing with this topic were examined, including adult and neonatal studies. RESULTS: One hundred eighteen references were included in the final review. Despite the widespread use of endotracheal suctioning, very little high-level evidence dealing with pediatric endotracheal suctioning exists. Studies of mechanically ventilated neonatal, pediatric, and adult patients have shown that suctioning causes a range of potentially serious complications. Current practice guidelines are not based on evidence from controlled clinical trials. There is no clear evidence that endotracheal suctioning improves respiratory mechanics, with most studies pointing to the detrimental effect it has on lung mechanics. Suctioning should be performed when obstructive secretions are present rather than routinely. There is no clear evidence for the superiority of closed- or open-system suctioning, nor is there clear evidence for appropriate vacuum pressures and suction catheter size. Sterility does not seem to be necessary when suctioning. Preoxygenation has short term benefits, but the longer-term impact is unknown. Routine saline instillation before suctioning should not be performed. Recruitment maneuvers performed after suctioning have not been shown to be useful as standard practice. CONCLUSIONS: Endotracheal suctioning is a procedure used regularly in the pediatric intensive care unit. Despite this, good evidence supporting its practice is limited. Further, controlled clinical studies are needed to develop evidence-based protocols for endotracheal suctioning of infants and children, and to examine the impact of different suctioning techniques on the duration of ventilatory support, incidence of nosocomial infection, and length of pediatric intensive care unit and hospital stay. PMID- 18679147 TI - The endotracheal tube air leak test does not predict extubation outcome in critically ill pediatric patients. AB - OBJECTIVE: Endotracheal tube air leak pressures are used to predict postextubation upper airway compromise such as stridor, upper airway obstruction, or risk of reintubation. To determine whether the absence of an endotracheal tube air leak (air leak test >/=30 cm H2O) measured during the course of mechanical ventilation predicts extubation failure in infants and children. DESIGN: Prospective, blinded cohort. SETTING: Multidisciplinary pediatric intensive care unit of a university hospital. PATIENTS: Patients younger than or equal to 18 yrs and intubated >/=24 hrs. INTERVENTIONS: The pressure required to produce an audible endotracheal tube air leak was measured within 12 hrs of intubation and extubation. Unless prescribed by the medical care team, patients did not receive neuromuscular blocking agents during air leak test measurements. MEASUREMENTS AND MAIN RESULTS: The need for reintubation (i.e., extubation failure) was recorded during the 24-hr postextubation period. Seventy-four patients were enrolled resulting in 59 observed extubation trials. The extubation failure rate was 15.3% (9 of 59). Seven patients were treated for postextubation stridor. Extubation failure was associated with a longer median length of ventilation, 177 vs. 78 hrs, p = 0.03. Extubation success was associated with the use of postextubation noninvasive ventilation (p = 0.04). The air leak was absent for the duration of mechanical ventilation (i.e., >/=30 cm H2O at intubation and extubation) in ten patients. Absence of the air leak did not predict extubation failure (negative predictive value 27%, 95% confidence interval 6-60). The air leak test was >/=30 cm H2O before extubation in 47% (28 of 59) of patients yet 23 patients extubated successfully (negative predictive value 18%). CONCLUSIONS: An endotracheal tube air leak pressure >/=30 cm H2O measured in the nonparalyzed patient before extubation or for the duration of mechanical ventilation was common and did not predict an increased risk for extubation failure. Pediatric patients who are clinically identified as candidates for an extubation trial but do not have an endotracheal tube air leak may successfully tolerate removal of the endotracheal tube. PMID- 18679148 TI - A prospective, randomized, controlled trial of noninvasive ventilation in pediatric acute respiratory failure. AB - OUTCOMES: To compare the benefits of noninvasive ventilation (NIV) plus standard therapy vs. standard therapy alone in children with acute respiratory failure; assess method effectiveness in improving gas exchange and vital signs; and assess method safety. DESIGN: Prospective, randomized, controlled study. SITE: Two pediatric intensive care units in Santiago, Chile, at Clinica Santa Maria and Clinica Davila, respectively. PATIENTS AND METHODS: Fifty patients with acute respiratory failure admitted to pediatric intensive care units were recruited; 25 patients were randomly allocated to noninvasive inspiratory positive airway pressure and expiratory positive airway pressure plus standard therapy (study group); the remaining 25 were given standard therapy (control group). Both groups were comparable in demographic terms. INTERVENTIONS AND MEASUREMENTS: The study group received NIV under inspiratory positive airway pressure ranging between 12 cm and 18 cm H2O and expiratory positive airway pressure between 6 cm and 12 cm H2O. Vital signs (cardiac and respiratory frequency), Po2, Pco2, pH, and Po2/Fio2 were recorded at the start and 1, 6, 12, 24, and 48 hrs into the study. RESULTS: Heart rate and respiratory rate improved significantly with NIV. Heart rate and respiratory rate were significantly lower after 1 hr of treatment compared with admission (p = 0.0009 and p = 0.004, respectively). The trend continued over time, heart rate being significantly lower than control after the first hour and heart rate after 6 hrs. With NIV, Po2/Fio2 improved significantly from the first hour. The endotracheal intubation was significantly lower (28%) in the NIV group than in the control group (60%; p = 0.045). CONCLUSIONS: NIV improves hypoxemia and the signs and symptoms of acute respiratory failure. NIV seems to afford these patients protection from endotracheal intubation. PMID- 18679149 TI - Single nucleotide polymorphisms in genes of circulatory homeostasis in surviving pediatric intensive care patients with meningococcal infection. AB - OBJECTIVE: In the course of a meningococcal infection, invasive and severe disease occurs in a restricted number of individuals. The predominant mechanism of death in case of meningococcal septic shock is circulatory failure. Inotropic requirements between patients vary widely. We investigated whether polymorphisms in genes regulating the hemodynamic response influence the amount of inotropics required or the susceptibility to severe meningococcal disease. DESIGN: Retrospective case control study. SETTING: Single-center pediatric intensive care unit (PICU). PATIENTS: Fifty-six cases (all consecutive patients admitted to the PICU between 1993 and 2001 with a proven meningococcal infection) and 136 controls. Patients were divided into two groups according to their inotropic requirements. INTERVENTION: DNA analysis was performed to determine the polymorphisms of the beta-adrenergic receptor gene-1, beta-adrenergic receptor gene-2, alpha-adducin, angiotensin converting enzyme, and angiotensin II type-1 receptor-1 genes. RESULTS: For the alpha-adducin gene a significant difference of the genotype distribution was found between the cases and controls. The odds ratio for admission to the PICU with meningococcal sepsis with or without meningitis, for carriers of the variant allele (Gly460Trp or Trp460Trp) was 2.1 (95% confidence interval 1.11-4.04; p < 0.02). Cases, homozygote for the wild type allele of the beta-1 adrenergic receptor at locus 389, were more likely to have a low pediatric risk of mortality score on admission (odds ratio 3.6, 95% confidence interval 1.11-11.76). No difference was found in the distribution of the beta-adrenergic receptor gene-1, beta-adrenergic receptor gene-2, angiotensin converting enzyme, and angiotensin II type-1 receptor-1 polymorphisms between the two groups of patients or between cases and controls. CONCLUSIONS: Among patients admitted to the PICU with a meningococcal infection, the variant allele of the alpha-adducin gene was more prevalent compared with controls. Patients with the variant allele of the beta-adrenergic receptor gene-1 at locus 389 were more likely to have a high pediatric risk of mortality score on admission. The mechanism and clinical relevance of these findings is unclear. PMID- 18679150 TI - The umbilical arterial catheter: a formula for improved positioning in the very low birth weight infant. AB - OBJECTIVE: : An umbilical arterial catheter is often used to monitor blood pressure and take frequent blood samples in the very low birth weight newborn infant requiring neonatal intensive care. Incorrect placement of the umbilical arterial catheter increases catheter complications, and adjustment of catheter position after radiograph increases infant handling and infection risk. Current methods overestimate insertion length in very low birth weight babies. We suggest a new formula for calculating insertion length that is more appropriate for today's neonatal intensive care population. The Umbilical Arterial Catheter Calculation Study, Australian Perinatal Trials Register PT0398, was set up to investigate this technical change. Our hypothesis was that the new formula would improve the siting of umbilical arterial catheters in very low birth weight infants. DESIGN: : Randomized control trial. SETTING: : Tertiary referral neonatal intensive care unit. PATIENTS: : All infants <1500 g who were free from major cardiovascular malformations and who received an umbilical arterial catheter for clinical reasons were eligible for the study. INTERVENTIONS: : Infants were randomized to current practice, using a nomogram derived from Dunn (control), or to the new formula: insertional length (cm) = (4 x birth weight [kg]) + 7. Primary outcome was correct catheter position (T6-10) on initial radiograph. MEASUREMENTS AND MAIN RESULTS: : Seventy-four randomized infants had catheters successfully inserted. There were no significant differences between the groups in birth weight, gestational age, or gender. There was a significant increase in correctly sited catheters (p = .003) with the new formula. Overinsertion of the umbilical arterial catheter was significantly less likely (p < .0001). Underinsertion was not significantly increased. Umbilical arterial catheter manipulation after radiograph was decreased from 50% to 5% (p = .007). There was no increase in adverse effects. CONCLUSIONS: : The use of the new formula results in better overall placement and in significantly less overinsertion of umbilical artery catheters in very low birth weight infants. PMID- 18679151 TI - Micafungin versus liposomal amphotericin B for pediatric patients with invasive candidiasis: substudy of a randomized double-blind trial. AB - BACKGROUND: Invasive candidiasis is increasingly prevalent in premature infants and seriously ill children, and pediatric data on available antifungal therapies are lacking. METHODS: We conducted a pediatric substudy as part of a double blind, randomized, multinational trial to compare micafungin (2 mg/kg) with liposomal amphotericin B (3 mg/kg) as first-line treatment of invasive candidiasis. Treatment success was defined as clinical and mycologic response at the