On what basis are medical cost-effectiveness thresholds set? Clashing opinions and an absence of data: a systematic review.
The amount a government should be willing to invest in adopting new medical treatments has long been under debate. With many countries using formal cost-effectiveness (C/E) thresholds when examining potential new treatments and ever-growing medical costs, accurately setting the level of a C/E threshold can be essential for an efficient healthcare system. The aim of this systematic review is to describe the prominent approaches to setting a C/E threshold, compile available national-level C/E threshold data and willingness-to-pay (WTP) data, and to discern whether associations exist between these values, gross domestic product (GDP) and health-adjusted life expectancy (HALE). This review further examines current obstacles faced with the presently available data. A systematic review was performed to collect articles which have studied national C/E thresholds and willingness-to-pay (WTP) per quality-adjusted life year (QALY) in the general population. Associations between GDP, HALE, WTP, and C/E thresholds were analyzed with correlations. Seventeen countries were identified from nine unique sources to have formal C/E thresholds within our inclusion criteria. Thirteen countries from nine sources were identified to have WTP per QALY data within our inclusion criteria. Two possible associations were identified: C/E thresholds with HALE (quadratic correlation of 0.63), and C/E thresholds with GDP per capita (polynomial correlation of 0.84). However, these results are based on few observations and therefore firm conclusions cannot be made. Most national C/E thresholds identified in our review fall within the WHO's recommended range of one-to-three times GDP per capita. However, the quality and quantity of data available regarding national average WTP per QALY, opportunity costs, and C/E thresholds is poor in comparison to the importance of adequate investment in healthcare. There exists an obvious risk that countries might either over- or underinvest in healthcare if they base their decision-making process on erroneous presumptions or non-evidence-based methodologies. The commonly referred to value of 100,000$ USD per QALY may potentially have some basis.


Human rights assessment in Parc Jean Marie Vincent, Port-au-Prince, Haiti.
Months after a 7.0 magnitude earthquake hit Port-au-Prince, Haiti, over one million remain homeless and living in spontaneous internally displaced person (IDP) camps. Billions of dollars from aid organizations and government agencies have been pledged toward the relief effort, yet many basic human needs, including food, shelter, and sanitation, continue to be unmet. The Sphere Project, "Humanitarian Charter and Minimum Standards in Disaster Response," identifies the minimum standards to be attained in disaster response. From a human rights perspective and utilizing key indicators from the Sphere Project as benchmarks, this article reports on an assessment of the living conditions approximately 12 weeks after the earthquake in Parc Jean Marie Vincent, a spontaneous IDP camp in Port-au-Prince. A stratified random sample of households in the camp, proportionate to the number of families living in each sector, was selected. Interview questions were designed to serve as "key indicators" for the Sphere Project minimum standards. A total of 486 interviews were completed, representing approximately 5% of households in each of the five sectors of the camp. Our assessment identified the relative achievements and shortcomings in the provision of relief services in Parc Jean Marie Vincent. At the time of this survey, the Sphere Project minimum standards for access to health care and quantity of water per person per day were being met. Food, shelter, sanitation, and security were below minimum accepted standard and of major concern. The formal assessment reported here was completed by September 2010, and is necessarily limited to conditions in Haiti before the cholera outbreak in October.


Post-assembly functionalization of supramolecular nanostructures with bioactive peptides and fluorescent proteins by native chemical ligation.
Post-assembly functionalization of supramolecular nanostructures has the potential to expand the range of their applications. We report here the use of the chemoselective native chemical ligation (NCL) reaction to functionalize self-assembled peptide amphiphile (PA) nanofibers. This strategy can be used to incorporate specific bioactivity on the nanofibers, and as a model, we demonstrate functionalization with the RGDS peptide following self-assembly. Incorporation of bioactivity is verified by the observation of characteristic changes in fibroblast morphology following NCL-mediated attachment of the signal to PA nanofibers. The NCL reaction does not alter the PA nanofiber morphology, and biotinylated RGDS peptide was found to be accessible on the nanofiber surface after ligation for binding with streptavidin-conjugated gold nanoparticles. In order to show that this strategy is not limited to short peptides, we utilized NCL to conjugate yellow fluorescent protein and/or cyan fluorescent protein to self-assembled PA nanofibers. Förster resonance energy transfer and fluorescence anisotropy measurements are consistent with the immobilization of the protein on the PA nanofibers. The change in electrophoretic mobility of the protein upon conjugation with PA molecules confirmed the formation of a covalent linkage. NCL-mediated attachment of bioactive peptides and proteins to self-assembled PA nanofibers allows the independent control of self-assembly and bioactivity while retaining the biodegradable peptide structure of the PA molecule and thus can be useful in tailoring design of biomaterials.


Disruption of mouse Slx4, a regulator of structure-specific nucleases, phenocopies Fanconi anemia.
The evolutionarily conserved SLX4 protein, a key regulator of nucleases, is critical for DNA damage response. SLX4 nuclease complexes mediate repair during replication and can also resolve Holliday junctions formed during homologous recombination. Here we describe the phenotype of the Btbd12 knockout mouse, the mouse ortholog of SLX4, which recapitulates many key features of the human genetic illness Fanconi anemia. Btbd12-deficient animals are born at sub-Mendelian ratios, have greatly reduced fertility, are developmentally compromised and are prone to blood cytopenias. Btbd12(-/-) cells prematurely senesce, spontaneously accumulate damaged chromosomes and are particularly sensitive to DNA crosslinking agents. Genetic complementation reveals a crucial requirement for Btbd12 (also known as Slx4) to interact with the structure-specific endonuclease Xpf-Ercc1 to promote crosslink repair. The Btbd12 knockout mouse therefore establishes a disease model for Fanconi anemia and genetically links a regulator of nuclease incision complexes to the Fanconi anemia DNA crosslink repair pathway.


The Renal Elimination Pathways of the Dabigatran Reversal Agent Idarucizumab and its Impact on Dabigatran Elimination.
Idarucizumab, a humanized monoclonal antibody fragment (Fab), provides rapid and sustained reversal of dabigatran-mediated anticoagulation. Idarucizumab and dabigatran are mainly eliminated via the kidneys. This analysis aimed to characterize the renal elimination of idarucizumab and investigate the influence of idarucizumab on the pharmacokinetics (PK) of dabigatran and vice versa. Studies were conducted in 5/6 nephrectomized rats, in human volunteers with and without renal impairment, and in a porcine liver trauma model. In both rats and humans, renal impairment increased idarucizumab exposure and initial half-life but did not affect its terminal half-life. Urinary excretion of unchanged idarucizumab increased with increasing idarucizumab dose, suggesting saturation of renal tubular reuptake processes at higher doses. The PK of idarucizumab was unaffected by dabigatran. In contrast, idarucizumab administration resulted in redistribution of dabigatran to the plasma, where it was bound and inactivated by idarucizumab. Urinary excretion of dabigatran after administration of idarucizumab was delayed, but total dabigatran excreted in urine was unaffected. Idarucizumab and dabigatran were eliminated together via renal pathways.


Aging affects the association between endothelial nitric oxide synthase gene polymorphism and acute myocardial infarction in the Korean male population.
The aging process affects responsiveness and other functions of endothelium and vascular smooth muscle cells, predisposing the old vessels to the development of atherosclerotic lesions. Endothelial nitric oxide synthase (ecNOS) gene polymorphisms were shown to affect the occurrence of acute myocardial infarction (AMI). We hypothesized that aging may affect the association between the ecNOS gene polymorphism and AMI. We investigated the age-related distribution of the ecNOS gene a/b polymorphism in 121 male AMI patients and 206 age-matched healthy male controls. The aa, ab and bb genotypes were found in 1, 49 and 156 cases among the control subjects and 5, 23 and 93 cases among the AMI patients, respectively. There was a significant correlation between the ecNOS polymorphism and AMI (p = 0.045). When the correlation was analyzed by age, the significance remained only in the group below the age of 51 (p = 0.009). The proportion of smokers was increased in the young patients when compared to the old patients (p = 0.033), indicating that smoking also has greater effect on the younger population. The incidences of hypertension and diabetes mellitus, however, were similar in both populations. Our work provides the first evidence that links ecNOS polymorphism to the risk of AMI in relation to age. Young persons who smoke or have ecNOSaa genotype may have an increased risk of developing AMI. The functional as well as structural changes associated with aging in the vascular endothelium may mask the effect of the ecNOS polymorphism in the development of AMI in old persons.


Clinical manifestations of food protein-induced enterocolitis syndrome.
To raise awareness among healthcare providers about the clinical and laboratory findings in acute and chronic food protein-induced enterocolitis syndrome (FPIES). FPIES can be caused by trivial exposure or rare foods. FPIES is a non-IgE-mediated reaction that usually presents with acute severe repetitive vomiting and diarrhea associated with lethargy, pallor, dehydration, and even hypovolemic shock. Manifestations resolve usually within 24-48 h of elimination of the causative food. In chronic cases, symptoms may include persistent diarrhea, poor weight gain, failure to thrive, and improvement may take several days after the food elimination. In the acute cases, laboratory evaluation may reveal thrombocytosis and neutrophilia, peaking about 6 h postingestion. Depending on the severity, metabolic acidosis and methemoglobinemia may occur. In chronic cases, anemia, hypoalbuminemia and eosinophilia may be seen. Radiologic evaluation or other procedures, such as endoscopy and gastric juice analysis may show nonspecific abnormal findings. The diagnosis is based on clinical manifestations. Further studies looking at the phenotypes of FPIES are needed to identify clinical subtypes, and to understand the predisposing factors for developing FPIES compared with immediate-type, IgE-mediated gastroenteropathies.


