Mega-dose vitamin C attenuated lung inflammation in mouse asthma model.
Asthma is a Th2-dependent disease mediated by IgE and Th2 cytokines, and asthmatic patients suffer from oxidative stresses from abnormal airway inflammation. Vitamin C is a micro-nutrient functioning as an antioxidant. When administered at a mega-dose, vitamin C has been reported to shift immune responses toward Th1. Thus, we tried to determine whether vitamin C exerted beneficial effects in asthma animal model. Asthma was induced in mice by sensitizing and challenging with ovalbumin. At the time of challenge, 3~5 mg of vitamin C was administered and the effects were evaluated. Vitamin C did not modulate Th1/Th2 balance in asthma model. However, it decreased airway hyperreactivity to methacholine, decreased inflammatory cell numbers in brochoalveolar lavage fluid, and moderate reduction of perivascular and peribronchiolar inflammatory cell infiltration. These results suggest that vitamin C administered at the time of antigen challenge exerted anti-inflammatory effects. Further studies based on chronic asthma model are needed to evaluate a long-term effect of vitamin C in asthma. In conclusion, even though vitamin C did not show any Th1/Th2 shifting effects in this experiment, it still exerted moderate anti-inflammatory effects. Considering other beneficial effects and inexpensiveness of vitamin C, mega-dose usage of vitamin C could be a potential supplementary modality for the management of asthma.


Anatomical and visual outcomes of three different scleral buckling techniques.
To compare the anatomical and visual outcomes of three different scleral buckling techniques and to explore the effect of cryotherapy and subretinal fluid drainage (SRFD) on outcomes of surgery. This retrospective study was performed on 111 eyes of 109 patients undergoing scleral buckling for rhegmatogenous retinal detachments (RRDs) by a single surgeon. Pre-, intra- and postoperative data were retrieved from hospital records. Buckles were radial in 27 (24.3%), circumferential (segmental) in 16 (14.4%) and encircling in 68 (61.3%) eyes. Anatomical and visual results were comparable with all three buckling techniques. Application of cryotherapy, the spot number, and SRFD did not affect anatomical and visual results. The only preoperative factor associated with poorer anatomical results was the presence of multiple retinal breaks (P=0.006). The following preoperative factors affected visual outcomes on univariate analysis: extent of retinal detachment (r=0.417, P=0.011) and relative afferent pupillary defect (r=0.423, P=0.02). Preoperative macular status (attached vs detached) also had a significant effect on visual outcomes (P<0.001). Based on multivariate analysis however, only preoperative macular status was significantly correlated with visual results (P=0.022). Silicone sponges placed for non-encircling surgery were removed due to ocular dysmotility in 4 (3.6%) eyes, cosmetic reasons in 3 (2.7%) cases and extrusion in 2 (1.8%) eyes. One encircling tire was also removed due to extrusion. Surgical technique and performing cryotherapy or SRFD do not seem to influence the anatomical and visual outcomes of scleral buckling. Postoperative complications seem to be more prevalent with non-encircling techniques.


Regulation of zebrafish skeletogenesis by ext2/dackel and papst1/pinscher.
Mutations in human Exostosin genes (EXTs) confer a disease called Hereditary Multiple Exostoses (HME) that affects 1 in 50,000 among the general population. Patients with HME have a short stature and develop osteochondromas during childhood. Here we show that two zebrafish mutants, dackel (dak) and pinscher (pic), have cartilage defects that strongly resemble those seen in HME patients. We have previously determined that dak encodes zebrafish Ext2. Positional cloning of pic reveals that it encodes a sulphate transporter required for sulphation of glycans (Papst1). We show that although both dak and pic are required during cartilage morphogenesis, they are dispensable for chondrocyte and perichondral cell differentiation. They are also required for hypertrophic chondrocyte differentiation and osteoblast differentiation. Transplantation analysis indicates that dak(-/-) cells are usually rescued by neighbouring wild-type chondrocytes. In contrast, pic(-/-) chondrocytes always act autonomously and can disrupt the morphology of neighbouring wild-type cells. These findings lead to the development of a new model to explain the aetiology of HME.


Complementary medicine for children and young people who have attention deficit hyperactivity disorder.
Despite effectiveness of medication in treating children and young people who have attention deficit hyperactivity disorder (ADHD), concerns about the effects of medication on children's developing brains, adverse side-effects, possibility of long-term use, and compliance issues have all contributed to the continuing search for alternative therapies. This article reviews the latest scientific evidence of the effectiveness and safety of these treatments in ADHD. Although there is evidence from a large randomized controlled study that neurofeedback has positive effects on reducing children's symptoms of ADHD, most recent randomized controlled trials have generally yielded negative results. Some positive results exist from a pilot study of repetitive transcranial magnetic stimulation. However, the sample size was far too small to enable any conclusions to be drawn about the evidence. Findings from the recent randomized controlled trials of supplements of essential fatty acids in children who have ADHD clearly demonstrated lack of superiority compared with placebo. Notwithstanding efforts made to increase the scientific rigor of previous studies, more recent studies have generally been unsuccessful in demonstrating adequate treatment effects of complementary medicine on children who have ADHD. Currently, there is no proof that complementary medicine provides a better alternative for children who have ADHD than treatments that are currently available within multimodal therapy.


A dynamic model of transmission and elimination of peste des petits ruminants in Ethiopia.
Peste des petits ruminants (PPR), a devastating viral disease of sheep and goats, has been targeted by the global community for eradication within the next 15 years. Although an efficacious attenuated live vaccine is available, the lack of knowledge about the transmission potential of PPR virus (PPRV) may compromise eradication efforts. By fitting a metapopulation model simulating PPRV spread to the results of a nationwide serological survey in Ethiopia, we estimated the level of viral transmission in an endemic setting and the vaccination coverage required for elimination. Results suggest that the pastoral production system as a whole acts as a viral reservoir, from which PPRV spills over into the sedentary production system, where viral persistence is uncertain. Estimated levels of PPRV transmission indicate that viral spread could be prevented if the proportion of immune small ruminants is kept permanently above 37% in at least 71% of pastoral village populations. However, due to the high turnover of these populations, maintaining the fraction of immune animals above this threshold would require high vaccine coverage within villages, and vaccination campaigns to be conducted annually. Adapting vaccination strategies to the specific characteristics of the local epidemiological context and small ruminant population dynamics would result in optimized allocation of limited resources and increase the likelihood of PPR eradication.


Mitochondrial long noncoding RNAs as blood based biomarkers for cardiac remodeling in patients with hypertrophic cardiomyopathy.
Hypertrophic cardiomyopathy (HCM) is a hereditary heart disease with a high risk for sudden cardiac death in young people. As a subtype, hypertrophic obstructive cardiomyopathy (HOCM) additionally has a left ventricular outflow gradient, showing stronger symptoms and requires a different treatment compared with hypertrophic nonobstructive cardiomyopathy (HNCM). In this study our aim was to investigate the regulation of mitochondrial and cardiac remodeling associated long noncoding RNAs (lncRNAs) in blood of patients affected with HOCM and HNCM. We included 28 HNCM, 57 HOCM, and 26 control inviduals. Already known mitochondrial and cardiac remodeling associated lncRNAs uc004cos.4, uc004coz.1, uc004cov.4, uc011mfi.2, uc022bqw.1, uc022bqs.1, and uc022bqu.1 were amplified in serum of these patients and correlated with clinical parameters. Long noncoding RNAs uc004cov.4 and uc022bqu.1 were significantly increased in patients with HOCM but not in patients with HNCM. With the use of receiver operator characteristic (ROC) curve analysis, lncRNAs uc004cov.4 and uc022bqu.1 were able to identify HOCM patients. In our study we evidenced that the specific mitochondrial long noncoding RNAs uc004cov.4 and uc022bqu.1 were upregulated in patients with HOCM and they were also able to identify HOCM and could be developed as useful clinical biomarkers in the future.


Which scoring method depicts spinal radiographic damage in early axial spondyloarthritis best? Five-year results from the DESIR cohort.
To compare the performance of different spinal radiographic damage scoring methods in patients with early axial spondyloarthritis (axSpA). Five-year spinal radiographs from the DESIR cohort were scored by three readers (averaged) for the calculation of the Stoke AS Spine Score (SASSS), modified Stoke Ankylosing Spondylitis Spine Score (mSASSS), Radiographic AS Spinal Score (RASSS), BASRI-spine and BASRI-total, and following the OMERACT filter, scores were compared according to truth, discrimination (reliability and sensitivity to change) and feasibility. The proportion of patients with a net change > smallest detectable change and >1 was calculated. The proportion of total variance explained by the patient (true variance) was calculated for the change scores as a measure of reliability, using analysis of variance. In total 699 patients were included. Five-year net changes > smallest detectable change (>1) were: RASSS 17% (17%), mSASSS 12% (12%), BASRI-spine and BASRI-total 12% (9%), SASSS 11% (11%). The mSASSS and the RASSS performed the best in terms of capturing the signal (positive change) related to noise (negative change). The proportion of variance explained by the patient was highest for the mSASSS and RASSS (85% for both 5-year progression scores vs 50-55% for other methods). The proportion of patient variance in the thoracic segment of the RASSS was unsatisfactory (46% for progression). The existing scoring methods to assess spinal radiographic damage performed well in early phases of axSpA. The mSASSS and RASSS captured most change. There was no clear gain in additionally scoring the thoracic spine for the RASSS. The mSASSS remains the most sensitive and valid scoring method in axSpA, including early phases of the disease.


Blastocystis hominis: pathogen or fellow traveler?
Blastocystis hominis, an intestinal organism of uncertain taxonomic position, has long been considered nonpathogenic. Some recent studies suggest, however, that it may be associated with diarrhea and may respond to treatment with iodoquinol or metronidazole. To investigate this possibility, we identified 148 persons whose stools contained this organism. Of this number, 32 had at least 6 stool examinations performed. Twenty-seven of the 32 persons were later found to have greater than or equal to 1 recognized pathogens--Entamoeba histolytica, Giardia lamblia or Dientamoeba fragilis--and, after receiving appropriate therapy, became asymptomatic. The B hominis infection, however, was unaffected by therapy. Five persons with only B. hominis infection were treated with iodoquinol without effect; these persons fulfilled the medical criteria for irritable bowel syndrome. We believe that when an apparently symptomatic B. hominis infection responds to therapy, the improvement probably represents elimination of some other undetected organism causing the infection.