end of therapy. Statistical analyses were descriptive, as the sample size meant that the study was not powered for hypothesis testing. RESULTS: One hundred six patients were included in the intent-to-treat population; and 98 patients-48 patients in the micafungin group and 50 patients in the liposomal amphotericin B group-in the modified intent-to-treat population. Baseline characteristics were balanced between treatment groups. Overall, 57 patients were <2 years old including 19 patients who were premature at birth; and 41 patients were 2 to <16 years old. Most patients (91/98, 92.9%) had candidemia, and 7/98 (7.1%) patients had other forms of invasive candidiasis. Treatment success was observed for 35/48 (72.9%) patients treated with micafungin and 38/50 (76.0%) patients treated with liposomal amphotericin B. The difference in proportions adjusted for neutropenic status was -2.4% [95% CI: (-20.1 to 15.3)]. Efficacy findings were consistent, independent of the neutropenic status, the age of the patient, and whether the patient was premature at birth. Both treatments were well tolerated, but with a lower incidence of adverse events that led to discontinuation in the micafungin group (2/52, 3.8%) compared with the liposomal amphotericin B group (9/54, 16.7%) (P = 0.05, Fisher exact test). CONCLUSIONS: Micafungin seems to be similarly effective and as safe as liposomal amphotericin B for the treatment of invasive candidiasis in pediatric patients. (ClinicalTrials.gov number, NCT00106288). PMID- 18679153 TI - Serious neurologic sequelae in cases of meningitis arising from infection by conjugate vaccine-related and nonvaccine-related serogroups of Streptococcus pneumoniae. AB - BACKGROUND: Introduction of the heptavalent conjugate vaccine for Streptococcus pneumoniae (PCV7) has led to a dramatic decline in meningitis by PCV7 serotypes, raising the possibility of similar trends by PCV7-related serogroups through cross-protection. A present concern, however, is of serotype replacement by pneumococci not related to PCV7 serogroups. If this occurs, there are currently few data to predict whether clinical outcomes will change substantially. METHODS: To address these questions, we analyzed medical records of 86 cases of pneumococcal meningitis treated at Nationwide Children's Hospital (1993-2004). Adverse neurologic sequelae and death were compared between cases with cerebrospinal fluid isolates characterized as vaccine-related serogroups serotypes belonging to PCV7 or related to PCV7 serogroups, and those designated nonvaccine serogroups-serotypes neither belonging to PCV7 nor related to PCV7 serogroups. Serotype 19A, because of recent reports of increased incidence, was subanalyzed separately. RESULTS: Thirty-six of 86 (42%) subjects had serious complications, including 6 who died. All 6 deaths occurred in patients with vaccine-related serogroups. Deafness was the most common complication, occurring in 26 (32.5%) survivors. There was no difference in the frequency of total complications between PCV7-related and non-PCV7 groups: 5 of 12 (42%) for non PCV7 serogroups versus 31 of 74 (42%) for PCV-related serogroups (OR: 1.0; 95% CI: 0.2-4.0). Serious outcomes occurred in 3 of 4 cases due to serogroup 19A. Non PCV7 serogroups increased slightly at the end of the study period. CONCLUSIONS: In children with pneumococcal meningitis, infections with non-PCV7 serogroups seem less likely to result in death. Among survivors, there is preliminary evidence of parity in neurologic sequelae between PCV7 and non-PCV7 serogroups. PMID- 18679152 TI - Morbidity and mortality among a cohort of human immunodeficiency virus type 1 infected and uninfected pregnant women and their infants from Malawi, Zambia, and Tanzania. AB - BACKGROUND: Morbidity and mortality patterns among pregnant women and their infants (before antiretroviral therapy was widely available) determines HIV-1 diagnostic, monitoring, and care interventions. METHODS: Data from mothers and their infants enrolled in a trial of antibiotics to reduce mother-to-child transmission of HIV-1 at 4 sub-Saharan African sites were analyzed. Women were enrolled during pregnancy and follow-up continued until the infants reached 12 months of age. We describe maternal and infant morbidity and mortality in a cohort of HIV-1-infected and HIV-1-uninfected mothers. Maternal and infant factors associated with mortality risk in the infants were assessed using Cox proportional hazard modeling. RESULTS: Among 2292 HIV-1-infected mothers, 166 (7.2%) had a serious adverse event (SAE) and 42 (1.8%) died, whereas no deaths occurred among the 331 HIV-1 uninfected mothers. Four hundred twenty-four (17.8%) of 2383 infants had an SAE and 349 (16.4%) died before the end of follow-up. Infants with early HIV-1 infection (birth to 4-6 weeks) had the highest mortality. Among infants born to HIV-1-infected women, maternal morbidity and mortality (P = 0.0001), baseline CD4 count (P = 0.0002), and baseline plasma HIV 1 RNA concentration (P < 0.0001) were significant predictors of infant mortality in multivariate analyses. CONCLUSIONS: The high mortality among infants with early HIV-1 infection supports access to HIV-1 diagnostics and appropriate early treatment for all infants of HIV-1-infected mothers. The significant association between stage of maternal HIV-1 infection and infant mortality supports routine CD4 counts at the time of prenatal HIV-1 testing. PMID- 18679154 TI - Extensive transmission of Mycobacterium tuberculosis among children on a school bus. AB - We report a tuberculosis outbreak in school children caused by exposure to a sputum-smear positive school bus driver. Exposed children were assessed using 2 versions of interferon-gamma release assays. Fifty-five percent (18/34) had a positive interferon-gamma release assays and 4 children developed tuberculosis. This suggests that transmission may be very efficient in this setting and highlights the need for early diagnosis. PMID- 18679155 TI - Neonatal meningoencephalitis caused by Bacillus cereus. AB - The classic organisms associated with central nervous system infection in the neonate are herpes simplex, Listeria monocytogenes, Escherichia coli, and Streptococcus agalactiae; we describe an unusual case of neonatal meningoencephalitis caused by Bacillus cereus. PMID- 18679156 TI - Insights into parturition biology from genetically altered mice. AB - With the growing frequency of preterm birth, increased effort has been made to elucidate the physiology of normal and aberrant parturition. As with many developmental processes, the study of genetically altered mice has led to an increased understanding of mechanisms controlling the maintenance and resolution of pregnancy. Studies in genetically altered mice have implicated critical roles for both prostaglandin synthesis and degradation in luteolysis and the progression of labor. The importance of local modulation of progesterone activity to cervical ripening has also been demonstrated. Although a decline in levels of serum progesterone is a part of normal labor initiation in mice but not humans, murine labor without progesterone withdrawal has been reported in some cases. These findings emphasize the importance of other components of the parturition cascade that are shared in mice and humans and highlights the importance of an increased understanding of the physiology of mouse parturition. PMID- 18679157 TI - Comparison of immunological status of African and European cord blood mononuclear cells. AB - The cellular aspects of the immunologic development of the fetus during pregnancy have been studied mainly in populations living in economically well developed countries, and there is no data concerning variation of the neonatal cellular immune system in geographically distinct areas with different environments. Here, we report a comparative immunologic marker analysis of the circulating mononuclear cell subsets in unstimulated cord blood of newborns from Gabon and Austria, assessing the activation and maturation status of T and B lymphocytes as well as antigen-presenting cells. Cells and markers hypothesized to be modulated by frequent exposure to microorganisms and parasites such as regulatory T cells and the expression of toll-like receptor 2 on antigen-presenting cells were also studied. We found marked differences in terms of expression of immunologic markers between the two populations, pointing to a comparatively enhanced maturation status of the neonatal immune system in general in the African setting. The observations suggest that environmental factors, including differential exposure to pathogens as well as nutritional differences, may have substantial impact on the development of the fetal immune system. PMID- 18679158 TI - Immunologic and biochemical factors of coincident celiac disease and type 1 diabetes mellitus in children. AB - The objective of the study was to investigate whether immunologic and biochemical events occurring in the course of type 1 diabetes mellitus might play a role in the development of the celiac disease. The study was carried out on 223 children with long-standing diabetes mellitus type 1 (DM1). All the patients had TSH, fT4, fT3, urinary albumin secretion rate, IgA, level of antigliadin antibodies (AGA) IgA and IgG, antitissue transglutaminase IgA antibodies, antiendomysium (EmA) IgA and IgG antibodies and antitireoglobulin antibodies, antithyroid peroxidase antibodies evaluated. Serum TNF-alpha, IL-6, and IL-10 levels were also measured. The group of children with coincident DM1 and celiac disease and without autoimmune thyroiditis was characterized by significantly higher glycosylated hemoglobin, higher serum TNF-alpha, IL-6 but lower serum IL-10 in relation to the remaining diabetic patients. A statistically significant positive correlation was observed between IgA-anti-tTG and serum TNF-alpha (R = 0.28, p = 0.026); between IgG AGA and serum IL-6 (R = 0.31, p = 0.023); and between glycosylated hemoglobin and IgA-anti-tTG (R = 0.21, p = 0.001) and IgA antiendomysium (R = 0.22, p = 0.001). Poor metabolic control, persistent elevated levels of proinflammatory cytokines, and decreased level of antiinflammatory cytokines occurring in the course of type 1 diabetes mellitus might influence the incidence of celiac disease. PMID- 18679159 TI - Preemptive morphine analgesia attenuates the long-term consequences of neonatal inflammation in male and female rats. AB - Despite mounting evidence on the importance of pain management in preterm infants, clinical use of analgesics in this population is limited. Our previous studies have shown that neonatal inflammation results in long-term alterations in adult somatosensory thresholds, characterized by decreased baseline nociceptive sensitivity, and enhanced hyperalgesia after a subsequent inflammatory insult. The present studies were conducted to determine whether preemptive morphine attenuates these negative consequences. At P0, pups received an injection of morphine sulfate before an intraplantar injection of 1% carrageenan. Control pups received either saline (SAL) followed by intraplantar carrageenan, morphine sulfate followed by intraplantar SAL, or SAL followed by intraplantar SAL. Preemptive morphine significantly attenuated neonatal injury-induced hypoalgesia in adolescence and adulthood. Similarly, morphine pretreated animals displayed significantly less hyperalgesia and recovered faster from a subsequent inflammatory