Goal-directed versus outcome-based financial incentives for weight loss among low-income patients with obesity: rationale and design of the Financial Incentives foR Weight Reduction (FIReWoRk) randomised controlled trial.
Obesity is a major public health challenge and exacerbates economic disparities through employment discrimination and increased personal health expenditures. Financial incentives for weight management may intensify individuals' utilisation of evidence-based behavioural strategies while addressing obesity-related economic disparities in low-income populations. Trials have focused on testing incentives contingent on achieving weight loss outcomes. However, based on social cognitive and self-determination theories, providing incentives for achieving intermediate behavioural goals may be more sustainable than incentivising outcomes if they enhance an individual's skills and self-efficacy for maintaining long-term weight loss. The objective of this paper is to describe the rationale and design of the Financial Incentives foR Weight Reduction study, a randomised controlled trial to test the comparative effectiveness and cost-effectiveness of two financial incentive strategies for weight loss (goal directed vs outcome based) among low-income adults with obesity, as well as compared with the provision of health behaviour change resources alone. We are recruiting 795 adults, aged 18-70 years with a body mass index ≥30 kg/m2, from three primary care clinics serving residents of socioeconomically disadvantaged neighbourhoods in New York City and Los Angeles. All participants receive a 1-year commercial weight loss programme membership, self-monitoring tools (bathroom scale, food journal and Fitbit Alta HR), health education and monthly check-in visits. In addition to these resources, those in the two intervention groups can earn up to $750 over 6 months for: (1) participating in an intensive weight management programme, self-monitoring weight and diet and meeting physical activity guidelines (goal-directed arm); or (2) a ≥1.5% to ≥5% reduction in baseline weight (outcome-based arm). To maximise incentive efficacy, we incorporate concepts from behavioural economics, including immediacy of payments and framing feedback to elicit regret aversion. We will use generalised mixed effect models for repeated measures to examine intervention effects on weight at 6, 9 and 12 months. Human research protection committees at New York University School of Medicine, University of California Los Angeles (UCLA) David Geffen School of Medicine and Olive-View-UCLA Medical Center granted ethics approval. We will disseminate the results of this research via peer-reviewed publications, conference presentations and meetings with stakeholders. NCT03157713.


Spectroscopy of element 115 decay chains.
A high-resolution α, x-ray, and γ-ray coincidence spectroscopy experiment was conducted at the GSI Helmholtzzentrum für Schwerionenforschung. Thirty correlated α-decay chains were detected following the fusion-evaporation reaction 48Ca + 243Am. The observations are consistent with previous assignments of similar decay chains to originate from element Z=115. For the first time, precise spectroscopy allows the derivation of excitation schemes of isotopes along the decay chains starting with elements Z>112. Comprehensive Monte Carlo simulations accompany the data analysis. Nuclear structure models provide a first level interpretation.


In vivo retinal imaging by optical coherence tomography.
We describe what are to our knowledge the first in vivo measurements of human retinal structure with optical coherence tomography. These images represent the highest depth resolution in vivo retinal images to date. The tomographic system, image-processing techniques, and examples of high-resolution tomographs and their clinical relevance are discussed.


Long-term maternal atherosclerotic morbidity in women with pre-eclampsia.
To investigate whether severe and recurrent pre-eclampsia increase the risk for long-term maternal atherosclerotic disease. A population-based study compared the incidence of long-term atherosclerotic morbidity in a cohort of women who delivered in the years 1988-2012. The exposure variable was pre-eclampsia. Mean follow-up duration was 11.2 years. Kaplan-Meier survival curves were used to estimate cumulative incidence of simple, complex (ie, angina pectoris and congestive heart failure, respectively) cardiovascular-related and renal-related hospitalisations. Cox proportional hazards models were used to estimate the adjusted HRs for cardiovascular and renal morbidity. During the study, 96 370 patients met the inclusion criteria; 7824 (8.1%) in patients who were diagnosed at least once with pre-eclampsia. Patients with pre-eclampsia had higher rates of cardiovascular morbidity including cardiac non-invasive (OR 1.4; 95% CI 1.1 to 1.7; p=0.005) and invasive diagnostic procedures (OR 1.7; 95% CI 1.2 to 2.3; p=0.001), simple (OR 1.5; 95% CI 1.2 to 1.8; p=0.001), as well as complex cardiovascular events (OR 2.4; 95% CI 2.2 to 2.8; p=0.001) and renal (OR 3.7; 95% CI 2.2 to 6.0; p=0.001) hospitalisations. A significant linear association was noted between the severity of pre-eclampsia (no pre-eclampsia, mild pre-eclampsia, severe pre-eclampsia and eclampsia) and cardiovascular (2.7% vs 4.5% vs 5.2% vs 5.7%, respectively; p=0.001), as well as renal disease (0.1% vs 0.2% vs 0.5% vs 1.1%, respectively; p=0.001). Likewise, a linear association was found between the number of previous pregnancies with pre-eclampsia (no pre-eclampsia, one event and ≥2 events of pre-eclampsia) and risk for future simple cardiovascular disease (1.2% vs 1.6% vs 2.2%, respectively; p=0.001), complex cardiovascular disease (1.3% vs 2.7% vs 4.6%, respectively; p=0.001) and total cardiovascular hospitalisations (2.7% vs 4.4% vs 6.0%, respectively; p=0.001). Using a Kaplan-Meier survival curve, patients with pre-eclampsia had significantly higher cumulative incidence of atherosclerotic-related hospitalisations. In a Cox proportional hazards model, adjusted for confounders such as maternal age, parity, diabetes mellitus and obesity, pre-eclampsia remained independently associated with atherosclerotic hospitalisations. Previous pregnancy with pre-eclampsia is an independent risk factor for long-term maternal atherosclerotic morbidity. The risk is more substantial for patients with severe and recurrent episodes of pre-eclampsia.


Involvement of phi29 DNA polymerase thumb subdomain in the proper coordination of synthesis and degradation during DNA replication.
Phi29 DNA polymerase achieves a functional coupling between its 3'-5' exonuclease and polymerization activities by means of important contacts with the DNA at both active sites. The placement and orientation of residues Lys538, Lys555, Lys557, Gln560, Thr571, Thr573 and Lys575 in a modelled phi29 DNA polymerase-DNA complex suggest a DNA-binding role. In addition, crystal structure of phi29 DNA polymerase-oligo (dT)5 complex showed Leu567, placed at the tip of the thumb subdomain, lying between the two 3'-terminal bases at the exonuclease site. Single replacement of these phi29 DNA polymerase residues by alanine was made, and mutant derivatives were overproduced and purified to homogeneity. The results obtained in the assay of their synthetic and degradative activities, as well as their coordination, allow us to propose: (1) a primer-terminus stabilization role at the polymerase active site for residues Lys538, Thr573 and Lys575, (2) a primer-terminus stabilization role at the exonuclease active site for residues Leu567 and Lys555 and (3) a primer-terminus binding role in both editing and polymerization modes for residue Gln560. The results presented here lead us to propose phi29 DNA polymerase thumb as the main subdomain responsible for the coordination of polymerization and exonuclease activities.


[The knowledge acquired during and adherence to a postgraduate course concerned with mental health aimed at school counselors].
To evaluate the academic performance in, and adherence to, a postgraduate course focused on promotion, prevention, identification, and early intervention with regard to the different mental problems and pathologies affecting children and adolescents. This was a before-and-after intervention study involving a postgraduate course. The population of school counselors from the Colombian regions of Boyacá and Cundinamarca were invited to participate in a postgraduate course that addressed mental health. Their knowledge was evaluated before and after having received it. The school counselors performed well on the academic test, with 48.5 % of the population obtaining a score of 4.0/5.0 before the intervention. After the course, 61.2 % achieved the same score. It was thus considered that the course functioned as a tool for improving knowledge about promotion, prevention, recognition, and early intervention with regard to mental health in educational institutions. There was a 90.6 % adherence to the 120-hour-long postgraduate course. The postgraduate course seemed to have a positive impact in terms of the knowledge acquired about mental health, and there was good adherence to the course.


Social and economic impact of diabetics in Bangladesh: protocol for a case-control study.
Diabetes affects both individuals and their families and has an impact on economic and social development of a country. Information on the availability, cost, and quality of medical care for diabetes is mostly not available for many low- and middle-income countries including Bangladesh. Complications from diabetes, which can be devastating, could largely be prevented by wider use of several inexpensive generic medicines, simple tests and monitoring and can be a cost saving intervention. This study will provide an in-depth and comprehensive picture of social and economic impacts of diabetes in Bangladesh and propose clear recommendations for improving prevention and management of diabetes. The objectives of the study are: 1) To study the association between diabetes and other health problems and its social impacts. 2) To estimate the economic impact of diabetes including total direct and indirect costs. 3) To measure the impact of diabetes on quality of life among diabetes patients in Bangladesh. 4) To study the impact of diabetes on the health care system This is a case-control study comparing cases with type 2 diabetes to controls without diabetes matched on age, sex and place of residence. 564 cases and 564 controls will be selected from the outpatient department of a tertiary hospital in Dhaka, Bangladesh. Data on socioeconomic status, health utility index, direct and indirect costs for diabetes, medication adherence, quality of life, treatment satisfaction, diet, physical activity, mental state examination, weight, height, hip and waist circumference, blood pressure, pulse, medication history, laboratory data and physical examination will be conducted. The primary outcome measures will be association between diabetes and other health problems, cost of diabetes, impact of diabetes on quality of life and secondary outcome measures are impact of diabetes on healthcare systems in Bangladesh. This study will provide an in-depth and comprehensive picture of social and economic impacts of diabetics in Bangladesh and propose clear recommendations for improving prevention and management of diabetics. It will help to develop programs and policies for better management of Diabetics and cost effective strategies in Bangladesh context.