[Assessment of a nutrition education intervention among construction workers].
Unhealthy lifestyles and overweight are common among blue collar workers. To evaluate a nutrition education intervention in construction workers to prevent chronic diseases. One hundred forty two participants were randomly assigned to an experimental group (n = 69) and or a control group (n = 73). The experimental group received nutrition education consisting in individual lifestyle counseling, group workshop, group counseling in healthy lifestyles, and environmental intervention. At baseline and after one year of intervention, clinical evaluations consisting in laboratory determinations, anthropometry and nutritional assessment evaluation were carried out. The experimental group experienced a significant decrease in waist circumference, total cholesterol, triglycerides, and an increase in HDL cholesterol. A significant reduction in total calorie intake, especially at the evening snack was observed during week-days. The same reduction in total calories occurred at dinner during the week-end. A significant increase was observed in the consumption of fruits, vegetables and fish. In the control group there was a significant increase in weight, waist circumference, body mass index, glucose and insulin, without changes in total calorie intake or healthy food consumption. High soft drink consumption did not change in both groups. In construction workers, integral nutrition intervention at the individual, group and environmental level had an impact in nutritional and biochemical indicators.


A Phase I clinical and pharmacological evaluation of sodium phenylbutyrate on an 120-h infusion schedule.
Sodium phenylbutyrate (PB) demonstrates potent differentiating capacity in multiple hematopoietic and solid tumor cell lines. We conducted a Phase I and pharmacokinetic study of PB by continuous infusion to characterize the maximum tolerated dose, toxicities, pharmacokinetics, and antitumor effects in patients with refractory solid tumors. Patients were treated with a 120-h PB infusion every 21 days. The dose was escalated from 150 to 515 mg/kg/day. Pharmacokinetics were performed during and after the first infusion period using a validated high-performance liquid chromatographic assay and single compartmental pharmacokinetic model for PB and its principal metabolite, phenylacetate. A total of 24 patients were enrolled on study, with hormone refractory prostate cancer being the predominant tumor type. All patients were evaluable for toxicity and response. A total of 89 cycles were administered. The dose-limiting toxicity (DLT) was neuro-cortical, exemplified by excessive somnolence and confusion and accompanied by clinically significant hypokalemia, hyponatremia, and hyperuricemia. One patient at 515 mg/kg/day and another at 345 mg/kg/day experienced this DLT. Toxicity resolved < or =12 h of discontinuing the infusion. Other toxicities were mild, including fatigue and nausea. The maximum tolerated dose was 410 mg/kg/day for 5 days. Pharmacokinetics demonstrated that plasma clearance of PB increased in a continuous fashion beginning 24 h into the infusion. In individuals whose V(max) for drug elimination was less than their drug-dosing rate, the active metabolite phenylacetate accumulated progressively. Plasma PB concentrations (at 410 mg/kg/day) remained above the targeted therapeutic threshold of 500 micromol/liter required for in vitro activity. The DLT in this Phase I study for infusional PB given for 5 days every 21 days is neuro-cortical in nature. The recommended Phase II dose is 410 mg/kg/day for 120 h.


Changes in volume of drinking after changes in alcohol taxes and travellers' allowances: results from a panel study.
The aim of this paper is to study short-term changes in alcohol consumption by subgroups of the population in Denmark, Finland and southern Sweden following large-scale decreases in alcohol taxation in Denmark and Finland and large increases in travellers' allowances in Finland and Sweden. General population random samples surveyed before and after the changes, using northern Sweden as a control site. Denmark, Finland, southern Sweden and northern Sweden. Respondents aged 16-69 years. Volume of drinking is the main measure reported. Changes are examined by gender, age, income and year 2003 consumption level. Consumption decreased or remained the same among women and men in all three study sites. Relative changes were similar across subgroups of age, gender and income in all countries. In absolute terms, there was a consistent differential change by age in Denmark, Finland and Southern Sweden, with the higher level of the young and lower level of the old converging. Women's and men's consumption converged in Finland and southern Sweden. The changes did not differ systematically by income. Changes were not larger among heavier drinkers. The results did not confirm expectations: an increase in consumption larger than that in the control site could not be shown in any of the countries or subgroups of the population. If there has been an effect -- as shown in aggregate data in Finland -- it seems to have been stronger among the old than the young and, in Finland and southern Sweden, among women rather than men.


Is chemotherapy-induced nausea and vomiting lower in smokers?
Chemotherapy-induced nausea and vomiting (CINV) is not understood completely. The aim of this study is to investigate the effects of smoking on CINV. 121 consecutive patients who received cisplatin (≥ 50 mg/m²) based chemotherapy were included in the study. The patients who reported motion sickness, pain, emesis during past pregnancy, emesis history of previous chemotherapy, with Karnofsky score < 70, peptic ulcer, gastroesophageal reflux, migraine, central nervous system metastasis and patients scheduled to receive radiation therapy were excluded from the study. A standard antiemetic treatment was given to all patients. The nausea and vomiting was assessed by the European Organization for the Research and Treatment of Cancer Quality of Life Questionnaire. After first cycle of chemotherapy, Grade 2 - 3 nausea and vomiting were questioned. Grade 2 - 3 nausea and vomiting was higher in non-smokers (p < 0.001). We found that nausea and vomiting in women is more frequent than in men (p < 0.001). In our study we found that smokers had a lower incidence of CINV. Further investigations are needed to confirm the findings of this pilot study.


Class IA PI3K inhibition inhibits cell growth and proliferation in mantle cell lymphoma.
Constitutive activation of the phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin signaling pathway preferentially occurs in aggressive blastoid variants of mantle cell lymphoma (MCL) and is implicated in the pathogenesis of this disease. In this study, we investigated the role of PI3K isoforms on proliferation of aggressive MCL cells. The changes in cell viability, cell cycle distribution and apoptosis induction by the PI3K isoform-selective inhibitors were evaluated. The molecular basis underlying the effects of the specific inhibition of PI3K isoforms was investigated by Western blot analysis. Our results demonstrated that a class IA PI3K isoform is most commonly involved in the constitutive activation of Akt in aggressive MCL. Treatment with a p110α isoform-specific inhibitor induced prominent cell cycle arrest followed by apoptosis through complete abolishment of phosphorylated (p)-Akt and its downstream targets. An inhibitor of isoform p110δ induced moderate cell cycle arrest with downregulation of p-Akt and p-S6K. A dual inhibitor of p110α and p110δ GDC-0941 caused more prominent cell growth inhibition compared to selective p110α or p110δ inhibitors. Inhibition of the class IB PI3K isoform p110γ did not cause cell cycle arrest or induce apoptosis in MCL cells. These findings suggest that the therapeutic ablation of class IA PI3K may be a promising strategy for the treatment of refractory, aggressive MCL.


pS2 transfection of murine adenocarcinoma cell line 410.4 enhances dispersed growth pattern in a 3-D collagen gel.
We describe the first model system employing human pS2 gene transfer and expression in a non-pS2-expressing cell line, mouse mammary adenocarcinoma 410.4, in order to analyse the potential effect of human trefoil peptide pS2 in glandular epithelium. Two selected clones, AA4 and AD4, were established and shown to have incorporated the pS2 cDNA sequence into the genome, express pS2 containing transcript and produce the pS2 peptide. When grown in 3-D collagen gels both transfectants show striking morphological changes compared to the vector control clone (VA5). VA5 forms large cohesive spherical aggregates with rare coarse spicular outgrowths, accompanied by prominent hyalinised extracellular matrix deposition. pS2 transfectants form poorly cohesive, stellate colonies with very little or no matrix deposition, radiating long cords composed of single elongated cells, an effect previously observed in other cell lines with hepatocyte growth factor. pS2 transfection had no demonstrable effect on proliferation and this is not a morphogenetic phenomenon, as tubulogenesis is not seen. Motility assays suggest that the pS2 'dispersant' effect in collagen gels is due to an increase in cell motility. There were no measurable alterations in either E-cadherin expression or E-cadherin-dependent cell-cell aggregation. pS2 may play a role in maintenance and restitution of mucosal integrity by accelerating migration/dispersion.


Background estimation in experimental spectra
A general probabilistic technique for estimating background contributions to measured spectra is presented. A Bayesian model is used to capture the defining characteristics of the problem, namely, that the background is smoother than the signal. The signal is allowed to have positive and/or negative components. The background is represented in terms of a cubic spline basis. A variable degree of smoothness of the background is attained by allowing the number of knots and the knot positions to be adaptively chosen on the basis of the data. The fully Bayesian approach taken provides a natural way to handle knot adaptivity and allows uncertainties in the background to be estimated. Our technique is demonstrated on a particle induced x-ray emission spectrum from a geological sample and an Auger spectrum from iron, which contains signals with both positive and negative components.


Variability over time and correlates of cholesterol and blood pressure in systemic lupus erythematosus: a longitudinal cohort study.
Total cholesterol (TC) and blood pressure (BP) are likely to take a dynamic course over time in patients with systemic lupus erythematosus (SLE). This would have important implications in terms of using single-point-in-time measurements of these variables to assess coronary artery disease (CAD) risk. The objective of this study was to describe and quantify variability over time of TC and BP among patients with SLE and to determine their correlates. Patients in the Toronto lupus cohort who had two or more serial measurements of TC and systolic and diastolic BP (SBP and DBP) were included in the analysis. Variability over time was described in terms of the proportion of patients whose TC and BP profile fluctuated between normal and elevated (TC>5.2 mmol/L; SBP>or=140 mm Hg or DBP>or=90 mm Hg), and also in terms of within- and between-patient variance quantified by using analysis of variance modeling. Generalized estimating equations (GEEs) were used to determine independent correlates of each of TC, SBP, and DBP, treated as continuous outcome variables. In total, 1,260 patients, comprising 26,267 measurements of each of TC, SBP, and DBP, were included. Mean+/-SD number of measurements per patient was 20.8+/-20. Mean+/-SD time interval between measurements was 5.4+/-9.7 months. Mean+/-SD time interval from the start to the end of the study was 9.3+/-8.5 years. Over time, 64.7% of patients varied between having normal and elevated cholesterol levels, whereas the status of 46.4% of patients varied between normotensive and hypertensive. By using analysis of variance (ANOVA), the within-patient percentage of total variance for each of TC, SBP, and DBP was 48.2%, 51.2%, and 63.9%, respectively. By using GEE, independent correlates of TC and BP included age, disease activity, and corticosteroids; antimalarial use was negatively correlated with TC (all P values<0.0001). TC and BP vary markedly over time in patients with SLE. This variability is due not only to lipid-lowering and antihypertensive medications, but also to disease- and treatment-related factors such as disease activity, corticosteroids, and antimalarials. The dynamic nature of TC and BP in SLE makes a compelling case for deriving summary measures that better capture cumulative exposure to these risk factors.