insult compared with controls. Neonatal morphine had no significant effect on morphine analgesia in adulthood. Interestingly, neonatally injured animals that did not receive morphine displayed a significant rightward shift in the morphine dose-response curve in the absence of peripheral inflammation. Together, these results demonstrate that preemptive morphine significantly attenuates the long-term behavioral impact of neonatal inflammatory injury. PMID- 18679160 TI - Campylobacter jejuni mediated disruption of polarized epithelial monolayers is cell-type specific, time dependent, and correlates with bacterial invasion. AB - The precise mechanism by which the most common cause of bacterial enterocolitis in humans, Campylobacter jejuni, perturbs the intestinal mucosa remains elusive. To define effects of C. jejuni infection on mucosal permeability, Madin-Darby canine kidney (MDCK)-I and T84 cell monolayers were infected with C. jejuni for up to 48 h. All three tested C. jejuni strains caused a 73-78% reduction in transepithelial electrical resistance (TER) in intestinal (T84) cell monolayers, whereas only one strain slightly reduced TER of MDCK-I cells by 25% after 48 h infection. Infection with C. jejuni strains also caused a 2.3-4.5-fold increase in dextran permeability, but only in T84 cells. C. jejuni infection of monolayers also caused morphologic changes in desmosomes, observed by transmission electron microscopy. The cell-type specificity, demonstrated by increased T84 monolayer permeability, correlated with higher bacterial invasion into these cells, relative to MDCK-I cells. In T84 cells, invasion and bacterial translocation preceded barrier disruption and inhibition of C. jejuni invasion using a pharmacological inhibitor of phosphoinositide 3-kinase, reduced the drop in TER. These findings suggest that C. jejuni disruption of monolayers is mediated by invasion, provide new insights into C. jejuni-host epithelial barrier interactions, and offer potential mechanisms of intestinal injury and chronic immune stimulation. PMID- 18679162 TI - Zebrafish as a developmental model organism for pediatric research. AB - Zebrafish has many advantages as a model of human pediatric research. Given the physical and ethical problems with performing experiments on human patients, biomedical research has focused on using model organisms to study biologic processes conserved between humans and lower vertebrates. The most common model organisms are small mammals, usually rats and mice. Although these models have significant advantages, they are also expensive to maintain, difficult to manipulate embryonically, and limited for large-scale genetic studies. The zebrafish model nicely complements these deficiencies in mammalian experimental models. The low cost, small size, and external development of zebrafish make it an excellent model for vertebrate development biology. Techniques for large-scale genome mutagenesis and gene mapping, transgenesis, protein overexpression or knockdown, cell transplantation and chimeric embryo analysis, and chemical screens have immeasurably increased the power of this model organism. It is now possible to rapidly determine the developmental function of a gene of interest in vivo, and then identify genetic and chemical modifiers of the processes involved. Discoveries made in zebrafish can be further validated in mammals. With novel technologies being regularly developed, the zebrafish is poised to significantly improve our understanding of vertebrate development under normal and pathologic conditions. PMID- 18679163 TI - Prenatal exposure to nicotine affects substance p and preprotachykinin-A mRNA levels in newborn rat. AB - Prenatal nicotine exposure influences neuronal development including effects on several neurotransmitter systems. It also attenuates the ventilatory response to hypoxia, known to require a functional substance P-ergic system. Previous studies have shown that nicotine increases the risk for sudden infant death syndrome (SIDS) by 4-fold, and that SIDS-victims have elevated brainstem levels of substance P. We, therefore, studied the effect of prenatal nicotine exposure on the levels of substance P-like immunoreactivity by RIA in the brain in newborn rat pups. The expression of the substance P precursor preprotachykinin A mRNA was also determined by real-time reverse transcriptase-polymerase chain reaction in carotid body, in petrosal/jugular and trigeminal ganglia, in cervical and lumbar dorsal root ganglia, and in the brainstem. We found that prenatal nicotine exposure increased levels of substance P-like immunoreactivity in the brainstem without changing levels in other parts of the brain or in the adrenals. Furthermore, mRNA levels were increased in the carotid bodies and in the petrosal ganglia, in contrast to the decreased levels in the cervical dorsal root ganglia. We conclude that nicotine causes alterations in the substance P-ergic system in the brainstem, possibly linked to the increased risk for SIDS after prenatal nicotine exposure. PMID- 18679164 TI - Neuroprotective effects of the nonpsychoactive cannabinoid cannabidiol in hypoxic ischemic newborn piglets. AB - To test the neuroprotective effects of the nonpsychoactive cannabinoid cannabidiol (CBD), piglets received i.v. CBD or vehicle after hypoxia-ischemia (HI: temporary occlusion of both carotid arteries plus hypoxia). Nonhypoxic ischemic sham-operated piglets remained as controls. Brain damage was studied by near-infrared spectroscopy (NIRS) and amplitude-integrated electroencephalography (aEEG) and by histologic assessment (Nissl and FluoroJadeB staining). In HI+vehicle, HI led to severe cerebral hemodynamic and metabolic impairment, as reflected in NIRS by an increase in total Hb index (THI) and a decrease in the fractional tissue oxygenation extraction (FTOE); in HI+CBD the increase of THI was blunted and FTOE remained similar to SHAM. HI profoundly decreased EEG amplitude, which was not recovered in HI+vehicle, indicating cerebral hypofunction; seizures were observed in all HI+vehicle. In HI+CBD, however, EEG amplitude recovered to 46.4 7.8% baseline and seizures appeared only in 4/8 piglets (both p < 0.05). The number of viable neurons decreased and that of degenerating neurons increased in HI+vehicle; CBD reduced both effects by more than 50%. CBD administration was free from side effects; moreover, CBD administration was associated with cardiac, hemodynamic, and ventilatory beneficial effects. In conclusion, administration of CBD after HI reduced short term brain damage and was associated with extracerebral benefits. PMID- 18679165 TI - The effect of fish oil supplementation on heart rate in healthy Danish infants. AB - Polyunsaturated n-3 fatty acids (n-3PUFA) may improve brain development and prevent cardiovascular disease. Heart rhythm is autonomically controlled and among the affected cardiovascular risk markers in adults. The aim of the study was to examine whether fish oil supplementation in late infancy could modify heart rate (HR) and heart rate variability (HRV). In a 2 x 2-intervention, 83 healthy Danish infants were randomized to +/- fish oil (3.4 +/- 1.1 mL/d) and cow's milk or infant formula from 9 to 12 mo of age. In 57 infants, 0.5-h ECG recordings were successfully obtained before and after the intervention and erythrocyte fatty acid composition was determined in 30 of these. Fish oil supplementation raised erythrocyte n-3PUFA content (p < 0.001). No significant group differences were seen in HR or HRV. However, a fish-oil x gender interaction was observed on mean RR interval (p = 0.001) with a 6% longer mean RR interval in fish-oil-supplemented boys (p = 0.007). Irrespective of gender, there was a positive association between the 9- and 12-mo changes in RR interval and erythrocyte n-3PUFA (p < 0.001). In infants with confirmed changes in erythrocyte n-3PUFA, mean RR interval was found to be longer (p = 0.011) in the fish-oil supplemented groups. The study suggests that fish oil may affect heart rhythm in infants similar to that observed in adults. This may imply low n-3PUFA-status in late infancy and n-3PUFA influence on CNS function. PMID- 18679166 TI - Sulfonylurea receptor 1 in the germinal matrix of premature infants. AB - Germinal matrix (GM) hemorrhage (GMH) is a major cause of mortality and of life long morbidity from cerebral palsy. GMH is typically preceded by hypoxic/ischemic events and is believed to arise from rupture of weakened veins in the GM. In the CNS, hypoxia/ischemia up-regulate sulfonylurea receptor 1 (SUR1)-regulated NCCa ATP channels in microvascular endothelium, with channel activation by depletion of ATP being responsible for progressive secondary hemorrhage. We hypothesized that this channel might be up-regulated in the GM of preterm infants at risk for GMH. Here, we studied expression of the regulatory subunit of the channel, SUR1, and its transcriptional antecedent, hypoxia inducible factor 1 (HIF1), in postmortem tissues of premature infants who either were at risk for or who sustained GMH. We found regionally specific up-regulation of HIF1 and of SUR1 protein and mRNA in GM tissues, compared with remote cortical tissues. Up regulation was prominent in most progenitor cells, whereas in veins, SUR1 was found predominantly in infants who had sustained GMH compared with those without hemorrhage. Our data suggest that the SUR1-regulated NCCa-ATP channel may be associated with GMH, and that pharmacological block of these channels could potentially reduce the incidence of this devastating complication of prematurity. PMID- 18679167 TI - Carbohydrate and lipid metabolism following infliximab therapy in pediatric Crohn's disease. AB - Improvements in insulin resistance after anti-TNF-alpha therapy have been reported in inflammatory conditions, although no changes were noted in adult patients with Crohn's disease. There is no information concerning insulin resistance and substrate metabolism in children with Crohn's disease after anti TNF-alpha therapy. Our aim was to describe changes in carbohydrate and lipid metabolism in children with active Crohn's disease after their initial dose of infliximab. Children with active Crohn's disease underwent measurement of plasma insulin and glucose just before and 2 wk after their initial infusion of infliximab, an anti-TNF-alpha antibody. In addition, resting energy expenditure, with determination of both carbohydrate and lipid oxidation rates, was determined. Measurements were conducted in both fasting and parenterally fed states. Despite no changes in resting energy expenditure, a significant reduction (p < 0.05) in RQ (5%) and carbohydrate oxidation rate (24%), with a corresponding increase in lipid oxidation rate (42%) was found during parenteral nutrition infusion. No differences in plasma insulin, glucose, and insulin resistance were noted when comparing pre- and postinfliximab measurements. PMID- 18679168 TI - Immune mediators in idiopathic nephrotic syndrome: evidence for a relation between interleukin 8 and proteinuria. AB - The pathogenesis of idiopathic nephrotic syndrome (INS) remains unknown. Several findings suggest a role for the immune system. This study aimed to evaluate immune mediators in INS by measuring plasma and urinary levels of transforming growth factor beta1 (TGF-beta1), monocyte chemoattractant protein-1 (MCP-1/CCL2), regulated on activation normal T-cell expressed and secreted (RANTES/CCL5) and IL 8 (IL-8/CXCL8) in pediatric patients with INS and in age-matched healthy controls. Patients were divided according to their response to corticosteroids: steroid-sensitive (SS, n = 8), or steroid-resistant (SR, n = 24). Immune mediators were also compared in regard with disease activity (relapse and remission). Immune mediators were measured by ELISA. Plasma TGF-beta1 levels in SR patients were approximately 2.8-fold higher than control values (p < 0.05). Urinary IL-8/CXCL8 was 2.9-fold higher in INS patients in relapse (proteinuria >100 mg/m2/24 h) when compared with patients in remission (p < 0.05), and levels had a positive correlation with individual proteinuria values (p < 0.05). Urinary IL-8/CXCL8 was significantly higher in relapsed SR than in SS patients in remission. No changes in MCP-1/CCL2 and RANTES/CCL5 levels were detected. Our findings suggest that IL-8/CXCL8 and TGF-beta1 are involved in the pathogenesis of INS: IL-8/CXCL8 associated with local changes in glomerular permeability and TGF-beta1 could be related to worse response to corticosteroids. PMID- 18679169 TI - Redressing the balance in autism research. PMID- 18679170 TI - Molecular mechanisms of mechanosensing and their roles in fungal contact sensing. AB - Numerous fungal species respond to contact with a surface by undergoing differentiation. Contact between plant pathogenic fungi and a surface results in the elaboration of the complex structures that enable invasion of the host plant, and for the opportunistic human pathogen Candida albicans, contact with a semi solid surface results in invasive growth into the subjacent material. The ability to sense contact with an appropriate surface therefore contributes to the ability of these fungi to cause disease in their respective hosts. This Review discusses molecular mechanisms of mechanosensitivity, the proteins involved, such as mechanosensitive ion channels, G-protein-coupled receptors and integrins, and their putative roles in fungal contact sensing. PMID- 18679172 TI - Protein-based organelles in bacteria: carboxysomes and related microcompartments. AB - Many bacteria contain intracellular microcompartments with outer shells that are composed of thousands of protein subunits and interiors that are filled with functionally related enzymes. These microcompartments serve as organelles by sequestering specific metabolic pathways in bacterial cells. The carboxysome, a prototypical bacterial microcompartment that is found in cyanobacteria and some chemoautotrophs, encapsulates ribulose-l,5-bisphosphate carboxylase/oxygenase (RuBisCO) and carbonic anhydrase, and thereby enhances carbon fixation by elevating the levels of CO2 in the vicinity of RuBisCO. Evolutionarily related, but functionally distinct, microcompartments are present in diverse bacteria. Although bacterial microcompartments were first observed more than 40 years ago, a detailed understanding of how they function is only now beginning to emerge. PMID- 18679173 TI - Evoking plasmin for beta-amyloid clearance. PMID- 18679171 TI - Revisiting the host as a growth medium. AB - The ability of the human body to play host to bacterial pathogens has been studied for more than 200 years. Successful pathogenesis relies on the ability to acquire the nutrients that are necessary for growth and survival, yet relatively little is understood about the in vivo physiology and metabolism of most human pathogens. This Review discusses how in vivo carbon sources can affect disease and highlights the concept that carbon metabolic pathways provide viable targets for antibiotic development. PMID- 18679174 TI - IFNgamma and B7-H1 in the immunology of mesenchymal stem cells. PMID- 18679176 TI - Pharmacoeconomics in the era of individualized medicine. PMID- 18679179 TI - Pharmacoeconomics is coming of age. PMID- 18679180 TI - Harnessing economic drivers for successful clinical implementation of pharmacogenetic testing. AB - Before pharmacogenetic (PGx) testing has a major impact on clinical practice, two levels of evidence must be generated. First, studies demonstrating the links between genetic variation and response to medications in defined populations are needed, along with development of valid tests to measure these specific variants. Second, studies should be conducted to evaluate whether PGx testing improves health outcomes for patients and whether the decision to test is cost-effective relative to usual care. This latter set of questions is typically of greatest relevance to clinicians and payers, the ultimate gatekeepers for the clinical integration of pharmacogenetics. To date, nearly all of the research efforts and funding for PGx have been focused on the first set of issues-getting the science right. However, now is the time to increase our research efforts on the second set of issues-to improve the PGx evidence base for both clinical and economic decision making. PMID- 18679181 TI - Nonprofit biomedical companies. AB - Nonprofit biomedical firms are an integrated market-based solution to improve incentives for investment in promising scientific areas that have high social value but minimal potential for profit. We briefly review the current market with an emphasis on the financing of innovative product development and propose ideas for new nonprofit companies centered on the health concerns of developed countries. We conclude with a suggestion that opportunities exist for nonprofit firms focused on cancer diagnostics, given the limitations of current financing incentives and ripe scientific opportunity. PMID- 18679182 TI - Using pharmacoeconomics to value pharmacotherapy. AB - Pharmacotherapeutic decisions are increasingly constrained in all clinical settings by the costs of drug treatment and medical care. Some biotech therapies (e.g., Avastin, Cerzyme, Herceptin, Gleevec, Erbitux) can cost from $10,000 to more than $100,000 per treatment episode. In 1996 the average drug treatment cost for advanced colon cancer was $500, and the average patient survival was 11 months. In 2006 the average drug treatment cost for such patients was $250,000, and the average patient survival was 24 months.(1) It is apparent that we are quickly arriving at a situation in which the determinants of medical decision making are not simply the clinical risks and benefits of treatment options but also how these are balanced against the economic costs of therapy. PMID- 18679183 TI - The European ACuteTox project: a modern integrative in vitro approach to better prediction of acute toxicity. AB - ACuteTox (optimization and prevalidation of an in vitro test strategy for predicting human acute toxicity) is an integrated European Union project under the Sixth Framework Programme with the aim of demonstrating that animal tests for acute systemic toxicity currently used for regulatory purposes could be replaced by a combination of alternative assays. Validated alternative test methods are urgently required for safety toxicology testing of chemicals, cosmetics, and drugs. PMID- 18679184 TI - Abstracts of the Shanghai International Symposium on Stem Cell Research. November 6-9, 2007. PMID- 18679363 TI - Degradation of BM SDF-1 by MMP-9: the role in G-CSF-induced hematopoietic stem/progenitor cell mobilization. AB - The major involvement of chemokines and proteolytic enzymes has recently been discovered in the mobilization process. Here, we report that the degradation of BM stromal cell-derived factor (SDF-1) by matrix metalloproteinase (MMP)-9 is important in G-CSF-mediated hematopoietic stem/progenitor cells (HSPCs) mobilization. In this study, the SDF-1 concentration in healthy donors BM plasma decreased significantly after 5 days of G-CSF administration, with no obvious change of SDF-1 in the peripheral blood. We also observed a similar result by immunohistochemical staining on the BM biopsy slides. In vitro, mobilized BM plasma exhibited decreased chemotactic effect on CD34(+) cells, compared with steady-state BM plasma. MMP-9 protein and mRNA increased dramatically in the BM plasma in accordance with SDF-1 decrease. MMP-9 enriched supernatant from HT1080 cell-conditioned medium upregulated CXCR4 expression and the migration of BM CD34(+) cells toward SDF-1. SDF-1 was a substrate for MMP-9, furthermore, SDF-1 also stimulated MMP-9 proteolytic enzyme activity of BM CD34(+) cells, which facilitate HSPCs migration. In BALB/c mice, HSPCs mobilization was significantly inhibited by anti-SDF-1 antibodies or MMP inhibitor, o-phenanthroline. In conclusion, the degradation of BM SDF-1 by MMP-9 is a vital step in mobilization. PMID- 18679365 TI - Myeloablative chemotherapy with autologous stem cell rescue for Ewing sarcoma. PMID- 18679366 TI - Impact of intensive PBSC mobilization therapy on outcomes following auto-SCT for non-Hodgkin's lymphoma. AB - The best method to mobilize PBSCs in patients with non-Hodgkin's Lymphoma (NHL) is uncertain. We hypothesized that PBSC mobilization using an intensive chemotherapy regimen would improve outcomes after autologous hematopoietic stem cell transplantation (ASCT) in NHL patients at high risk for relapse. Fifty NHL patients were prospectively allocated to intense mobilization with high-dose etoposide plus either high-dose cytarabine or CY if they were 'high risk' for relapse, whereas 30 patients were allocated to nonintense mobilization with CY if they were 'standard risk' (all patients, +/-rituximab). All intensely mobilized patients were hospitalized compared with one-third of nonintensely mobilized patients. The EFS after ASCT was the same between the two groups, but overall survival (OS) was better for intensely mobilized patients (<0.01), including the diffuse large B-cell subgroup (P<0.04). We conclude that the intense mobilization of PBSCs in patients with NHL is more efficient than nonintense mobilization, but with greater toxicity. The equalization of EFS and superiority of OS in patients intensely mobilized to those nonintensely mobilized suggests that a treatment strategy using intensive chemotherapy for mobilization may be improving NHL outcomes after ASCT. PMID- 18679367 TI - Acute appendicitis in adult neutropenic patients with hematologic malignancies. PMID- 18679368 TI - Human herpesvirus 6-associated limbic encephalitis in adult recipients of unrelated umbilical cord blood transplantation. PMID- 18679369 TI - Hematopoietic cell transplantation for hemophagocytic lymphohistiocytosis: a journey of a thousand miles begins with a single (big) step. AB - Hemophagocytic lymphohistiocytosis (HLH) is a rare, highly fatal disorder of uncontrolled inflammation, usually affecting infants. Significant progress in the treatment of this disorder has been achieved during the last decade, and outcomes for larger series of patients have been reported in recent years. Although medical therapy has advanced, hematopoietic cell transplantation remains the only curative therapy for patients with the familial form of this disorder. Unfortunately, these patients have demonstrated relatively poor post-transplant outcomes for a nonmalignant disorder, with approximately 30% mortality in the first 100 days. Early deaths were attributable to infection, GVHD, and unusually high rates of primary nonengraftment, venoocclusive disease and pneumonitis. In addition, a significant number of deaths were due to HLH reactivation, a unique complication seen in this patient group. In contrast, late complications were relatively infrequent and essentially all patients