Epitope-specific regulation of immunoglobulin class switching in mice immunized with malarial merozoite surface proteins.
Antibodies that bind to Fc receptors and activate complement are implicated in the efficient control of pathogens, but the processes that regulate their induction are still not well understood. To investigate antigen-dependent factors that regulate class switching, we have developed an in vivo model of class switching to immunoglobulin G2b (IgG2b) using the malaria antigen Plasmodium falciparum merozoite surface protein 2 (MSP2). C57BL/6 mice were immunized with recombinant proteins representing discrete domains of MSP2, and a T-cell epitope (C8) was identified within the conserved C terminus of the protein that preferentially induces IgG2b antibodies. The ability of C8 to induce IgG2b is ablated in both homozygous gamma interferon-negative and interleukin 10-negative mice. The IgG2b-inducing properties of C8 override the IgG1-inducing properties of both the fusion protein partner, glutathione S-transferase, and the adjuvant. Furthermore, when attached to other proteins that normally induce IgG1 responses, C8 induces a switch to IgG2b secretion. This is the first description of a defined T-cell epitope that drives specific IgG2b subclass switching, and our data offer proof of the concept that chimeric vaccines incorporating specific T-cell "switch epitopes" might be used to enhance qualitative aspects of the antibody response.


Human and mouse macrophage-inducible C-type lectin (Mincle) bind Candida albicans.
Candida albicans is a causative agent in mycoses of the skin, oral cavity, and gastrointestinal tract. Identification of receptors, and their respective ligands, that are engaged by immune cells when in contact with C. albicans is crucial for understanding inflammatory responses leading to invasive candidiasis. Mincle is a recently identified macrophage-expressed receptor that is important for host responses to C. albicans. The carbohydrate-recognition domain of human and mouse Mincle were expressed, purified under denaturing conditions, and successfully refolded. In addition to oligomers, there are isolatable monomeric and dimeric forms of the protein that occur under two different buffer solutions. The human and mouse homologues bound yeast extract, and the isolated dimeric and monomeric species also demonstrated the recognition of whole C. albicans yeast cells. The data are indicative of several functional states mediating the interaction of Mincle and yeast at the surface of the macrophage.


Factors preventing the performance of oxygen isotope ratios as indicators of grain yield in maize.
This paper provides new insights into source-sink relationships and transpiration processes which will eventually help to interpret δ (18) O as a genotype selection and ecophysiological tool for maize adaptation to drought. Oxygen isotope composition (δ(18)O) has been proposed as a phenotyping tool to integrate leaf transpiration in C4 crops, such as maize. Within this context we hypothesize that δ(18)O in leaves may reflect primarily environmental and genetic variability in evaporative processes, but that this signal may become dampened in transit from source to sink tissues. The aim of this study was to assess the relative importance of transpirative or translocation-related factors affecting δ(18)O in plant tissues of maize. We performed two water regime experiments, one with two varieties under semi-controlled conditions, and another in the field with 100 genotypes during two consecutive years. The δ(18)O in organic matter at the leaf base was strongly correlated with the δ(18)O in stem water, indicating that it could be a good proxy for source water in extensive samplings. Compared to leaves, we observed an (18)O depletion in silks and grains, but not in stem-soluble organic matter. We interpret this as evidence of exchange with unenriched water from source to sink, but mainly occurring within sink tissues. Although grain yield (GY) and physiological variables did not show clear intra-trial patterns against δ(18)O, the only tissues that correlated with GY in the linear regression approach were that of silks, giving an insight of evapotranspirative demand during female flowering and thus of potential maize lines that are better adapted to drought. This finding will eventually help to interpret δ(18)O as a genotype selection and ecophysiological tool for the adaption of maize and other crops to drought, offering insight into source-sink relationships and transpiration processes.


How orchids concentrate? The relationship between physiological stress reactivity and cognitive performance during infancy and early childhood.
The Autonomic Nervous System (ANS) is involved both in higher-order cognition such as attention and learning, and in responding to unexpected, threatening events. Increased ANS reactivity may confer both superior short-term cognitive performance, and heightened long-term susceptibility to adverse events. Here, we evaluate this hypothesis within the Differential Susceptibility Theory (DST) framework. We hypothesise that individuals with increased reactivity may show heightened biological sensitivity to context, conferring both positive (development-enhancing) effects (superior attention and learning) and negative (risk-promoting) effects (increased sensitivity to unsupportive environments). First, we examine how ANS reactivity relates to early cognitive performance. We hypothesise that increased phasic ANS reactivity, observed at lower tonic (pre-stimulus) ANS activity, is associated with better attention and learning. We conclude that the evidence is largely in support. Second we discuss whether ANS reactivity to 'positive', attention-eliciting and to 'negative', aversive stimuli is a one-dimensional construct; and evaluate evidence for how the real-world environment influences physiological stress over short and long time-frames. We identify three areas where the evidence is currently inconclusive.


Prevalence of known and unknown primary tumor sites in spinal metastasis patients.
A retrospective study. THREE OBJECTIVES HAVE BEEN DESIGNATED FOR THIS STUDY: (1) to determine the prevalence of identifiable and non-identifiable primary tumor sites in patients with spinal metastasis, (2) to identify the most common site of the known primary tumor sites, and (3) to identify the factors associated with survival time. The spine is the third most common metastatic site for several primary visceral carcinomas. The primary tumor site could not be identified in 15% to 20% of patients who had been diagnosed of with a skeletal metastasis. Most of the previous studies on skeletal metastasis have not been limited to spinal metastasis alone. Between January 2007 and July 2011 reviews were done for 82 patients with spinal metastasis who had not received a previous diagnosis of carcinoma. The assessment parameters included the following: general demographic data, Karnofsky score, Frankel score, number of spinal vertebra affected, region of the spine affected by metastasis, other skeletal metastasis site, visceral metastasis, known or unknown primary sites of metastasis, histological cell type of metastasis, and the survival period. The log-rank test and Cox proportional hazard model were used to study the survival analysis. Of the 82 patients included in the study, 56 were male. The mean age was 57 years. 86.6% had a known primary carcinoma site while the remaining 13.4% had none. The two most common known carcinoma sites were the lung and biliary systems. Among the 11 unknown primary sites, the most common histological finding was adenocarcinoma. The mean survival period was 8.7 ± 11.7 months. The survival analysis revealed two statistically significant factors: the primary tumor site's aggressiveness (P<0.005) and the presence of visceral metastasis (P<0.05). The prevalence of identifiable primary site was 86.6% and the most common site was the lungs followed by the biliary system. The primary carcinoma site's aggressiveness and the presence of visceral metastasis were the factors associated with patient survival.


Structure, mechanism and catalytic duality of thiamine-dependent enzymes.
Thiamine is an essential cofactor that is required for processes of general metabolism amongst all organisms, and it is likely to have played a role in the earliest stages of the evolution of life. Here, we review from a structural perspective the enzymatic mechanisms that involve this cofactor. We explore asymmetry within homodimeric thiamine diphosphate (ThDP)-dependent enzyme structures and discuss how this may be correlated with the kinetic properties of half-of-the-sites reactivity, and negative cooperativity. It is likely these structural and kinetic hallmarks may arise through reciprocal coupling of active sites. This mode of communication between distant active sites is not unique to ThDP-dependent enzymes, but is widespread in other classes of oligomeric enzyme. Thus, it appears likely to be a general phenomenon reflecting a powerful mechanism of accelerating the rate of a chemical pathway. Finally, we speculate on the early evolutionary history of the cofactor and its ancient association with protein and RNA.


Dramatic Response of BRAF V600E Mutant Papillary Craniopharyngioma to Targeted Therapy.
We recently reported that BRAF V600E is the principal oncogenic driver of papillary craniopharyngioma, a highly morbid intracranial tumor commonly refractory to treatment. Here, we describe our treatment of a man age 39 years with multiply recurrent BRAF V600E craniopharyngioma using dabrafenib (150mg, orally twice daily) and trametinib (2mg, orally twice daily). After 35 days of treatment, tumor volume was reduced by 85%. Mutations that commonly mediate resistance to MAPK pathway inhibition were not detected in a post-treatment sample by whole exome sequencing. A blood-based BRAF V600E assay detected circulating BRAF V600E in the patient's blood. Re-evaluation of the existing management paradigms for craniopharyngioma is warranted, as patient morbidity might be reduced by noninvasive mutation testing and neoadjuvant-targeted treatment.


HLA-A11-restricted epitope polymorphism among Epstein-Barr virus strains in the highly HLA-A11-positive Chinese population: incidence and immunogenicity of variant epitope sequences.
An individual's CD8(+)-cytotoxic-T-lymphocyte (CTL) response to Epstein-Barr virus (EBV) latent cycle antigens focuses on a small number of immunodominant epitopes often presented by just one of the available HLA class I alleles; for example, HLA-A11-positive Caucasians frequently respond to two immunodominant HLA A11 epitopes, IVTDFSVIK (IVT) and AVFDRKSDAK (AVF), within the nuclear antigen EBNA3B. Here, we reexamine the spectrum of EBV strains present in the highly HLA-A11-positive Chinese population for sequence changes in these epitopes relative to the Caucasian type 1 prototype strain B95.8. The IVT epitope was altered in 61 of 64 Chinese type 1 viruses, with four different sequence variants being observed, and the AVF epitope was altered in 46 cases with six different sequence variants; by contrast, all 10 Chinese type 2 viruses retained the prototype 2 epitope sequences. All but one of the type 1 epitope variants were poorly recognized by IVT- or AVF-specific CTLs in pulse-chase assays of peptide-mediated target cell lysis. More importantly, we screened HLA-A11-positive Chinese donors carrying viruses with known epitope mutations for evidence of epitope-specific CTL memory by enzyme-linked immunospot assays: none of the type 1 variants tested, nor the type 2 prototype, appeared to be immunogenic in vivo. The data remain consistent with the possibility that, during virus-host coevolution, pressure from the host CTL-mediated immune response has given A11 epitope-loss viruses a selective advantage.