Factors impacting antimicrobial resistance in the South East Asian food system and potential places to intervene: A participatory, one health study.
With AMU projected to increase, South East Asia (SEA) is at high risk of experiencing disproportionate health, social, and economic burdens due to antimicrobial resistance (AMR). Our objective was to identify factors influencing AMR in SEA's food system and places for intervention by integrating the perspectives of experts from the region to inform policy and management decisions. We conducted two 6.5 h workshops and two 90-min interviews involving 18 AMR and other disciplinary experts from human, animal, and environment sectors who brainstormed the factors influencing AMR and identified leverage points (places) for intervention. Transcripts and workshop materials were coded for factors and their connections and transcribed into a causal loop diagram (CLD). Thematic analysis described AMR dynamics in SEA's food system and leverage points for intervention. The CLD and themes were confirmed via participant feedback. Participants constructed a CLD of AMR in the SEA food system that contained 98 factors interlinked by 362 connections. CLD factors reflected eight sub-areas of the SEA food system (e.g., government). Seven themes [e.g., antimicrobial and pesticide use and AMR spread (n = 40 quotes)], six "overarching factors" that impact the entire AMR system [e.g., the drive to survive (n = 12 quotes)], and 10 places for intervention that target CLD factors (n = 5) and overarching factors (n = 2) emerged from workshop discussions. The participant derived CLD of factors influencing AMR in the SEA food system demonstrates that AMR is a product of numerous interlinked actions taken across the One Health spectrum and that finding solutions is no simple task. Developing the model enabled the identification of potentially promising leverage points across human, animal, and environment sectors that, if comprehensively targeted using multi-pronged interventions, could evoke system wide changes that mitigate AMR. Even targeting some leverage points for intervention, such as increasing investments in research and capacity building, and setting and enforcing regulations to control antimicrobial supply, demand, and use could, in turn, shift mindsets that lead to changes in more difficult to alter leverage points, such as redefining the profit-driven intent that drives system behavior in ways that transform AMU and sustainably mitigate AMR.


Factors associated with commencing smoking in 12-year-old students in Catalonia (Spain): a cross-sectional population-based study.
Over the last decade notable progress has been made in developed countries on monitoring smoking although experimenting with cigarettes and smoking in young people remains a serious public health problem. This paper reports a cross-sectional study at the beginning of the 3-year follow-up community study TA_BES. The aim was to study the prevalence of smoking in addition to determining predictive factors for when smoking commences in a representative population of 12-year-old first year compulsory secondary education students. Twenty-nine secondary schools (N = 29) from an area of Catalonia participated in the study. In these schools 2245 students answered a questionnaire to study the attitudes, behaviors, and tobacco consumption in the subject's surrounding circle and family in relation to smoking; carbon monoxide measurements were taken by means of co-oximetry on 2 different occasions. A smoker was defined as a student who had smoked daily or occasionally in the last 30 days. For non-smokers the criteria of not considering was set up for those who answered that in the future they would not be smokers and considering those who answered that they did not rule out becoming a smoker in the future. Among the total 2245 students included in the analysis 157(7%) were classified as smokers. Among non-smokers we differentiated between those not considering smoking 1757 (78.3%) and those considering smoking 288 (12.8%).Age is among the factors related to commencing smoking. The risk of becoming a smoker increases 2.27 times/year. The influence of the group of friends with a very high risk for boys OR 149.5 and lower, albeit high, in girls OR 38.1. Tobacco consumption of parents produces different effects in young people. A smoking father does not produce alterations in the smoking behavior of young people. However having a smoking mother or former smoking is a risk factor for boys and a protective factor for girls.We detected a gradual risk of becoming a smoker by means of the co-oximetry test. A boy/girl with a test between 6 p.p.m and 10 p.p.m increased the probability of smoking by 2.29 and co-oximetry values > 10 p.p.m multiplied the risk 4 times over. Results indicate that the age of commencing smoking is maintained in spite of prevalence having decreased in the last few years. The risk factors identified should be used to involve families and the educational community by offering them tobacco weaning programmes.


A Bayesian parametric approach to handle missing longitudinal outcome data in trial-based health economic evaluations.
Trial-based economic evaluations are typically performed on cross-sectional variables, derived from the responses for only the completers in the study, using methods that ignore the complexities of utility and cost data (e.g. skewness and spikes). We present an alternative and more efficient Bayesian parametric approach to handle missing longitudinal outcomes in economic evaluations, while accounting for the complexities of the data. We specify a flexible parametric model for the observed data and partially identify the distribution of the missing data with partial identifying restrictions and sensitivity parameters. We explore alternative non-ignorable missingness scenarios through different priors for the sensitivity parameters, calibrated on the observed data. Our approach is motivated by, and applied to, data from a trial assessing the cost-effectiveness of a new treatment for intellectual disability and challenging behaviour.


Plasmodium falciparum dihydroartemisinin-piperaquine failures in Cambodia are associated with mutant K13 parasites presenting high survival rates in novel piperaquine in vitro assays: retrospective and prospective investigations.
The declining efficacy of dihydroartemisinin-piperaquine against Plasmodium falciparum in Cambodia, along with increasing numbers of recrudescent cases, suggests resistance to both artemisinin and piperaquine. Available in vitro piperaquine susceptibility assays do not correlate with treatment outcome. A novel assay using a pharmacologically relevant piperaquine dose/time exposure was designed and its relevance explored in retrospective and prospective studies. The piperaquine survival assay (PSA) exposed parasites to 200 nM piperaquine for 48 hours and monitored survival 24 hours later. The retrospective study tested 32 culture-adapted, C580Y-K13 mutant parasites collected at enrolment from patients treated with a 3-day course of dihydroartemisinin-piperaquine and having presented or not with a recrudescence at day 42 (registered ACTRN12615000793516). The prospective study assessed ex vivo PSA survival rate alongside K13 polymorphism of isolates collected from patients enrolled in an open-label study with dihydroartemisinin-piperaquine for uncomplicated P. falciparum malaria in Cambodia (registered ACTRN12615000696594). All parasites from recrudescent cases had in vitro or ex vivo PSA survival rates ≥10%, a relevant cut-off value for piperaquine-resistance. Ex vivo PSA survival rates were higher for recrudescent than non-recrudescent cases (39.2% vs. 0.17%, P <1 × 10(-7)). Artemisinin-resistant K13 mutants with ex vivo PSA survival rates ≥10% were associated with 32-fold higher risk of recrudescence (95% CI, 4.5-224; P = 0.0005). PSA adequately captures the piperaquine resistance/recrudescence phenotype, a mainstay to identify molecular marker(s) and evaluate efficacy of alternative drugs. Combined ex vivo PSA and K13 genotyping provides a convenient monitor for both artemisinin and piperaquine resistance where dihydroartemisinin-piperaquine is used.


Streptococcus mutans and Streptococcus sanguinis Expression of Competition-Related Genes, Under Sucrose.
Streptococcus mutans synthesizes 3 glucosyltransferases (Gtfs) associated with cariogenic biofilms, while commensal Streptococcus sanguinis produces only one; gtfP and hydrogen peroxide (H2O2) by SpxB. The aim was to test the hypothesis that under a sucrose-induced cariogenic challenge, the expression of competition-related genes is differentially regulated depending on whether S. sanguinis or S. mutans primarily colonize enamel. Dual-species biofilms of S. sanguinis and S. mutans were formed under different colonization sequences on enamel slabs and exposed to 10% sucrose for 5 min, 3×/day for 5 days. Biofilms were analyzed for the transcriptional response of competition-related genes encoding gtfB, gtfC, and gtfD for S. mutans and gtfP and spxB for S. sanguinis. In addition, acidogenicity (pH) and viable cells in each of the conditions were determined. For all the genes, a downregulation was observed during simultaneous colonization by both bacterial species. In contrast, gtfB was upregulated when S. sanguinis was the first colonizer (p < 0.05). Both gtfC and gtfD were upregulated during sequential inoculation with S. sanguinis as the first colonizer. An eleven-fold upregulation of gtfP was observed in biofilms with S. mutans as initial colonizer (p < 0.05), with a moderate increase in spxB expression. The lowest pH values and viable cells of S. sanguinis were observed when S. mutans first colonized the enamel slabs, compared to the other conditions (p < 0.05). Demanding sucrose-challenged oral environment requires increased expression of virulence traits to effectively compete and thrive in the dental biofilm, especially when the competitor has already colonized the ecological niche.


Optogenetic and electrical microstimulation systematically bias visuospatial choice in primates.
Optogenetics is a recently developed method in which neurons are genetically modified to express membrane proteins sensitive to light, enabling precisely targeted control of neural activity [1-3]. The temporal and spatial precision afforded by neural stimulation by light holds promise as a powerful alternative to current methods of neural control, which rely predominantly on electrical and pharmacological methods, in both research and clinical settings [4, 5]. Although the optogenetic approach has been widely used in rodent and other small animal models to study neural circuitry [6-8], its functional application in primate models has proven more difficult. In contrast to the relatively large literature on the effects of cortical electrical microstimulation in perceptual and decision-making tasks [9-13], previous studies of optogenetic stimulation in primates have not demonstrated its utility in similar paradigms [14-18]. In this study, we directly compare the effects of optogenetic activation and electrical microstimulation in the lateral intraparietal area during a visuospatial discrimination task. We observed significant and predictable biases in visual attention in response to both forms of stimulation that are consistent with the experimental modulation of a visual salience map. Our results demonstrate the power of optogenetics as a viable alternative to electrical microstimulation for the precise dissection of the cortical pathways of high-level processes in the primate brain.


Template-Free 3D Microprinting of Metals Using a Force-Controlled Nanopipette for Layer-by-Layer Electrodeposition.
A novel 3D printing method for voxel-by-voxel metal printing is presented. Hollow atomic force microscopy (AFM) cantilevers are used to locally supply metal ions in an electrochemical cell, enabling a localized electroplating reaction. By exploiting the deflection feedback of these probes, electrochemical 3D metal printing is, for the first time, demonstrated in a layer-by-layer fashion, enabling the fabrication of arbitrary-shaped geometries.