Retrospective study of triclosan and methyl-triclosan residues in fish and suspended particulate matter: results from the German Environmental Specimen Bank.
A retrospective monitoring of triclosan (TCS; period 1994-2003 and 2008) and its potential transformation product methyl-triclosan (MTCS; period 1994-2008) was performed using archived fish samples from German rivers (16 sites, including Elbe and Rhine). At four of these sites suspended particulate matter (SPM) was also investigated covering the period 2005-2007. Samples were analyzed by GC/MS, either directly (MTCS) or after derivatization (TCS). TCS burdens of fish muscle tissue ranged from <0.2-3.4 ng g(-1) ww (wet weight; corresponding to <2-69 ng g(-1) lw, lipid weight) without apparent concentration trends over time. MTCS was detected at considerably higher concentrations in fish ranging from 1.0-33 ng g(-1) ww (47-1010 ng g(-1) lw) and increased until about 2003-2005. Thereafter, concentrations generally were lower, although at some sites single higher values were observed in recent years. In SPM, decreasing MTCS concentrations in the range 1-4 ng g(-1) dry weight were detected while TCS was always below the limit of quantification. Assuming that MTCS concentrations are correlated to TCS consumption, the observed decrease in MTCS levels may be partly a result of the voluntary renunciation of TCS use in detergents for, e.g., laundry or dishwashing declared by a manufacturers' association in 2001. Because of a lack of ecotoxicity studies for MTCS, a QSAR-derived predicted no effect concentration (PNEC) was compared to averaged ambient water concentrations of fish which were calculated from maximum tissue residues by applying an appropriate bioconcentration factor from literature. Since these calculated water concentrations were below the PNEC it is assumed that MTCS alone poses no immediate risk to aquatic organism. The conversion to a PNEC for SPM organisms and comparison with detected SPM levels of MTCS also revealed no risk.


Bone morphogenetic protein-6 enhances gonadotrophin-dependent progesterone and inhibin secretion and expression of mRNA transcripts encoding gonadotrophin receptors and inhibin/activin subunits in chicken granulosa cells.
The aims were to examine ovarian expression of bone morphogenetic protein (BMP) ligands/receptor mRNAs in the chicken and to test the hypothesis that theca-derived BMP(s) modulates granulosa cell function in a paracrine manner. RT-PCR revealed expression of multiple BMPs in granulosa and theca cells from pre hierarchical and preovulatory follicles with greater expression in theca cells; both cell types expressed BMP receptors-IA, -IB and -II consistent with tissue responsiveness. Preovulatory granulosa cells (F1, F2 and F3/4) were cultured with BMP-6 (expressed by theca but not granulosa) in the presence/absence of LH, FSH or 8-Br-cAMP. BMP-6 increased 'basal' and gonadotrophin-induced inhibin-A and progesterone secretion by each cell type but did not enhance the effect of 8-Br-cAMP. This indicates that the observed synergism between BMP-6 and gonadotrophin might involve BMP-induced up-regulation of gonadotrophin receptors. In support of this, BMP-6 alone increased LH-receptor (LHR) mRNA in F1 cells and FSH-receptor (FSHR) mRNA in F1, F2 and F3/4 cells. BMP-6 also enhanced LH/FSH-induced LHR transcript amount in each cell type but did not raise FSHR transcript amounts above those induced by BMP-6 alone. To further explore BMP-6 action on inhibin-A secretion, we quantified inhibin/activin subunits (alpha, beta(A), beta(B)) mRNAs. Consistent with its effect on inhibin-A secretion, BMP-6 enhanced 'basal' expression of alpha- and beta(A)-subunit mRNA in F1, F2 and F3/4 cells, and beta(B)-subunit mRNA in F3/4 cells. BMP-6 markedly enhanced FSH/LH-induced expression of alpha-subunit in all follicles and FSH-induced beta(A)-subunit in F2 and F3/4 follicles but not in F1 follicles. Neither BMP-6 alone, nor FSH/LH alone, affected 'basal' beta(B) mRNA abundance. However, co-treatment with gonadotrophin and BMP-6 greatly increased beta(B)-subunit expression, the response being lowest in F1 follicles and greatest in F3/4 follicles. Collectively, these results support the hypothesis that intraovarian BMPs of thecal origin have a paracrine role in modulating granulosa cell function in the chicken in a preovulatory stage-dependent manner.


Patterns of endogenous steroids in apathetic refugee children are compatible with long-term stress.
During the last few years, a number of children of asylum applicants in Sweden developed an apathetic or unconscious state. The syndrome was perceived as new, and various explanations were advanced such as factitious disorder, intoxication, or stress. Considering a potential association between traumatic stress and regulation of steroids biosynthesis, this study explored whether changes in concentrations of endogenous steroids were associated with the above syndrome. Eleven children were recruited in the study. Concentrations of steroids in blood samples were determined using high sensitivity liquid chromatography tandem mass spectrometry methods. Symptoms were assessed with a clinical rating scale developed for the study. Steroid concentrations were measured at the entry into study and after recovery; and concentrations were evaluated for the association with the symptoms in apathetic children. Cortisol and cortisone concentrations at baseline were negatively associated with duration of the symptoms from entry into the study to clinical recovery. Higher concentrations of pregnanes (pregnenolone, 17-OH-pregnenolone, and dehydroepiandrosterone) were observed in the symptomatic state and the concentrations decreased after the recovery. Pattern of low cortisol concentrations found in apathetic children is consistent with long-term stress. An increase of upstream steroid metabolites (pregnanes) was found to be associated with the symptomatic state.


Is England facing an opioid epidemic?
The opioid crisis in the United States (US) is one of the most high-profile public health scandals of the 21st century with millions of people unknowingly becoming dependent on opioids. The United Kingdom (UK) had the world's highest rate of opioid consumption in 2019, and opiate-related drug poisoning deaths have increased by 388% since 1993 in England and Wales. This article explores the epidemiological definitions of public health emergencies and epidemics in the context of opioid use, misuse, and mortality in England, to establish whether England is facing an opioid crisis.


Multimode Interference of Bloch Surface Electromagnetic Waves.
Integrated photonics aims at on-chip controlling light in the micro- and nanoscale ranges utilizing the waveguide circuits, which include such basic elements as splitters, multiplexers, and phase shifters. Several photonic platforms, including the well-developed silicon-on-insulator and surface-plasmon polaritons ones, operate well mostly in the IR region. However, operating in the visible region is challenging because of the drawbacks originating from absorption or sophisticated fabrication technology. Recently, a new promising all-dielectric platform based on Bloch surface electromagnetic waves (BSWs) in multilayer structures and functioning in the visible range has emerged finding a lot of applications primarily in sensing. Here, we show the effect of multimode interference (MMI) of BSWs and propose a method for implementing the advanced integrated photonic devices on the BSW platform. We determine the main parameters of MMI effect and demonstrate the operation of Mach-Zehnder interferometers with a predefined phase shift proving the principle of MMI BSW-based photonics in the visible spectrum. Our research will be useful for further developing a versatile toolbox of the BSW platform devices which can be essential in integrated photonics, lab-on-chip, and sensing applications.


Full-electron ligand-to-ligand charge transfer in a compact Re(I) complex.
Ligand-to-ligand charge transfer (LLCT) states in transition metal complexes are often characterized by fractional electron transfer due to coupling of the LLCT state with many other states via the metal. We designed and characterized a compact Re(I) complex that displays essentially full-electron charge transfer in the LLCT excited state. The complex, [Re(DCEB)(CO)3(L)](+) (DCEB = 4,4'-dicarboxyethyl-2,2'-bipyridine), referred to as ReEBA, features two redox active ligands with strong electron accepting (DCEB) and electron donating (L is 3-dimethylaminobenzonitrile (3DMABN)) properties. The lowest energy excited state formed with a ca. 10 ps time constant and was characterized as the full-electron 3DMABN → DCEB LLCT state using time-resolved infrared spectroscopy (TRIR), transient absorption spectroscopy, and DFT computations. Analysis of a range of vibrational modes helped to assign the charge transfer characteristics of the complex. The LLCT state lifetime in ReEBA shows a strong dependence on the solvent polarity and features solvent dependent frequency shifts for several vibrational reporters. The formation of a full-electron LLCT state (∼92%) was enabled by tuning the redox properties of the electron accepting ligand (DCEB) and simultaneously decoupling the redox active group of the electron donating ligand (3DMABN) from the metal center. This strategy is generally applicable for designing compact transition metal complexes that have full-electron LLCT states.


Inflammatory responses of Nile tilapia Oreochromis niloticus to Streptococcus agalactiae: effects of vaccination and yeast diet supplement.
This study evaluated the effects of dietary supplementation with 0.3% Saccharomyces cerevisiae yeast cell wall and of vaccination against Streptococcus agalactiae on the cellular component of acute inflammation induced in the coelomic cavity of Nile tilapia Oreochromis niloticus and on survival of the fish after challenge. A total of 84 tilapia of mean (±SD) weight 125.0 ± 1.5 g were distributed among twelve 310 l fiberglass tanks according to a 2 × 2 × 3 factorial design in the following manner: with and without supplementation; 2 stimulations (oily solution without S. agalactiae vaccine and vaccination); 15 d later all fish were intracoelomically challenged with 108 CFU ml-1 of a homologous strain of S. agalactiae, and evaluated after 6, 24 and 48 h, with 7 replicates. The fish received the non-supplemented or supplemented diet for a total of 77 d. The vaccination was performed on the 60th day, intracoelomically, as a single injection of 0.5 ml of the vaccine containing 108 CFU ml-1. Fifteen days later, all the fish were challenged with S. agalactiae by means of an intracoelomic inoculation of 108 CFU ml-1. No mortality was observed among the supplemented fish. The fish that were fed the non-supplemented diet and immunized with the bacterium presented a mortality rate of 28.5%. Among the non-supplemented and non-immunized fish, the mortality rate was 38.09%. Supplementation, in both vaccinated and non-vaccinated fish, induced larger accumulations of thrombocytes, lymphocytes and macrophages at the inflammatory focus. The results suggest that supplementation with 0.3% yeast cell wall, in both vaccinated and non-vaccinated fish, improved the inflammatory response of the fish and protected against the challenge. Vaccination increased the defense response, but the effect was stronger when associated with supplementation with S. cerevisiae.


Moving Well-Being Well: Investigating the maturation of fundamental movement skill proficiency across sex in Irish children aged five to twelve.
Fundamental movement skills (FMS) are the basic building blocks of more advanced, complex movements required to participate in physical activity. This study examined FMS proficiency across the full range of Irish primary school children (n = 2098, 47% girls, age range 5-12 years). Participants were assessed using the Test of Gross Motor Development, 3rd edition (TGMD-3), Victorian Fundamental Movement skills manual, and the balance subtest from the Bruininks-Oseretsky Test of Motor Proficiency 2 (BOT-2). Independent sample t-tests and a one way between groups ANOVA with planned comparisons were used analyse sex and age differences. Mastery or near mastery of skills ranged from 16% for overhand throw, to 75.3% for run. Girls scored significantly higher than boys in the locomotor and balance subtests with the boys outperforming the girls in object control skills. Improvements in ability can be seen over time (F(8,1968) = 70.18, p < 0.001), with significant increases in FMS proficiency seen up to the age of 10, after which proficiency begins to decline. The findings demonstrate the low levels of FMS proficiency amongst Irish primary school children, the differences between sex that exist, and highlights the need for more programmes that focus on developing these FMS at an early age.