Survival of dairy cows after surgery to correct abomasal displacement: 2. Association of clinical and laboratory parameters with survival in cows with left abomasal displacement.
This study examines the association of pre-surgical clinical parameters in cows with left displacement of the abomasum (LDA) with the probability of cure. Data from 466 cases of LDA in which omentopexy was performed were analysed. Cows were classified in three categories according to the outcome of treatment. Cows in the first category were cured, i.e. sent back to the farm for further milk production. Cows in the second category were sent for slaughter because of an unfavourable prognosis concerning productivity. Cows in the third category either died or were salvaged because of poor prognosis concerning survival. Factors were examined for their possible influence on the outcome of treatment by comparing the values of successfully treated cows to those cows that either died or had to be sent for slaughter. Factors that were associated with a favourable prognosis were a short duration of disease, an undisturbed general condition, good appetite, normal faeces, a higher body weight, lower haematocrit, haemoglobin and erythrocyte counts, lower urea, AST and bilirubin, and higher sodium, potassium and chloride concentrations compared with cows with an unfavourable prognosis. Haemoconcentration was associated with higher AST, bilirubin and urea and with low sodium, potassium and chloride. Acetonuria was associated with increased AST and bilirubin and low urea. Low sodium and potassium concentrations were associated with an impaired general condition. It is concluded that a thorough clinical examination with special emphasis on general condition, liver function and dehydration status is of great importance in determining the prognosis of abdominal surgery in cows with LDA.


Universality, maximum radiation, and absorption in high-energy collisions of black holes with spin.
We explore the impact of black hole spins on the dynamics of high-energy black hole collisions. We report results from numerical simulations with γ factors up to 2.49 and dimensionless spin parameter χ=+0.85, +0.6, 0, -0.6, -0.85. We find that the scattering threshold becomes independent of spin at large center-of-mass energies, confirming previous conjectures that structure does not matter in ultrarelativistic collisions. It has further been argued that in this limit all of the kinetic energy of the system may be radiated by fine tuning the impact parameter to threshold. On the contrary, we find that only about 60% of the kinetic energy is radiated for γ=2.49. By monitoring apparent horizons before and after scattering events we show that the "missing energy" is absorbed by the individual black holes in the encounter, and moreover the individual black-hole spins change significantly. We support this conclusion with perturbative calculations. An extrapolation of our results to the limit γ→∞ suggests that about half of the center-of-mass energy of the system can be emitted in gravitational radiation, while the rest must be converted into rest-mass and spin energy.


Retrospective injury epidemiology of one hundred one competitive Oceania power lifters: the effects of age, body mass, competitive standard, and gender.
The injury epidemiology of competitive power lifters was investigated to provide a basis for injury prevention initiatives in power lifting. Self-reported retrospective injury data for 1 year and selected biographical and training information were obtained via a 4-page injury survey from 82 men and 19 women of varying ages (Open and Masters), body masses (lightweight and heavyweight), and competitive standards (national and international). Injury was defined as any physical damage to the body that caused the lifter to miss or modify one or more training sessions or miss a competition. A total of 118 injuries, which equated to 1.2 +/- 1.1 injuries per lifter per year and 4.4 +/- 4.8 injuries per 1,000 hours of training, were reported. The most commonly injured body regions were the shoulder (36%), lower back (24%), elbow (11%), and knee (9%). More injuries appeared to be of a sudden (acute) (59%) rather than gradual (chronic) nature (41%). National competitors had a significantly greater rate of injury (5.8 +/- 4.9 per 1,000 hours) than international competitors (3.6 +/- 3.6 per 1,000 hours). The relative proportion of injuries at some body regions varied significantly as a function of competitive standard and gender. No significant differences in injury profile were seen between Open and Masters or between lightweight and heavyweight lifters. Power lifting appears to have a moderately low risk of injury, regardless of the lifter's age, body mass, competitive standard, or gender, compared with other sports. Future research should utilize a prospective cohort or case-controlled design to examine the effect of a range of other intrinsic and extrinsic factors on injury epidemiology and to assess the effects of various intervention strategies.


Laparoscopic antrectomy for the treatment of type I gastric carcinoid tumors.
While the optimal treatment for type I gastric carcinoid tumors remains controversial, there is evidence to suggest that in multifocal disease, antrectomy may not only control local disease but also may lead to enterochromaffin-like cell (ECL) hyperplasia regression compared to medical and endoscopic treatments. A single institution retrospective review of eight consecutive patients with multifocal type I gastric carcinoid tumor patients with no evidence of metastatic disease was performed from 2005 to 2006. All of these patients underwent laparoscopic antrectomy with Billroth II reconstruction. Patients' preoperative gastrin, chromogranin A levels, and biopsy and surgical specimen slides were compared with postoperative laboratory and biopsy slides. Pathology slides were reanalyzed by a blinded pathologist from our institution for evidence of tumor and ECL hyperplasia regression. All patients tolerated the procedure well with no reoperations or mortalities. Six of eight patient complained of mild reflux which was treated medically. One of eight had a mild wound infection which resolved with a course of cephalexin. Gastrin levels significantly decreased (98.9%) in all patients (P = 0.001). Furthermore, chromogranin A levels also significantly decreased (81.4%). Eight of eight patients showed no evidence of carcinoid tumor after surgery at mean biopsy follow-up of 17 mo (range 2-35 mo), however there was ECL hyperplasia after resection. Four of eight patients (50%) showed regression of ECL hyperplasia on postop biopsy, while the remaining four of eight showed no evidence of regression. This is the largest case series to investigate the surgical, clinical, and histologic outcomes of laparoscopic antrectomy in type I gastric carcinoid. Our data suggest that laparoscopic antrectomy is a safe and minimally invasive approach to treat nonmetastatic type I gastric carcinoid. All patients had no evidence of gross or microscopic disease at follow-up biopsy and almost half had regression of ECL hyperplasia at follow-up suggesting that antrectomy may be sufficient to prevent tumor recurrence. However, continued regular endoscopic surveillance and medical follow-up of patients with ECL hyperplasia are recommended.


Planar Laser-Induced Fluorescence fuel concentration measurements in isothermal Diesel sprays.
This paper presents a complete methodology to perform fuel concentration measurements of Diesel sprays in isothermal conditions using the Planar Laser-Induced Fluorescence (PLIF) technique. The natural fluorescence of a commercial Diesel fuel is used with an excitation wavelength of 355 nm. The correction and calibration procedures to perform accurate measurements are studied. These procedures include the study of the fluorescence characteristics of the fuel as well as the correction of the laser sheet non-homogeneities and the losses due to Mie scattering, absorption and autoabsorption. The results obtained are compared with theoretical models and other experimental techniques.


In vitro-in vivo correlation of parenteral risperidone polymeric microspheres.
The objective of the present study was to determine whether an in vitro-in vivo correlation (IVIVC) can be established for polymeric microspheres that are equivalent in formulation composition but prepared with different manufacturing processes. Risperidone was chosen as a model therapeutic and poly(lactic-co-glycolic acid) (PLGA) with similar molecular weight as that used in the commercial product Risperdal® Consta® was used to prepare risperidone microspheres. Various manufacturing processes were investigated to produce the risperidone microspheres with similar drug loading (approx. 37%) but distinctly different physicochemical properties (e.g. porosity, particle size and particle size distribution). In vitro release of the risperidone microspheres was investigated using different release testing methods (such as sample-and-separate and USP apparatus 4). In vivo pharmacokinetic profiles of the risperidone microsphere formulations following intramuscular administration were determined using a rabbit model. Furthermore, the obtained pharmacokinetic profiles were deconvoluted using the Loo-Riegelman method and the calculated in vivo release was compared with the in vitro release of these microspheres. Level A IVIVCs were established and validated for the compositionally equivalent risperidone microspheres based on the in vitro release data obtained using USP apparatus 4. The developed IVIVCs demonstrated good predictability and were robust. These results showed that the developed USP apparatus 4 method was capable of discriminating PLGA microspheres that are equivalent in formulation composition but with manufacturing differences and predicting their in vivo performance in the investigated animal model.


Can enhanced screening of men with a history of prior syphilis infection stem the epidemic in men who have sex with men? A mathematical modelling study.
The aim of this study is to determine the transmission impact of using prior syphilis infection to guide a focused syphilis screening intervention among men who have sex with men (MSM). We parameterised a deterministic model of syphilis transmission in MSM to reflect the 2011-2015 syphilis outbreak in Winnipeg, Canada. Enhanced screening of 75% of men with prior syphilis every 3 months (A) was compared with distributing equivalent number tests to all MSM (B) or those with the highest partner number (C). We compared early syphilis incidence, diagnoses and prevalence after 10 years, relative to a base case of 30% of MSM screened annually. Strategy A was expected to avert 52% of incident infections, 44% of diagnosed cases and reduce early syphilis prevalence by 89%. Strategy B had the least impact. Strategy C was most effective, averting 59% of incident cases. When screening frequency was semiannual or annual, strategy A was the most effective. Enhanced screening of MSM with prior syphilis may efficiently reduce transmission, especially when identification of high-risk men via self-reported partner numbers or high-frequency screening is difficult to achieve.


The role of pregnancy-specific beta-1 glycoprotein (SP1) in assessing the human blastocyst quality in vitro.
The advent of new culture techniques resulting in more than 60% of embryos developing in vitro to the blastocyst stage suggests that blastocyst transfer in humans with its potential to increase in-vitro fertilization success rates could be a feasible option. Blastocyst quality markers, however, are required to ensure that an increase in numbers is not achieved at the expense of lowering quality. We have previously reported a morphology based method for grading blastocysts. The current study sought to determine whether the secretion of pregnancy-specific beta-1-glycoprotein (SP1) (a trophoblast product known to be associated with fetal well-being) by blastocysts of differing quality was reflected in the morphological grading. SP1 concentrations were measured in the culture medium of grade 1 (n = 19), grade 2 (n = 4) and grade 3 (n = 4) blastocysts as well as vacuolated morulae (n = 6) daily from day 1 to day 14. Cumulative SP1 concentrations secreted by blastocysts were significantly higher than those secreted by vacuolated morulae. However, SP1 levels could not distinguish between blastocysts of differing morphological grades. The inconsistent pattern of secretion suggests that at this early stage of development, SP1 secretion is probably not sufficiently defined to allow any differences in levels to be reflected by the morphological grading.