Baseline characteristics of the infant aphakia treatment study population: predicting recognition acuity at 4.5 years of age.
To identify patient baseline characteristics that predict recognition acuity at 4.5 years of age in the Infant Aphakia Treatment Study, a study of patients with monocular infantile cataracts. We analyzed baseline characteristics of the 114 infants enrolled in the Infant Aphakia Treatment Study to determine which were most predictive of visual outcome at 4.5 years of age. All infants underwent cataract surgery between 1 and 7 months of age. Monocular acuity was assessed at 4.5 years of age by a traveling examiner using the Amblyopia Treatment Study HOTV protocol. Age at cataract surgery was weakly associated with visual acuity (Spearman rank correlation coefficient = 0.19, P = 0.041) with median visual acuity better among the younger patients (28-48 days: 0.50 logMAR, 49-210 days: 1.10 logMAR, P = 0.046). Patients from families with private insurance had significantly better median visual acuity (0.60 vs. 1.40 logMAR, P = 0.0004). No other baseline characteristic revealed a significant bivariate relationship with visual acuity. A multiple linear regression relating visual acuity to all baseline characteristics demonstrated that only the availability of private insurance was statistically significant, accounting for 12% of the variance. This analysis concurs with previous studies that early surgery is important for good visual outcomes in patients with unilateral infantile cataracts. The fact that only one baseline variable (private insurance) contributed to the multivariate analysis, accounting for 12% of the variance, suggests that predicting visual outcome for these patients is complicated at best, and cannot be estimated from baseline characteristics alone. (ClinicalTrials.gov number, NCT00212134.).


Photocatalytic H2 production from water with rhenium and cobalt complexes.
Photocatalytic hydrogen production in pure water for three component systems using a series of rhenium-based photosensitizers (PS) and cobalt-based water reduction catalysts (WRC), with triethanolamine (TEOA) as an irreversible electron donor, is described. Besides the feasibility of this reaction in water, key findings are reductive quenching of the excited state of the PS by TEOA (k(q) = 5-8 × 10(7) M(-1) s(-1); Φ(cage) = 0.75) and subsequent transfer of an electron to the WRC (k(Co(III)) = 1.1 × 10(9) M(-1) s(-1)). Turnover numbers in rhenium (TON(Re), H/Re) above 500 were obtained, whereas TON(Co) (H(2)/Co) did not exceed 17. It is shown that the cobalt-based WRC limits long-term performance. Long-term performance critically depends on pH and the type of WRC used but is unaffected by the type of PS or the concentration of WRC. A quantum yield of 30% was obtained (H/photon).


Prevalence and Outcomes of Opioid Use Disorder in Pediatric Surgical Patients: A Retrospective Cohort Study.
Chronic opioid use among adolescents is a leading preventable public health problem in the United States. Consequently, a sizable proportion of surgical patients in this age group may have a comorbid opioid use disorder (OUD). No previously published study has examined the prevalence of OUD and its impact on postoperative morbidity or mortality in the adolescent surgical population. Our objective was to investigate the prevalence of comorbid OUD and its association with surgical outcomes in a US adolescent surgical population. We hypothesized that OUD among adolescent surgical patients is on an upward trajectory and that the presence of OUD is associated with higher risk of postoperative morbidity or mortality. Using the pediatric health information system, we performed a 1:1 propensity score-matched, retrospective cohort study of adolescents (10-18 years of age) undergoing inpatient surgery between 2004 and 2019. The primary outcome was inpatient mortality. The secondary outcomes were surgical complications and postoperative infection. We also evaluated indicators of resource utilization, including mechanical ventilation, intensive care unit (ICU) admission, and postoperative length of stay (LOS). Of 589,098 adolescents, 563 (0.1%) were diagnosed with comorbid OUD (563 were matched on OUD). The prevalence of OUD in adolescents undergoing surgery increased from 0.4 per 1000 cases in 2004 to 1.6 per 1000 cases in 2019, representing an average annual percent change (AAPC) of 9.7% (95% confidence interval [CI], 5.7-13.9; P value < .001). The overall postoperative mortality rate was 0.50% (n = 2941). On univariable analysis, mortality rate was significantly higher in adolescents with comorbid OUD than those without comorbid OUD (3.37% vs 0.50%; P < .001). Among propensity-matched pairs, comorbid OUD diagnosis was associated with an estimated 57% relative increase in the risk of surgical complications (adjusted relative risk [aRR], 1.57; 95% CI, 1.24-2.00; P < .001). The relative risk of postoperative infection was 2-fold higher in adolescents with comorbid OUD than in those without OUD (aRR, 2.02; 95% CI, 1.62-2.51; P < .001). Adolescents with comorbid OUD had an increased risk of ICU admission, mechanical ventilation, and extended postoperative LOS. OUD is becoming increasingly prevalent in adolescents presenting for surgery. Comorbid OUD is an important determinant of surgical complications, postoperative infection, and resource utilization, underscoring the need to consider OUD as a critical, independent risk factor for postsurgical morbidity.


Structural similarities and functional differences clarify evolutionary relationships between tRNA healing enzymes and the myelin enzyme CNPase.
Eukaryotic tRNA splicing is an essential process in the transformation of a primary tRNA transcript into a mature functional tRNA molecule. 5'-phosphate ligation involves two steps: a healing reaction catalyzed by polynucleotide kinase (PNK) in association with cyclic phosphodiesterase (CPDase), and a sealing reaction catalyzed by an RNA ligase. The enzymes that catalyze tRNA healing in yeast and higher eukaryotes are homologous to the members of the 2H phosphoesterase superfamily, in particular to the vertebrate myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase). We employed different biophysical and biochemical methods to elucidate the overall structural and functional features of the tRNA healing enzymes yeast Trl1 PNK/CPDase and lancelet PNK/CPDase and compared them with vertebrate CNPase. The yeast and the lancelet enzymes have cyclic phosphodiesterase and polynucleotide kinase activity, while vertebrate CNPase lacks PNK activity. In addition, we also show that the healing enzymes are structurally similar to the vertebrate CNPase by applying synchrotron radiation circular dichroism spectroscopy and small-angle X-ray scattering. We provide a structural analysis of the tRNA healing enzyme PNK and CPDase domains together. Our results support evolution of vertebrate CNPase from tRNA healing enzymes with a loss of function at its N-terminal PNK-like domain.


Prolongation of electrode lifetime in biofuel cells by periodic enzyme renewal.
Enzymatically catalyzed biofuel cells show unique specificity and promise high power densities, but suffer from a limited lifetime due to enzyme deactivation. In the present work, we demonstrate a novel concept to extend the lifetime of a laccase-catalyzed oxygen reduction cathode in which we decouple the electrode lifetime from the limited enzyme lifetime by a regular resupply of fresh enzymes. Thereto, the adsorption behavior of laccase from Trametes versicolor to buckypaper electrode material, as well as its time-dependent deactivation characteristics, has been investigated. Laccase shows a Langmuir-type adsorption to the carbon nanotube-based buckypaper electrodes, with a mean residence time of 2 days per molecule. In a citrate buffer of pH 5, laccase does not show any deactivation at room temperature for 2 days and exhibits a half-life of 9 days. In a long-term experiment, the laccase electrodes were operated at a constant galvanostatic load. The laccase-containing catholyte was periodically exchanged against a freshly prepared one every second day to provide sufficient active enzymes in the catholyte for the replacement of desorbed inactive enzymes. Compared to a corresponding control experiment without catholyte exchange, this procedure resulted in a 2.5 times longer cathode lifetime of 19 ± 9 days in which the electrode showed a potential above 0.744 V vs. normal hydrogen electrode at 110 μA cm(-2). This clearly indicates the successful exchange of molecules by desorption and re-adsorption and is a first step toward the realization of a self-regenerating enzymatic biofuel cell in which enzyme-producing microorganisms are integrated into the electrode to continuously resupply fresh enzymes.


Separated carbon nanotube macroelectronics for active matrix organic light-emitting diode displays.
Active matrix organic light-emitting diode (AMOLED) display holds great potential for the next generation visual technologies due to its high light efficiency, flexibility, lightweight, and low-temperature processing. However, suitable thin-film transistors (TFTs) are required to realize the advantages of AMOLED. Preseparated, semiconducting enriched carbon nanotubes are excellent candidates for this purpose because of their excellent mobility, high percentage of semiconducting nanotubes, and room-temperature processing compatibility. Here we report, for the first time, the demonstration of AMOLED displays driven by separated nanotube thin-film transistors (SN-TFTs) including key technology components, such as large-scale high-yield fabrication of devices with superior performance, carbon nanotube film density optimization, bilayer gate dielectric for improved substrate adhesion to the deposited nanotube film, and the demonstration of monolithically integrated AMOLED display elements with 500 pixels driven by 1000 SN-TFTs. Our approach can serve as the critical foundation for future nanotube-based thin-film display electronics.


Inter-brain synchronization during social interaction.
During social interaction, both participants are continuously active, each modifying their own actions in response to the continuously changing actions of the partner. This continuous mutual adaptation results in interactional synchrony to which both members contribute. Freely exchanging the role of imitator and model is a well-framed example of interactional synchrony resulting from a mutual behavioral negotiation. How the participants' brain activity underlies this process is currently a question that hyperscanning recordings allow us to explore. In particular, it remains largely unknown to what extent oscillatory synchronization could emerge between two brains during social interaction. To explore this issue, 18 participants paired as 9 dyads were recorded with dual-video and dual-EEG setups while they were engaged in spontaneous imitation of hand movements. We measured interactional synchrony and the turn-taking between model and imitator. We discovered by the use of nonlinear techniques that states of interactional synchrony correlate with the emergence of an interbrain synchronizing network in the alpha-mu band between the right centroparietal regions. These regions have been suggested to play a pivotal role in social interaction. Here, they acted symmetrically as key functional hubs in the interindividual brainweb. Additionally, neural synchronization became asymmetrical in the higher frequency bands possibly reflecting a top-down modulation of the roles of model and imitator in the ongoing interaction.