Vulnerability factors in OCD symptoms: cross-cultural comparisons between Turkish and Canadian samples.
Recent findings have suggested some potential psychological vulnerability factors for development of obsessive-compulsive (OC) symptoms, including cognitive factors of appraisal and thought control, religiosity, self-esteem and personality characteristics such as neuroticism. Studies demonstrating these associations usually come from Western cultures, but there may be cultural differences relevant to these vulnerability factors and OC symptoms. The present study examined the relationship between putative vulnerability factors and OC symptoms by comparing non-clinical samples from Turkey and Canada, two countries with quite different cultural characteristics. The findings revealed some common correlates such as neuroticism and certain types of metacognition, including appraisals of responsibility/threat estimation and perfectionism/need for certainty, as well as thought-action fusion. However, culture-specific factors were also indicated in the type of thought control participants used. For OC disorder symptoms, Turkish participants were more likely to utilize worry and thought suppression, while Canadian participants tended to use self-punishment more frequently. The association with common factors supports the cross-cultural validity of some factors, whereas unique factors suggest cultural features that may be operative in cognitive processes relevant to OC symptoms.


Synergistic effects of colchicine combined with atorvastatin in rats with hyperlipidemia.
Inflammation and endothelial dysfunction is implicated in the atherosclerosis initiation and progression in the setting of hyperlipidemia. Colchicine is a potent anti-inflammatory agent and whether colchicine combined with atorvastatin has synergistic effects on inflammation amelioration and endothelial function improvement is unknown. Hyperlipidemic rat model was produced by high-fat and high-cholesterol diet for 6 weeks. Rats with normal diet were served as shame group. In hyperlipidemic group, normal saline, atorvastatin (10 mg/kg body weight/day), colchicines (0.5 mg/kg body weight/day), or atorvastatin combined with colchicines (same dosages) were prescribed for 2 weeks. Serum levels of lipid profile, C-reactive protein (CRP), liver enzyme, lipoprotein associated phospholipase A2 (Lp-PLA2) and nitric oxide (NO) production were serially assessed. Before the beginning of the study, all laboratory variables were comparable among each group. After 6 weeks of hyperlipidemic model production, serum levels of cholesterols, CRP and Lp-PLA2 were significantly increased when compared to sham group, whereas NO production was reduced. With 2 weeks of colchicine therapy, serum levels of CRP and Lp-PLA2 were decreased and NO production was enhanced in the colchicine group in a lipid-lowering independent manner. Added colchicine into atorvastatin therapy further improved NO production and decreased CRP and Lp-PLA2 levels, indicating a potential synergism of colchicine and atorvastatin. Colchicine combined with atorvastatin may have stronger protective effects on improving endothelial function and ameliorating inflammation in rats with hyperlipidemia.


Interaction-Temporal GCN: A Hybrid Deep Framework For Covid-19 Pandemic Analysis.
The Covid-19 pandemic is still spreading around the world and seriously imperils humankind's health. This swift spread has caused the public to panic and look to scientists for answers. Fortunately, these scientists already have a wealth of data-the Covid-19 reports that each country releases, reports with valuable spatial-temporal properties. These data point toward some key actions that humans can take in their fight against Covid-19. Technically, the Covid-19 records can be described as sequences, which represent spatial-temporal linkages among the data elements with graph structure. Therefore, we propose a novel framework, the Interaction-Temporal Graph Convolution Network (IT-GCN), to analyze pandemic data. Specifically, IT-GCN introduces ARIMA into GCN to model the data which originate on nodes in a graph, indicating the severity of the pandemic in different cities. Instead of regular spatial topology, we construct the graph nodes with the vectors via ARIMA parameterization to find out the interaction topology underlying in the pandemic data. Experimental results show that IT-GCN is able to capture the comprehensive interaction-temporal topology and achieve well-performed short-term prediction of the Covid-19 daily infected cases in the United States. Our framework outperforms state-of-art baselines in terms of MAE, RMSE and MAPE. We believe that IT-GCN is a valid and reasonable method to forecast the Covid-19 daily infected cases and other related time-series. Moreover, the prediction can assist in improving containment policies.


Protein Semisynthesis Provides Access to Tau Disease-Associated Post-translational Modifications (PTMs) and Paves the Way to Deciphering the Tau PTM Code in Health and Diseased States.
The microtubule-associated protein Tau plays a central role in neurodegeneration and is a leading therapeutic target for the treatment of Alzheimer's disease (AD). Several lines of evidence suggest that post-translational modifications (PTMs) regulate the function(s) of Tau, including its subcellular localization, clearance, aggregation, toxicity, and pathological spreading. However, the lack of tools and methodologies that allow site-specific introduction of PTMs in Tau have limited our ability to dissect the role of PTMs in regulating Tau functions in health and disease. To facilitate deciphering the Tau PTM code, we have developed, for the first time, semisynthetic strategies that allow for the site-specific introduction of single or multiple physiological or disease-associated PTMs that occur within residues 246-441 of Tau, which includes the microtubule-binding domain (MTBD). As a proof of concept, we produced unmodified Tau and three Tau variants with single or multiple disease-associated PTMs that were not previously accessible as homogeneously modified proteins, AcK280, pY310, and pS396/pS404. We then focused on investigating the effect of acetylation at lysine 280 (AcK280) on the structure, aggregation, and microtubule binding properties of Tau. Our results show that site-specific acetylation at K280 significantly enhances the aggregation rate of Tau and impairs microtubule assembly. Surprisingly, compared with unmodified Tau, which forms long and flexible filaments, AcK280 Tau forms predominantly globular oligomers and short fibrils (<200 nm) that exhibit a reduced propensity to assemble into long filaments. These findings are consistent with the increased aggregation propensity and pathogenicity of this mutant in animal models of AD and suggest that acetylation at this residue might enhance the seeding capacity or formation of toxic Tau species in vivo. Beyond acetylation and phosphorylation, the development of this semisynthetic strategy provides new opportunities to investigate other types of Tau PTMs and to study the cross-talk between PTMs that occurs within residues 246-441, which were previously inaccessible, thereby paving the way to deciphering the Tau PTM code in health and disease.


Effect of stratified culture compared to confluent culture in monolayer on proliferation and differentiation of human articular chondrocytes.
With conventional tissue culture of cells, it is generally assumed that when the available 2D substrate is fully occupied, growth ceases or is greatly reduced.However, in nature wound repair mostly involves proliferation of cells that are attracted to the defect site in a 3D environment.Hence, proliferation continues in 3D until the defect site is filled with cells contributing to repair tissue. With this in mind,we examined the growth behavior of human articular chondrocytes during stratified culture as opposed to routine culture to confluency. Additionally, we studied the influence of growth factors on proliferation during stratified culture and differentiation thereafter. Chondrocytes were cultured in monolayer on tissue culture plastic to confluency or stratified for an additional 7 days. Culture medium was based on DMEM with 10% serum and either supplemented with high concentrations of nonessential amino acids (NEAA) and ascorbic acid (AsAP), or instead with basic fibroblastic growth factor (bFGF), platelet-derived growth factor (PDBF-BB), and/or transforming growth factor beta1 (TGF-beta). After expansion, cells were harvested, counted, and their differentiation capacity was examined in pellet culture assay. It was shown that chondrocytes, cultured stratified proliferate exponentially for up to an additional 4 days and that cell yield increased 5-fold. Furthermore, during stratified culture the number of cells increased further in the presence of bFGF, PDBF-BB, and TGFbeta1 or high concentrations of NEAA and AsAP. Depending on donor variation and factors supplemented the cell yield ranged from 0.06 up to 1.1 million cells/cm2 at the second passage. During stratified culture in the presence of either bFGF and PDGF or high concentrations of NEAA and AsAP, exponential growth continued for up to 7 days. Finally, cells maintained their differentiation capacity when cultured stratified with or without growth factors (bFGF, TGF-beta, and PDGF), but not when cultured with high levels of AsAP and NEAA. In contrast to other 3D culture techniques like microcarrier or suspension culture, nutrient consumption remained the same as with conventional expansion. Because this allows culturing of clinically relevant amounts of chondrocytes without increasing the amount of serum, chondrocytes can be fully expanded in the presence autologous serum, avoiding the risk of viral and/or prion disease transmission associated with the use of animal-derived serum or serum replacers with animal-derived constituents.


The efficacy of novel anatomical sites for the assessment of muscle oxygenation during central hypovolemia.
Muscle tissue oxygenation (SmO2) can track central blood volume loss associated with hemorrhage. Traditional peripheral measurement sites (e.g., forearm) may not be practical due to excessive movement or injury (e.g., amputation). The aim of this study was to evaluate the efficacy of three novel anatomical sites for the assessment of SmO2 under progressive central hypovolemia. 10 male volunteers were exposed to stepwise prone lower body negative pressure to decrease central blood volume, while SmO2 was assessed at four sites-the traditional site of the flexor carpi ulnaris (ARM), and three novel sites not previously investigated during lower body negative pressure, the deltoid, latissimus dorsi, and trapezius. SmO2 at the novel sites was compared to the ARM sensor and to stroke volume responses. A reduction in SmO2 was detected by the ARM sensor at the first level of lower body negative pressure (-15 mmHg; P = 0.007), and at -30 (the deltoid), -45 (latissimus dorsi), and -60 mmHg lower body negative pressure (trapezius) at the novel sites (P ≤ 0.04). SmO2 responses at all novel sites were correlated with responses at the ARM (R ≥ 0.89), and tracked the reduction in stroke volume (R ≥ 0.87); the latissimus dorsi site exhibited the strongest linear correlations (R ≥ 0.96). Of the novel sensor sites, the latissimus dorsi exhibited the strongest linear associations with SmO2 at the ARM, and with reductions in central blood volume. These findings have important implications for detection of hemorrhage in austere environments (e.g., combat) when use of a peripheral sensor may not be ideal, and may facilitate incorporation of these sensors into uniforms.