GI-RADS reporting system for ultrasound evaluation of adnexal masses in clinical practice: a prospective multicenter study.
To assess the clinical usefulness of a structured reporting system based on ultrasound findings for management of adnexal masses. This was a prospective multicenter study comprising 432 adnexal masses in 372 women (mean age, 44.0 (range, 13-78) years) over a 36-month period. Ninety-three (25%) women were postmenopausal and 279 (75%) women were premenopausal. Patients were evaluated with transvaginal ultrasound by one of three examiners expert in gynecological ultrasound. Reporting was provided to referring clinicians according to the Gynecologic Imaging Report and Data System (GI-RADS) classification. A predetermined management protocol was offered to referral clinicians. It was suggested that patients classified as GI-RADS 2 be managed with follow-up scan, patients classified as GI-RADS 3 undergo laparoscopic surgery and patients classified as GI-RADS 4 or 5 be referred to a gynecologic oncologist. Definitive histologic diagnosis was available in 370 cases and 62 additional cases were considered as benign because of spontaneous resolution during follow-up. These outcomes were used as the gold standard for calculating the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (LR+) and negative likelihood ratio (LR-) of GI-RADS classification for identifying adnexal masses at high risk of malignancy, considering GI-RADS 4 and 5 as being malignant. Of the 432 tumors, 112 were malignant and 320 benign. The GI-RADS classification rate was as follows: GI-RADS 2, 92 (21%) cases; GI-RADS 3, 184 (43%) cases; GI-RADS 4, 40 (9%) cases; GI-RADS 5, (27%) 116 cases. Sensitivity for this system was 99.1% (95% CI, 95.1-99.8%), specificity was 85.9% (95% CI, 81.7-89.3%), LR+ was 7.05 (95% CI, 5.37-9.45) and LR- was 0.01 (95% CI, 0.001-0.07). PPV and NPV were 71.1% and 99.6%, respectively. The GI-RADS reporting system performed well in identifying adnexal masses at high risk of malignancy and seems to be useful for clinical decision-making.


Demonstrating compliance with protection objectives for non-human biota within post-closure safety cases for radioactive waste repositories.
Over recent years, a number of approaches have been developed that enable the calculation of dose rates to animals and plants following the release of radioactivity to the environment. These approaches can be used to assess the potential impacts of activities that may release radioactivity to the environment, such as the operation of waste repositories. A number of national and international studies have identified screening criteria to indicate those assessment results below which further consideration is not generally required. However no internationally agreed criteria are currently available and consistency in criteria between countries has not been achieved. Furthermore, since screening criteria are not intended to be applied as limits, it is clear that they cannot always form a sufficient basis for assessing the adequacy of protection afforded. Typically, exceeding a screening value leads to a regulatory requirement to undertake a further, more detailed assessment. It does not, per se, imply that there is inadequate protection of the organism types at the specific site under assessment. Therefore, there is a need to develop a more structured approach to dealing with situations in which current screening criteria are exceeded. As a contribution to the developing international discussions, and as an interim measure for application where assessments are required currently, a two-tier, three zone framework is proposed here, relevant to the long term assessment of potential impacts from the deep disposal of radioactive wastes. The purpose of the proposed framework is to promote a proportionate and risk-based approach to the level of effort required in undertaking and interpreting an assessment.


Quick measurement of protein sulfhydryls with Ellman's reagent and with 4,4'-dithiodipyridine.
Since its introduction in 1959, Ellman's reagent (5,5'-dithio-bis(2-nitrobenzoic acid)) has been the favorite reagent for spectrophotometric measurement of protein sulfhydryls. Meanwhile however, evidence has accumulated that many protein sulfhydryls give an incomplete reaction with Ellman's reagent, even during prolonged assay times. In the present study, the kinetic problem was solved by including cystamine as a "mediator" between the protein sulfhydryl and Ellman's reagent, as previously applied in an enzymatic thiol assay [9]. As an alternative, 4,4'-dithiodipyridine (DTDP) was used in place of Ellman's reagent. Due to its small size, amphiphilic nature, and lack of charge, DTDP quickly reacts with poorly accessible protein sulfhydryls, without any catalysis by cystamine. The DTDP method and the Ellman/cystamine method were both optimized for maximal sensitivity, minimal sample consumption (detection limit 0.2 nmol mL(-1), determination limit 0.6 nmol mL(-1)), and minimal assay time (5 min). In validation experiments, both methods gave identical results and the measured sulfhydryls/protein matched the expected values. Electronic supplementary material to this paper can be obtained by using the Springer Link server located at http://dx.doi.org/10.1007/s00216-002-1347-2.


Warfarin-cranberry juice interaction.
To report a case of warfarin-cranberry juice interaction, which resulted in an international normalized ratio (INR) elevation on 2 separate occasions. A 46-year-old female was receiving a total weekly dose of 56 mg of warfarin. During the 4 months prior to the incident INR, her average INR was 2.0, with a range of 1.6-2.2, while taking the same weekly dose of warfarin. Her INR increased to 4.6 after drinking approximately 1.5 quarts (1420 mL) of cranberry juice cocktail daily for 2 days. Her INR 14 days later without cranberry juice cocktail consumption was 2.3. For the next 3 months, while taking warfarin 56 mg per week, her average INR was 2.1, with a range of 1.4-2.5. At a subsequent visit, after drinking approximately 2 quarts (1893 mL) of cranberry juice cocktail daily for 3-4 days, her INR had increased to 6.5. Her INR after holding warfarin for 3 days was 1.86. Her INR 7 days after resuming the weekly dose of warfarin 56 mg was 3.2. During both of the elevated INR episodes, no other factors were identified that would have resulted in an elevated INR, such as drug, herbal, disease, or other food interactions. An objective causality assessment revealed the interaction was highly probable. Warfarin is the most commonly used anticoagulant for chronic therapy. There have been several case reports of cranberry juice or cranberry sauce potentiating the effects of warfarin by elevating the INR; however, clinical trials evaluating this interaction have failed to demonstrate a significant effect on an INR. Our case report describes INR elevations in a patient previously stable on warfarin after ingestion of cranberry juice cocktail daily for several days. This elevation occurred on 2 separate occasions, which distinguishes our case from other published literature.


Does Ureaplasma spp. cause chronic lung disease of prematurity: ask the audience?
Ureaplasma has long been implicated in the pathogenesis of both preterm labour and neonatal morbidity, particularly chronic lung disease of prematurity (CLD), but despite numerous studies, reviews and meta-analyses, its exact role remains unclear. Many papers call for a definitive randomised control trial to determine if eradication of pulmonary Ureaplasma decreases the rates of CLD but few address in detail the obstacles to an adequately powered clinical trial. We review the evidence for Ureaplasma as a causative agent in CLD, asking why a randomised control trial has not been performed. We surveyed the opinions of senior neonatologists in the UK on whether they felt that there was sufficient evidence for Ureaplasma either causing or not causing CLD and whether a definitive trial was needed, as well as their views on the design of such a trial. Additionally, we ascertained current practice with respect to Ureaplasma detection in preterm neonates in the UK. There is clear support for an adequately powered randomised controlled clinical trial by senior neonatologists in the UK. There are no reasons why a definitive trial cannot be conducted especially as the appropriate samples, and methods to culture or identify the organism by PCR are already available.


Delineation of clinical conditions: conceptual models of 'physical' and 'mental disorders.
The dysfunctional consequences of the Cartesian dichotomy have been enhanced by the power of biomedical technology. Technical virtuosity reifies the mechanical model and widens the gap between what patients seek and doctors provide. Patients suffer 'illnesses'; doctors diagnose and treat 'diseases'. Illnesses are experiences of discontinuities in states of being and perceived role performances. Diseases, in the scientific paradigm of modern medicine, are abnormalities in the function and/or structure of body organs and systems. Traditional healers also redefine illness as disease: because they share symbols and metaphors consonant with lay beliefs, their healing rituals are more responsive to the psychosocial context of illness. Psychiatric disorders offer an illuminating perspective on the basic medical dilemma. The paradigms for psychiatric practice include multiple and ostensibly contradictory models: organic, psychodynamic, behavioural and social. This mélange of concepts stems from the fact that the fundamental manifestations of psychosis are disordered behaviours. The psychotic patient remains a person; his self-concept and relationships with others are cental to the therapeutic encounter, whatever pharmacological adjuncts are employed. The same truths hold for all patients. The social matrix determines when and how the patient seeks what kind of help, his 'compliance' with the recommended regimen and, to a significant extent, the functional outcome. When physicians dismiss illness because ascertainable 'disease' is absent, they fail to meet their socially assigned responsibility. It is essential to reintegrate 'scientific' and 'social' concepts of disease and illness as a basis for a functional system of medical research and care.


Experimental evaluation of iterative reconstruction versus filtered back projection for 3D [15O]water PET activation studies using statistical parametric mapping analysis.
Iterative reconstructions are increasingly used for clinical PET studies owing to the better noise properties compared with filtered backprojection (FBP). The purpose of the present study was to compare ordered subsets expectation maximization (OSEM) iterative reconstruction with FBP as input for statistical parametric mapping (SPM) analysis of PET activation studies. Two phantom studies were performed simulating both motor and cognitive tasks and acquiring data with both high and low statistics. In contrast to clinical studies, where no a priori information is known, phantom studies allow for an accurate and detailed comparison between different reconstruction techniques. The significance of "activations" during "tasks" was determined using SPM99 software. Using region of interest analysis of SPM results, it was found that the maximum and average t values within each hot spot of the phantom were higher for OSEM than for FBP. In addition, OSEM4 x 16 (4 iterations, 16 subsets) produced fewer false-positive voxels than FBP, OSEM1 x 16 and OSEM2 x 16. In conclusion, for PET activation studies use of OSEM4 x 16 seems to give the best tradeoff between signal detection and noise reduction.


Relatedness and inbreeding avoidance: counterploys in the communally nesting acorn woodpecker.
Acorn woodpeckers (Melanerpes formicivorus) live in family groups within which more than one female may lay eggs communally in a single nest. Communally nesting females are usually closely related and share evenly in nesting activities. Although birds of either sex may breed in their natal territory, reproductive inhibition of offspring by the presence of their parent of the opposite sex and dispersal by unisexual sibling units ensure that inbreeding between close relatives is rare.