Effect of saffron supplementation on symptoms of depression and anxiety: a systematic review and meta-analysis.
Saffron (Crocus sativus L.) has gained interest as a potential treatment in psychiatry. This systematic review and meta-analysis sought to investigate the effect of saffron supplementation, as both an adjunctive therapy and monotherapy, on symptoms of depression and anxiety in clinical and general populations compared with pharmacotherapy or placebo. Using the PRISMA guidelines, a systematic literature review of randomized controlled trials was conducted. A meta-analysis was conducted to determine treatment effect. Risk of bias was assessed using the Jadad scale. Twenty-three studies were included. Saffron had a large positive effect size when compared with placebo for depressive symptoms (g = 0.99, P < 0.001) and anxiety symptoms (g = 0.95, P < 0.006). Saffron also had a large positive effect size when used as an adjunct to antidepressants for depressive symptoms (g = 1.23, P = 0.028). Egger's regression test found evidence of publication bias. Saffron could be an effective intervention for symptoms of depression and anxiety; however, due to evidence of publication bias and lack of regional diversity, further trials are required. PROSPERO registration CRD42017070060.


The safety, tolerability, and efficacy of a liquid formulation of deferiprone in young children with transfusional iron overload.
Limited data are available on the use of deferiprone in children younger than 10 years of age. This study evaluated the safety and efficacy of a new liquid formulation of deferiprone for the treatment of transfusional iron overload in children 1-10 years old. One hundred children (91 thalassemia major, 8 Hb E-β thalassemia, and 1 sickle cell disease) were enrolled for a 6-month treatment with deferiprone (50 to 100 mg/kg/d). The safety profile was similar to or better than that reported in earlier studies with deferiprone tablets in older children and adults. No unexpected adverse reactions were observed. Gastrointestinal intolerance (GI) was observed in 11% and an increased serum ALT in 12% of the children. Both events were transient. Mild neutropenia, observed in 6% of patients, did not progress to agranulocytosis and resolved despite continuous deferiprone treatment. Two patients experienced agranulocytosis that resolved without complications upon discontinuation of therapy. Deferiprone use was associated with a significant decline in mean serum ferritin level from 2532±1463 μg/L at baseline to 2176±1144 μg/L (P<0.0005). The results of this study show a favorable benefit/risk ratio of deferiprone oral solution for the treatment of young children with transfusional iron overload.


[Sociocultural factors and physical activity level in early adolescence].
To describe the level of physical activity in adolescents born in 1993 in the city of Pelotas, Brazil, and to evaluate the effect of sociocultural variables on those levels. A combined ethno-epidemiological methodology was employed. In the epidemiological study, 4 452 adolescents born in 1993 were interviewed. Physical activity level was evaluated by means of a questionnaire. Adolescents with fewer than 300 minutes of physical activity per week were classified as sedentary. The ethnographic study included 69 adolescents randomly selected from among all cohort participants. On average, three in-depth interviews were carried out (about one every 6 months), with mothers and adolescents being interviewed separately. The prevalence of a sedentary lifestyle was 48.7% (95%CI: 46.5 to 50.8) in boys and 67.5% (95%CI: 65.6 to 69.5) in girls (P < 0.001). The independent variable presenting the strongest association with physical activity level was the weekly frequency with which the adolescent met friends outside of school. The ethnographic study showed that boys have more social and family support to engage in physical activities in adolescence, but that many parents associate poor school performance with the amount of time spent outside the home. This study suggests that physical activity is often viewed as competing with family values in adolescence. This perspective should be explored in future studies and discussed with families so that physical activity may be adopted as a cultural norm.


Models for Trypanosoma evansi (surra), its control and economic impact on small-hold livestock owners in the Philippines.
Simple demographic and infectious disease models of buffaloes and other domestic hosts for animal trypanosomosis (surra) caused by Trypanosoma evansi were developed. The animal models contained deterministic and stochastic elements and were linked to simulate the benefit of control regimes for surra in village domestic animal populations in Mindanao, Philippines. The impact of the disease on host fertility and mortality were key factors in determining the economic losses and net-benefit from the control regimes. If using a high (99%) efficacy drug in surra-moderate to high risk areas, then treating all animals twice each year yielded low prevalence in 2 years; targeted treatment of clinically sick animals, constantly monitored (monthly), required 75% fewer treatments but took longer to reach a low prevalence than treating all animals twice each year. At high drug efficacy both of these treatment strategies increased the benefit over untreated animals by 81%. If drug efficacy declined then the benefit obtained from twice yearly treatment of all animals declined rapidly compared with regular monitoring and targeting treatment to clinically sick animals. The current control regimen applied in the Philippines of annual sero-testing for surra and only treating sero-positive animals provided the lowest net-benefit of all the control options simulated and would not be regarded as effective control. The total net-benefit from effective surra control for a typical village in a moderate/high risk area was 7.9 million pesos per annum (US $158,000). The value added to buffaloes, cattle, horses, goats/sheep and pigs as a result of this control was US $88, $84, $151, $7, $114 per animal/year, respectively.


Transgenic reexpression of GLUT1 or GLUT2 in pancreatic beta cells rescues GLUT2-null mice from early death and restores normal glucose-stimulated insulin secretion.
GLUT2-null mice are hyperglycemic, hypoinsulinemic, hyperglucagonemic, and glycosuric and die within the first 3 weeks of life. Their endocrine pancreas shows a loss of first phase glucose-stimulated insulin secretion (GSIS) and inverse alpha to beta cell ratio. Here we show that reexpression by transgenesis of either GLUT1 or GLUT2 in the pancreatic beta cells of these mice allowed mouse survival and breeding. The rescued mice had normal-fed glycemia but fasted hypoglycemia, glycosuria, and an elevated glucagon to insulin ratio. Glucose tolerance was, however, normal. In vivo, insulin secretion assessed following hyperglycemic clamps was normal. In vitro, islet perifusion studies revealed that first phase of insulin secretion was restored as well by GLUT1 or GLUT2, and this was accompanied by normalization of the glucose utilization rate. The ratio of pancreatic insulin to glucagon and volume densities of alpha to beta cells were, however, not corrected. These data demonstrate that 1) reexpression of GLUT1 or GLUT2 in beta cells is sufficient to rescue GLUT2-null mice from lethality, 2) GLUT1 as well as GLUT2 can restore normal GSIS, 3) restoration of GSIS does not correct the abnormal composition of the endocrine pancreas. Thus, normal GSIS does not depend on transporter affinity but on the rate of uptake at stimulatory glucose concentrations.


A high precision feature based on LBP and Gabor theory for face recognition.
How to describe an image accurately with the most useful information but at the same time the least useless information is a basic problem in the recognition field. In this paper, a novel and high precision feature called BG2D2LRP is proposed, accompanied with a corresponding face recognition system. The feature contains both static texture differences and dynamic contour trends. It is based on Gabor and LBP theory, operated by various kinds of transformations such as block, second derivative, direct orientation, layer and finally fusion in a particular way. Seven well-known face databases such as FRGC, AR, FERET and so on are used to evaluate the veracity and robustness of the proposed feature. A maximum improvement of 29.41% is achieved comparing with other methods. Besides, the ROC curve provides a satisfactory figure. Those experimental results strongly demonstrate the feasibility and superiority of the new feature and method.


QEEG guided neurofeedback therapy in personality disorders: 13 case studies.
According to DSM-IV, personality disorder constitutes a class only when personality traits are inflexible and maladaptive and cause either significant functional impairment or subjective distress. Classical treatment of choice for personality disorders has been psychotherapy and/or psychopharmacotherapy. Our study is to determine if subjects with antisocial personality disorders will benefit from quantitative EEG (qEEG) guided neurofeedback treatment. Thirteen subjects (9 male, 4 female) ranged in age from 19 to 48 years. All the subjects were free of medications and illicit drugs. We excluded subjects with other mental disorders by clinical assessment. Psychotherapy or psychopharmacotherapy or any other treatment model was not introduced to any of the subjects during or after neurofeedback treatment. For the subject who did not respond to neurofeedback, training was applied with 38 sessions of LORETA neurofeedback training without success. Evaluation measures included qEEG analysis with Nx Link data base, MMPI, T.O.V.A tests and SA-45 questionaries at baseline, and at the end of neurofeedback treatment. Lexicor qEEG signals were sampled at 128 Hz with 30 minutes-neurofeedback sessions completed between 80-120 sessions depending on the case, by Biolex neurofeedback system. At baseline and after every 20 sessions, patients were recorded with webcam during the interview. Twelve out of 13 subjects who received 80-120 sessions of neurofeedback training showed significant improvement based on SA-45 questionaries, MMPI, T.O.V.A. and qEEG/Nx Link data base (Neurometric analysis) results, and interviewing by parent/family members. Neurofeedback can change the view of psychiatrists and psychologists in the future regarding the treatment of personality disorders. This study provides the first evidence for positive effects of neurofeedback treatment in antisocial personality disorders. Further study with controls is warranted.


Hepatitis C virus nonstructural protein specific T cells are associated with virological responses to combination therapy in chronic HCV patients.
Virus-specific T-cell responses play a major role in antiviral immune response. However, the effect of hepatitis C virus (HCV)-specific T-cell responses on combination therapy still remains controversial. To identify the association between HCV-specific T cell responses and efficiency of combination therapy. To address this issue, a longitudinal analysis of HCV-specific T-cell responses to overlapping peptides covering HCV-nonstructural protein (NS) was performed using ELISpot assay in 48 chronically infected HCV-1b patients during combination treatment with peginterferon-alfa and ribavirin. Fifty-two percent of chronic HCV patients showed detectable HCV-NS3, NS4 or NS5A specific T-cell responses before therapy, with NS3 appearing to be the most immunodominant protein followed by NS5A and NS4. In addition, the percentage of patients responding to peptide stimulation was higher in patients with sustained virological response (SVR) when compared with those without SVR. Dynamics of HCV-NS-specific T-cell responses were further analysed; we found that HCV-specific T-cell responses maintained higher levels at 12 weeks into treatment in patients with SVR. In contrast, HCV-specific T-cell responses in patients without SVR declined significantly at 4 weeks into treatment and maintained low levels at 12 weeks. We found that the HCV-specific T-cell responses were associated with good viral control in patients with combination therapy.