Magnetic apatite for structural insights on the plasma membrane.
The iron oxide-hydroxyapatite (FeOxHA) nanoparticles reported here differ from those reported before by their advantage of homogeneity and simple preparation; moreover, the presence of carboxymethyldextran (CMD), together with hydroxyapatite (HA), allows access to the cellular membrane, which makes our magnetic apatite unique. These nanoparticles combine magnetic behavior, Raman label ability and the property of interaction with the cellular membrane; they therefore represent an interesting material for structural differentiation of the cell membrane. It was observed by Raman spectroscopy, scanning electron microscopy (SEM) and fluorescence microscopy that FeOxHA adheres to the plasma membrane and does not penetrate the membrane. These insights make the nanoparticles a promising material for magnetic cell sorting, e.g. in microfluidic device applications.


Urinary soluble CD163 and monocyte chemoattractant protein-1 in the identification of subtle renal flare in anti-neutrophil cytoplasmic antibody-associated vasculitis.
Prior work has shown that urinary soluble CD163 (usCD163) displays excellent biomarker characteristics for detection of active renal vasculitis using samples that included new diagnoses with highly active renal disease. This study focused on the use of usCD163 in the detection of the more clinically relevant state of mild renal flare and compared results of usCD163 testing directly to testing of urinary monocyte chemoattractant protein-1 (uMCP-1). Patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV, n = 88) were identified within a serially sampled, longitudinal and multicentre cohort. Creatinine-normalized usCD163 and uMCP-1 levels were measured by enzyme-linked immunosorbent assay and, both alone and in combination, were compared between times of active renal AAV and during remission and/or active non-renal AAV. Samples from 320 study visits included times of active renal vasculitis (n = 39), remission (n = 233) and active extrarenal vasculitis (n = 48). Median creatinine levels were 0.9 mg/dL [interquartile range (IQR) 0.8-1.2] in remission and 1.4 mg/dL (IQR 1.0-1.8) during renal flare. usCD163 levels were higher in patients with active renal vasculitis compared with patients in remission and those with active extrarenal vasculitis, with median values of 162 ng/mmol (IQR 79-337), 44 (17-104) and 38 (7-76), respectively (P < 0.001). uMCP-1 levels were also higher in patients with active renal vasculitis compared with patients in remission and those with active extrarenal vasculitis, with median values of 10.6 pg/mmol (IQR 4.6-23.5), 4.1 (2.5-8.4) and 4.1 (1.9-6.8), respectively (P < 0.001). The proposed diagnostic cut-points for usCD163 and uMCP-1 were 72.9 ng/mmol and 10.0 pg/mmol, respectively. usCD163 and uMCP-1 levels were marginally correlated (r2 = 0.11, P < 0.001). Combining novel and existing biomarkers using recursive tree partitioning indicated that elevated usCD163 plus either elevated uMCP-1 or new/worse proteinuria improved the positive likelihood ratio (PLR) of active renal vasculitis to 19.2. A combination of usCD163 and uMCP-1 measurements appears to be useful in identifying the diagnosis of subtle renal vasculitis flare.


lumi: a pipeline for processing Illumina microarray.
Illumina microarray is becoming a popular microarray platform. The BeadArray technology from Illumina makes its preprocessing and quality control different from other microarray technologies. Unfortunately, most other analyses have not taken advantage of the unique properties of the BeadArray system, and have just incorporated preprocessing methods originally designed for Affymetrix microarrays. lumi is a Bioconductor package especially designed to process the Illumina microarray data. It includes data input, quality control, variance stabilization, normalization and gene annotation portions. In specific, the lumi package includes a variance-stabilizing transformation (VST) algorithm that takes advantage of the technical replicates available on every Illumina microarray. Different normalization method options and multiple quality control plots are provided in the package. To better annotate the Illumina data, a vendor independent nucleotide universal identifier (nuID) was devised to identify the probes of Illumina microarray. The nuID annotation packages and output of lumi processed results can be easily integrated with other Bioconductor packages to construct a statistical data analysis pipeline for Illumina data. The lumi Bioconductor package, www.bioconductor.org


Membrane-associated carbonic anhydrase in cultured rabbit nonpigmented ciliary epithelium.
To measure the activity of membrane-associated carbonic anhydrase (CA) in cultured rabbit nonpigmented epithelial (NPE) cells, determine its identity and its sensitivity to extracellular trypsin, and compare the ability of acetazolamide and a cell-impermeant dextran-bound CA inhibitor to change cytoplasmic pH. Studies were conducted using a cell line derived from rabbit NPE. The cells were lysed and separated into soluble and insoluble fractions by differential centrifugation. CA activity in these fractions was determined using a CO2 hydration assay. In studies with intact cells, a membrane-impermeable high-molecular-weight dextran-bound inhibitor (DBI) was synthesized and used to selectively bind and inhibit the extracellular-facing membrane-bound CA. Measurements of CA activity in intact red blood cells were conducted to confirm DBI remains extracellular. Acetazolamide, a membrane-permeable CA inhibitor, was used to inhibit total CA activity. Intracellular pH was determined using the pH-dependent absorbance of the fluorescent dye BCECF-AM. A low-speed pellet enriched with plasma membrane material accounted for 22.3 +/- 6.1% (n = 18) of the total CA activity in the cultured NPE. When intact cells were exposed to trypsin-EDTA, a 28% reduction of membrane-associated CA activity was observed; DBI inhibited this CA activity loss. Cytosolic CA activity was inhibited by 0.2% sodium dodecyl sulfate (SDS). In contrast, membrane-associated CA was SDS resistant, a characteristic of the CA-IV isozyme. By Western blot, CA-IV immunoreactive polypeptide was detected in the cultured cells and also in native rabbit and porcine ciliary epithelium. Inhibition of total CA activity with acetazolamide and inhibition of extracellular-facing membrane-associated CA with DBI caused an identical intracellular pH decrease in intact NPE cells. Expression of the CA-IV isozyme could account for the significant fraction of CA activity in the cultured NPE, which is membrane associated and SDS resistant. Sensitivity to tryptic hydrolysis suggests the membrane-associated CA partially faces extracellularly. As judged by responses to an extracellular CA inhibitor, the membrane-associated CA has a functional role in maintaining cytoplasmic pH.


Mammillothalamic functional connectivity and memory function in Wernicke's encephalopathy.
There is still debate over the neural mechanisms underlying pathogenic and even recovery processes of Wernicke's encephalopathy. Therefore, we attempted to validate the usefulness of resting-state functional connectivity analysis in assessing memory function and its neural correlation with the mammillothalamic tract in patients recovering from Wernicke's encephalopathy. Seven chronic alcoholics recovering from Wernicke's encephalopathy, 14 alcoholic comparisons without Wernicke's encephalopathy, and 14 healthy comparisons underwent functional connectivity MRI scans, as well as verbal and non-verbal memory tests after at least a 1 month abstinence from alcohol. Resting-state functional connectivity strength between the anterior thalamus and the mammillary bodies was investigated by calculating temporal correlations in magnetic resonance signal levels between the two regions during a 5-min passive viewing task. The mean values of the functional connectivity strength between the left anterior thalamus and the ipsilateral mammillary body differed significantly between Wernicke's encephalopathy patients and healthy comparisons (P = 0.014). This connectivity strength in alcoholic comparisons fell between those of the former two groups, with a significant difference from that of healthy comparisons (P = 0.038). In addition, the strength of this left-sided functional connectivity significantly correlated with delayed verbal recall scores (r = 0.771, P = 0.042) and verbal recognition score (r = 0.825, P = 0.022) in patients with Wernicke's encephalopathy. Our findings indicate that memory function in patients recovering from Wernicke's encephalopathy parallels the level of the mammillothalamic functional connectivity; this supports the usefulness of resting-state functional connectivity analysis as a practical alternative to pathological examination of the mammillothalamic tract in living patients with Wernicke's encephalopathy.


The serine/threonine phosphatase inhibitor, calyculin A, inhibits and dissociates macrophage responses to lipopolysaccharide.
LPS-stimulated macrophages (M phi) produce inflammatory mediators that are largely responsible for the pathophysiology associated with septic shock. M phi respond to LPS with rapid protein phosphorylation and dephosphorylation on serine, threonine, and tyrosine residues. If these events are critical for the cellular response to LPS, the kinases and/or phosphatases involved may be vulnerable targets for pharmacologic intervention. Recent studies demonstrated that tyrosine kinase inhibitors block LPS-induced tyrosine phosphorylation of MAP kinases as well as TNF-alpha and IL-1 beta production. To investigate a role for serine/threonine phosphatases, we evaluated the effect of calyculin A, a potent serine/threonine phosphatase inhibitor, on LPS stimulation of murine M phi. Pretreatment of M phi with calyculin A inhibited LPS-induced expression of six immediate-early genes: TNF-alpha, IL-1 beta, IFN-beta, IP-10, IRF-1, and TNFR-2. Calyculin A added 1.5 h after LPS treatment greatly reduced accumulation of IP-10, IRF-1, and TNFR-2 mRNA, but not TNF-alpha, IL-1 beta, and IFN-beta mRNA. Calyculin A, in the absence or presence of LPS, resulted in sustained tyrosine phosphorylation of the MAP kinases. These findings suggest that an "early" serine/threonine phosphatase activity is essential for LPS stimulation of M phi and that the activation of MAP kinases is not sufficient for the induction of these immediate-early genes. The requirement for a "late" phosphatase activity for expression of a subset of LPS-inducible genes dissociates at least two regulatory pathways in LPS signal transduction.


The effect of neoadjuvant chemotherapy and chemoradiotherapy on exercise capacity and outcome following upper gastrointestinal cancer surgery: an observational cohort study.
In 2014 approximately 21,200 patients were diagnosed with oesophageal and gastric cancer in England and Wales, of whom 37 % underwent planned curative treatments. Potentially curative surgical resection is associated with significant morbidity and mortality. For operable locally advanced disease, neoadjuvant chemotherapy (NAC) improves survival over surgery alone. However, NAC carries the risk of toxicity and is associated with a decrease in physical fitness, which may in turn influence subsequent clinical outcome. Lower levels of physical fitness are associated with worse outcome following major surgery in general and Upper Gastrointestinal Surgery (UGI) surgery in particular. Cardiopulmonary exercise testing (CPET) provides an objective assessment of physical fitness. The aim of this study is to test the hypothesis that NAC prior to upper gastrointestinal cancer surgery is associated with a decrease in physical fitness and that the magnitude of the change in physical fitness will predict mortality 1 year following surgery. This study is a multi-centre, prospective, blinded, observational cohort study of participants with oesophageal and gastric cancer scheduled for neoadjuvant cancer treatment (chemo- and chemoradiotherapy) and surgery. The primary endpoints are physical fitness (oxygen uptake at lactate threshold measured using CPET) and 1-year mortality following surgery; secondary endpoints include post-operative morbidity (Post-Operative Morbidity Survey (POMS)) 5 days after surgery and patient related quality of life (EQ-5D-5 L). The principal benefits of this study, if the underlying hypothesis is correct, will be to facilitate better selection of treatments (e.g. NAC, Surgery) in patients with oesophageal or gastric cancer. It may also be possible to develop new treatments to reduce the effects of neoadjuvant cancer treatment on physical fitness. These results will contribute to the design of a large, multi-centre trial to determine whether an in-hospital exercise-training programme that increases physical fitness leads to improved overall survival. ClinicalTrials.gov NCT01325883 - 29(th) March 2011.