Wallerian degeneration of the corticofugal tracts in chronic stroke: a pilot study relating diffusion tensor imaging, transcranial magnetic stimulation, and hand function.
This study tested the hypothesis that degeneration of the corticofugal tracts (CFTs) is related to poor functional outcome in the upper limb after stroke. . The authors used diffusion tensor imaging to determine the degree of white matter integrity of the CFT (FA(AH/UH)) in chronic stroke patients and controls. The functional integrity of the corticospinal pathway was examined using transcranial magnetic stimulation. Recruitment curves and silent period duration were measured. The relationship between FA(AH/UH) and functional outcome of the upper limb was also assessed using a battery of upper limb function tests. In patients, FA( AH/UH) correlated positively with the slope of recruitment curves (RC(AH/UH)) and grip strength. FA(AH/UH) also correlated negatively with the silent period duration (SP(AH/UH)). According to the FA(AH/UH), patients were also classified into groups with minimal or extensive Wallerian degeneration (WD). Patients with more extensive WD had poorer grip strength, dexterity, and range of movement. The authors' results suggest that structural and functional integrity of the CFT is essential to maintain function of the upper limb after stroke.


Symptom Interference and Relation between the Domains of Quality of Life among Cancer Patients of Tertiary Care Hospital.
Quality of life (QOL) should be considered as the main outcome measure for patient with advanced cancer. QOL of cancer patients is affected by several factors. The objective of the study was to assess the symptom interference and the relation between the domains of QOL among cancer patients. An exploratory survey conducted among 768 patients diagnosed as Stage III or IV of cancer of breast/cervix/head and neck/gastrointestinal tract/lung/colorectal. Data were collected using pretested reliable structured interview questionnaire. Out of 768 cancer patients, symptoms burden was interfered completely in their relationship with others among 445 (57.9%), sleep among 491 (63.9%), and QOL among 433 (56.4%). The domains of QOL were general well-being, physical well-being, psychological well-being, familial relationship, sexual and personal abilities, cognitive well-being, economic well-being, informational support, doctors' cooperation, and body image. Since the data were not following the normal distribution, Spearman rho was computed to assess the relationship between the domains. The data revealed that there is a positive statistically significant correlation between the domains of general well-being of the participants with physical wellbeing (r = 0.265, P = 0.001), psychological well-being (r = 0.195, P = 0.001), sexual and personal abilities (r = 0.278, P = 0.001), and body image (r = 0.168, P = 0.001). The study inferred that cancer patients with good cognitive well-being and high economic status had a sense of positive body image. Cancer patients have poor QOL in physical and psychological domains. QOL of patients is affected by the symptoms suffering during the treatment and diagnosis. Symptoms need to be managed effectively to improve the QOL of cancer patients.


COL4A2 in the tissue-specific extracellular matrix plays important role on osteogenic differentiation of periodontal ligament stem cells.
Periodontal ligament stem cells (PDLSCs) can repair alveolar bone defects in periodontitis in a microenvironment context-dependent manner. This study aimed to determine whether different extracellular matrices (ECMs) exert diverse effects on osteogenic differentiation of PDLSCs and accurately control alveolar bone defect repair. Methods: The characteristics of PDLSCs and bone marrow mesenchymal stem cells (BMSCs) with respect to surface markers and multi-differentiation ability were determined. Then, we prepared periodontal ligament cells (PDLCs)-derived and bone marrow cells (BMCs)-derived ECMs (P-ECM and B-ECM) and the related decellularized ECMs (dECMs). Transmission electron microscopy (TEM), scanning electron microscopy (SEM), atomic force microscopy (AFM), and protein mass spectrometry were used to distinguish the ECMs. The expression of Type IV collagen A2 (COL4A2) in the ECMs was inhibited by siRNA or activated by lentiviral transduction of relevant cells. The stemness, proliferation, and differentiation of PDLSCs were determined in vitro in different dECMs. For the in vivo analysis, different dECMs under the regulation of COL4A2 mixed with PDLSCs and Bio-Oss bone powder were subcutaneously implanted into immunocompromised mice or in defects in rat alveolar bone. The repair effects were identified by histological or immunohistochemical staining and micro-CT. Results: B-dECM exhibited more compact fibers than P-dECM, as revealed by TEM, SEM, and AFM. Protein mass spectrometry showed that COL4A2 was significantly increased in B-dECM compared with P-dECM. PDLSCs displayed stronger proliferation, stemness, and osteogenic differentiation ability when cultured on B-dECM than P-dECM. Interestingly, B-dECM enhanced the osteogenic differentiation of PDLSCs to a greater extent than P-dECM both in vitro and in vivo, whereas downregulation of COL4A2 in B-dECM showed the opposite results. Furthermore, the classical Wnt/β-catenin pathway was found to play an important role in the negative regulation of osteogenesis through COL4A2, confirmed by experiments with the Wnt inhibitor DKK-1 and the Wnt activator Wnt3a. Conclusion: These findings indicate that COL4A2 in the ECM promotes osteogenic differentiation of PDLSCs through negative regulation of the Wnt/β-catenin pathway, which can be used as a potential therapeutic strategy to repair bone defects.


Auto-context and its application to high-level vision tasks and 3D brain image segmentation.
The notion of using context information for solving high-level vision and medical image segmentation problems has been increasingly realized in the field. However, how to learn an effective and efficient context model, together with an image appearance model, remains mostly unknown. The current literature using Markov Random Fields (MRFs) and Conditional Random Fields (CRFs) often involves specific algorithm design in which the modeling and computing stages are studied in isolation. In this paper, we propose a learning algorithm, auto-context. Given a set of training images and their corresponding label maps, we first learn a classifier on local image patches. The discriminative probability (or classification confidence) maps created by the learned classifier are then used as context information, in addition to the original image patches, to train a new classifier. The algorithm then iterates until convergence. Auto-context integrates low-level and context information by fusing a large number of low-level appearance features with context and implicit shape information. The resulting discriminative algorithm is general and easy to implement. Under nearly the same parameter settings in training, we apply the algorithm to three challenging vision applications: foreground/background segregation, human body configuration estimation, and scene region labeling. Moreover, context also plays a very important role in medical/brain images where the anatomical structures are mostly constrained to relatively fixed positions. With only some slight changes resulting from using 3D instead of 2D features, the auto-context algorithm applied to brain MRI image segmentation is shown to outperform state-of-the-art algorithms specifically designed for this domain. Furthermore, the scope of the proposed algorithm goes beyond image analysis and it has the potential to be used for a wide variety of problems for structured prediction problems.


A descriptive analysis of internal and external loads for elite-level tennis drills.
Planning tennis sessions accentuating physical development requires an understanding of training load (TL). The aims were to describe the external and internal TL of drills and analyze relationships between ratings of perceived exertion (RPE), TL, and other measures. Fourteen elite-level junior tennis athletes completed 259 individual drills. Six coaches helped devise classifications for all drills: recovery/defensive, open pattern, accuracy, 2-on-1 open, 2-on-1 net play, closed technical, point play, and match play. Notational analysis on stroke and error rates was performed postsession. Drill RPE and mental exertion were collected postdrill, while heart rate (HR) was recorded continuously. Recovery/defensive, open pattern, and point play were significantly greater than closed technical drills (P < .05) for RPE and mental exertion, as were accuracy drills and match play (P < .05). Recovery/defensive, open-pattern, accuracy, and 2-on-1 open drills had higher stroke rates than match play (P < .05). Error rates of closed technical drills were significantly higher than for open pattern, 2-on-1 drills, point play, and match play (P < .05). No HR differences were observed (P > .05) between categories. Substantial correlations existed for drill RPE and TL with mental exertion (r > .62) for several categories. TL was substantially correlated with total strokes (r > .65), while HR and stroke and error rates were in slight to moderate agreement with RPE and TL (r < .51). Recovery/defensive drills are highest in physiological stress, making them ideal for maximizing physicality. Recovery/defensive drills compromised training quality, eliciting high error rates. In contrast, 2-on-1 net-play drills provided the lowest error rates, potentially appropriate for error-amelioration practice. Open-pattern drills were characterized by significantly higher stroke rates, suggesting congruence with high-repetition practice. Finally, with strong relationships between physical and mental perception, mental exertion may complement currently used monitoring strategies (TL and RPE).


Impaired skeletal muscle microvascular function and increased skeletal muscle oxygen consumption in severe falciparum malaria.
Organ dysfunction and tissue hypoxia in severe falciparum malaria result from an imbalance between oxygen delivery and demand. In severe malaria, microvascular obstruction from parasite sequestration decreases oxygen delivery. However, host microvascular function (defined as the capacity to increase oxygen delivery in response to ischemia) and oxygen consumption have not been assessed. We used near-infrared resonance spectroscopy to measure thenar muscle microvascular function (StO(2)recov) and oxygen consumption (VO(2)) in 36 adults in Papua, Indonesia, with severe malaria, 33 with moderately severe malaria (MSM), 24 with severe sepsis, and 36 healthy controls. In the severe malaria group, the StO(2)recov of 2.7%/second was 16% and 22% lower than that in the MSM group (3.1%/second) and control group (3.5%/second), respectively (P < .001), and comparable to that in the severe sepsis group (2.5%/second). In the severe malaria group, StO(2)recov was inversely correlated with lactate level (r = -0.63; P < .001) and predicted death (area under the receiver operating characteristic curve, 0.71 [95% confidence interval {CI}, .51-.92]), with each percentage decrease associated with an increased odds of mortality (odds ratio, 2.49 [95% CI, 1.05-6.2]). Conversely, VO(2) increased in the severe malaria group by 18%, compared with levels in the control and severe sepsis groups (P < .001), and was associated with parasite biomass (r = 0.49; P = .04). Impaired microvascular function is associated with increased mortality among individuals with severe malaria, while oxygen consumption is increased. Tissue hypoxia may result not only from microvascular obstruction, but also from impaired ability of the microvasculature to match oxygen delivery to increased oxygen demand.