Near miss experiences of transport and recreational cyclists in New South Wales, Australia. Findings from a prospective cohort study.
This paper investigates events in which cyclists perceive a cycling crash is narrowly avoided (henceforth, a near miss). A cohort of 2038 adult transport and recreational cyclists from New South Wales (Australia) provided self-reported prospectively collected data from cycling diaries to allow the calculation of an exposure-based rate of near misses and investigation of near miss circumstances. During 25,971days of cycling, 3437 near misses were reported. For a given time cycling, cyclists who rode mainly for transport (compared with those who rode mainly for recreation), and cyclists with less experience (compared to those with more experience) were more likely to report a near miss; older cyclists (60+ years) were less likely to report a near miss than younger cyclists (25-59 years). Where type of near miss was recorded, 72.0% involved motor vehicles, 10.9% involved pedestrians and 6.9% involved other cyclists. Results indicate some similarities between near misses and crashes reported by this cohort during the same reporting period. A bias toward reporting near misses with motor vehicles was suggested, which likely reflects cyclists' perceptions that crashes involving motor vehicles are particularly serious, and highlights their impact on perceived safety. Given the relative rarity of crashes, and the limited breadth and depth of administrative data, collection of near miss data may contribute to our understanding of cycling safety by increasing the volume and detail of information available for analysis. Addressing the causes of near misses may offer an opportunity to improve both perceived and actual safety for cyclists.


Association between depression and mortality in older adults: the Cardiovascular Health Study.
Studies of the association between depressive symptoms and mortality in elderly populations have yielded contradictory findings. To address these discrepancies, we test this association using the most extensive array of sociodemographic and physical health control variables ever studied, to our knowledge, in a large population-based sample of elderly individuals. To examine the relation between baseline depressive symptoms and 6-year all-cause mortality in older persons, systematically controlling for sociodemographic factors, clinical disease, subclinical disease, and health risk factors. A total of 5201 men and women aged 65 years and older from 4 US communities participated in the study. Depressive symptoms and 4 categories of covariates were assessed at baseline. The primary outcome measure was 6-year mortality. Of the 5201 participants, 984 (18.9%) died within 6 years. High baseline depressive symptoms were associated with a higher mortality rate (23.9%) than low baseline depression scores (17.7%) (unadjusted relative risk [RR], 1.41; 95% confidence interval [CI], 1.22-1.63). Depression was also an independent predictor of mortality when controlling for sociodemographic factors (RR, 1.43; 95% CI, 1.23-1.66), prevalent clinical disease (RR, 1.25; 95% CI, 1.07-1.45), subclinical disease indicators (RR, 1.35; 95% CI, 1.15-1.58), or biological or behavioral risk factors (RR, 1.42; 95% CI, 1.22-1.65). When the best predictors from all 4 classes of variables were included as covariates, high depressive symptoms remained an independent predictor of mortality (RR, 1.24; 95% CI, 1.06-1.46). High levels of depressive symptoms are an independent risk factor for mortality in community-residing older adults. Motivational depletion may be a key underlying mechanism for the depression-mortality effect.


Differential ergoline and ergopeptine binding to 5-hydroxytryptamine2A receptors: ergolines require an aromatic residue at position 340 for high affinity binding.
In this paper we show that a highly conserved aromatic residue, phenylalanine at the 340-position, is essential for ergoline binding to 5-hydroxytryptamine2A receptors. We hypothesized that F340 was essential for a specific aromatic-aromatic interaction (e.g., pi-pi or hydrophobic) between the phenyl moiety of F340 and the aromatic ring of the ergoline nucleus. To test this hypothesis, eight point mutations of adjacent (F340 and F339) and nonadjacent (F125) phenylanaines were made, using conservative (phenylalanine to tyrosine) and nonconservative (phenylalanine to leucine, alanine, or serine) substitutions. The binding affinities of all of the tested simple ergolines were greatly reduced by specific mutations of F340 in which aromatic-aromatic interactions (e.g., F340A and F340L) were abolished, but they were unaffected when the replacement residue was aromatic (e.g., F340Y). In contrast, the binding affinities of four ergopeptines (bromocryptine, ergocryptine, ergocornine, and ergotamine) were relatively unaffected by the F340L substitution. Neither ergoline nor ergopeptine affinities were consistently altered by F339 mutations. These results support the notion that aromatic-aromatic interactions (either pi-pi of hydrophobic) with F340 are essential for the binding of simple ergolines but not ergopeptines to 5-hydroxytryptamine2A receptors. Our findings support models of ergoline and ergopeptine binding to serotonin receptors that suggest that the nature of the substituent at the 8-position of the ergoline nucleus may give rise to different modes of binding for the two classes of agents, particularly with respect to the phenyl ring of F340.


Agonist binding, agonist affinity and agonist efficacy at G protein-coupled receptors.
Measurements of affinity and efficacy are fundamental for work on agonists both in drug discovery and in basic studies on receptors. In this review I wish to consider methods for measuring affinity and efficacy at G protein coupled receptors (GPCRs). Agonist affinity may be estimated in terms of the dissociation constant for agonist binding to a receptor using ligand binding or functional assays. It has, however, been suggested that measurements of affinity are always contaminated by efficacy so that it is impossible to separate the two parameters. Here I show that for many GPCRs, if receptor/G protein coupling is suppressed, experimental measurements of agonist affinity using ligand binding (K(obs)) provide quite accurate measures of the agonist microscopic dissociation constant (KA). Also in pharmacological functional studies, good estimates of agonist dissociation constants are possible. Efficacy can be quantitated in several ways based on functional data (maximal effect of the agonist (E(max)), ratio of agonist dissociation constant to concentration of agonist giving half maximal effect in functional assay (K(obs)/EC50), a combined parameter E(max)K(obs)/EC50). Here I show that E(max)K(obs)/EC50 provides the best assessment of efficacy for a range of agonists across the full range of efficacy for full to partial agonists. Considerable evidence now suggests that ligand efficacy may be dependent on the pathway used to assess it. The efficacy of a ligand may, therefore, be multidimensional. It is still, however, necessary to have accurate measures of efficacy in different pathways.


The INTERVAL trial to determine whether intervals between blood donations can be safely and acceptably decreased to optimise blood supply: study protocol for a randomised controlled trial.
Ageing populations may demand more blood transfusions, but the blood supply could be limited by difficulties in attracting and retaining a decreasing pool of younger donors. One approach to increase blood supply is to collect blood more frequently from existing donors. If more donations could be safely collected in this manner at marginal cost, then it would be of considerable benefit to blood services. National Health Service (NHS) Blood and Transplant in England currently allows men to donate up to every 12 weeks and women to donate up to every 16 weeks. In contrast, some other European countries allow donations as frequently as every 8 weeks for men and every 10 weeks for women. The primary aim of the INTERVAL trial is to determine whether donation intervals can be safely and acceptably decreased to optimise blood supply whilst maintaining the health of donors. INTERVAL is a randomised trial of whole blood donors enrolled from all 25 static centres of NHS Blood and Transplant. Recruitment of about 50,000 male and female donors started in June 2012 and was completed in June 2014. Men have been randomly assigned to standard 12-week versus 10-week versus 8-week inter-donation intervals, while women have been assigned to standard 16-week versus 14-week versus 12-week inter-donation intervals. Sex-specific comparisons will be made by intention-to-treat analysis of outcomes assessed after two years of intervention. The primary outcome is the number of blood donations made. A key secondary outcome is donor quality of life, assessed using the Short Form Health Survey. Additional secondary endpoints include the number of 'deferrals' due to low haemoglobin (and other factors), iron status, cognitive function, physical activity, and donor attitudes. A comprehensive health economic analysis will be undertaken. The INTERVAL trial should yield novel information about the effect of inter-donation intervals on blood supply, acceptability, and donors' physical and mental well-being. The study will generate scientific evidence to help formulate blood collection policies in England and elsewhere. Current Controlled Trials ISRCTN24760606, 25 January 2012.


N-acetylcysteine blocks SOS induction and mutagenesis produced by fluoroquinolones in Escherichia coli.
Fluoroquinolones such as ciprofloxacin induce the mutagenic SOS response and increase the levels of intracellular reactive oxygen species (ROS). Both the SOS response and ROS increase bacterial mutagenesis, fuelling the emergence of resistant mutants during antibiotic treatment. Recently, there has been growing interest in developing new drugs able to diminish the mutagenic effect of antibiotics by modulating ROS production and the SOS response. To test whether physiological concentrations of N-acetylcysteine, a clinically safe antioxidant drug currently used in human therapy, is able to reduce ROS production, SOS induction and mutagenesis in ciprofloxacin-treated bacteria without affecting antibiotic activity. The Escherichia coli strain IBDS1 and its isogenic mutant deprived of SOS mutagenesis (TLS-) were treated with different concentrations of ciprofloxacin, N-acetylcysteine or both drugs in combination. Relevant parameters such as MICs, growth rates, ROS production, SOS induction, filamentation and antibiotic-induced mutation rates were evaluated. Treatment with N-acetylcysteine reduced intracellular ROS levels (by ∼40%), as well as SOS induction (by up to 75%) and bacterial filamentation caused by subinhibitory concentrations of ciprofloxacin, without affecting ciprofloxacin antibacterial activity. Remarkably, N-acetylcysteine completely abolished SOS-mediated mutagenesis. Collectively, our data strongly support the notion that ROS are a key factor in antibiotic-induced SOS mutagenesis and open the possibility of using N-acetylcysteine in combination with antibiotic therapy to hinder the development of antibiotic resistance.


Influence of neighbourhood ethnic density, diet and physical activity on ethnic differences in weight status: a study of 214,807 adults in Australia.
We investigated whether ethnic and country of birth differences in adult Body Mass Index (BMI) were associated with differences in diet, physical activity and ethnic density (the percentage of an ethnic group within the neighbourhood environment). A sample of 214,807 adults living in Australia was extracted from the 45 and Up Study. Analyses comprised multilevel modelling of BMI for 38 ethnic and country of birth groups. Physical activity was ascertained using the Active Australia Survey. Dietary measures included self-reported consumption of fruit, vegetables, meat and cheese. Ethnic density was objectively measured using 2006 Australian Census data. Possible confounders included age, gender, household income, educational qualifications, economic status, couple status, language, duration of residence, neighbourhood affluence and remoteness. Compared to Australian-born Australians (age-gender adjusted mean BMI = 27.1, 95%CI 27.1, 27.2), overseas-born groups often had lower mean BMI, especially the Chinese born in China (23.2, 23.0, 23.4). Exceptions included the Italians (BMI = 28.1), Greeks (28.5), Maltese (27.6), Lebanese (28.4) and Croatians (27.8) born in their ethnic-country of origin. Regardless of birthplace, BMI was lower for the English, Scottish, and Chinese, but higher for Italians and Greeks. Some ethnic differences reflected the 'healthy migrant' hypothesis, whereas others did not. These differences were only partially attenuated by controls for portions of fruit and vegetables, meat and cheese, frequency of participation in physical activity, and other explanatory variables. Ethnic density was associated with lower BMI for the English and Irish (p < 0.05), regardless of whether they were born in the UK, Ireland, or Australia. Ethnic differences in adult weight status in Australia do not appear to be fully explained by conventional risk factors. For some groups, but not all, living among others of the same ethnic group may proxy unmeasured health-promoting factors and these contexts, along with other factors that harm health (e.g. racial discrimination) warrant further investigation.


