Domain-Targeted Metabolomics Delineates the Heterocycle Assembly Steps of Colibactin Biosynthesis.
Modular polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs) comprise giant multidomain enzymes responsible for the "assembly line" biosynthesis of many genetically encoded small molecules. Site-directed mutagenesis, protein biochemical, and structural studies have focused on elucidating the catalytic mechanisms of individual multidomain proteins and protein domains within these megasynthases. However, probing their functions at the cellular level typically has invoked the complete deletion (or overexpression) of multidomain-encoding genes or combinations of genes and comparing those mutants with a control pathway. Here we describe a "domain-targeted" metabolomic strategy that combines genome editing with pathway analysis to probe the functions of individual PKS and NRPS catalytic domains at the cellular metabolic level. We apply the approach to the bacterial colibactin pathway, a genotoxic NRPS-PKS hybrid pathway found in certain Escherichia coli. The pathway produces precolibactins, which are converted to colibactins by a dedicated peptidase, ClbP. Domain-targeted metabolomics enabled the characterization of "multidomain signatures", or functional readouts of NRPS-PKS domain contributions to the pathway-dependent metabolome. These multidomain signatures provided experimental support for individual domain contributions to colibactin biosynthesis and delineated the assembly line timing events of colibactin heterocycle formation. The analysis also led to the structural characterization of two reactive precolibactin metabolites. We demonstrate the fate of these reactive intermediates in the presence and absence of ClbP, which dictates the formation of distinct product groups resulting from alternative cyclization cascades. In the presence of the peptidase, the reactive intermediates are converted to a known genotoxic scaffold, providing metabolic support of our mechanistic model for colibactin-induced genotoxicity. Domain-targeted metabolomics could be more widely used to characterize NRPS-PKS pathways with unprecedented genetic and metabolic precision.


Propranolol in the prevention of the first hemorrhage from esophagogastric varices: A multicenter, randomized clinical trial. The Boston-New Haven-Barcelona Portal Hypertension Study Group.
To assess the effectiveness of propranolol in the prevention of initial variceal hemorrhage, a double-blind, randomized trial was carried out in three centers. Patients with cirrhosis (78% alcoholic), hepatic venous pressure gradients greater than 12 mm Hg and endoscopically proven esophageal varices were randomly assigned to propranolol (51 patients) or placebo (51 patients). Of the 102 patients, 58% were Child's class A, 34% were Child's class B and 8% were Child's class C. Daily dosage was determined by the administration of progressively increasing doses of propranolol with the hepatic vein catheter in place to achieve a 25% decrease in hepatic venous pressure gradient, a decrease in hepatic venous pressure gradient to less than 12 mm Hg or a decrease in resting heart rate to less than 55 beats/min. During a mean follow-up period of 16.3 mo, 11 patients in the placebo group (22%) bled from esophageal varices compared with 2 in the propranolol group (4%) during a mean period of 17.1 mo (p less than 0.01). Three additional patients (6%) in the placebo group bled from portal hypertensive gastropathy compared with none in the propranolol group. Propranolol appeared effective in preventing bleeding from large varices. Eleven deaths (22%) occurred in the placebo group compared with eight deaths (16%) in the propranolol group (NS). The mean dose of propranolol was 132 mg/day, and the median dose was 80 mg/day. Using a compliance index (pill count, clinic attendance, alcohol and propranolol levels and alcohol history), 81% of the propranolol patients and 77% of the placebo patients were considered compliant. Complications severe enough to require cessation of therapy occurred in eight patients (16%) in the propranolol group and four in the placebo group (8%) (NS). We conclude that propranolol effectively prevents the first variceal hemorrhage in patients with alcoholic cirrhosis and large esophageal varices but does not improve survival.


Systematic review on hormone receptor testing in breast cancer.
Assessment of hormone receptors (estrogen and progesterone) helps to direct therapy for women with breast cancer. Immunohistochemistry is most commonly used to assess hormone receptor status and it is essential that these tests are performed accurately and reliably within and across laboratories. The overall purpose of this guideline is to improve the quality and accuracy of hormone receptor testing and its utility as a prognostic and predictive marker for invasive and in situ breast cancer. Medline, EMBASE, the Cochrane Database of Systematic Reviews, and abstracts from the San Antonio Breast Cancer Symposium were searched. An environmental scan of the internet and of international guideline developers and key organizations was performed. Preanalytic elements such as the collection, fixation, and storage of samples, and analytic elements such as selection of antibodies and scoring methods that seem to offer the best results for immunohistochemical assessment of hormone receptors are presented. Proficiency testing or quality assurance of immunohistochemistry is described.