An Exclusion Zone for Ca2+ Channels around Docked Vesicles Explains Release Control by Multiple Channels at a CNS Synapse.
The spatial arrangement of Ca2+ channels and vesicles remains unknown for most CNS synapses, despite of the crucial importance of this geometrical parameter for the Ca2+ control of transmitter release. At a large model synapse, the calyx of Held, transmitter release is controlled by several Ca2+ channels in a "domain overlap" mode, at least in young animals. To study the geometrical constraints of Ca2+ channel placement in domain overlap control of release, we used stochastic MCell modelling, at active zones for which the position of docked vesicles was derived from electron microscopy (EM). We found that random placement of Ca2+ channels was unable to produce high slope values between release and presynaptic Ca2+ entry, a hallmark of domain overlap, and yielded excessively large release probabilities. The simple assumption that Ca2+ channels can be located anywhere at active zones, except below a critical distance of ~ 30 nm away from docked vesicles ("exclusion zone"), rescued high slope values and low release probabilities. Alternatively, high slope values can also be obtained by placing all Ca2+ channels into a single supercluster, which however results in significantly higher heterogeneity of release probabilities. We also show experimentally that high slope values, and the sensitivity to the slow Ca2+ chelator EGTA-AM, are maintained with developmental maturation of the calyx synapse. Taken together, domain overlap control of release represents a highly organized active zone architecture in which Ca2+ channels must obey a certain distance to docked vesicles. Furthermore, domain overlap can be employed by near-mature, fast-releasing synapses.


CMEIAS color segmentation: an improved computing technology to process color images for quantitative microbial ecology studies at single-cell resolution.
Quantitative microscopy and digital image analysis are underutilized in microbial ecology largely because of the laborious task to segment foreground object pixels from background, especially in complex color micrographs of environmental samples. In this paper, we describe an improved computing technology developed to alleviate this limitation. The system's uniqueness is its ability to edit digital images accurately when presented with the difficult yet commonplace challenge of removing background pixels whose three-dimensional color space overlaps the range that defines foreground objects. Image segmentation is accomplished by utilizing algorithms that address color and spatial relationships of user-selected foreground object pixels. Performance of the color segmentation algorithm evaluated on 26 complex micrographs at single pixel resolution had an overall pixel classification accuracy of 99+%. Several applications illustrate how this improved computing technology can successfully resolve numerous challenges of complex color segmentation in order to produce images from which quantitative information can be accurately extracted, thereby gain new perspectives on the in situ ecology of microorganisms. Examples include improvements in the quantitative analysis of (1) microbial abundance and phylotype diversity of single cells classified by their discriminating color within heterogeneous communities, (2) cell viability, (3) spatial relationships and intensity of bacterial gene expression involved in cellular communication between individual cells within rhizoplane biofilms, and (4) biofilm ecophysiology based on ribotype-differentiated radioactive substrate utilization. The stand-alone executable file plus user manual and tutorial images for this color segmentation computing application are freely available at http://cme.msu.edu/cmeias/ . This improved computing technology opens new opportunities of imaging applications where discriminating colors really matter most, thereby strengthening quantitative microscopy-based approaches to advance microbial ecology in situ at individual single-cell resolution.


Droplet sorting based on the number of encapsulated particles using a solenoid valve.
Droplet microfluidics provides a high-throughput platform for screening subjects and conditions involved in biology. Droplets with encapsulated beads and cells have been increasingly used for studying molecular and cellular biology. Droplet sorting is needed to isolate and analyze the subject of interest during such screening. The vast majority of current sorting techniques use fluorescence intensity emitted by each droplet as the only criterion. However, due to the randomness and imperfections in the encapsulation process, typically a mixed population of droplets with an uneven number of encapsulated particles results and is used for screening. Thus droplet sorting based on the number of encapsulated particles becomes necessary for isolating or enriching droplets with a specific occupancy. In this work, we developed a fluorescence-activated microfluidic droplet sorter that integrated a simple deflection mechanism based on the use of a solenoid valve and a sophisticated signal processing system with a microcontroller as the core. By passing droplets through a narrow interrogation channel, the encapsulated particles were detected individually. The microcontroller conducted the computation to determine the number of encapsulated particles in each droplet and made the sorting decision accordingly that led to actuation of the solenoid valve. We tested both fluorescent beads and stained cells and our results showed high efficiency and accuracy for sorting and enrichment.


GRADE guidelines 6. Rating the quality of evidence--imprecision.
GRADE suggests that examination of 95% confidence intervals (CIs) provides the optimal primary approach to decisions regarding imprecision. For practice guidelines, rating down the quality of evidence (i.e., confidence in estimates of effect) is required if clinical action would differ if the upper versus the lower boundary of the CI represented the truth. An exception to this rule occurs when an effect is large, and consideration of CIs alone suggests a robust effect, but the total sample size is not large and the number of events is small. Under these circumstances, one should consider rating down for imprecision. To inform this decision, one can calculate the number of patients required for an adequately powered individual trial (termed the "optimal information size" [OIS]). For continuous variables, we suggest a similar process, initially considering the upper and lower limits of the CI, and subsequently calculating an OIS. Systematic reviews require a somewhat different approach. If the 95% CI excludes a relative risk (RR) of 1.0, and the total number of events or patients exceeds the OIS criterion, precision is adequate. If the 95% CI includes appreciable benefit or harm (we suggest an RR of under 0.75 or over 1.25 as a rough guide) rating down for imprecision may be appropriate even if OIS criteria are met.


The Contribution of Genetic Risk and Lifestyle Factors in the Development of Adult-Onset Inflammatory Bowel Disease: A Prospective Cohort Study.
The joint associations across genetic risk, modifiable lifestyle factors, and inflammatory bowel disease (IBD) remains unclear. Genetic susceptibility to Crohn's disease (CD) and ulcerative colitis (UC) was estimated by polygenic risk scores and further categorized into high, intermediate, and low genetic risk categories. Weighted healthy lifestyle scores were constructed based on 5 common lifestyle factors and categorized into favorable (4 or 5 healthy lifestyle factors), intermediate (3 healthy lifestyle factors), and unfavorable (0-2 healthy lifestyle factors) groups. Cox proportional hazards regression model was used to estimate the hazard ratios (HR) and 95% confidence interval (CI) for their associations. During the 12-year follow-up, 707 cases with CD and 1576 cases with UC were diagnosed in the UK Biobank cohort. Genetic risk and unhealthy lifestyle categories were monotonically associated with CD and UC risk with no multiplicative interaction between them. The HR of CD and UC were 2.24 (95% CI 1.75-2.86) and 2.15 (95% CI 1.82-2.53) for those with a high genetic risk, respectively. The HR of CD and UC for individuals with an unfavorable lifestyle were 1.94 (95% CI 1.61-2.33) and 1.98 (95% CI 1.73-2.27), respectively. The HR of individuals with a high genetic risk but a favorable lifestyle (2.33, 95% CI 1.58-3.44 for CD, and 2.05, 95% CI 1.58-2.66 for UC) were reduced nearly by half, compared with those with a high genetic risk but an unfavorable lifestyle (4.40, 95% CI 2.91-6.66 for CD and 4.44, 95% CI 3.34-5.91 for UC). Genetic and lifestyle factors were independently associated with susceptibility to incident CD and UC. Adherence to a favorable lifestyle was associated with a nearly 50% lower risk of CD and UC among participants at a high genetic risk.


Molecular characterization of sour orange (Citrus aurantium) accessions and their relatives using SSR and SRAP markers.
Citrus production with its many varieties is of importance since it provides economically important products for Turkish exports. Sour orange is a rootstock commonly used for propagating the different scion varieties. Knowledge of the genetic diversity of the rootstock accessions would be useful in order to improve citrus breeding programs. We studied genetic relationships and diversity of 51 accessions of sour orange (Citrus aurantium) and their relatives using SSR (simple sequence repeat) and SRAP (sequence-related amplified polymorphism) molecular markers. Twenty-one SRAP primer combinations were tested on these accessions and relatives, producing 167 polymorphic fragments, with a mean of 8.0 and a mean polymorphism information content value of 0.47. Seventeen SSR primers also produced 30 polymorphic fragments, with a mean of 1.4 per primer and a mean polymorphism information content value of 0.39. The unweighted pair-group method with arithmetic average analysis using combined SSR and SRAP data showed a similarity range from 0.71 to 1.00 among the accessions. In the cluster analysis, sour orange relatives were indicated as a separate group from sour orange. 'Macrophylla' and 'Mexican lime' were the accessions most distinct (0.71) from the others. We conclude that genetic diversity in these sour orange accessions is lower and some of them were identical.


Androgen receptor as potential therapeutic target in metastatic endometrial cancer.
The expression and involvement of estrogen (ER) and progesterone receptor (PR) is extensively studied in endometrial cancer. Androgen receptor (AR) is a hormone receptor less studied in female cancers, and we here aim to investigate the expression level of AR in endometrial cancer precursor lesions, primary tumors and metastases, and its potential as therapeutic target. Expression of AR was observed in 93% of hyperplasias, but only in 41% of non-endometrioid tumors. Compared to estrogen and progesterone receptor AR is more commonly expressed in metastatic lesions, and AR status is discordant in primary and metastatic lesions in a large proportion of cases. AR protein level was significantly associated with survival (P < 0.001), and a calculated AR to ERα ratio identified a subgroup of patients with particular poor outcome. The anti-androgen enzalutamide may have a growth inhibitory effect in endometrial cancer cells based on experiments with primary endometrial tumor cells. 718 primary endometrial cancers and 298 metastatic lesions (from 142 patients) were investigated for expression of AR in relation to survival, clinical and histopathological data. Protein levels were investigated by immunohistochemistry and reverse phase protein array; mRNA levels by DNA oligonucleotide microarray. The effect of androgen stimulation and inhibition was tested on primary endometrial tumor cells. A large proportion of metastatic endometrial cancer lesions express AR, which may be a potential target in these patients. Treatment targeting AR may be of particular benefit in patients with high AR levels compared to ERα levels.


Deconstructing crop processes and models via identities.
This paper is part review and part opinion piece; it has three parts of increasing novelty and speculation in approach. The first presents an overview of how some of the major crop simulation models approach the issue of simulating the responses of crops to changing climatic and weather variables, mainly atmospheric CO2 concentration and increased and/or varying temperatures. It illustrates an important principle in models of a single cause having alternative effects and vice versa. The second part suggests some features, mostly missing in current crop models, that need to be included in the future, focussing on extreme events such as high temperature or extreme drought. The final opinion part is speculative but novel. It describes an approach to deconstruct resource use efficiencies into their constituent identities or elements based on the Kaya-Porter identity, each of which can be examined for responses to climate and climatic change. We give no promise that the final part is 'correct', but we hope it can be a stimulation to thought, hypothesis and experiment, and perhaps a new modelling approach.