Deoxyguanosine is a TLR7 agonist.
Toll-like receptor 7 (TLR7) is an innate immune sensor for single-strand RNA (ssRNA). Recent structural analysis revealed that TLR7 has an additional binding site for nucleosides such as guanosine, and is activated when both guanosine and ssRNA bind. The nucleoside binding site also accommodates imidazoquinoline derivatives such as R848, which activate TLR7 in the absence of ssRNA. Here, we report that deoxyguanosine (dG) triggered cytokine production in murine bone marrow derived macrophages and plasmacytoid dendritic cells, as well as in human peripheral blood mononuclear cells, including type I interferons and pro-inflammatory factors such as TNF and IL-6. This signalling activity of dG was dependent on TLR7 and its adaptor MyD88 and did not require amplification via the type I interferon receptor. dG-triggered cytokine production required endosomal maturation but did not depend on the concurrent provision of RNA. We conclude that dG induces an inflammatory response through TLR7 and propose that dG is an RNA-independent TLR7 agonist.


Novel heterocyclic trans olefin analogues of N-{4-[4-(2,3-dichlorophenyl)piperazin-1-yl]butyl}arylcarboxamides as selective probes with high affinity for the dopamine D3 receptor.
Dopamine D3 receptor subtypes have been hypothesized to play a pivotal role in modulating the reinforcing and drug-seeking effects induced by cocaine. However, definitive pharmacological investigations have been hampered by the lack of highly D3 receptor selective compounds that can be used in vivo. To address this problem, the potent and D3-receptor-selective antagonist NGB 2904 (1, 9H-fluorene-2-carboxylic acid {4-[(2,3-dichlorophenyl)-piperazin-1-yl]-butyl}-amide, Ki (hD3) = 2.0 nM, Ki (hD2L) = 112 nM, D2/D3 selectivity ratio of 56) was chosen as a lead structure for chemical modification in an attempt to reduce its high lipophilicity (c log D = 6.94) while optimizing D3 receptor binding affinity and D2/D3 selectivity. A series of >30 novel analogues were synthesized, and their binding affinities were evaluated in competition binding assays in HEK 293 cells transfected with either D2(L), D3, or D4 human dopamine receptors using the high affinity, selective D2-like receptor antagonist (125)I-IABN. Structural diversity in the aryl amide end of the molecule was found to have a major influence on (sub)nanomolar D3 receptor affinity and D2/D3 selectivity, which was optimized using a more rigid trans-butenyl linker between the aryl amide and the piperazine. Several analogues demonstrated superior D3 receptor binding affinities and selectivities as compared to the parent ligand. Compound 29 (N-{4-[4-(2,3-dichlorophenyl)-piperazin-1-yl]-trans-but-2-enyl}-4-pyridine-2-yl-benzamide) displayed the most promising pharmacological profile (Ki (hD3) = 0.7 nM, Ki (hD2L) = 93.3 nM, D2/D3 selectivity ratio of 133). In addition, this ligand inhibited quinpirole stimulation of mitogenesis at human dopamine D3 receptors transfected into Chinese hamster ovary (CHO) cells, with an EC50 value of 3.0 nM. Compound 29 was a nearly 5 times more potent antagonist at the D3 receptor than 1 (EC50 = 14.4 nM). Moreover, a decrease in c log D value of approximately 2 orders of magnitude was determined for this novel D3-receptor-preferring ligand, compared to 1. In summary, chemical modification of 1 has resulted in compounds with high affinity and selectivity for D3 receptors. The most promising candidate, compound 29, is currently being evaluated in animal models of cocaine abuse and will provide an important tool with which to elucidate the role of D3 receptors in drug reinforcement in vivo.


Comparison of one- and two-stage basilic vein transposition for arterio-venous fistula formation in haemodialysis patients: preliminary results.
This study aimed to compare the results of one-and two-stage basilic vein transposition (BVT) in haemodialysis patients. This was a non-randomised, retrospective study between January 2007 and January 2012 on 96 patients who were diagnosed with end-stage renal failure (ESRF) (54 males, 42 females; mean age 43.6 ± 14 years) and underwent one- or two-stage BVT in our clinic. All patients who were not eligible for a native radio-cephalic or brachio-cephalic arterio-venous fistula (AVF) were scheduled for one- or two-stage BVT after arterial (brachial, radial and ulnar) and venous (basilic and cephalic) Doppler ultrasonography. Patients were retrospectively divided into two groups: group 1, basilic vein diameter > 3 mm and patients who underwent one-stage BVT; and group 2, basilic vein diameter < 3 mm and patients who underwent two-stage BVT. In group 1, the basilic vein with a single incision was anastomosed to the brachial artery, followed by superficialisation. In group 2, the basilic vein was anastomosed to the brachial artery and they underwent the superficialisation procedure one month postoperatively. Fistula maturation and postoperative complications were assessed. The mean diameter of the basilic vein was statistically significantly higher in group 1 (3.46 ± 0.2 mm) than in group 2 (2.79 ± 0.1 mm) (p < 0.05). In terms of postoperative complications, thrombosis, haemorrhage and haematoma were significantly higher in group 1 (34, 36 and 17%, respectively) than in group 2 (23, 14 and 6%, respectively) (p < 0.05). The rate of fistula maturation was significantly lower in group 1 (66%), compared to group 2 (77%) (p < 0.05).Time to fistula maturation was significantly shorter in group 1 (mean 41 ± 14 days), compared to group 2 (mean 64 ± 28 days) (p < 0.05). Two-stage BVT was superior to one-stage BVT due to its lower rate of postoperative complications and higher fistula maturation, despite its disadvantage of late fistula use. Although the diameter of the basilic vein was larger in patients who underwent one-stage BVT, we observed that one-stage BVT was disadvantageous in terms of postoperative complications and fistula maturation.


Learning and removing cast shadows through a multidistribution approach.
Moving cast shadows are a major concern for foreground detection algorithms. The processing of foreground images in surveillance applications typically requires that such shadows be identified and removed from the detected foreground. This paper presents a novel pixel-based statistical approach to model moving cast shadows of nonuniform and varying intensity. This approach uses the Gaussian mixture model (GMM) learning ability to build statistical models describing moving cast shadows on surfaces. This statistical modeling can deal with scenes with complex and time-varying illumination, including light saturated areas, and prevent false detection in regions where shadows cannot be detected. The proposed approach can be used with pixel-based descriptions of shadowed surfaces found in the literature. It significantly reduces their false detection rate without increasing the missed detection rate. Results obtained with different scene types and shadow models show the robustness of the approach.


Progression free survival rate at 9 and 18 weeks predict overall survival in patients with malignant pleural mesothelioma: an individual patient pooled analysis of 10 European Organisation for Research and Treatment of Cancer Lung Cancer Group studies and an independent study validation.
Response criteria have always been difficult to apply to malignant pleural mesothelioma (MPM), due to its unique pattern of growth. We developed some models to show that progression free survival rate (PFSR) could be a better predictor of overall survival (OS) than the response rate (RR) in MPM patients. The results were validated independently in the European Organisation for Research and Treatment of Cancer (EORTC) 08052, a phase II study in MPM. Individual patient data from 10 EORTC-Lung Cancer Group (LCG) studies of first-line chemotherapy in MPM were pooled. Response to therapy was assessed according to World Health Organisation (WHO) criteria in all except the two most recent trials, which used Response Evaluation Criteria in Solid Tumours (RECIST). Landmark analyses (LA) at 9 weeks and 18 weeks after registration/randomisation were performed to assess the association between PFSR and OS. Independent validation of the results was conducted in EORTC 08052 study (82 patients) employing the same LA. All 10 studies (N=523 patients) were included in the LA of PFSR at 9 and 18 weeks (PFSR-9 and PFSR-18). PFSR-9 and PFSR-18 were confirmed as predictors of OS, with hazard ratio (HR) of 0.37 (95% confidence interval (CI), 0.30-0.47) and 0.50 (0.38-0.65) and C-index of 0.62 and 0.58, respectively. In the validation study, 28.4% achieved CR/PR and 77.8% had disease control (CR/PR/SD) as their best overall response. PFSR-9 and PFSR-18 weeks were both strongly correlated with OS (HR of 0.35 [80% CI, 0.25-0.49] and 0.46 (0.32-0.67) and C-index of 0.66 and 0.60, respectively). PFSR-18 was strongly correlated and discriminated patients with better OS from the poorer prognosis patients. An earlier end-point, PFSR-9 was also strongly correlated to OS with better discriminating capacity. The results were independently validated.


Oral spore-based probiotic supplementation was associated with reduced incidence of post-prandial dietary endotoxin, triglycerides, and disease risk biomarkers.
To determine if 30-d of oral spore-based probiotic supplementation could reduce dietary endotoxemia. Apparently healthy men and women (n = 75) were screened for post-prandial dietary endotoxemia. Subjects whose serum endotoxin concentration increased by at least 5-fold from pre-meal levels at 5-h post-prandial were considered "responders" and were randomized to receive either placebo (rice flour) or a commercial spore-based probiotic supplement [Bacillus indicus (HU36), Bacillus subtilis (HU58), Bacillus coagulans, and Bacillus licheniformis, and Bacillus clausii] for 30-d. The dietary endotoxemia test was repeated at the conclusion of the supplementation period. Dietary endotoxin (LAL) and triglycerides (enzymatic) were measured using an automated chemistry analyzer. Serum disease risk biomarkers were measured using bead-based multiplex assays (Luminex and Milliplex) as secondary, exploratory measures. Data were statistically analyzed using repeated measures ANOVA and a P < 0.05. We found that spore-based probiotic supplementation was associated with a 42% reduction in endotoxin (12.9 ± 3.5 vs 6.1 ± 2.6, P = 0.011) and 24% reduction in triglyceride (212 ± 28 vs 138 ± 12, P = 0.004) in the post-prandial period Placebo subjects presented with a 36% increase in endotoxin (10.3 ± 3.4 vs 15.4 ± 4.1, P = 0.011) and 5% decrease in triglycerides (191 ± 24 vs 186 ± 28, P = 0.004) over the same post-prandial period. We also found that spore-based probiotic supplementation was associated with significant post-prandial reductions in IL-12p70 (24.3 ± 2.2 vs 21.5 ± 1.7, P = 0.017) and IL-1β (1.9 ± 0.2 vs 1.6 ± 0.1, P = 0.020). Compared to placebo post supplementation, probiotic subject had less ghrelin (6.8 ± 0.4 vs 8.3 ± 1.1, P = 0.017) compared to placebo subjects. The key findings of the present study is that oral spore-based probiotic supplementation reduced symptoms indicative of "leaky gut syndrome".