Use of computed tomography scout film and Hounsfield unit of computed tomography scan in predicting the radio-opacity of urinary calculi in plain kidney, ureter and bladder radiographs.
The objective of this study is to determine the diagnostic utility of computed tomography (CT)- scout film with an optimal non-contrast helical CT scan Hounsfield unit (HU) in predicting the appearance of urinary calculus in the plain kidneys, ureter, urinary bladder (KUB)-radiograph. A prospective cross-sectional study was executed and data were collected from June 2007 to June 2012 at a tertiary hospital. The included subjects were diagnosed to have <10mm urolithiasis with non-contrast helical CT scan and KUB X-ray, which were carried out on the same day. Both KUB radiographs and CT-scout film were read by two qualified radiologists with inter-observer standardization prior to the study. Urolithiasis characteristics such as stone location, CT attenuation value, CT-scout film and KUB radiograph appearance were recorded independently by two observers. Univariate logistic analysis with receiver operating characteristic curve was generated to determine the best cut-off HU value of urolithiases not identified in CT-scout film, but determined radio-opaque in KUB X-ray. Subsequently, its sensitivity, specificity, predictive values and likelihood ratios were calculated. Statistical significance was set at P value of 0.05 or less. Two hundred and three valid cases were included. 73 out of 75 CT-scout film detected urolithiasis were identified on plain radiograph and determined as radio-opaque. The determined best cut off value of HU utilized for prediction of radiographic characteristics was 630HU at which urinary calculi were not seen at CT-scout film and were KUB X-ray radio-opaque. The set HU cut-off was established of ideal accuracy with an overall sensitivity of 82.2%, specificity of 96.9% and a positive predictive value of 96.5% and negative predictive value of 83.5%. Urolithiases identified on the CT-scout film were also seen as radiopaque on the KUB radiograph while those stones not visible on the CT-scout film, but above the optimal HU cut-off value of 630 are also likely to be radiopaque.


Inverse source problem and minimum-energy sources.
We present a new linear inversion formalism for the scalar inverse source problem in three-dimensional and one-dimensional (1D) spaces, from which a number of previously unknown results on minimum-energy (ME) sources and their fields readily follow. ME sources, of specified support, are shown to obey a homogeneous Helmholtz equation in the interior of that support. As a consequence of that result, the fields produced by ME sources are shown to obey an iterated homogeneous Helmholtz equation. By solving the latter equation, we arrive at a new Green-function representation of the field produced by a ME source. It is also shown that any square-integrable (L2), compactly supported source that possesses a continuous normal derivative on the boundary of its support must possess a nonradiating (NR) component. A procedure based on our results on the inverse source problem and ME sources is described to uniquely decompose an L2 source of specified support and its field into the sum of a radiating and a NR part. The general theory that is developed is illustrated for the special cases of a homogeneous source in 1D space and a spherically symmetric source.


Fluid-structure coupled biotransport processes in aortic valve disease.
Biological transport processes near the aortic valve play a crucial role in calcific aortic valve disease initiation and bioprosthetic aortic valve thrombosis. Hemodynamics coupled with the dynamics of the leaflets regulate these transport patterns. Herein, two-way coupled fluid-structure interaction (FSI) simulations of a 2D bicuspid aortic valve and a 3D mechanical heart valve were performed and coupled with various convective mass transport models that represent some of the transport processes in calcification and thrombosis. Namely, five different continuum transport models were developed to study biochemicals that originate from the blood and the leaflets, as well as residence-time and flow stagnation. Low-density lipoprotein (LDL) and platelet activation were studied for their role in calcification and thrombosis, respectively. Coherent structures were identified using vorticity and Lagrangian coherent structures (LCS) for the 2D and 3D models, respectively. A very close connection between vortex structures and biochemical concentration patterns was shown where different vortices controlled the concentration patterns depending on the transport mechanism. Additionally, the relationship between leaflet concentration and wall shear stress was revealed. Our work shows that blood flow physics and coherent structures regulate the flow-mediated biological processes that are involved in aortic valve calcification and thrombosis, and therefore could be used in the design process to optimize heart valve replacement durability.