Maintenance of p53 alterations throughout breast cancer progression.
Overexpression of the nuclear phosphoprotein p53 is one of the most common abnormalities in primary human cancer and appears to be due to point mutation within a highly conserved region of the p53 gene which then encodes for a mutant, more stable protein. In this study different stages of breast cancer progression were examined, from in situ to metastatic disease, to determine at what stage mutational activation occurs and whether it is maintained during tumor progression. Two (13%) of 15 pure intraductal tumors expressed high levels of p53 in all malignant epithelial cells. Sequencing of p53 mRNA from one of these tumors demonstrated a nucleotide substitution altering the amino acid composition of the protein. Six (17%) of 35 specimens which contained both in situ and invasive disease expressed high levels of p53. All malignant epithelial cells in these 6 cases stained positively and in no specimen did one component express different levels of the protein than the other growth phase. Sequence analysis of a tissue with significant amounts of both in situ and invasive disease revealed only a single point mutation, without evidence of wild-type nucleotide at the site of substitution, suggesting that p53 mRNA from each component of the tumor contained the same nucleotide substitution. Eleven (50%) of 22 pairs of primary tumors and their lymph node metastases expressed elevated levels of p53, and in each case, expression levels were identical in the primary and secondary sites. Identical mutations were found in the p53 mRNA from two paired primary and metastatic sites. Therefore, mutation within a highly conserved region of the p53 gene leading to overexpression of the protein product can occur in the earliest recognized phase of breast cancer and this alteration is maintained during progression from intraductal to infiltrating carcinoma. Mutations are also conserved during the process of metastatic spread.


Senescence-related changes in the responsiveness to ouabain of canine Purkinje fibers.
Clinical studies have suggested that old adults are more susceptible to the toxic effects of digitalis than young adults. To test whether this reflects an age-related change in responsiveness to digitalis of cardiac fibers, standard microelectrode techniques were used to study the effects of ouabain (O) on Purkinje fibers (PF) from 5- and 10-year-old beagles. PF driven at cycle length = 600 msec were superfused with O, 2 x 10(-7) M, containing 3% [3H]O for 25 min. PF-O uptake was the same at both ages. Control maximum diastolic potential, action potential amplitude, upstroke velocity (Vmax) and duration to 50% repolarization (APD50) did not differ significantly between the two age groups. However, the effects of O on action potential characteristics and on delayed after depolarizations in PF from the 10 year old PF were significantly greater than that on the 5 year old PF. Additional experiments were done in which we superfused PF from adult and old animals with O for long periods to determine the time-response relationships for ouabain effects on maximum diastolic potential, action potential amplitude, Vmax and APD50. The rate of change was 2 to 3 times greater in the 10 than the 5 year old PF. After 55 min of O, only 2 of 7 10-year-old PF remained excitable, compared with 6 of 6 5-year-old PF. In conclusion, PF from old dogs show greater responsiveness to the toxic effects of O than those of younger adults, explaining in part the clinical observation of increased digitalis toxicity with age.


Modulation of the immunological synapse: a key to HIV-1 pathogenesis?
AIDS is the result of a constant struggle between the lentivirus HIV and the immune system. Infection with HIV interferes directly with the function of CD4(+) T cells and manipulates the host immune response to the virus. Recent studies indicate that the viral protein Nef, a central player in HIV pathogenesis, impairs the ability of infected lymphocytes to form immunological synapses with antigen-presenting cells and affects T-cell-receptor-mediated stimulation. An integrative picture of the abnormal behaviour of HIV-infected lymphocytes is therefore emerging. We propose that modulating lymphocyte signalling, apoptosis and intracellular trafficking ensures efficient spread of the virus in the hostile environment of the immune system.


Horizontal gene transfer and gene dosage drives adaptation to wood colonization in a tree pathogen.
Some of the most damaging tree pathogens can attack woody stems, causing lesions (cankers) that may be lethal. To identify the genomic determinants of wood colonization leading to canker formation, we sequenced the genomes of the poplar canker pathogen, Mycosphaerella populorum, and the closely related poplar leaf pathogen, M. populicola. A secondary metabolite cluster unique to M. populorum is fully activated following induction by poplar wood and leaves. In addition, genes encoding hemicellulose-degrading enzymes, peptidases, and metabolite transporters were more abundant and were up-regulated in M. populorum growing on poplar wood-chip medium compared with M. populicola. The secondary gene cluster and several of the carbohydrate degradation genes have the signature of horizontal transfer from ascomycete fungi associated with wood decay and from prokaryotes. Acquisition and maintenance of the gene battery necessary for growth in woody tissues and gene dosage resulting in gene expression reconfiguration appear to be responsible for the adaptation of M. populorum to infect, colonize, and cause mortality on poplar woody stems.


Synergy and remarkable specificity of antimicrobial peptides in vivo using a systematic knockout approach.
Antimicrobial peptides (AMPs) are host-encoded antibiotics that combat invading microorganisms. These short, cationic peptides have been implicated in many biological processes, primarily involving innate immunity. In vitro studies have shown AMPs kill bacteria and fungi at physiological concentrations, but little validation has been done in vivo. We utilized CRISPR gene editing to delete all known immune-inducible AMPs of Drosophila, namely: 4 Attacins, 4 Cecropins, 2 Diptericins, Drosocin, Drosomycin, Metchnikowin and Defensin. Using individual and multiple knockouts, including flies lacking all 14 AMP genes, we characterize the in vivo function of individual and groups of AMPs against diverse bacterial and fungal pathogens. We found that Drosophila AMPs act primarily against Gram-negative bacteria and fungi, contributing either additively or synergistically. We also describe remarkable specificity wherein certain AMPs contribute the bulk of microbicidal activity against specific pathogens, providing functional demonstrations of highly specific AMP-pathogen interactions in an in vivo setting.


SOX6 suppresses cyclin D1 promoter activity by interacting with beta-catenin and histone deacetylase 1, and its down-regulation induces pancreatic beta-cell proliferation.
Sex-determining region Y-box (SOX) 6 negatively regulates glucose-stimulated insulin secretion from beta-cells and is a down-regulated transcription factor in the pancreatic islet cells of hyperinsulinemic obese mice. To determine the contribution of SOX6 to insulin resistance, we analyzed the effects of SOX6 on cell proliferation. Small interfering RNA-mediated attenuation of SOX6 expression stimulated the proliferation of insulinoma INS-1E and NIH-3T3 cells, whereas retroviral overexpression resulted in inhibition of cell growth. Quantitative real time-PCR analysis revealed that the levels of cyclin D1 transcripts were markedly decreased by SOX6 overexpression. Luciferase-reporter assay with beta-catenin showed that SOX6 suppresses cyclin D1 promoter activities. In vitro binding experiments showed that the LZ/Q domain of SOX6 physically interacts with armadillo repeats 1-4 of beta-catenin. Furthermore, chromatin immunoprecipitation assay revealed that increased SOX6 expression significantly reduced the levels of acetylated histones H3 and H4 at the cyclin D1 promoter. By using a histone deacetylase (HDAC) inhibitor and co-immunoprecipitation analysis, we showed that SOX6 suppressed cyclin D1 activities by interacting withbeta-catenin and HDAC1. The data presented suggest that SOX6 may be an important factor in obesity-related insulin resistance.


Piscivore efficiency and refuging prey: the importance of predator search mode.
In predator-prey interactions, the efficiency of the predator is dependent on characteristics of both the predator and the prey, as well as the structure of the environment. In a field enclosure experiment, we tested the effects of a prey refuge on predator search mode, predator efficiency and prey behaviour. Replicated enclosures containing young of the year (0+) and 1-year-old (1+) perch were stocked with 3 differentially sized individuals of either of 2 piscivorous species, perch (Perca fluviatilis), pike (Esox lucius) or no piscivorous predators. Each enclosure contained an open predator area with three small vegetation patches, and a vegetated absolute refuge for the prey. We quantified the behaviour of the predators and the prey simultaneously, and at the end of the experiment the growth of the predators and the mortality and habitat use of the prey were estimated. The activity mode of both predator species was stationary. Perch stayed in pairs in the vegetation patches whereas pike remained solitary and occupied the corners of the enclosure. The largest pike individuals stayed closest to the prey refuge whereas the smallest individuals stayed farthest away from the prey refuge, indicating size-dependent interference among pike. Both size classes of prey showed stronger behavioural responses to pike than to perch with respect to refuge use, distance from refuge and distance to the nearest predator. Prey mortality was higher in the presence of pike than in the presence of perch. Predators decreased in body mass in all treatments, and perch showed a relatively stronger decrease in body mass than pike during the experiment. Growth differences of perch and pike, and mortality differences of prey caused by predation, can be explained by predator morphology, predator attack efficiency and social versus interference behaviour of the predators. These considerations suggest that pike are more efficient piscivores around prey refuges such as the littoral zones of lakes, whereas perch have previously been observed to be more efficient in open areas, such as in the pelagic zones of lakes.


Epidemiology of Lung Cancer.
Lung cancer continues to be one of the most common causes of cancer death despite understanding the major cause of the disease: cigarette smoking. Smoking increases lung cancer risk 5- to 10-fold with a clear dose-response relationship. Exposure to environmental tobacco smoke among nonsmokers increases lung cancer risk about 20%. Risks for marijuana and hookah use, and the new e-cigarettes, are yet to be consistently defined and will be important areas for continued research as use of these products increases. Other known environmental risk factors include exposures to radon, asbestos, diesel, and ionizing radiation. Host factors have also been associated with lung cancer risk, including family history of lung cancer, history of chronic obstructive pulmonary disease and infections. Studies to identify genes associated with lung cancer susceptibility have consistently identified chromosomal regions on 15q25, 6p21 and 5p15 associated with lung cancer risk. Risk prediction models for lung cancer typically include age, sex, cigarette smoking intensity and/or duration, medical history, and occupational exposures, however there is not yet a risk prediction model currently recommended for general use. As lung cancer screening becomes more widespread, a validated model will be needed to better define risk groups to inform screening guidelines.