Is the involvement of opinion leaders in the implementation of research findings a feasible strategy?
There is only limited empirical evidence about the effectiveness of opinion leaders as health care change agents. To test the feasibility of identifying, and the characteristics of, opinion leaders using a sociometric instrument and a self-designating instrument in different professional groups within the UK National Health Service. Postal questionnaire survey. All general practitioners, practice nurses and practice managers in two regions of Scotland. All physicians and surgeons (junior hospital doctors and consultants) and medical and surgical nursing staff in two district general hospitals and one teaching hospital in Scotland, as well as all Scottish obstetric and gynaecology, and oncology consultants. Using the sociometric instrument, the extent of social networks and potential coverage of the study population in primary and secondary care was highly idiosyncratic. In contrast, relatively complex networks with good coverage rates were observed in both national specialty groups. Identified opinion leaders were more likely to have the expected characteristics of opinion leaders identified from diffusion and social influence theories. Moreover, opinion leaders appeared to be condition-specific. The self-designating instrument identified more opinion leaders, but it was not possible to estimate the extent and structure of social networks or likely coverage by opinion leaders. There was poor agreement in the responses to the sociometric and self-designating instruments. The feasibility of identifying opinion leaders using an off-the-shelf sociometric instrument is variable across different professional groups and settings within the NHS. Whilst it is possible to identify opinion leaders using a self-designating instrument, the effectiveness of such opinion leaders has not been rigorously tested in health care settings. Opinion leaders appear to be monomorphic (different leaders for different issues). Recruitment of opinion leaders is unlikely to be an effective general strategy across all settings and professional groups; the more specialised the group, the more opinion leaders may be a useful strategy.


Cruciform DNA structure underlies the etiology for palindrome-mediated human chromosomal translocations.
There is accumulating evidence to suggest that palindromic AT-rich repeats (PATRRs) represent hot spots of double-strand breakage that lead to recurrent chromosomal translocations in humans. As a mechanism for such rearrangements, we proposed that the PATRR forms a cruciform structure that is the source of genomic instability. To test this hypothesis, we have investigated the tertiary structure of a cloned PATRR. We have observed that a plasmid containing this PATRR undergoes a conformational change, causing temperature-dependent mobility changes upon agarose gel electrophoresis. The mobility shift is observed in physiologic salt concentrations and is most prominent when the plasmid DNA is incubated at room temperature prior to electrophoresis. Analysis using two-dimensional gel electrophoresis indicates that the mobility shift results from the formation of a cruciform structure. S1 nuclease and T7 endonuclease both cut the plasmid into a linear form, also suggesting cruciform formation. Furthermore, anti-cruciform DNA antibody reduces the electrophoretic mobility of the PATRR-containing fragment. Finally, we have directly visualized cruciform extrusions from the plasmid DNA with the size expected of hairpin arms using atomic force microscopy. Our data imply that for human chromosomes, translocation susceptibility is mediated by PATRRs and likely results from their unstable conformation.


Functional effects of GRM1 suppression in human melanoma cells.
Ectopic expression of a neuronal receptor, metabotropic glutamate receptor 1 (Grm1), in melanocytes has been implicated in melanoma development in mouse models. The human relevance of this receptor's involvement in melanoma pathogenesis was shown by detecting GRM1 expression in subsets of human melanomas, an observation lacking in benign nevi or normal melanocytes. Grm1-transformed mouse melanocytes and a conditional Grm1 transgenic mouse model confirmed a requirement for sustained expression of Grm1 for the maintenance of transformed phenotypes in vitro and tumorigenicity in vivo. Here, we investigate if continued GRM1 expression is also required in human melanoma cell lines by using two inducible, silencing RNA systems: the ecdysone/Ponasterone A and tetracycline on/off approaches to regulate GRM1 expression in the presence of each inducer. Various in vitro assays were conducted to assess the consequences of a reduction in GRM1 expression on cell proliferation, apoptosis, downstream targeted signaling pathways, and in vivo tumorigenesis. We showed that suppression of GRM1 expression in several human melanoma cell lines resulted in a reduction in the number of viable cells and a decrease in stimulated mitogen-activated protein kinase (MAPK) and PI3K/AKT and suppressed tumor progression in vivo. These results reinforce earlier observations where a reduction in cell growth in vitro and tumorigenesis in vivo were correlated with decreased GRM1 activities by pharmacologic inhibitors of the receptor, supporting the notion that GRM1 plays a role in the maintenance of transformed phenotypes in human melanoma cells in vitro and in vivo and could be a potential therapeutic target for the treatment of melanoma.


A Non-Reciprocal Autosomal Translocation 64,XX, t(4;10)(q21;p15) in an Arabian Mare with Repeated Early Embryonic Loss.
Balanced autosomal translocations are a known cause for repeated early embryonic loss (REEL) in horses. In most cases, carriers of such translocations are phenotypically normal, but the chromosomal aberration negatively affects gametogenesis giving rise to both genetically balanced and unbalanced gametes. The latter, if involved in fertilization, result in REEL, whereas gametes with the balanced form of translocation will pass the defect into next generation. Therefore, in order to reduce the incidence of REEL, identification of translocation carriers is critical. Here, we report about a phenotypically normal 3-year-old Arabian mare that had repeated resorption of conceptuses prior to day 45 of gestation and was diagnosed with REEL. Conventional and molecular cytogenetic analyses revealed that the mare had normal chromosome number 64,XX but carried a non-mosaic and non-reciprocal autosomal translocation t(4;10)(q21;p15). This is a novel translocation described in horses with REEL and the first such report in Arabians. Previous cases of REEL due to autosomal translocations have exclusively involved Thoroughbreds. The findings underscore the importance of routine cytogenetic screening of breeding animals.


Character independent font recognition on a single Chinese character.
A novel algorithm for font recognition on a single unknown Chinese character, independent of the identity of the character, is proposed in this paper. We employ a wavelet transform on the character image and extract wavelet features from the transformed image. After a Box-Cox transformation and LDA (Linear Discriminant Analysis) process, the discriminating features for font recognition are extracted and classified through a MQDF (Modified Quadric Distance Function) classifier with only one prototype for each font class. Our experiments show that our algorithm can achieve a recognition rate of 90.28 percent on a single unknown character and 99.01 percent if five characters are used for font recognition. Compared with existing methods, all of which are based on a text block, our method can provide a higher recognition rate and is more flexible and robust, since it is based on a single unknown character. Additionally, our method demonstrates that it is possible to extract subtle yet discriminative signals embedded in a much larger noisy background.


Consent for long-term storage of blood samples by Indigenous Australian research participants: the DRUID Study experience.
Little is known about the characteristics of people who do and do not agree to the long-term storage and use of their biological materials, or about potential biases that may be introduced as a result of differential consent. More specifically, concerns about tissue storage and use are especially relevant among population groups for whom blood and other biological materials are culturally significant, such as Indigenous Australians. Using data from a 2003-2005 study of 1,004 Indigenous Australians, we examined participants' choices regarding long-term storage of excess blood for possible use in future studies. Overall, 55% of participants agreed to long-term storage. Among 854 participants with a fasting blood sample and completed questionnaire, consent for storage was more likely among those aged 45+ years than those 15-44 (odds ratio (OR) = 1.55, 95% confidence interval (CI): 1.14, 2.11), and was similar for males and females. After adjustment for age and other covariates using logistic regression, consent was more likely for never smokers than current smokers (OR = 1.48, 95% CI: 1.04, 2.10), those reporting any non-Indigenous grandparent(s) (OR = 2.07, 95% CI: 1.50, 2.85), and those whose consent form was administered/witnessed by an Indigenous staff member (OR 1.43, 95% CI: 1.05, 1.94). Consent for long-term storage was associated with only small differences (generally less than +/- 5%) in the results of assays performed on all participants' blood samples as part of the baseline health examination. These data show that consent for blood storage among these research participants was neither rare nor universal. It was associated with some socio-demographic/cultural factors but not with blood biochemistry. Decisions about requesting or giving consent for storage and later use of tissue samples must recognize a number of important, and potentially competing, ethical and logistical considerations.


Lack of familial aggregation of Parkinson disease and Alzheimer disease.
To investigate the risk of Alzheimer disease (AD) in first-degree relatives of patients with Parkinson disease (PD) compared with first-degree relatives of controls. Case-control study, family history method, and reconstructed cohort approach. Probands with PD without dementia and control probands, matched by age strata, sex, and ethnicity, were examined in person and enrolled without knowledge of family history of PD and other neurological disorders. Disease status in first-degree relatives of probands with PD and control probands was ascertained through a structured family history interview administered to the proband and a second informant (self-report or another informant). Cox proportional hazards models with double-censoring techniques for missing information on age of onset of AD were used to analyze the risk of AD in first-degree relatives of patients with PD compared with first-degree relatives of controls. Four hundred eighty-seven probands with PD and 409 control probands provided family history information on 4819 first-degree relatives older than 30 years (2534 relatives of probands with PD and 2285 relatives of control probands). One hundred thirteen first-degree relatives (2.3%; 61 relatives [2.4%] of patients with PD and 52 relatives [2.3%] of controls) were diagnosed with AD. The risk of AD was not increased in relatives of patients with PD compared with relatives of controls (hazard ratio, 1.1; 95% confidence interval, 0.7-1.6; P =.65). Similarly, no significantly increased risk of AD was observed when comparing relatives of patients with early-onset (< or =50 years) and late-onset (>50 years) PD with relatives of controls. The lack of familial aggregation of PD and AD does not support the hypothesis of major shared genetic contributions to the etiology of the 2 most common neurodegenerative disorders.