Ezrin has a COOH-terminal actin-binding site that is conserved in the ezrin protein family.
Ezrin, previously also known as cytovillin, p81, and 80K, is a cytoplasmic protein enriched in microvilli and other cell surface structures. Ezrin is postulated to have a membrane-cytoskeleton linker role. Recent findings have also revealed that the NH2-terminal domain of ezrin is associated with the plasma membrane and the COOH-terminal domain with the cytoskeleton (Algrain, M., O. Turunen, A. Vaheri, D. Louvard, and M. Arpin. 1993. J. Cell Biol. 120: 129-139). Using bacterially expressed fragments of ezrin we now demonstrate that ezrin has an actin-binding capability. We used glutathione-S-transferase fusion proteins of truncated ezrin in affinity chromatography to bind actin from the cell extract or purified rabbit muscle actin. We detected a binding site for filamentous actin that was localized to the COOH-terminal 34 amino acids of ezrin. No binding of monomeric actin was detected in the assay. The region corresponding to the COOH-terminal actin-binding site in ezrin is highly conserved in moesin, actin-capping protein radixin and EM10 protein of E. multilocularis, but not in merlin/schwannomin. Consequently, this site is a potential actin-binding site also in the other members of the protein family. Furthermore, the actin-binding site in ezrin shows sequence homology to the actin-binding site in the COOH terminus of the beta subunit of the actin-capping protein CapZ and one of the potential actin-binding sites in myosin heavy chain. The actin-binding capability of ezrin supports its proposed role as a membrane-cytoskeleton linker.


Complications Following Enucleations and Subsequent Oculoplastic Surgeries.
To analyze the complications and subsequent type and frequency of oculoplastic surgeries after enucleation in adult patients. The authors conducted a retrospective case note review of adult patients who underwent enucleation followed by placement of an alloplastic implant wrapped in donor sclera between 2001 and 2013. The data collected included patient demographics, surgical indication, implant size, postoperative complications, and subsequent oculoplastic surgical procedures. The authors included 186 patients who underwent enucleation during the study period. Malignancy was the leading cause for this operation (79.6%) followed by a blind painful eye (12.4%). Most postoperative complications were managed conservatively with an adjustment of the size of the ocular prosthesis. In most cases, the 20-mm and 22-mm implants were used, and implant size ranged from 16 to 22 mm. There was no correlation between implant size and complication rate. Twenty-six patients required subsequent surgery after enucleation (14%). In total, 9.7% (18 patients of 186) patients underwent eyelid surgery after enucleation, most frequently for blepharoptosis (7%). The interval between enucleation and eyelid surgeries was, on average, 1.9 years. Less frequently, surgery is needed for socket repair for anterior surface breakdown (1.6%), and the interval between enucleation and socket surgery was 0.9 years. The most frequent complications following enucleation were blepharoptosis and enophthalmos with a deep upper eyelid sulcus. About 15% of patients required subsequent oculoplastic procedures after, on average, 2 years, while surgery in the early postoperative phase was rarely indicated.


Distinct populations of innate CD8+ T cells revealed in a CXCR3 reporter mouse.
CXCR3, expressed mainly on activated T and NK cells, is implicated in a host of immunological conditions and can contribute either to disease resolution or pathology. We report the generation and characterization of a novel CXCR3 internal ribosome entry site bicistronic enhanced GFP reporter (CIBER) mouse in which enhanced GFP expression correlates with surface levels of CXCR3. Using CIBER mice, we identified two distinct populations of innate CD8(+) T cells based on constitutive expression of CXCR3. We demonstrate that CXCR3(+) innate CD8(+) T cells preferentially express higher levels of Ly6C and CD122, but lower levels of CCR9 compared with CXCR3(-) innate CD8(+) T cells. Furthermore, we show that CXCR3(+) innate CD8(+) T cells express higher transcript levels of antiapoptotic but lower levels of proapoptotic factors, respond more robustly to IL-2 and IL-15, and produce significantly more IFN-γ and granzyme B. Interestingly, CXCR3(+) innate CD8(+) T cells do not respond to IL-12 or IL-18 alone, but produce significant amounts of IFN-γ on stimulation with a combination of these cytokines. Taken together, these findings demonstrate that CXCR3(+) and CXCR3(-) innate CD8(+) T cells are phenotypically and functionally distinct. These newly generated CIBER mice provide a novel tool for studying the role of CXCR3 and CXCR3-expressing cells in vivo.


2D nanostructures for water purification: graphene and beyond.
Owing to their atomically thin structure, large surface area and mechanical strength, 2D nanoporous materials are considered to be suitable alternatives for existing desalination and water purification membrane materials. Recent progress in the development of nanoporous graphene based materials has generated enormous potential for water purification technologies. Progress in the development of nanoporous graphene and graphene oxide (GO) membranes, the mechanism of graphene molecular sieve action, structural design, hydrophilic nature, mechanical strength and antifouling properties and the principal challenges associated with nanopore generation are discussed in detail. Subsequently, the recent applications and performance of newly developed 2D materials such as 2D boron nitride (BN) nanosheets, graphyne, molybdenum disulfide (MoS2), tungsten chalcogenides (WS2) and titanium carbide (Ti3C2Tx) are highlighted. In addition, the challenges affecting 2D nanostructures for water purification are highlighted and their applications in the water purification industry are discussed. Though only a few 2D materials have been explored so far for water treatment applications, this emerging field of research is set to attract a great deal of attention in the near future.


Gestational Age-Dependent Increase of Survival Motor Neuron Protein in Umbilical Cord-Derived Mesenchymal Stem Cells.
Spinal muscular atrophy (SMA) is the most common genetic neurological disease leading to infant death. It is caused by loss of survival motor neuron (SMN) 1 gene and subsequent reduction of SMN protein in motor neurons. Because SMN is ubiquitously expressed and functionally linked to general RNA metabolism pathway, fibroblasts (FBs) are most widely used for the assessment of SMN expression in SMA patients but usually isolated from skin biopsy samples after the onset of overt symptoms. Although recent translational studies of SMN-targeted therapies have revealed the very limited time window for effective SMA therapies during perinatal period, the exact time point when SMN shortage became evident is unknown in human samples. In this study, we analyzed SMN mRNA and protein expression during perinatal period by using umbilical cord-derived mesenchymal stem cells (UC-MSCs) obtained from preterm and term infants. UC-MSCs were isolated from 16 control infants delivered at 22-40 weeks of gestation and SMA fetus aborted at 19 weeks of gestation (UC-MSC-Control and UC-MSC-SMA). FBs were isolated from control volunteer and SMA patient (FB-Control and FB-SMA). SMN mRNA and protein expression in UC-MSCs and FBs was determined by RT-qPCR and Western blot. UC-MSC-Control and UC-MSC-SMA expressed the comparable level of MSC markers on their cell surface and were able to differentiate into adipocytes, osteocytes, and chondrocytes. At steady state, SMN mRNA and protein expression was decreased in UC-MSC-SMA compared to UC-MSC-Control, as observed in FB-SMA and FB-Control. In response to histone deacetylase inhibitor valproic acid, SMN mRNA and protein expression in UC-MSC-SMA and FB-SMA was increased. During perinatal development from 22 to 40 weeks of gestation, SMN mRNA and protein expression in UC-MSC-Control was positively correlated with gestational age. UC-MSCs isolated from 17 fetus/infant of 19-40 weeks of gestation are expressed functional SMN mRNA and protein. SMN mRNA and protein expression in UC-MSCs is increased with gestational age during perinatal development.


Genetically encoding unnatural amino acids for cellular and neuronal studies.
Proteins participate in various biological processes and can be harnessed to probe and control biological events selectively and reproducibly, but the genetic code limits the building block to 20 common amino acids for protein manipulation in living cells. The genetic encoding of unnatural amino acids will remove this restriction and enable new chemical and physical properties to be precisely introduced into proteins. Here we present new strategies for generating orthogonal tRNA-synthetase pairs, which made possible the genetic encoding of diverse unnatural amino acids in different mammalian cells and primary neurons. Using this new methodology, we incorporated unnatural amino acids with extended side chains into the K+ channel Kv1.4, and found that the bulkiness of residues in the inactivation peptide is essential for fast channel inactivation, a finding that had not been possible using conventional mutagenesis. This technique will stimulate and facilitate new molecular studies using tailored unnatural amino acids for cell biology and neurobiology.


Post-mortem stability of RNA in skeletal muscle and adipose tissue and the tissue-specific expression of myostatin, perilipin and associated factors in the horse.
Obesity, a major concern for equine welfare, is highly prevalent in the leisure horse population. Skeletal-muscle and adipose tissues are important determinants of maintenance energy requirements. The myostatin and perilipin pathways play key roles in the regulation of muscle mass and lipolysis respectively and have both been associated with obesity predisposition in other mammalian species. High quality samples, suitable for molecular biology, are an essential prerequisite for detailed investigations of gene and protein expression. Hence, this study has evaluated a) the post-mortem stability of RNA extracted from skeletal-muscle and adipose-tissues collected under commercial conditions and b) the tissue-specific presence of myostatin, the moystatin receptor (activin receptor IIB, ActRIIB), follistatin and perilipin, genes and proteins across a range of equine tissues. Objectives were addressed using tissues from 7 Thoroughbred horses presented for slaughter at a commercial abattoir; a) samples were collected at 7 time-points from Masseter muscle and perirenal adipose from 5 minutes to 6 hours post-mortem. Extracted RN was appraised by Optical Density analysis and agarose-gel electrophoresis. b) Quantitative real time PCR and Western Blotting were used to evaluate gene and protein expression in anatomically-defined samples collected from 17 tissues (6 organs, 4 skeletal muscles and 7 discrete adipose depots). The results indicate that, under the present collection conditions, intact, good quality RNA could be extracted from skeletal-muscle for up to 2 hours post-mortem. However, RNA from adipose tissue may be more susceptible to degradation/contamination and samples should be collected no later than 30 minutes post-mortem. The data also show that myostatin and ActRIIB genes and proteins were almost exclusively expressed in skeletal muscle. The follistatin gene showed a more diverse gene expression profile, with expression evident in several organs, adipose tissue depots and skeletal muscles. Perilipin gene and protein were almost exclusively expressed by adipose tissue.