Continental-scale, seasonal movements of a heterothermic migratory tree bat.
Long-distance migration evolved independently in bats and unique migration behaviors are likely, but because of their cryptic lifestyles, many details remain unknown. North American hoary bats (Lasiurus cinereus cinereus) roost in trees year-round and probably migrate farther than any other bats, yet we still lack basic information about their migration patterns and wintering locations or strategies. This information is needed to better understand unprecedented fatality of hoary bats at wind turbines during autumn migration and to determine whether the species could be susceptible to an emerging disease affecting hibernating bats. Our aim was to infer probable seasonal movements of individual hoary bats to better understand their migration and seasonal distribution in North America. We analyzed the stable isotope values of non-exchangeable hydrogen in the keratin of bat hair and combined isotopic results with prior distributional information to derive relative probability density surfaces for the geographic origins of individuals. We then mapped probable directions and distances of seasonal movement. Results indicate that hoary bats summer across broad areas. In addition to assumed latitudinal migration, we uncovered evidence of longitudinal movement by hoary bats from inland summering grounds to coastal regions during autumn and winter. Coastal regions with nonfreezing temperatures may be important wintering areas for hoary bats. Hoary bats migrating through any particular area, such as a wind turbine facility in autumn, are likely to have originated from a broad expanse of summering grounds from which they have traveled in no recognizable order. Better characterizing migration patterns and wintering behaviors of hoary bats sheds light on the evolution of migration and provides context for conserving these migrants.


Developing a new national MDMA policy: Results of a multi-decision multi-criterion decision analysis.
Ecstasy (3,4-methylenedioxymethamphetamine (MDMA)) has a relatively low harm and low dependence liability but is scheduled on List I of the Dutch Opium Act ('hard drugs'). Concerns surrounding increasing MDMA-related criminality coupled with the possibly inappropriate scheduling of MDMA initiated a debate to revise the current Dutch ecstasy policy. An interdisciplinary group of 18 experts on health, social harms and drug criminality and law enforcement reformulated the science-based Dutch MDMA policy using multi-decision multi-criterion decision analysis (MD-MCDA). The experts collectively formulated policy instruments and rated their effects on 25 outcome criteria, including health, criminality, law enforcement and financial issues, thematically grouped in six clusters. The experts scored the effect of 22 policy instruments, each with between two and seven different mutually exclusive options, on 25 outcome criteria. The optimal policy model was defined by the set of 22 policy instrument options which gave the highest overall score on the 25 outcome criteria. Implementation of the optimal policy model, including regulated MDMA sales, decreases health harms, MDMA-related organised crime and environmental damage, as well as increases state revenues and quality of MDMA products and user information. This model was slightly modified to increase its political feasibility. Sensitivity analyses showed that the outcomes of the current MD-MCDA are robust and independent of variability in weight values. The present results provide a feasible and realistic set of policy instrument options to revise the legislation towards a rational MDMA policy that is likely to reduce both adverse (public) health risks and MDMA-related criminal burden.


Histone H2A C-terminus regulates chromatin dynamics, remodeling, and histone H1 binding.
The tails of histone proteins are central players for all chromatin-mediated processes. Whereas the N-terminal histone tails have been studied extensively, little is known about the function of the H2A C-terminus. Here, we show that the H2A C-terminal tail plays a pivotal role in regulating chromatin structure and dynamics. We find that cells expressing C-terminally truncated H2A show increased stress sensitivity. Moreover, both the complete and the partial deletion of the tail result in increased histone exchange kinetics and nucleosome mobility in vivo and in vitro. Importantly, our experiments reveal that the H2A C-terminus is required for efficient nucleosome translocation by ISWI-type chromatin remodelers and acts as a novel recognition module for linker histone H1. Thus, we suggest that the H2A C-terminal tail has a bipartite function: stabilisation of the nucleosomal core particle, as well as mediation of the protein interactions that control chromatin dynamics and conformation.