with durable engraftment remained in remission indefinitely. In this review, we will discuss recent progress in the transplant management of patients with HLH and potential future strategies, including the use of reduced intensity conditioning regimens. PMID- 18679370 TI - Outcome after reduced-intensity conditioning allogeneic SCT for AML in first complete remission: comparison of two regimens. PMID- 18679371 TI - Occurrence of donor-derived CLL 8 years after sibling donor SCT for CML. PMID- 18679372 TI - Predictors of outcome following myeloablative allo-SCT for therapy-related myelodysplastic syndrome and AML. AB - Administration of alkylating agents (Alk), topoisomerase II inhibitors (Topo II) and radiotherapy (RT) can result in therapy-related myelodysplastic syndrome or acute myelogenous leukaemia (t-MDS/t-AML), the optimal treatment for which is allo-SCT. A retrospective review was performed of 24 patients who underwent related- or unrelated-donor SCT for t-MDS/t-AML at our institution. Eight patients remain alive and in continuous remission (median follow-up 54 months (range, 12-161)) with estimated 5-year EFS being 30% (95% confidence intervals 16 58%). Corresponding actuarial risks of relapse and non-relapse mortality (NRM) are 39% (19-60%) and 30% (13-50%), respectively. EFS was 40% in Alk/RT-related t MDS/t-AML and 11% in Topo II-related t-MDS/t-AML (P=0.05), with an increased risk of relapse in the latter (56 vs 29%, respectively (P=0.05)). In multivariate analysis, development of acute GVHD (P=0.009) and Topo II-related t-MDS/t-AML (P=0.018) were associated with inferior EFS. Patients with acute GVHD had an increased risk of NRM (P=0.03) whereas risk of relapse was higher for patients of advanced age (P=0.046) and for patients who underwent bone marrow (vs blood) SCT (P=0.032). Allo-SCT can result in long-term survival for individuals with t-MDS/t AML although outcome in Topo II-related t-MDS/t-AML patients remains suboptimal. PMID- 18679373 TI - Imatinib mesylate versus allogeneic BMT for patients with chronic myeloid leukemia in first chronic phase. AB - Imatinib mesylate (IM) is now first-line treatment for CML. To study the results of treatment with IM after IFN failure/intolerance versus allogeneic BMT (allo BMT), we retrospectively analyzed 264 patients treated for CML in first chronic phase in three different institutions. Over a 6-year period (2001-2006), 174 patients received IM after failure of or intolerance to IFN. During the same period of time, 90 patients received an allo-BMT from an HLA-matched sibling (n=83) or an unrelated donor (n=7). The IM group was older (41 versus 33 years, P<0.001). Five-year EFS was 62% among patients receiving IM and 52% among patients undergoing allo-BMT (P=0.0002). OS at 5 years was 93% for IM-treated patients and 59% for patients undergoing allo-BMT (P<0.0001). Allo-BMT cannot be considered as first-line treatment for CML patients in first chronic phase. PMID- 18679374 TI - Neuropsychological outcome in very young hematopoietic SCT recipients in relation to pretransplant conditioning. AB - Stem cell transplantation involves conditioning with TBI and/or intensive chemotherapy, which may cause long-term neuropsychological deficits, particularly in children treated at a very young age. The aim of this study was to investigate whether very young children who receive chemotherapy-based conditioning only (BUCY) may have a more favorable neuropsychological outcome than children conditioned with TBI-CY. Twenty-two children who underwent allogeneic SCT at 0.4 3.6 years of age were subject to an extensive neuropsychological assessment at an average of 6.5 years post-therapy. The test results of 10 children exposed to BU were compared to the results of 12 children who had received TBI. Ten of them had received single-dose TBI, whereas two had received fractionated TBI. The BU group performed at age level on verbal measures, but tended to score below age level in the executive and visuo-spatial domains (P<0.01). By comparison, children treated with TBI had more pervasive neuropsychological impairments, including motor deficits (P<0.01) and varying degrees of perceptual (P<0.05), executive and cognitive (P<0.05) problems. In conclusion, children transplanted at a very young age had a more favorable neuropsychological development if conditioned with BUCY than if conditioned with single-dose TBI-CY. PMID- 18679376 TI - The Nrf2 transcription factor protects from toxin-induced liver injury and fibrosis. AB - The liver is frequently exposed to insults, including toxic chemicals and alcohol, viral infection or metabolic overload. Although it can fully regenerate after acute injury, chronic liver damage causes liver fibrosis and cirrhosis, which can result in complete liver failure. In this study, we demonstrate that the NF-E2-related factor 2 (Nrf2) transcription factor protects the liver from acute and chronic toxin-mediated damage. Repair of the liver injury that occurs after a single treatment with the hepatotoxin carbon tetrachloride (CCl(4)) was severely delayed in Nrf2-deficient mice. The defect in repair was accompanied by an enhanced and prolonged inflammatory and profibrotic response. After long-term CCl(4) treatment, liver fibrosis was strongly aggravated in the Nrf2 knockout mice and inflammation was enhanced. We demonstrate that these abnormalities are at least in part due to the reduced expression of known and novel Nrf2 target genes in hepatocytes, which encode enzymes involved in the detoxification of CCl(4) and its metabolites. These results suggest that activation of Nrf2 may be a novel strategy to prevent or ameliorate toxin-induced liver injury and fibrosis. PMID- 18679375 TI - Hematopoietic SCT from partially HLA-mismatched (HLA-haploidentical) related donors. AB - Hematopoietic SCT from a partially HLA-mismatched (HLA-haploidentical) first degree relative offers the benefits of rapid and near universal donor availability but also the risks that result from traversing the HLA barrier; namely, graft failure, severe GVHD and prolonged immunodeficiency. Improvements over the last 10 years in conditioning regimens, graft engineering and pharmacological immunoprophylaxis of GVHD have substantially reduced the morbidity and mortality of HLA-haploidentical SCT. Highly immunosuppressive but nonmyeloablative conditioning extends the availability of HLA-haploidentical SCT to elderly hematologic malignancy patients lacking HLA-matched donors and permits recovery of autologous hematopoiesis in the event of graft failure. Current regimens for HLA-haploidentical SCT are associated with a 2-year non-relapse mortality of 20+/-5%, relapse of 35+/-15% and overall survival of 50+/-20%. Major developmental areas include harnessing natural killer cell alloreactivity to reduce the risk of disease relapse and improving immune reconstitution by delayed infusions of lymphocytes selectively depleted of alloreactive cells. Hematologic malignancy patients who lack suitably matched related or unrelated donors can now be treated with HLA-haploidentical related donor or unrelated umbilical cord blood SCT. Future clinical trials will assess the relative risks and benefits of these two graft sources. PMID- 18679378 TI - Generation of endogenous hydrogen sulfide by cystathionine gamma-lyase limits renal ischemia/reperfusion injury and dysfunction. AB - The generation of endogenous hydrogen sulfide may either limit or contribute to the degree of tissue injury caused by ischemia/reperfusion. A total of 74 male Wistar rats were used to investigate the effects of endogenous and exogenous hydrogen sulfide in renal ischemia/reperfusion. Administration of the irreversible cystathionine gamma-lyase (CSE) inhibitor, dL-propargylglycine, prevented the recovery of renal function after 45 min ischemia and 72 h reperfusion. The hydrogen sulfide donor sodium hydrosulfide attenuated the (renal, tubular, and glomerular) dysfunction and injury caused by 45 min ischemia and 6 h reperfusion. Western blot analysis of kidneys taken at 30 min reperfusion showed that sodium hydrosulfide significantly attenuated phosphorylation of mitogen-activated protein kinases (p-38, c-JUN N-terminal protein kinase 1/2, and extracellular signal-regulated kinase 1/2) and activation of nuclear factor kappaB. At 6 h reperfusion, sodium hydrosulfide significantly attenuated the histological score for acute tubular necrosis, the activation of caspase-3 and Bid, the decline in the expression of anti-apoptotic Bcl-2, and the expression of nuclear factor-kappaB-dependent proteins (inducible nitric oxide synthase, cyclo oxygenase-2, and intercellular adhesion molecule-1). These findings suggest that (1) the synthesis of endogenous hydrogen sulfide by CSE is essential to protect the kidney against ischemia/reperfusion injury and dysfunction and aids in the recovery of renal function following ischemia/reperfusion, (2) hydrogen sulfide generated by sodium hydrosulfide reduces ischemia/reperfusion injury and dysfunction, and morphological changes of the kidney, and (3) the observed protective effects of hydrogen sulfide are due to both anti-apoptotic and anti inflammatory effects. PMID- 18679379 TI - Liver injury caused by antibodies against dengue virus nonstructural protein 1 in a murine model. AB - Clinical manifestations of severe dengue diseases include thrombocytopenia, vascular leakage, and liver damage. Evidence shows that hepatic injury is involved in the pathogenesis of dengue infection; however, the mechanisms are not fully resolved. Our previous in vitro studies suggested a mechanism of molecular mimicry in which antibodies directed against dengue virus (DV) nonstructural protein 1 (NS1) cross-reacted with endothelial cells and caused inflammatory activation and apoptosis. In this study, the pathogenic effects of anti-DV NS1 antibodies were further examined in a murine model. We found, in liver sections, that anti-DV NS1 antibodies bound to naive mouse vessel endothelium and the binding activity was inhibited by preabsorption of antibodies with DV NS1. Active immunization with DV NS1 resulted in antibody deposition to liver vessel endothelium, and also apoptotic cell death of liver endothelium. Liver tissue damage was observed in DV NS1-immunized mice by histological examination. The serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were increased in mice either actively immunized with DV NS1 protein or passively immunized with antibodies obtained from DV NS1-immunized mice. Furthermore, histological examination revealed mononuclear phagocyte infiltration and cell apoptosis in mice passively immunized with antibodies obtained from mice immunized with DV NS1. Increased AST and ALT levels were observed in mice passively immunized with purified immunoglobulin G (IgG) from dengue patients compared with normal control human IgG-immunized mice. The increased AST and ALT levels were inhibited when dengue patient serum IgG was preabsorbed with DV NS1. In conclusion, active immunization with DV NS1 protein causes immune-mediated liver injury in mice. Passive immunization provides additional evidence that anti DV NS1 antibodies may play a role in liver damage, which is a pathologic manifestation in dengue virus disease. PMID- 18679377 TI - Placental expression of ceruloplasmin in pregnancies complicated by severe preeclampsia. AB - There is consensus that ischemia/reperfusion injury associated with preeclampsia (PE) promotes