Mesenchymal stem cell therapy for liver disease: current status and future perspectives.
Liver transplantation is the gold standard for the treatment of end-stage liver disease. However, a shortage of donor organs, high cost, and surgical complications limit the use of this treatment. Cellular therapies using hepatocytes, hematopoietic stem cells, bone marrow mononuclear cells, and mesenchymal stem cells (MSCs) are being investigated as alternative treatments to liver transplantation. The purpose of this review is to describe studies using MSC transplantation for liver diseases based on the reported literature and to discuss prospective research designed to improve the efficacy of MSC therapy. MSCs have several properties that show potential to regenerate injured tissues or organs, such as homing, transdifferentiation, immunosuppression, and cellular protective capacity. Additionally, MSCs can be noninvasively isolated from various tissues and expanded ex vivo in sufficient numbers for clinical evaluation. Currently, there is no approved MSC therapy for the treatment of liver disease. However, MSC therapy is considered a promising alternative treatment for end-stage liver diseases and is reported to improve liver function safely with no side effects. Further robust preclinical and clinical studies will be needed to improve the therapeutic efficacy of MSC transplantation.


Specificity between lactobacilli and hymenopteran hosts is the exception rather than the rule.
Lactobacilli (Lactobacillales: Lactobacillaceae) are well known for their roles in food fermentation, as probiotics, and in human health, but they can also be dominant members of the microbiota of some species of Hymenoptera (ants, bees, and wasps). Honey bees and bumble bees associate with host-specific lactobacilli, and some evidence suggests that these lactobacilli are important for bee health. Social transmission helps maintain associations between these bees and their respective microbiota. To determine whether lactobacilli associated with social hymenopteran hosts are generally host specific, we gathered publicly available Lactobacillus 16S rRNA gene sequences, along with Lactobacillus sequences from 454 pyrosequencing surveys of six other hymenopteran species (three sweat bees and three ants). We determined the comparative secondary structural models of 16S rRNA, which allowed us to accurately align the entire 16S rRNA gene, including fast-evolving regions. BLAST searches and maximum-likelihood phylogenetic reconstructions confirmed that honey and bumble bees have host-specific Lactobacillus associates. Regardless of colony size or within-colony oral sharing of food (trophallaxis), sweat bees and ants associate with lactobacilli that are closely related to those found in vertebrate hosts or in diverse environments. Why honey and bumble bees associate with host-specific lactobacilli while other social Hymenoptera do not remains an open question. Lactobacilli are known to inhibit the growth of other microbes and can be beneficial whether they are coevolved with their host or are recruited by the host from environmental sources through mechanisms of partner choice.


Impacts of compression on crystallization behavior of freeze-dried amorphous sucrose.
An amorphous matrix comprised of sugar molecules is used as excipient and stabilizing agent for labile ingredients in the pharmaceutical industry. The amorphous sugar matrix is often compressed into a tablet form to reduce the volume and improve handling. Herein, the effect of compression on the crystallization behavior of an amorphous sucrose matrix was investigated. Amorphous sucrose samples were prepared by freeze-drying and compressed under different conditions, followed by analyses by differential scanning calorimetry, isothermal crystallization tests, X-ray powder diffractometry, Fourier transform infrared spectroscopy (FTIR), and gas pycnometry. The compressed sample had a lower crystallization temperature and a shorter induction period for isothermal crystallization, indicating that compression facilitates the formation of the critical nucleus of a sucrose crystal. Based on FTIR and molecular dynamics simulation results, the conformational distortion of sucrose molecules due to the compression appears to contribute to the increase in the free energy of the system, which leads to the facilitation of critical nucleus formation. An isothermal crystallization test indicated an increase in the growth rate of sucrose crystals by the compression. This can be attributed to the transformation of the microstructure from porous to nonporous, as the result of compression.


Designing porous electronic thin-film devices: band offsets and heteroepitaxy.
Increasing numbers of electrically active porous framework materials are being reported, with conductivities that make them attractive for technological applications. As design strategies for efficient carrier transport emerge, the next challenge is to incorporate the materials into a functioning device. In thin-film devices interface effects are of critical importance to overall function. In this article we present a method to identify compatible materials combinations to achieve mechanically robust, electronically optimal pairings. The computational screening is based on a two-step procedure: (i) matching of lattice constants to ensure interfaces with minimal epitaxial strain and therefore maximal mechanical and chemical stability; (ii) matching of absolute electron energies to construct energy-band-alignment diagrams, which can be used to screen for particular electronic applications. We apply the methodology to search for zeolitic imidazolate framework (ZIF) type materials that are compatible with native metal electrodes. The procedure allows us to predict simple routes for electrochemical deposition of ZIFs for application as conductive porous electrodes.


Important parameters affecting efficiency of protein refolding by reversed micelles.
Refolding of denatured RNase A as a model of inclusion bodies was performed by reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) in isooctane. In the novel refolding process, a solid-liquid extraction was utilized as an alternative to the ordinary protein extraction by reversed micelles based on a liquid-liquid extraction. First, the effects of operational parameters such as concentration of AOT, W(o) (= [H(2)O]/[AOT]), and pH were examined on the solubilization of solid denatured proteins into a reversed micellar solution. The solubilization was facilitated by a high AOT concentration, a high W(o) value, and a high pH in water pools. These conditions are favorable for the dispersion of the solid protein aggregates in an organic solvent. Second, the renaturation of the denatured RNase A solubilized into the reversed micellar solution was conducted by addition of glutathione as a redox reagent. A complete renaturation of RNase A was accomplished by adjusting the composition of the redox reagent even at a high protein concentration in which protein aggregation would usually occur in aqueous media. In addition, the renaturation rates were improved by optimizing water content (W(o)) and the pH of water pools in reversed micelles. Finally, the recovery of renatured RNase A from the reversed micellar solution was performed by adding a polar organic solvent such as acetone into the reversed micellar solution. This precipitation method was effective for recovering proteins from reversed micellar media without any significant reduction in enzymatic activity.


Factors determining cardiac hypertrophy in hypertensive patients with or without peripheral vascular disease.
1. Coronary ischaemic disease and congestive heart failure are the principal causes of mortality in patients with peripheral vascular disease. Whether cardiac hypertrophy is present and even more pronounced in peripheral vascular disease than in other populations has never been explored.2.Twenty-five hypertensive patients were investigated, 11 without and 14 with peripheral vascular disease, matched for age, sex, mean arterial pressure and antihypertensive drug treatment. Cardiac mass was determined using echocardiography together with measurement of systemic blood pressure, ratio between ankle systolic pressure (ASP) and brachial systolic pressure (BSP), and standard biochemical parameters including natriuresis per 24 h.3. At the same mean arterial pressure, patients with peripheral vascular disease had a significantly higher cardiac mass (157+/-12 versus 116+/-6 g/m2; P<0.01), pulse pressure (81+/-5 versus 55+/-4 mmHg; P<0.01) and natriuresis (180+/-17 versus 144+/-6 mmol/24h; P<0. 01) than controls. Using univariate correlations, cardiac mass was positively associated with pulse pressure, mean arterial pressure and natriuresis, and negatively with the ASP/BSP ratio. On the basis of multivariate regression analysis, only natriuresis was positively correlated to cardiac mass.4. Patients with peripheral vascular disease develop a higher degree of cardiac hypertrophy in comparison with hypertensive subjects with the same level of mean arterial pressure. Sodium intake rather than mechanical factors seems to be the major modulating factor which influences the degree of cardiac hypertrophy.


Favorable Effects of Endoscopic Ligation Combined with Drugs on Rebleeding and Mortality in Cirrhotic Patients: A Network Meta-Analysis.
To assess the effects of combination therapies (endoscopic plus drug[s], drug combinations) on variceal/any-cause rebleeding and mortality among cirrhotic patients with one previous episode of variceal hemorrhage. We searched PubMed, Embase, Cochrane Library, and Web of Science for eligible studies. We included 26 randomized controlled trials involving 2,536 adults using OR to measure the effects. Endoscopic variceal ligation (EVL) plus nadolol ranked first for reducing recurrent bleeds. Both EVL + nadolol and EVL + drugs (nadolol, sucralfate) decreased the risk of any-cause rebleeding than EVL alone (OR 0.34, 95% CI 0.12-0.97; OR 0.40, 95% CI 0.18-0.88, respectively). Meanwhile, EVL + drugs ranked first lowering mortality rates (P-score >0.85) with a marginal superiority over EVL alone (OR 0.52, 95% CI 0.26-1.01). Beta-blockers with isosorbide mononitrate (ISMN) also reached a marginal superiority (OR 0.78, 95% CI 0.56-1.09) for improving mortality. Key Messages: Our findings indicated that EVL + nadolol might be the preferred choice to cirrhotic patients with one previous episode of variceal hemorrhage for preventing rebleeding. EVL + nadolol + sucralfate and beta-blockers + ISMN may be potential alternatives to improve mortality. Further, well-controlled studies are warranted to compare the promising combination therapies.


A Simple and Easy Process for the Determination of Estimated Plasma Glucose Level in Patients Presenting to Hospital: An Example of Multicentric Data Mining.
The aim of the present study was to determine the relation between the simultaneous fasting plasma glucose level and HbA1c in a large population of patients presenting to the hospital, based on various measurement methods available for HbA1c. HbA1c levels of 162,210 patients presenting to various hospitals and laboratories were measured based on seven different systems, and at the same time, eAG levels were calculated based on HbA1c levels. The correlation coefficients (r) between serum plasma glucose and HbA1c levels were found to be 0.809, 0.774, 0.779, 0.817, 0.704, 0.796, and 0.747 in Bio-Rad Variant II, Tosoh G8, ADAMS A1c, Trinity Boronate Affinity, Chromsystems HPLC, Roche Tina-quant, and Abbott Architect, respectively. The concordance correlation coefficients between the eAG levels as calculated with the formulas provided in the text and the eAG levels as calculated according to NGSP directions (where eAG = (28.7*HbA1c) - 46.7) were found to be between 0.9339 and 0.9866. Despite the progress made for the standardization of HbA1c measurements, the relation between serum glucose and HbA1c still demonstrated certain discrepancies pertaining to the differences in measurement methodologies. As a conclusion, each laboratory could determine different eAG levels depending on the data originated by their individual analyzer.