A Comparison of Pig Farmers' and Veterinarians' Perceptions and Intentions to Reduce Antimicrobial Usage in Six European Countries.
Antimicrobial (AM) resistance is an increasing problem in human and veterinary medicine. To manage this problem, the usage of AM should be reduced in pig farming, as well as in other areas. It is important to investigate the factors that influence both pig farmers' and veterinarians' intentions to reduce AM usage, which is a prerequisite for developing intervention measures. We conducted a mail survey among pig farmers (N = 1,294) and an online survey among veterinarians (N = 334) in Belgium, Denmark, France, Germany, Sweden and Switzerland. The farmers' survey assessed the perceived risks and benefits of and need for AM usage; the intention to reduce AM usage; farmers' efficacy (i.e. perception of their ability to reduce AM usage); support from their veterinarian; and the future reduction potential of AM usage. Additionally, self-reported reduction behaviours, the perceived farmers' barriers to reduce AM usage and relationships with farmers were assessed in the veterinarians' survey. The results showed that farmers and veterinarians had similar perceptions of the risks and benefits of AM usage. Veterinarians appeared to be more optimistic than pig farmers about reducing AM usage in pig farming. Farmers believed that their efficacy over AM reduction was relatively high. Farmers' intention to reduce AM usage and veterinarians' self-reported reduction behaviours were mainly associated with factors concerning the feasibility of reducing AM usage. To promote prudent AM usage, pig farmers should learn and experience how to reduce usage by applying alternative measures, whereas veterinarians should strengthen their advisory role and competencies to support and educate farmers.


Effect of alternative NAD+-regenerating pathways on the formation of primary and secondary aroma compounds in a Saccharomyces cerevisiae glycerol-defective mutant.
Saccharomyces cerevisiae maintains a redox balance under fermentative growth conditions by re-oxidizing NADH formed during glycolysis through ethanol formation. Excess NADH stimulates the synthesis of mainly glycerol, but also of other compounds. Here, we investigated the production of primary and secondary metabolites in S. cerevisiae strains where the glycerol production pathway was inactivated through deletion of the two glycerol-3-phosphate dehydrogenases genes (GPD1/GPD2) and replaced with alternative NAD(+)-generating pathways. While these modifications decreased fermentative ability compared to the wild-type strain, all improved growth and/or fermentative ability of the gpd1Δgpd2Δ strain in self-generated anaerobic high sugar medium. The partial NAD(+) regeneration ability of the mutants resulted in significant amounts of alternative products, but at lower yields than glycerol. Compared to the wild-type strain, pyruvate production increased in most genetically manipulated strains, whereas acetate and succinate production decreased in all strains. Malate production was similar in all strains. Isobutanol production increased substantially in all genetically manipulated strains compared to the wild-type strain, whereas only mutant strains expressing the sorbitol producing SOR1 and srlD genes showed increases in isoamyl alcohol and 2-phenyl alcohol. A marked reduction in ethyl acetate concentration was observed in the genetically manipulated strains, while isobutyric acid increased. The synthesis of some primary and secondary metabolites appears more readily influenced by the NAD(+)/NADH availability. The data provide an initial assessment of the impact of redox balance on the production of primary and secondary metabolites which play an essential role in the flavour and aroma character of beverages.


Spatial configuration and composition of charge modulates transport into a mucin hydrogel barrier.
The mucus barrier is selectively permeable to a wide variety of molecules, proteins, and cells, and establishes gradients of these particulates to influence the uptake of nutrients, the defense against pathogens, and the delivery of drugs. Despite its importance for health and disease, the criteria that govern transport through the mucus barrier are largely unknown. Studies with uniformly functionalized nanoparticles have provided critical information about the relevance of particle size and net charge for mucus transport. However, these particles lack the detailed spatial arrangements of charge found in natural mucus-interacting substrates, such as certain viruses, which may have important consequences for transport through the mucus barrier. Using a novel, to our knowledge, microfluidic design that enables us to measure real-time transport gradients inside a hydrogel of mucins, the gel-forming glycoprotein component of mucus, we show that two peptides with the same net charge, but different charge arrangements, exhibit fundamentally different transport behaviors. Specifically, we show that certain configurations of positive and negative charges result in enhanced uptake into a mucin barrier, a remarkable effect that is not observed with either charge alone. Moreover, we show that the ionic strength within the mucin barrier strongly influences transport specificity, and that this effect depends on the detailed spatial arrangement of charge. These findings suggest that spatial charge distribution is a critical parameter to modulate transport through mucin-based barriers, and have concrete implications for the prediction of mucosal passage, and the design of drug delivery vehicles with tunable transport properties.