Visual appearance of a virtual upper limb modulates the temperature of the real hand: a thermal imaging study in Immersive Virtual Reality.
To explore the link between Sense of Embodiment (SoE) over a virtual hand and physiological regulation of skin temperature, 24 healthy participants were immersed in virtual reality through a Head Mounted Display and had their real limb temperature recorded by means of a high-sensitivity infrared camera. Participants observed a virtual right upper limb (appearing either normally, or with the hand detached from the forearm) or limb-shaped non-corporeal control objects (continuous or discontinuous wooden blocks) from a first-person perspective. Subjective ratings of SoE were collected in each observation condition, as well as temperatures of the right and left hand, wrist and forearm. The observation of these complex, body and body-related virtual scenes resulted in increased real hand temperature when compared to a baseline condition in which a 3d virtual ball was presented. Crucially, observation of non-natural appearances of the virtual limb (discontinuous limb) and limb-shaped non-corporeal objects elicited high increase in real hand temperature and low SoE. In contrast, observation of the full virtual limb caused high SoE and low temperature changes in the real hand with respect to the other conditions. Interestingly, the temperature difference across the different conditions occurred according to a topographic rule that included both hands. Our study sheds new light on the role of an external hand's visual appearance and suggests a tight link between higher-order bodily self-representations and topographic regulation of skin temperature.


Translating comparative effectiveness of depression medications into practice by comparing the depression medication choice decision aid to usual care: study protocol for a randomized controlled trial.
Comparative effectiveness research (CER) documents important differences in antidepressants in terms of efficacy, safety, cost, and burden to the patient. Decision aids can adapt this evidence to help patients participate in making informed choices. In turn, antidepressant therapy will more likely reflect patients' values and context, leading to improved adherence and mood outcomes. The objective of this study is to develop the Depression Medication Choice decision aid for use during primary care encounters, and to test its efficacy by conducting a clustered practical randomized trial comparing the decision aid to usual depression care in primary care practices.We will use a novel practice-based, patient-centered approach based on participatory action research that involves a multidisciplinary team of designers, investigators, clinicians, patient representatives, and other stakeholders for the development of the decision aid. We will then conduct a clustered practical randomized trial enrolling clinicians and their patients (n = 300) with moderate to severe depression from rural, suburban and inner city primary care practices (n = 10). The intervention will consist of the use of the depression medication choice decision aid during the clinical encounter. This trial will generate preliminary evidence of the relative impact of the decision aid on patient involvement in decision making, decision making quality, patient knowledge, and 6-month measures of medication adherence and mental health compared to usual depression care. Upon completion of the proposed research, we will have developed and evaluated the efficacy of the decision aid depression medication choice as a novel translational tool for CER in depression treatment, engaged patients with depression in their care, and refined the process by which we conduct practice-based trials with limited research footprint. Clinical Trials.gov: NCT01502891.


Sequencing and de novo analysis of a coral larval transcriptome using 454 GSFlx.
New methods are needed for genomic-scale analysis of emerging model organisms that exemplify important biological questions but lack fully sequenced genomes. For example, there is an urgent need to understand the potential for corals to adapt to climate change, but few molecular resources are available for studying these processes in reef-building corals. To facilitate genomics studies in corals and other non-model systems, we describe methods for transcriptome sequencing using 454, as well as strategies for assembling a useful catalog of genes from the output. We have applied these methods to sequence the transcriptome of planulae larvae from the coral Acropora millepora. More than 600,000 reads produced in a single 454 sequencing run were assembled into approximately 40,000 contigs with five-fold average sequencing coverage. Based on sequence similarity with known proteins, these analyses identified approximately 11,000 different genes expressed in a range of conditions including thermal stress and settlement induction. Assembled sequences were annotated with gene names, conserved domains, and Gene Ontology terms. Targeted searches using these annotations identified the majority of genes associated with essential metabolic pathways and conserved signaling pathways, as well as novel candidate genes for stress-related processes. Comparisons with the genome of the anemone Nematostella vectensis revealed approximately 8,500 pairs of orthologs and approximately 100 candidate coral-specific genes. More than 30,000 SNPs were detected in the coral sequences, and a subset of these validated by re-sequencing. The methods described here for deep sequencing of the transcriptome should be widely applicable to generate catalogs of genes and genetic markers in emerging model organisms. Our data provide the most comprehensive sequence resource currently available for reef-building corals, and include an extensive collection of potential genetic markers for association and population connectivity studies. The characterization of the larval transcriptome for this widely-studied coral will enable research into the biological processes underlying stress responses in corals and evolutionary adaptation to global climate change.


Small molecule modulator of aggrephagy regulates neuroinflammation to curb pathogenesis of neurodegeneration.
Plethora of efforts fails to yield a single drug to reverse the pathogenesis of Parkinson's disease (PD) and related α-synucleopathies. Using chemical biology, we identified a small molecule inhibitor of c-abl kinase, PD180970 that could potentially clear the toxic protein aggregates. Genetic, molecular, cell biological and immunological assays were performed to understand the mechanism of action. In vivo preclinical disease model of PD was used to assess its neuroprotection efficacy. In this report, we show the ability of a small molecule inhibitor of tyrosine kinases, PD180970, to induce autophagy (cell lines and mice midbrain) in an mTOR-independent manner and ameliorate the α-synuclein mediated toxicity. PD180970 also exerts anti-neuroinflammatory potential by inhibiting the release of proinflammatory cytokines such as IL-6 (interleukin-6) and MCP-1 (monocyte chemoattractant protein-1) through reduction of TLR-4 (toll like receptor-4) mediated NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) activation. In vivo studies show that PD180970 is neuroprotective by degrading the toxic protein oligomers through induction of autophagy and subsiding the microglial activation. These protective mechanisms ensure the negation of Parkinson's disease related motor impairments. FUND: This work was supported by Wellcome Trust/DBT India Alliance Intermediate Fellowship (500159-Z-09-Z), DST-SERB grant (EMR/2015/001946), DBT (BT/INF/22/SP27679/2018) and JNCASR intramural funds to RM, and SERB, DST (SR/SO/HS/0121/2012) to PAA, and DST-SERB (SB/YS/LS-215/2013) to JPC and BIRAC funding to ETA C-CAMP.


Characterization of vanM carrying clinical Enterococcus isolates and diversity of the suppressed vanM gene cluster.
Here we report the prevalence of the suppressed vanM gene cluster as a reservoir of vancomycin resistance genes. Among 1284 clinical isolates of enterococci from four hospitals in Hangzhou, China, 55 isolates of Enterococcus faecium and one isolate of Enterococcus faecalis were screened positive for the vanM genotype. Antimicrobial susceptibility testing showed that 55 of the 56 vanM-positive isolates were susceptible to vancomycin and teicoplanin. Most of them (54/56) belonged to the main epidemic lineage CC17, mostly the ST78 type. The vanM gene clusters in the 55 vancomycin-susceptible isolates showed sequence diversity owing to different insertion locations of IS1216E. The vanM transposons could be classified into five types and they all carried two or more IS1216E elements, leading to complete or partial deletions of vanR, vanS, or vanX. Quantitative reverse transcription polymerase chain reaction showed that the expression level of vanM was significantly lower in the vancomycin-susceptible isolates than in the vancomycin-resistant isolate. Considering the prevalence of the vanM genotype and the potential for conversion to a resistant phenotype, vanM might act as an important determinant of glycopeptide resistance in the future. It is essential to strengthen the surveillance of vanM-containing enterococci to control the dissemination of vancomycin resistance.


FSTL1 Promotes Metastasis and Chemoresistance in Esophageal Squamous Cell Carcinoma through NFκB-BMP Signaling Cross-talk.
Esophageal squamous cell carcinoma (ESCC) has a generally poor prognosis, and molecular markers to improve early detection and predict outcomes are greatly needed. Here, we report that the BMP-binding follistatin-like protein FSTL1 is overexpressed in ESCCs, where it correlates with poor overall survival. Genetic amplification of FSTL1 or chromosome 3q, where it is located, occurred frequently in ESCC, where FSTL1 copy number correlated positively with higher FSTL1 protein expression. Elevating FSTL1 levels by various means was sufficient to drive ESCC cell proliferation, clonogenicity, migration, invasion, self-renewal, and cisplatin resistance in vitro and tumorigenicity and distant metastasis in vivo Conversely, FSTL1 attenuation by shRNA or neutralizing antibody elicited the opposite effects in ESCC cells. mRNA profiling analyses suggested that FSTL1 drives ESCC oncogenesis and metastasis through various pathways, with deregulation of NFκB and BMP signaling figuring prominently. Cross-talk between the NFκB and BMP pathways was evidenced by functional rescue experiments using inhibitors of NFκB and TLR4. Our results establish the significance of FSTL1 in driving oncogenesis and metastasis in ESCC by coordinating NFκB and BMP pathway control, with implications for its potential use as a diagnostic or prognostic biomarker and as a candidate therapeutic target in this disease setting. Cancer Res; 77(21); 5886-99. ©2017 AACR.


Parents' and professionals' perceptions on causes and treatment options for Autism Spectrum Disorders (ASD) in a multicultural context on the Kenyan Coast.
To explore parents' and professionals' perceived causes and treatment of Autism Spectrum Disorders (ASD) on the Kenyan Coast. In-depth interviews and focus group discussions using guiding questions were utilized in data collection. One hundred and three participants, who included parents of children with ASD, special needs teachers, clinicians, and social workers from diverse cultural background, participated in this study. The interviews and focus groups were recorded, transcribed verbatim and then translated to English. Themes were generated using content analysis. Preternatural causes were mentioned and included evil spirits, witchcraft, and curses. Biomedical causes comprised infections, drug abuse, birth complications, malnutrition, and genetic related problems. Treatment varied from traditional and spiritual healing to modern treatment in health facilities, and included consultations with traditional healers, offering prayers to God, and visits to hospitals. The results suggest that regardless of cultural backgrounds, people on the Kenyan Coast have similar views on perceived causes and treatment of ASD. These findings provide valuable conceptual understanding for professionals when planning and implementing community based rehabilitation interventions targeting children with ASD within a local context.