Symbolic representation of recurrent neural network dynamics.
Simple recurrent error backpropagation networks have been widely used to learn temporal sequence data, including regular and context-free languages. However, the production of relatively large and opaque weight matrices during learning has inspired substantial research on how to extract symbolic human-readable interpretations from trained networks. Unlike feedforward networks, where research has focused mainly on rule extraction, most past work with recurrent networks has viewed them as dynamical systems that can be approximated symbolically by finite-state machine (FSMs). With this approach, the network's hidden layer activation space is typically divided into a finite number of regions. Past research has mainly focused on better techniques for dividing up this activation space. In contrast, very little work has tried to influence the network training process to produce a better representation in hidden layer activation space, and that which has been done has had only limited success. Here we propose a powerful general technique to bias the error backpropagation training process so that it learns an activation space representation from which it is easier to extract FSMs. Using four publicly available data sets that are based on regular and context-free languages, we show via computational experiments that the modified learning method helps to extract FSMs with substantially fewer states and less variance than unmodified backpropagation learning, without decreasing the neural networks' accuracy. We conclude that modifying error backpropagation so that it more effectively separates learned pattern encodings in the hidden layer is an effective way to improve contemporary FSM extraction methods.


Child abuse as an international issue.
This paper provides a background and suggests a strategy for an international approach to policy development concerning child abuse. First, child abuse is defined in a way that makes it applicable across cultures and national boundaries as that portion of harm to children that results from human action that is proscribed, proximate and preventable. A number of other dimensions, such as the degree of social sanction or social censure, are outlined that also affect the likelihood that given harm will be regarded as child abuse. Cross-cultural research also reveals that certain categories of children--such as those in poor health, females, unwanted children and those born under difficult circumstances or with disvalued traits or under conditions of rapid socioeconomic change--are more vulnerable to maltreatment in many countries. The paper argues for a two-pronged international strategy that first urges individual countries to make a priority of the particular types of abuse that are in most urgent need of attention in their society as well as participating at the same time in a concerted international focus on three widely occurring forms of child abuse: parental child battering, selective neglect, and sexual abuse.


Ocular manifestations of a metastatic adenocarcinoma in a horse.
A 10-year-old German Warmblood gelding was referred to the Equine Department of the Vetsuisse Faculty, University of Zurich, Switzerland, for an iris mass OD, lethargy, intermittent fever, and coughing. Ophthalmic examination revealed a 7 × 9 mm raised, fleshy, whitish to pinkish, vascularized iris mass at the 2 o`clock position OD. Fundic examination showed multifocal round, brown to black, slightly raised lesions with indistinct margins and a surrounding hyperreflective zone OU. Physical examination revealed a temperature of 39.2 °C, sinus tachycardia, preputial and ventral edema, and an enlarged right mandibular lymph node. Results of a complete blood count and plasma biochemical profile showed mild anemia, leukocytosis, and thrombocytopenia. Severe splenopathy, moderate splenomegaly, and severe pulmonary pathology with nodules and large areas of consolidated lung parenchyma were observed on abdominal ultrasound and thoracic radiographs, respectively. Fine needle aspirates of the enlarged mandibular lymph node showed malignant epithelial neoplastic cells. The horse was euthanized because of the poor prognosis and subsequently underwent postmortem examination. Macroscopic necropsy and histopathology revealed an adenocarcinoma of suspected pulmonary origin with involvement of eyes, heart, liver, kidneys, spleen, diaphragm, skeletal muscles, mandibular, pulmonary, and internal iliac lymph nodes. Metastatic adenocarcinoma should be considered as a differential diagnosis in horses with iris masses, multifocal chorioretinal infiltrates, and clinical signs that conform to a paraneoplastic syndrome.


Cellular Ca2+ Signals Generate Defined pH Signatures in Plants.
Ca2+ play a key role in cell signaling across organisms. The question of how a simple ion can mediate specific outcomes has spurred research into the role of Ca2+ signatures and their encoding and decoding machinery. Such studies have frequently focused on Ca2+ alone and our understanding of how Ca2+ signaling is integrated with other responses is poor. Using in vivo imaging with different genetically encoded fluorescent sensors in Arabidopsis (Arabidopsis thaliana) cells, we show that Ca2+ transients do not occur in isolation but are accompanied by pH changes in the cytosol. We estimate the degree of cytosolic acidification at up to 0.25 pH units in response to external ATP in seedling root tips. We validated this pH-Ca2+ link for distinct stimuli. Our data suggest that the association with pH may be a general feature of Ca2+ transients that depends on the transient characteristics and the intracellular compartment. These findings suggest a fundamental link between Ca2+ and pH dynamics in plant cells, generalizing previous observations of their association in growing pollen tubes and root hairs. Ca2+ signatures act in concert with pH signatures, possibly providing an additional layer of cellular signal transduction to tailor signal specificity.


Robot Guided 'Pen Skill' Training in Children with Motor Difficulties.
Motor deficits are linked to a range of negative physical, social and academic consequences. Haptic robotic interventions, based on the principles of sensorimotor learning, have been shown previously to help children with motor problems learn new movements. We therefore examined whether the training benefits of a robotic system would generalise to a standardised test of 'pen-skills', assessed using objective kinematic measures [via the Clinical Kinematic Assessment Tool, CKAT]. A counterbalanced, cross-over design was used in a group of 51 children (37 male, aged 5-11 years) with manual control difficulties. Improved performance on a novel task using the robotic device could be attributed to the intervention but there was no evidence of generalisation to any of the CKAT tasks. The robotic system appears to have the potential to support motor learning, with the technology affording numerous advantages. However, the training regime may need to target particular manual skills (e.g. letter formation) in order to obtain clinically significant improvements in specific skills such as handwriting.


Platelets and the innate immune system: mechanisms of bacterial-induced platelet activation.
It has become clear that platelets are not simply cell fragments that plug the leak in a damaged blood vessel; they are, in fact, also key components in the innate immune system, which is supported by the presence of Toll-like receptors (TLRs) on platelets. As the cells that respond first to a site of injury, they are well placed to direct the immune response to deal with any resulting exposure to pathogens. The response is triggered by bacteria binding to platelets, which usually triggers platelet activation and the secretion of antimicrobial peptides. The main platelet receptors that mediate these interactions are glycoprotein (GP)IIb-IIIa, GPIbα, FcγRIIa, complement receptors, and TLRs. This process may involve direct interactions between bacterial proteins and the receptors, or can be mediated by plasma proteins such as fibrinogen, von Willebrand factor, complement, and IgG. Here, we review the variety of interactions between platelets and bacteria, and look at the potential for inhibiting these interactions in diseases such as infective endocarditis and sepsis.


Interaction between retinoid acid receptor-related orphan receptor alpha (RORA) and neuropeptide S receptor 1 (NPSR1) in asthma.
Retinoid acid receptor-related Orphan Receptor Alpha (RORA) was recently identified as a susceptibility gene for asthma in a genome-wide association study. To investigate the impact of RORA on asthma susceptibility, we performed a genetic association study between RORA single nucleotide polymorphisms (SNPs) in the vicinity of the asthma-associated SNP (rs11071559) and asthma-related traits. Because the regulatory region of a previously implicated asthma susceptibility gene, Neuropeptide S receptor 1 (NPSR1), has predicted elements for RORA binding, we hypothesized that RORA may interact biologically and genetically with NPSR1. 37 RORA SNPs and eight NPSR1 SNPs were genotyped in the Swedish birth cohort BAMSE (2033 children) and the European cross-sectional PARSIFAL study (1120 children). Seven RORA SNPs confined into a 49 kb region were significantly associated with physician-diagnosed childhood asthma. The most significant association with rs7164773 (T/C) was driven by the CC genotype in asthma cases (OR = 2.0, 95%CI 1.36-2.93, p = 0.0003 in BAMSE; and 1.61, 1.18-2.19, p = 0.002 in the combined BAMSE-PARSIFAL datasets, respectively), and strikingly, the risk effect was dependent on the Gln344Arg mutation in NPSR1. In cell models, stimulation of NPSR1 activated a pathway including RORA and other circadian clock genes. Over-expression of RORA decreased NPSR1 promoter activity further suggesting a regulatory loop between these genes. In addition, Rora mRNA expression was lower in the lung tissue of Npsr1 deficient mice compared to wildtype littermates during the early hours of the light period. We conclude that RORA SNPs are associated with childhood asthma and show epistasis with NPSR1, and the interaction between RORA and NPSR1 may be of biological relevance. Combinations of common susceptibility alleles and less common functional polymorphisms may modify the joint risk effects on asthma susceptibility.


Inter-residue coupling contributes to high-affinity subtype-selective binding of α-bungarotoxin to nicotinic receptors.
The crystal structure of a pentameric α7 ligand-binding domain chimaera with bound α-btx (α-bungarotoxin) showed that of the five conserved aromatic residues in α7, only Tyr¹⁸⁴ in loop C of the ligand-binding site was required for high-affinity binding. To determine whether the contribution of Tyr¹⁸⁴ depends on local residues, we generated mutations in an α7/5HT(3A) (5-hydroxytryptamine type 3A) receptor chimaera, individually and in pairs, and measured ¹²⁵I-labelled α-btx binding. The results show that mutations of individual residues near Tyr¹⁸⁴ do not affect α-btx affinity, but pairwise mutations decrease affinity in an energetically coupled manner. Kinetic measurements show that the affinity decreases arise through increases in the α-btx dissociation rate with little change in the association rate. Replacing loop C in α7 with loop C from the α-btx-insensitive α2 or α3 subunits abolishes high-affinity α-btx binding, but preserves acetylcholine-elicited single channel currents. However, in both the α2 and α3 construct, mutating either residue that flanks Tyr¹⁸⁴ to its α7 counterpart restores high-affinity α-btx binding. Analogously, in α7, mutating both residues that flank Tyr¹⁸⁴ to the α2 or α3 counterparts abolishes high-affinity α-btx binding. Thus interaction between Tyr¹⁸⁴ and local residues contributes to high-affinity subtype-selective α-btx binding.