Calponin distribution in human ciliary muscle and other anterior segment tissues.
Calponins are a family of actin-binding proteins known to regulate aortic and tracheal smooth muscle contraction. This investigation was undertaken to assess the presence, subtype, and distribution of calponin proteins in human ciliary muscle, iris, and other anterior segment tissues as well as expression in ciliary muscle cells in vitro. The distribution of calponin immunoreactivity was assessed in paraffin sections of human anterior segment tissue. Human ciliary muscle proteins were analyzed by polyacrylamide gel electrophoresis and Western blotting. The regulation of calponin expression was compared with alpha-sm-actin expression in preconfluent and postconfluent ciliary muscle cell cultures by immunocytochemistry. To determine total cell counts, the cultures were counter-stained with ethidium homodimer. As control specimens, expression of calponin and alpha-sm-actin also was assessed in human Tenon fibroblast cultures. Strong calponin immunoreactivity was present in ciliary muscle, iris dilator and sphincter muscles, and blood vessel smooth muscle. Fine immunostained strands also were observed in the scleral spur. This distribution was similar to alpha-sm-actin. Western blotting showed a single band of calponin with a molecular weight of 32 kDa. In the cultured ciliary muscle cells, calponin stained straight cable-like fibers running parallel along the long axis of the cells. Although the proportion of calponin immunoreactive cells was reduced substantially in preconfluent cultures, virtually all cells were stained in confluent primary through fourth-passage cultures. Cultured human Tenon fibroblasts did not show either calponin or alpha-sm-actin immunoreactivity. Calponin is expressed in human ciliary muscle, iris smooth muscles, blood vessel smooth muscle, as well as within the scleral spur. In addition, calponin is expressed by ciliary smooth muscle cells in vitro. The role of calponin in contraction of these tissues should be investigated.


Highly parallel direct RNA sequencing on an array of nanopores.
Sequencing the RNA in a biological sample can unlock a wealth of information, including the identity of bacteria and viruses, the nuances of alternative splicing or the transcriptional state of organisms. However, current methods have limitations due to short read lengths and reverse transcription or amplification biases. Here we demonstrate nanopore direct RNA-seq, a highly parallel, real-time, single-molecule method that circumvents reverse transcription or amplification steps. This method yields full-length, strand-specific RNA sequences and enables the direct detection of nucleotide analogs in RNA.


Theory of mind in nonhuman primates.
Since the BBS article in which Premack and Woodruff (1978) asked "Does the chimpanzee have a theory of mind?," it has been repeatedly claimed that there is observational and experimental evidence that apes have mental state concepts, such as "want" and "know." Unlike research on the development of theory of mind in childhood, however, no substantial progress has been made through this work with nonhuman primates. A survey of empirical studies of imitation, self-recognition, social relationships, deception, role-taking, and perspective-taking suggests that in every case where nonhuman primate behavior has been interpreted as a sign of theory of mind, it could instead have occurred by chance or as a product of nonmentalistic processes such as associative learning or inferences based on nonmental categories. Arguments to the effect that, in spite of this, the theory of mind hypothesis should be accepted because it is more parsimonious than alternatives or because it is supported by convergent evidence are not compelling. Such arguments are based on unsupportable assumptions about the role of parsimony in science and either ignore the requirement that convergent evidence proceed from independent assumptions, or fail to show that it supports the theory of mind hypothesis over nonmentalist alternatives. Progress in research on theory of mind requires experimental procedures that can distinguish the theory of mind hypothesis from nonmentalist alternatives. A procedure that may have this potential is proposed. It uses conditional discrimination training and transfer tests to determine whether chimpanzees have the concept "see." Commentators are invited to identify flaws in the procedure and to suggest alternatives.


Real-time high resolution 3D imaging of the lyme disease spirochete adhering to and escaping from the vasculature of a living host.
Pathogenic spirochetes are bacteria that cause a number of emerging and re-emerging diseases worldwide, including syphilis, leptospirosis, relapsing fever, and Lyme borreliosis. They navigate efficiently through dense extracellular matrix and cross the blood-brain barrier by unknown mechanisms. Due to their slender morphology, spirochetes are difficult to visualize by standard light microscopy, impeding studies of their behavior in situ. We engineered a fluorescent infectious strain of Borrelia burgdorferi, the Lyme disease pathogen, which expressed green fluorescent protein (GFP). Real-time 3D and 4D quantitative analysis of fluorescent spirochete dissemination from the microvasculature of living mice at high resolution revealed that dissemination was a multi-stage process that included transient tethering-type associations, short-term dragging interactions, and stationary adhesion. Stationary adhesions and extravasating spirochetes were most commonly observed at endothelial junctions, and translational motility of spirochetes appeared to play an integral role in transendothelial migration. To our knowledge, this is the first report of high resolution 3D and 4D visualization of dissemination of a bacterial pathogen in a living mammalian host, and provides the first direct insight into spirochete dissemination in vivo.