both placental damage and the release of factors leading to maternal endothelium dysfunction, a hallmark of this potentially life-threatening syndrome. These factors include plasminogen activator inhibitor-1 (PAI-1) and soluble fms-like tyrosine kinase-1 (sFlt-1). The goal of this study was to further characterize placental factors involved in the pathophysiology of PE. Thus, DNA microarray gene profiling was utilized to identify mRNA differentially regulated in placentas from women with severe PE compared to both preterm (PC) and term control (TC) groups. Microarray studies detected an upregulation of mRNA for ceruloplasmin, a copper-containing iron transport protein with antioxidant ferroxidase properties, in PE compared to PC and TC placentas, respectively. Quantitative real-time PCR confirmed these results by demonstrating significant increases in ceruloplasmin mRNA in PE vs PC and TC placentas. Supporting previous reports, the expression of sFlt-1 and PAI-1 were also upregulated in PE placentas. Immunohistochemistry localized ceruloplasmin to the intervillous space in PE and PC placentas, whereas stronger syncytial staining was noted in PE. Western blotting confirmed a significant increase in ceruloplasmin levels in placental tissue in PE compared to PC groups. PCR identified the presence of mRNA for ceruloplasmin in primary cultures of syncytiotrophoblasts, but not villous derived fibroblasts, suggesting that syncytium is the site of ceruloplasmin synthesis in placenta. Hypoxic treatment (1% O(2)) of syncytiotrophoblasts enhanced levels of ceruloplasmin mRNA approximately 25-fold, a significantly greater upregulation than that noted for PAI-1 and sFlt-1, suggesting that enhanced ceruloplasmin expression is a sensitive marker of syncytial hypoxia. We suggest that syncytial ceruloplasmin and its associated ferroxidase activity, induced by the hypoxia accompanying severe PE, is important in an endogenous cellular program to mitigate the damaging effects of subsequent reperfusion injury at this site. PMID- 18679380 TI - Which patients with venous thromboembolism are at risk for fatal pulmonary embolism? AB - In a registry of 15,520 patients treated for symptomatic deep vein thrombosis or pulmonary embolism (PE), the 90-day mortality was 8.65% and death was attributed to PE in 1.68% of patients (19.4% of all deaths). Multivariate analysis defined five simple predictors of death from PE during the first 3 months after presentation. The odds ratio for fatal PE was raised to 5.4 by initially nonmassive symptomatic PE (compared with deep vein thrombosis and no symptoms of PE), to 17.5 by initially massive PE (systolic blood pressure below 90 mmHg), 4.9 by immobility as a result of neurological disease, 2.5 by age over 75 years, and 2.0 by the presence of cancer. Of all the deaths from PE, 75% occurred within 12 days of presentation and 50% occurred within 5 days. These results reinforce previous observations that also linked symptomatic PE, massive PE, old age, and cancer to a raised likelihood of death from PE despite appropriate therapy. PMID- 18679381 TI - From randomized trials to registry studies: translating data into clinical information. AB - All clinicians face the challenge of practicing evidence-based medicine and are confronted with data from a variety of studies, ranging from prospective randomized and registry studies to retrospective analyses. Unfortunately, the data frequently provide conflicting recommendations. How then should one interpret the information so that study findings can be applied directly in patient care? To evaluate the relevance of the abundance of studies published and how they apply to an individual patient, physicians must understand subtle nuances of study design and their effect on the interpretation of the results. In this Review, we examine the strengths and weaknesses of different study designs with the aim of providing the reader with a greater understanding how best to apply study results in the clinical setting. PMID- 18679382 TI - Bioabsorbable drug-eluting stents: the future of coronary angioplasty? AB - In the ABSORB trial, the feasibility of using a fully bioabsorbable everolimus eluting stent (BVS; Abbott Vascular, Santa Clara, CA) in patients with ischemia and a single de novo coronary lesion was investigated. Some important concerns about the efficacy of this device were raised by this study. The in-stent late loss with the BVS (0.44 mm) was comparable to that seen with some drug-eluting stents currently on the market, and the late lumen loss was primarily due to reduction in stent area. Technical improvements aimed at definitively preventing late vessel recoil and negative remodeling, as well as clarification of the behavior of these devices with longer follow-up, are needed before fully bioabsorbable drug-eluting stents can be considered a therapeutic option for patients with coronary artery disease. PMID- 18679383 TI - Management of asymptomatic severe aortic stenosis: is aortic valve replacement indicated? PMID- 18679384 TI - Is activation of coronary venous cells the key to cardiac regeneration? PMID- 18679385 TI - Poor outcomes in diabetes care: is medication nonadherence or lack of treatment intensification to blame? PMID- 18679386 TI - What is the best strategy to combat low bone mineral density in functional hypothalamic amenorrhea? PMID- 18679387 TI - Does glycemic status at admission predict mortality in noncritically ill hospitalized patients? PMID- 18679388 TI - Esophageal cancer: epidemiology, pathogenesis and prevention. AB - Esophageal cancer is highly aggressive and is a common cancer both worldwide and in the US. In the past two decades, the incidence and mortality of esophageal cancer in the US have both increased, where as the incidence and mortality of other cancers have decreased. Although esophageal squamous cell carcinoma and esophageal adenocarcinoma differ in their histology and epidemiologic distribution, some of their risk factors (e.g. dietary deficiencies and tobacco) and underlying mechanisms of carcinogenesis are the same. Intensive research into risk factors combined with the ability to identify precursor lesions (e.g.squamous dysplasia in esophageal squamous cell carcinoma and Barrett's esophagus in esophageal adenocarcinoma) has paved the way for studies of chemoprevention for esophageal cancer, some of which have shown promising results. PMID- 18679389 TI - Efficient diagnosis of suspected functional bowel disorders. AB - Functional bowel disorders (FBDs) are common disorders that are characterized by various combinations of abdominal pain and/or discomfort, bloating and changes in bowel habits. At present, diagnosing FBDs often incurs considerable health-care costs, partly because unnecessary investigations are performed. Patients are currently diagnosed as having an FBD on the basis of a combination of typical symptoms, normal physical examination and the absence of alarm features indicative of an organic gastrointestinal disease. Basic laboratory investigations, such as a complete blood count, measurement of the erythrocyte sedimentation rate and serological tests for celiac disease, are useful in the initial evaluation. No further investigations are needed for most patients who have typical symptoms and no alarm symptoms. The most important alarm symptoms include signs of gastrointestinal bleeding, symptom onset above 50 years of age, a family history of colorectal cancer, documented weight loss and nocturnal symptoms. The presence of alarm symptoms obviously does not exclude an FBD, but further investigation is needed before confirmation of the diagnosis. For patients with predominant and severe diarrhea, a more thorough diagnostic work-up should normally be considered, including colonoscopy with colonic biopsies and a test for bile-acid malabsorption. PMID- 18679390 TI - Immunosuppression minimization protocols: how should they be monitored? AB - This Practice Point commentary discusses the experience of Shapiro et al. with renal allograft recipients at a single center who underwent significant minimization of immunosuppressive treatment with alemtuzumab induction and tacrolimus monotherapy (weaned to three times weekly by 1 year after transplantation). Donor-specific antibodies (DSAs) against human leukocyte antigen were monitored after transplantation in some patients. Although 44% of patients did not develop acute rejection or DSAs and had excellent 2-year graft survival (96%), spaced weaning was not attempted in 20% of patients, generally those with early rejection and poor allograft function. During tapering, 20% of patients developed acute rejection and 15% developed DSAs. Two-year graft survival was 63% in patients for whom weaning was not attempted and 78% in patients who experienced acute rejection. Alemtuzumab preconditioning with tacrolimus monotherapy and spaced weaning increased the risk of acute rejection and development of DSAs. Patients should be carefully selected for this protocol and monitored closely for DSAs. Results of a long-term controlled trial that includes protocol biopsies are required to reach more-definitive conclusions. PMID- 18679391 TI - Implications of ethnicity for the treatment of hypertensive kidney disease, with an emphasis on African Americans. AB - The recognition of chronic kidney disease (CKD) as an important public health issue has fostered an increasing number of strategies to increase CKD awareness and to reduce both the prevalence and the complications of CKD. Despite these advances, end-stage renal disease (ESRD) and cardiovascular events remain the major complications of CKD. Although the ESRD epidemic is attributed in greater part to the increasing rate of diabetes, hypertension remains the second most common reported cause of ESRD and is present in approximately 90% of cases of diabetes-related ESRD. The disproportionately high prevalence of hypertension in ethnic minorities, as well as the difficulty of achieving adequate blood-pressure control in these populations, contributes substantially to the high rate of CKD progression and complications in these groups. Although the role of hypertension as a primary cause of CKD is debated, hypertension is commonly recognized as the most important CKD progression factor. Important differences have been reported in the degree and likelihood of blood-pressure response to antihypertensive medications between ethnic groups, but ethnicity seems to be less important as a determinant of clinical outcomes. In this Review we examine key ethnic variations in hypertensive CKD in terms of pathophysiology, response to antihypertensive therapy, clinical outcomes, and evidence-based recommendations for blood-pressure control, with an emphasis on African Americans. PMID- 18679392 TI - A case of cocaine-induced basilar artery thrombosis. AB - BACKGROUND: A 40-year-old man presented to an emergency department with slurred speech, diplopia and agitation several hours after cocaine use. His level of consciousness rapidly dropped in the hours following presentation. INVESTIGATIONS: Physical examination, CT scan of the head, cerebral angiography. DIAGNOSIS: Cocaine-induced basilar artery thrombosis. MANAGEMENT: Intra-arterial thombolysis, mechanical clot aspiration, intravenous abciximab, neurointensive care support, rehabilitation at a specialist stroke unit. PMID- 18679393 TI - The rationale for sentinel-node biopsy in primary melanoma. PMID- 