Techniques for radiofrequency ablation of head and neck tumors.
To describe the various techniques that have been developed for application of radiofrequency ablation in the palliative treatment of recurrent and advanced head and neck malignant tumors. Case series with a follow-up of 3 to 18 months. Faculty practice, research protocol, tertiary care academic medical center. Fifteen treatments were administered over a 3-year period to 12 patients with advanced and/or recurrent tumors. Eleven tumors were squamous cell carcinomas of the oral cavity, oropharynx, and maxillary sinus, and 1 tumor was a medullary thyroid carcinoma. Three of the 12 patients were treated on 2 separate occasions. Patients were selected as a referred sample and sent specifically for treatment with radiofrequency ablation because they were not candidates for the standard curative options of radiation or surgery. No patient refused enrollment, nor were any patients withdrawn because of adverse events. Radiofrequency ablation is a method of localized hyperthermia resulting in tissue necrosis. Ninety to 150 W of energy is applied, achieving intratumoral temperatures of 60 to 110 degrees C for 5 to 15 minutes per ablation. Techniques have been developed to apply radiofrequency ablation under direct vision, endoscopically, percutaneously, and with ultrasound and computed tomographic guidance. The radiofrequency ablation probe was accurately placed and treatment administered on 15 occasions. No perioperative deaths occurred. One patient suffered a stroke. Subjective patient improvement was reported with regard to pain (n=9), appearance (n=3), and function (n=4).


Expression and function of conserved nuclear receptor genes in Caenorhabditis elegans.
The Caenorhabditis elegans genome encodes 284 nuclear receptor (NR) genes. Among these 284 NR genes are 15 genes conserved among the Metazoa. Here, we analyze the expression and function of eight heretofore uncharacterized conserved C. elegans NR genes. Reporter gene analysis demonstrates that these genes have distinct expression patterns and that a majority of the C. elegans cell types express a conserved NR gene. RNA interference with NR gene function resulted in visible phenotypes for three of the genes, revealing functions in various processes during postembryonic development. Five of the conserved NR genes are orthologs of NR genes that function during molting and metamorphosis in insects. Functional studies confirm a role for most of these 'ecdysone cascade' NR orthologs during the continuous growth and dauer molts. Transcript levels for these genes fluctuate in a reiterated pattern during the molting cycles, reminiscent of the expression hierarchy observed in the insect ecdysone response. Together, these analyses provide a foundation for further dissecting the role of NRs in nematode development as well as for evaluating conservation of NR functions among the Metazoa.


Continuing efficacy of milnacipran following long-term treatment in fibromyalgia: a randomized trial.
Previous studies of long-term treatment response in fibromyalgia and other chronic pain states have generally been limited to approximately one year, leaving questions about the longer-term durability of response. The purpose of this study was to demonstrate continuing efficacy of milnacipran by characterizing changes in pain and other fibromyalgia symptoms after discontinuing long-term treatment. The mean length of milnacipran treatment at the time of randomized withdrawal was 36.1 months from initial exposure to milnacipran (range, 17.9 to 54.4 months). After completing a long-term, open-label, lead-in study of milnacipran (which followed varying periods of exposure in previous studies), adult patients with fibromyalgia entered the four-week open-label period of the current study for evaluation of ongoing treatment response. After the four-week period to confirm new baseline status, 151 patients taking milnacipran ≥100 mg/day and reporting ≥50% improvement from pre-milnacipran exposure in Visual Analogue Scale (VAS) pain scores were classified as responders. These responders entered the 12-week, double-blind withdrawal period in which they were randomized 2:1 to continue milnacipran or switched to placebo. The prespecified primary parameter was loss of therapeutic response (LTR), defined as increase in VAS pain score to <30% reduction from pre-milnacipran exposure or worsening of fibromyalgia requiring alternative treatment. Adverse events and vital signs were also monitored. Time to LTR was shorter in patients randomized to placebo than in patients continuing milnacipran (P < 0.001). Median time to LTR was 56 days with placebo and was not calculable for milnacipran, because less than half of the latter group of patients lost therapeutic response by study end. Additionally, 81% of patients continuing on milnacipran maintained clinically meaningful pain response (≥30% improvement from pre-milnacipran exposure), compared with 58% of patients switched to placebo (sensitivity analysis II; P < 0.001). The incidences of treatment-emergent adverse events were 58% and 47% for placebo and milnacipran, respectively. Mean decreases in blood pressure and heart rate were found in both groups, with greater decreases for patients switched to placebo. Continuing efficacy of milnacipran was demonstrated by the loss of effect following withdrawal of treatment in patients who received an average of three years of milnacipran treatment. ClinicalTrials.gov: NCT01014585.


Surgery in the patient with liver disease.
Liver dysfunction is a prominent entity in Western medicine that has historically affected patients suffering from chronic viral or alcoholic hepatitis. The incidence of these conditions has not changed dramatically in recent years but the overall number of patients with liver dysfunction has increased considerably with the emergence of the obesity epidemic. Nonalcoholic fatty liver disease (NAFLD) has become increasingly recognized as the most common cause of chronic liver disease in the United States. Although the rate of progression of NAFLD to overt cirrhosis is low, the high prevalence of this condition, combined with the moderate degree of liver dysfunction it engenders, has resulted in a significant increase in the number of patients with liver disease that can be encountered by a surgical practice. Any degree of clinically evident liver disease in a prospective surgical patient should raise concern for the entire surgical team. This particularly applies to intraabdominal surgery whereby the presence of hepatomegaly, portal hypertension, variceal bleeding, and ascites can turn even the most routine operation into a morbid and life-threatening procedure. Nonabdominal surgery avoids some of the technical challenges presented by liver disease but the anesthetic management of a cirrhotic patient still makes any operation potentially more dangerous. In this article, approaches to minimize the risk when surgery becomes necessary in the presence of liver disease are discussed.


Foot posture as a risk factor for lower limb overuse injury: a systematic review and meta-analysis.
Static measures of foot posture are regularly used as part of a clinical examination to determine the need for foot level interventions. This is based on the premise that pronated and supinated foot postures may be risk factors for or associated with lower limb injury. This systematic review and meta-analysis investigates foot posture (measured statically) as a potential risk factor for lower limb overuse injuries. A systematic search was performed using Medline, CINAHL, Embase, SportDiscus in April 2014, to identify prospective cohort studies that investigated foot posture and function as a risk factor for lower limb overuse injury. Eligible studies were classified based on the method of foot assessment: (i) static foot posture assessment; and/or (ii) dynamic foot function assessment. This review presents studies evaluating static foot posture. The methodological quality of included studies was evaluated by two independent reviewers, using an adapted version of the Epidemiological Appraisal Instrument (EAI). Where possible, effects were expressed as standardised mean differences (SMD) for continuous scaled data, and risk ratios (RR) for nominal scaled data. Meta-analysis was performed where injuries and outcomes were considered homogenous. Twenty-one studies were included (total n = 6,228; EAI 0.8 to 1.7 out of 2.0). There was strong evidence that a pronated foot posture was a risk factor for medial tibial stress syndrome (MTSS) development and very limited evidence that a pronated foot posture was a risk factor for patellofemoral pain development, although associated effect sizes were small (0.28 to 0.33). No relationship was identified between a pronated foot posture and any other evaluated pathology (i.e. foot/ankle injury, bone stress reactions and non-specific lower limb overuse injury). This systematic review identified strong and very limited evidence of small effect that a pronated foot posture is a risk factor for MTSS and patellofemoral pain respectively. Evaluation of static foot posture should be included in a multifactorial assessment for both MTSS and patellofemoral pain, although only as a part of the potential injury risk profile. Whilst the included measures are clinically applicable, further studies are required to determine their relationship with dynamic foot function.


Internal Morphology-Controllable Self-Assembly in Poly(Ionic Liquid) Nanoparticles.
Precise control of the interior and outer shapes of polymer nanoparticles has found broad interest in nanosciences, for example, in fundamental studies of their physical properties, colloidal behavior, and corresponding applications. Realizing such control below the 50 nm scale (i.e., a size scale close to individual polymer chains) requires accurate manipulation of polymerization techniques and a judicious choice of the chemical structure in monomers and/or polymers. Here, we constructed a series of well-defined sub-50 nm homopolymer nanoparticles with controllable shape and highly ordered, complex internal structures with sub-5 nm domain spacings, starting from 1-vinyl-1,2,4-triazolium-type ionic liquids in a one-pot dispersion polymerization. With cryogenic electron microscopy and tomography, a morphological evolution of particle shape and interior at this extremely small size end, unusual for polymer colloids, was identified and investigated in detail.


Macrophage NCOR1 protects from atherosclerosis by repressing a pro-atherogenic PPARγ signature.
Nuclear receptors and their cofactors regulate key pathophysiological processes in atherosclerosis development. The transcriptional activity of these nuclear receptors is controlled by the nuclear receptor corepressors (NCOR), scaffolding proteins that form the basis of large corepressor complexes. Studies with primary macrophages demonstrated that the deletion of Ncor1 increases the expression of atherosclerotic molecules. However, the role of nuclear receptor corepressors in atherogenesis is unknown. We generated myeloid cell-specific Ncor1 knockout mice and crossbred them with low-density lipoprotein receptor (Ldlr) knockouts to study the role of macrophage NCOR1 in atherosclerosis. We demonstrate that myeloid cell-specific deletion of nuclear receptor corepressor 1 (NCOR1) aggravates atherosclerosis development in mice. Macrophage Ncor1-deficiency leads to increased foam cell formation, enhanced expression of pro-inflammatory cytokines, and atherosclerotic lesions characterized by larger necrotic cores and thinner fibrous caps. The immunometabolic effects of NCOR1 are mediated via suppression of peroxisome proliferator-activated receptor gamma (PPARγ) target genes in mouse and human macrophages, which lead to an enhanced expression of the CD36 scavenger receptor and subsequent increase in oxidized low-density lipoprotein uptake in the absence of NCOR1. Interestingly, in human atherosclerotic plaques, the expression of NCOR1 is reduced whereas the PPARγ signature is increased, and this signature is more pronounced in ruptured compared with non-ruptured carotid plaques. Our findings show that macrophage NCOR1 blocks the pro-atherogenic functions of PPARγ in atherosclerosis and suggest that stabilizing the NCOR1-PPARγ binding could be a promising strategy to block the pro-atherogenic functions of plaque macrophages and lesion progression in atherosclerotic patients.