Reticularis thalami neurons revisited: activity changes during shifts in states of vigilance.
This study tested the hypothesis that inhibitory actions are exerted by reticularis thalami (RE) neurons upon thalamocortical neurons. The RE neurons were recorded in the rostral pole and lateral districts of the nucleus, and were activated monosynaptically by cortical volleys. Thalamocortical neurons were identified antidromically in intralaminar and ventrolateral nuclei. During sleep with EEG synchronization, prolonged spike barrages of RE neurons extended over the whole spindle sequences. This result suggests that RE neurons are depolarized throughout spindle oscillations, whereas thalamocortical neurons show, simultaneously, long hyperpolarizations and short rebounds. During waking, parallelism rather than reciprocity was found between RE and thalamocortical neurons. Spontaneous discharge rates almost doubled in RE neurons on arousal from sleep, and the probability of cortically evoked short-latency discharges increased. The increase in spontaneous firing rates of RE neurons during natural arousal is consistent with their short-latency synaptic excitation by stimulating the rostral brain stem reticular formation after chronic degeneration of passing fibers. We suggest that RE cells inhibit GABAergic local-circuit cells, in addition to inhibiting thalamocortical neurons, and that different ratios of inhibitory effects are exerted by RE neurons upon these two cell classes during waking and sleep. We further suggest that, upon arousal, disinhibition of thalamocortical neurons (via the local-circuit neurons) outweighs direct inhibition of the thalamocortical neurons.


Effects of Cannabinoids on T-cell Function and Resistance to Infection.
This review examines the effects of cannabinoids on immune function, with a focus on effects on T-cells, as well as on resistance to infection. The paper considers the immune modulating capacity of marijuana, of ∆(9)-THC extracted from the marijuana plant, and synthetic cannabinoids. Of particular interest are synthetic compounds that are CB2 receptor (CB2R) selective agonists. As the CB2R is principally expressed on cells of the immune system, agonists that target this receptor, and not CB1 (which is mainly expressed on neurons), have the possibility of altering immune function without psychoactive effects. The overall conclusion of the studies discussed in this review is that cannabinoids that bind to the CB2 receptor, including ∆(9)-THC and CB2 selective agonists are immunosuppressive. The studies provide objective evidence for potentially beneficial effects of marijuana and ∆(9)-THC on the immune system in conditions where it is desirable to dampen immune responses. Evidence is also reviewed supporting the conclusion that these same compounds can sensitize to some infections through their immunosuppressive activities, but not to others. An emerging area of investigation that is reviewed is evidence to support the conclusion that CB2 selective agonists are a new class of immunosuppressive and anti-inflammatory compounds that may have exceptional beneficial effects in a variety of conditions, such as autoimmune diseases and graft rejection, where it is desirable to dampen the immune response without psychoactive effects.


A more responsive, multi-pronged strategy is needed to strengthen HIV healthcare for men who have sex with men in a decentralized health system: qualitative insights of a case study in the Kenyan coast.
HIV healthcare services for men who have sex with men (MSM) in Kenya have not been openly provided because of persistent stigma and lack of healthcare capacity within Kenya's decentralized health sector. Building on an evaluation of a developed online MSM sensitivity training programme offered to East and South African healthcare providers, this study assessed views and responses to strengthen HIV healthcare services for MSM in Kenya. The study was conducted between January and July 2017 in Kilifi County, coastal Kenya. Seventeen policymakers participated in an in-depth interview and 59 stakeholders, who were purposively selected from three key groups (i.e. healthcare providers, implementing partners and members of MSM-led community-based organizations) took part in eight focus group discussions. Discussions aimed to understand gaps in service provision to MSM from different perspectives, to identify potential misconceptions, and to explore opportunities to improve MSM HIV healthcare services. Interviews and focus group discussions were recorded, transcribed verbatim and analysed using Braun and Clarke's thematic analysis. Participants' responses revealed that all key groups navigated diverse challenges related to MSM HIV health services. Specific challenges included priority-setting by county government staff; preparedness of leadership and management on MSM HIV issues at the facility level; data reporting at the implementation level and advocacy for MSM health equity. Strong power inequities were observed between policy leadership, healthcare providers and MSM, with MSM feeling blamed for their sexual orientation. MSM agency, as expressed in their actions to access HIV services, was significantly constrained by county context, but can potentially be improved by political will, professional support and a human rights approach. To strengthen HIV healthcare for MSM within a decentralized Kenyan health system, a more responsive, multi-pronged strategy adaptable and relevant to MSM's healthcare needs is required. Continued engagement with policy leadership, collaboration with health facilities, and partnerships with different community stakeholders are critical to improve HIV healthcare services for MSM.