18679394 TI - Treatment of Hodgkin lymphoma: the past, present, and future. AB - Significant advances in the biology and treatment of Hodgkin lymphoma (HL) have been accomplished over the past decades. In a landmark study, DeVita and colleagues showed that half of patients with advanced-stage HL experienced long term disease-free survival following treatment with a four-drug chemotherapy regimen. Subsequent reports and randomized clinical trials conducted over the past 40 years have defined prognostic categories and refined the treatment options for patients with early-stage and advanced-stage HL. New treatment concepts and regimens have continued to increase the cure rate of HL, while other analyses have documented the acute and long-term morbid and potentially fatal side effects of HL therapy. Increased knowledge of HL biology has been gained, in particular, much has been learnt about the genetic and phenotypic characteristics of malignant cells and the varied oncogenic signaling pathways involved in HL. Continued translational research is needed to improve the long-term survival and to lessen the toxicities associated with therapy. Furthermore, continued clinical trial involvement by oncologists and patients is imperative to further advance the field of HL. PMID- 18679395 TI - Does administration of transdermal dihydrotestosterone gel before hypospadias repair improve postoperative outcomes? AB - In an effort to improve the outcome of hypospadias repair, Kaya et al. performed a prospective, randomized study of transdermal dihydrotestosterone (DHT) gel as a preoperative adjunct to surgery. They found that patients who received DHT gel had fewer complications and better cosmesis than patients who underwent surgery alone. The authors attribute the differences between the two groups to an increased blood flow produced by the gel. The series of patients was small, and a larger sample size might have produced different results. Some of the outcomes measured were subjective, and the methods used to assess these outcomes were not clear. If a larger study confirms the results obtained by Kaya and colleagues, transdermal DHT gel might prove to be a valuable adjunct to hypospadias surgery. PMID- 18679396 TI - A case of acute renal failure caused by bilateral, multifocal malacoplakia lesions of the bladder and ureters. AB - BACKGROUND: A 51-year-old woman with a history of hypertension and a 5-year history of recurrent perimenopausal Escherichia coli urinary tract infections presented with fatigue. She had a history of occasional urinary frequency, but denied gross hematuria, flank pain, abdominal pain, immunodeficiency or immunosuppression. INVESTIGATIONS: Basic metabolic profile, ultrasonography, cystoscopy, retrograde pyelography, pathologic evaluation of resected lesions from the bladder, ureteroscopy, pathologic evaluation of resected lesions from the ureters, and MRI. DIAGNOSIS: Acute renal failure caused by bilateral, multifocal malacoplakia lesions of the bladder and ureters. MANAGEMENT: Endoscopic resection and laser ablation, ureteral stent placement, and medical treatment with fluoroquinolones, vitamin C and bethanechol. PMID- 18679397 TI - Do topical NSAIDs work? PMID- 18679398 TI - Somatic pharmacogenomics in cancer. AB - Many of the initial examples of the clinical utility of pharmacogenetics were elucidated in the field of oncology. Those examples were largely based on the existence of germline genetic variation that influences the metabolism of cytotoxic drugs. However, with the development of kinase inhibitors, drugs designed to preferentially target altered proteins driving oncogenesis, pharmacogenetics in cancer has shifted to understanding the somatic differences that determine response to these targeted agents. It is becoming increasingly clear that understanding the molecular genetics of cancer will lead to the further development of targeted therapeutics. Therefore, it is imperative that pharmacogenomics researchers understand the motivations and challenges of developing targeted therapies to treat cancer as a paradigm for personalized medicine. However, much of the discussion in the pharmacogenomics community in cancer is still largely focused on the germline variants as predictors of drug toxicity. In light of that fact, this review presents a detailed discussion of the development of commonly used targeted therapies for the treatment of hematological and solid tumors, the somatic mutations that determine response to those therapies, and the mechanisms of drug resistance. PMID- 18679399 TI - Low-echoic lesions underneath the skin in subjects with spinal-cord injury. AB - STUDY DESIGN: Criterion standard and survey cases. OBJECTIVES: To assess the utility of ultrasonography for detecting deep tissue injury or incipient pressure ulcers and to determine the patterns of development of pressure ulcers in subjects with chronic spinal-cord injury (SCI). SETTING: Ambulatory setting at public hospital. METHODS: The subjects were 43 men with SCI between C5 and L1 (age: 42.6+/-11.6 years, mean+/-s.d.). A total of 129 areas (sacral region and bilateral ischial regions in each subject) were examined by inspection, palpation and ultrasonography. RESULTS: Of the 129 areas, 112 were normal by inspection, palpation and ultrasound imaging. Nine areas were abnormal on ultrasonography alone and six were lesion positive by palpation and ultrasonography. Only two areas were abnormal by all three methods. Ultrasonography always detected a heterogeneous pattern and low-echoic areas directly adjacent to the bone. CONCLUSIONS: Our results indicated that low-echoic lesions, signaling deep tissue injuries or early pressure ulcers, originated in areas near the bone and extended toward the epidermis. The results suggest that ultrasonography is a useful tool for the early detection of deep tissue injuries or pressure ulcers. PMID- 18679400 TI - Profile of spinal injuries in Lagos, Nigeria. AB - STUDY DESIGN: Retrospective study. OBJECTIVE: To describe the pattern and outcome of spinal cord injuries in Lagos, Nigeria. SETTING: Nigeria, Lagos. METHODS: This retrospective review of cases was conducted at Lagos University Teaching Hospital (LUTH), Lagos, between January 1992 and December 2006. RESULTS: A total of 468 cases of spinal cord injury over the 15-year period were studied. Three hundred and twelve of them (66.2%) were aged 40 years and below, with the peak age incidence as 21-30 years. There were 328 males (70.1%) and 140 females (29.9%) showing a male to female ratio of 2.34:1. Road traffic accident accounted for 362 (77.4%) patients. At presentation, 430 (91.9%) of the injuries were of ASIA (American Spinal Injury Association) class A. Lumbar spine was affected in 278 (59.4%) of patients, whereas cervical spine accounted for 142 (30.3%). The most frequently seen complication was bedsore, seen in 163 (59.9%) of 272 patients that had complications. On discharge, 230 patients (49.1%) were in ASIA class A and 34 (7.5%) in ASIA E. Eighty-two patients (17.5%) died. CONCLUSION: Spinal injuries in Lagos occurred mostly in young adult males affecting mainly the lumbar spine. Traffic-related incidents were the leading cause and bedsore the most common complication. PMID- 18679401 TI - Cost-effectiveness of transanal irrigation versus conservative bowel management for spinal cord injury patients. AB - STUDY DESIGN: Cost-effectiveness analysis following international guidelines and taking the societal viewpoint. OBJECTIVES: To estimate the cost-effectiveness of transanal irrigation using a self-administered irrigation system when compared with conservative bowel management. SETTING: A randomized clinical trial was conducted at five spinal centres situated in Denmark, Germany, Italy, United Kingdom and Sweden. Estimates of resources and unit costs were made for the German health care system. METHODS: Efficacy outcomes were drawn from a randomized controlled trial conducted in 2003-2005. Adult spinal cord-injured patients with neurogenic bowel dysfunction were randomized to 10 weeks with either transanal irrigation using Peristeen Anal Irrigation or to conservative bowel management. Costs were calculated based on results from the clinical trial and on 24 interviews conducted in Germany. Unit costs were obtained from the Federal Statistical Office Germany and product list prices. RESULTS: When comparing outcome measures at termination, transanal irrigation significantly reduced symptoms of neurogenic bowel dysfunction. Product-related costs were higher for transanal irrigation using the self-administered system; however, costs for a carer to help with bowel management and changes/washing due to leakage were lower. For transanal irrigation, costs associated with urinary tract infections and patient time spent were reduced. Thus, the total cost to society is lower when patients use transanal irrigation. The results were shown to be robust in the sensitivity analysis. CONCLUSION: Transanal irrigation using a self administered system reduces symptoms of neurogenic bowel dysfunction and results in a lower total cost to society than conservative bowel management. SPONSORSHIP: The study was supported by Coloplast A/S. PMID- 18679402 TI - Unmyelinated fibers in human spinal ventral roots: C4 to S2. AB - STUDY DESIGN: Histological examination of human spinal ventral roots. OBJECTIVES: To determine the proportion of unmyelinated fibers in human ventral roots from the 4th cervical (C4) to 2nd sacral (S2) segment, and to evaluate differences in the proportions of unmyelinated fibers between the cervical, thoracic, lumbar and sacral segments, and between autonomic and other segments. SETTING: University Teaching Hospital, Busan, Korea. METHODS: Eight embalmed adult human cadavers (six males and two females; mean age 56.3 years) were collected. The ventral root samples were obtained by transverse cuts of the ventral roots within 1 cm proximal to the medial portion of the dorsal root ganglion from the C4 to S2 segment. The number of unmyelinated and myelinated fibers was counted in four fields, and the mean number of unmyelinated fibers was calculated. The percentage of unmyelinated fibers was calculated from the ratio of unmyelinated fibers to total fibers (myelinated fibers+unmyelinated fibers). RESULTS: The mean percentages of unmyelinated axons in cervical (C4-C8), thoracic (T1-T12), lumbar (L1-L5) and sacral (S1-S2) ventral roots were 16.3, 21.4, 17.8 and 20.7%, respectively. The percentage of unmyelinated fibers in thoracic ventral roots was higher than that for other segments (P<0.001). There was no significant difference in proportions of unmyelinated fibers between the sympathetic segments (T11-L2), parasympathetic segments (S2) and the other segments (C4-T10 and L3-S1) (P=0.1784). CONCLUSIONS: Approximately 20% of human spinal ventral root fibers were unmyelinated. The proportion of unmyelinated fibers was highest in the thoracic segments. PMID- 18679403 TI - Contrasting alteration patterns of different cartilage plates in knee articular cartilage after spinal cord injury in rats. AB - STUDY DESIGN: Experimental, controlled trial, animal study. OBJECTIVE: To assess morphologic changes in different cartilage plates after spinal cord injury and identify the localization of these alterations. SETTING: Saitama, Japan. METHODS: A total of 16 Wistar rats were used. Eight rats underwent a spinal cord injury and eight rats had no intervention as