Modeling and enhanced sampling of molecular systems with smooth and nonlinear data-driven collective variables.
Collective variables (CVs) are low-dimensional representations of the state of a complex system, which help us rationalize molecular conformations and sample free energy landscapes with molecular dynamics simulations. Given their importance, there is need for systematic methods that effectively identify CVs for complex systems. In recent years, nonlinear manifold learning has shown its ability to automatically characterize molecular collective behavior. Unfortunately, these methods fail to provide a differentiable function mapping high-dimensional configurations to their low-dimensional representation, as required in enhanced sampling methods. We introduce a methodology that, starting from an ensemble representative of molecular flexibility, builds smooth and nonlinear data-driven collective variables (SandCV) from the output of nonlinear manifold learning algorithms. We demonstrate the method with a standard benchmark molecule, alanine dipeptide, and show how it can be non-intrusively combined with off-the-shelf enhanced sampling methods, here the adaptive biasing force method. We illustrate how enhanced sampling simulations with SandCV can explore regions that were poorly sampled in the original molecular ensemble. We further explore the transferability of SandCV from a simpler system, alanine dipeptide in vacuum, to a more complex system, alanine dipeptide in explicit water.


Effects of cinacalcet on atherosclerotic and nonatherosclerotic cardiovascular events in patients receiving hemodialysis: the EValuation Of Cinacalcet HCl Therapy to Lower CardioVascular Events (EVOLVE) trial.
Premature cardiovascular disease limits the duration and quality of life on long-term hemodialysis. The objective of this study was to define the frequency of fatal and nonfatal cardiovascular events attributable to atherosclerotic and nonatherosclerotic mechanisms, risk factors for these events, and the effects of cinacalcet, using adjudicated data collected during the EValuation of Cinacalcet HCl Therapy to Lower CardioVascular Events (EVOLVE) Trial. EVOLVE was a randomized, double-blind, placebo-controlled clinical trial that randomized 3883 hemodialysis patients with moderate to severe secondary hyperparathyroidism to cinacalcet or matched placebo for up to 64 months. For this post hoc analysis, the outcome measure was fatal and nonfatal cardiovascular events reflecting atherosclerotic and nonatherosclerotic cardiovascular diseases. During the trial, 1518 patients experienced an adjudicated cardiovascular event, including 958 attributable to nonatherosclerotic disease. Of 1421 deaths during the trial, 768 (54%) were due to cardiovascular disease. Sudden death was the most frequent fatal cardiovascular event, accounting for 24.5% of overall mortality. Combining fatal and nonfatal cardiovascular events, randomization to cinacalcet reduced the rates of sudden death and heart failure. Patients randomized to cinacalcet experienced fewer nonatherosclerotic cardiovascular events (adjusted relative hazard 0.84, 95% CI 0.74 to 0.96), while the effect of cinacalcet on atherosclerotic events did not reach statistical significance. Accepting the limitations of post hoc analysis, any benefits of cinacalcet on cardiovascular disease in the context of hemodialysis may result from attenuation of nonatherosclerotic processes. Unique identifier: NCT00345839. URL: ClinicalTrials.gov.


Bistable chimera attractors on a triangular network of oscillator populations.
We study a triangular network of three populations of coupled phase oscillators with identical frequencies. The populations interact nonlocally, in the sense that all oscillators are coupled to one another, but more weakly to those in neighboring populations than to those in their own population. This triangular network is the simplest discretization of a continuous ring of oscillators. Yet it displays an unexpectedly different behavior: in contrast to the lone stable chimera observed in continuous rings of oscillators, we find that this system exhibits two coexisting stable chimeras. Both chimeras are, as usual, born through a saddle-node bifurcation. As the coupling becomes increasingly local in nature they lose stability through a Hopf bifurcation, giving rise to breathing chimeras, which in turn get destroyed through a homoclinic bifurcation. Remarkably, one of the chimeras reemerges by a reversal of this scenario as we further increase the locality of the coupling, until it is annihilated through another saddle-node bifurcation.