control. The cartilage alterations of the knee joint were evaluated with radiography and histomorphometric analysis. To quantify cartilage alterations, we selected the histologic characteristics: thickness of the articular cartilage, number of chondrocytes, matrix staining to toluidine blue as a reflection of proteoglycan content and surface irregularity. RESULTS: No differences in knee joints were found between the groups by radiography. In the medial knee joint, cartilage thickness of spinal-cord-injured knees increased at the anterior femoral region and decreased at the tibial and posterior femoral regions; however, in the lateral knee, that of spinal cord injuries did not change compared with control knees. Spinal cord injuries decreased the number of chondrocytes, especially at the anterior femoral regions. Matrix staining increased partially at the tibial regions. Surface irregularity of spinal-cord-injured knees was comparable to that of control knees in all cartilage plates. CONCLUSION: The present findings exhibit characteristics of the cartilage after spinal cord injury. These alterations were different in nature between the medial and lateral regions. Future studies should assess separately different cartilage plates, to overestimate these severities when the changes at the medial knee were examined and to underestimate when the changes at the lateral knee were examined. PMID- 18679404 TI - Rectum perforation during transanal irrigation: a case story. AB - STUDY DESIGN: Case report. OBJECTIVES: Report a case of rectum perforation during transanal irrigation (TAI). SETTING: Clinic for Spinal Cord Injuries, and Departments of Gastroenterological Surgery and Radiology. CASE REPORT: A 54-year old woman with spinal cord lesion for 35 years emptied for years her bowel using oral laxative. This became more difficult and took more than 2 h three times a week with reflex stimulation after a chlysma. She wanted to try TAI, and went through the procedure with a nurse one time. The next time she performed the TAI by herself without difficulty. Two hours later she started shivering with a temperature at 38.3 degrees C with normal blood pressure (BP). At rectal exploration, a spoonful fresh blood was found. After another 2 h, she became septic and was transferred to a gastroenterological surgical department. An abdominal X-ray gave no suspicion of free air in the abdomen. Sigmoideoscopy showed 3-5 cm oral to the dentate line a 1 x 1 cm transmural circular lesion. A colon X-ray with water contrast showed a perforation of approximately 2 x 0.5 cm. CONCLUSION: Even in experienced individuals who are proper trained, TAI can cause rectal perforation, which always have to be born in mind. PMID- 18679405 TI - Motor scores on the functional independence measure after pediatric spinal cord injury. AB - STUDY DESIGN: Retrospective descriptive analysis. OBJECTIVES: The purpose of this study was to report the functional ability of children with spinal cord injury (SCI) as recorded on motor items of the functional independence measure (FIM) and to examine the factors associated with FIM motor admission and post-discharge gain scores. METHODS: Scores on FIM motor items were analyzed from 941 children (age range: 0-21 years; mean: 13 years 4 months; s.d.: 4 years 8 months) admitted in acute-to-chronic time periods post-SCI to Shriners Hospitals for Children (USA). FIM motor scores at admission and gains at discharge were examined along with neurological level, completeness of injury, age, etiology of injury, and length of time between injury and admission and admission and discharge. RESULTS: The FIM motor scores at admission were negatively correlated with age, neurological level and completeness of injury. Gain in FIM motor scores was significant across neurological levels, and was associated with lower admission FIM motor scores, lower neurological level, incomplete injury, traumatic injury and less time between injury and admission. CONCLUSIONS: The motor function of children after pediatric SCI depends on neurological level and completeness of injury, among other factors. FIM motor scores can improve with intervention even several years after the injury. PMID- 18679406 TI - The implications of gene-environment interactions in depression: will cause inform cure? AB - In a number of human diseases, including depression, interactions between genetic and environmental factors have been identified in the absence of direct genotype disorder associations. The lack of genes with major direct pathogenic effect suggests that genotype-specific vulnerabilities are balanced by adaptive advantages and implies aetiological heterogeneity. A model of depression is proposed that incorporates the interacting genetic and environmental factors over the life course and provides an explanatory framework for the heterogeneous aetiology of depression. Early environmental influences act on the genome to shape the adaptability to environmental changes in later life. The possibility is explored that genotype- and epigenotype-related traits can be harnessed to develop personalized therapeutic interventions. As diagnosis of depression alone is a weak predictor of response to specific treatments, aetiological subtypes can be used to inform the choice between treatments. As a specific application of this notion, a hypothesis is proposed regarding relative responsiveness of aetiological subtypes of depression to psychological treatment and antidepressant medication. Other testable predictions are likely to emerge from the general framework of interacting genetic, epigenetic and environmental mechanisms in depression. PMID- 18679407 TI - Dendritic cells and C-type lectin receptors: coupling innate to adaptive immune responses. AB - Dendritic cells (DCs) have an important function in the initiation and differentiation of immune responses, linking innate information to tailored adaptive responses. Depending on the pathogen invading the body, specific immune responses are built up that are crucial for eliminating the pathogen from the host. Host recognition of invading microorganisms relies on evolutionarily ancient, germline-encoded pattern recognition receptors (PRRs) that are highly expressed on the cell surface of DCs, of which the Toll-like receptors (TLRs) are well characterized and recognize bacterial or viral components. Moreover, they bind a variety of self-proteins released from damaged tissues including several heat-shock proteins. The membrane-associated C-type lectin receptors (CLRs) recognize glycan structures expressed by host cells of the immune system or on specific tissues, which upon recognition allow cellular interactions between DCs and other immune or tissue cells. In addition, CLRs can function as PRRs. In contrast to TLRs, CLRs recognize carbohydrate structures present on the pathogens. Modification of glycan structures on pathogens to mimic host glycans can thereby alter CLR interactions that subsequently modifies DC-induced polarization. In this review, we will discuss in detail how specific glycosylation of antigens can dictate both the innate and adaptive interactions that are mediated by CLRs on DCs and how this balances immune activation and inhibition of DC function. PMID- 18679408 TI - The increase of fatty acid-binding protein aP2 in overweight and obese children: interactions with dietary fat and impact on measures of subclinical inflammation. AB - BACKGROUND: In adults, circulating aP2 may link obesity, inflammation and the metabolic syndrome, but there are few data in children. Experimental models support that dietary factors, particularly dietary fat, may be major determinants of phenotype. OBJECTIVE: The aim of this study was to investigate, in normal, overweight and obese children, the relationships among aP2, the metabolic syndrome, inflammation and diet. DESIGN: This was a cross-sectional study conducted in Northern Switzerland. SUBJECTS: Subjects for this study were 6- to 14-year-old, prepubertal and early pubertal, normal weight, overweight and obese children (n=124). MAIN OUTCOME MEASURES: Body mass index (BMI), body fat percent, waist-to-hip ratio, blood pressure, circulating aP2, fasting insulin, C-reactive protein (CRP), plasma lipids and dietary intakes of macro- and micronutrients were determined. RESULTS: Circulating aP2 markedly increased with increasing central and total adiposity, and predicted measures of insulin resistance. Independent of BMI standard deviation scores and puberty, aP2 correlated with intake of the antioxidant vitamins A, C and E as well as circulating concentrations of CRP, leptin and low-density lipoprotein cholesterol. Children with lower aP2 concentrations consuming high-fat diets did not show an increase in fasting insulin or CRP, whereas those with higher aP2 concentrations showed marked increases in these measures with high intakes of fat or saturated fat. CONCLUSIONS: Increased central and overall adiposity in children are associated with higher circulating aP2 concentrations. In children with high dietary intakes of total fat and saturated fat, but not those with low intakes, higher aP2 concentrations are associated with measures of insulin resistance and inflammation. PMID- 18679411 TI - Vitamin B6 status improves in overweight/obese women following a hypocaloric diet rich in breakfast cereals, and may help in maintaining fat-free mass. AB - OBJECTIVE: To analyze the changes in vitamin B6 status in women following slightly hypocaloric diets based on the relative increase consumption of foods whose intakes are below those recommended, and to study how these changes influence the proportion of fat-free mass. DESIGN: Intervention study of two slightly hypocaloric diets: diet V (increased consumption of vegetables), or diet C (increased consumption of cereals, especially breakfast cereals). SUBJECTS: A total of 49 women with a body mass index (BMI) of 25-35 kg/m2. MEASUREMENTS: Dietetic, anthropometric and biochemical data were collected at the start of the study and at 2 and 6 weeks. RESULTS: Both the C and V subjects showed a reduction in their energy intake, body weight, BMI and fat mass. Pyridoxine intake increased in both groups and plasma pyridoxal phosphate (PLP) levels increased only with diet C. An association was found between the increase in plasma PLP at 6 weeks and the increase in pyridoxine intake (r=0.451; P<0.01). Multiple regression analysis showed a positive association between the increase in PLP at the end of the study and the increases in the pyridoxine intake, B6 density or B6/protein ratio. At the end of the study, and only in those women whose PLP levels were increased, the higher the increase in PLP level, the higher the increase in fat-free mass percentage (r=0.4426, P<0.05). CONCLUSIONS: Interventions aimed at weight control should also try to maintain or improve nutritional status. A diet rich in cereals (especially fortified breakfast cereals) appears to be useful in improving vitamin B6 status. Such an improvement could help maintain fat-free mass during periods of weight loss. PMID- 18679412 TI - Egg breakfast enhances weight loss. AB - OBJECTIVE: To test the hypotheses that an egg breakfast, in contrast to a bagel breakfast matched for energy density and total energy, would enhance weight loss in overweight and obese participants while on a reduced-calorie weight loss diet. SUBJECTS: Men and women (n=152), age 25-60 years, body mass index (BMI) >or=25 and