Antihypertensive effect of mitochondria-targeted proxyl nitroxides.
Superoxide ( [Formula: see text] ) has been implicated in the pathogenesis of many human diseases including hypertension. Mitochondria-targeted superoxide scavenger mitoTEMPO reduces blood pressure; however, the structure-functional relationships in antihypertensive effect of mitochondria-targeted nitroxides remain unclear. The nitroxides are known to undergo bioreduction into hydroxylamine derivatives which reacts with [Formula: see text] with much lower rate. The nitroxides of pyrrolidine series (proxyls) are much more resistant to bioreduction compared to TEMPOL derivatives suggesting that mitochondria-targeted proxyls can be effective antioxidants with antihypertensive activity. In this work we have designed and studied two new pyrrolidine mitochondria targeted nitroxides: 3-[2-(triphenyphosphonio)acetamido]- and 3-[2-(triphenyphosphonio) acetamidomethyl]-2,2,5,5-tetramethylpyrrolidine-1-oxyl (mCP2) and (mCP1). These new mitochondria targeted nitroxides have 3- to 7-fold lower rate constants of the reaction with O2(-•) compared with mitoTEMPO; however, the cellular bioreduction of mCP1 and mCP2 was 3- and 2-fold slower. As a consequence incubation with cells afforded much higher intracellular concentration of mCP1 and mCP2 nitroxides compared to mitoTEMPO nitroxide. This has compensated for the difference in the rate of O2(-•) scavenging and all nitroxides similarly protected mitochondrial respiration in H2O2 treated endothelial cells. Treatment of hypertensive mice with mCP1 and mCP2 (1.4mg/kg/day) after onset of angiotensin II-induced hypertension significantly reduced blood pressure to 133±5mmHg and 129±6mmHg compared to 163±5mmHg in mice infused with angiotensin II alone. mCP1 and mCP2 reduced vascular O2(-•) and prevented decrease of endothelial nitric oxide production. These data indicate that resistance to bioreduction play significant role in antioxidant activity of nitroxides. Studies of nitroxide analogs such as mCP1 and mCP2 may help in optimization of chemical structure of mitochondria-targeted nitroxides for improved efficacy and pharmacokinetics of these drugs in treatment of hypertension and many other conditions including atherosclerosis, diabetes and degenerative neurological disorders in which mitochondrial oxidative stress seems to play a role.


Metabolic disturbances in diseases with neurological involvement.
Degeneration of specific neuronal populations and progressive nervous system dysfunction characterize neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease. These findings are also reported in inherited diseases such as phenylketonuria and glutaric aciduria type I. The involvement of mitochondrial dysfunction in these diseases was reported, elicited by genetic alterations, exogenous toxins or buildup of toxic metabolites. In this review we shall discuss some metabolic alterations related to the pathophysiology of diseases with neurological involvement and aging process. These findings may help identifying early disease biomarkers and lead to more effective therapies to improve the quality of life of the patients affected by these devastating illnesses.


Family Features of Social Withdrawal Syndrome (Hikikomori).
Background: Family may play an important role in the origin, maintenance, and treatment of people with social withdrawal. The aim of this study is to analyze family factors related to social withdrawal syndrome. Methods: Socio-demographic, clinical, and family data, including family psychiatric history, dysfunctional family dynamics, and history of family abuse were analyzed in 190 cases of social withdrawal with a minimum duration of 6 months that started an at-home treatment program. Data were analyzed at baseline and at 12 months. Results: In 36 cases (18%) neither the patient nor the family allowed at home evaluation and treatment by the Crisis Resolution Home Treatment (CRHT) team. Patients had high rates of dysfunctional family dynamics (n = 115, 61.5%), and family psychiatric history (n = 113, 59.3%), especially maternal affective (n = 22, 42.9%), and anxiety disorders (n = 11 20.4%). There was a non-negligible percentage of family maltreatment in childhood (n = 35, 20.7%) and single-parent families (n = 66, 37.8%). Most of the cases lived with their families (n = 135, 86%), had higher family collaboration in the therapeutic plan (n = 97, 51.9%) and families were the ones to detect patient isolation and call for help (n = 140, 73.7%). Higher social withdrawal severity (as defined by at least one of: early age of onset, no family collaboration, lack of insight, higher CGSI score, and higher Zarit score), was associated with family psychiatric history, dysfunctional family dynamics, and family abuse history. All of these predictive variables were highly correlated one to each other. Conclusions: There is a high frequency of family psychiatric history, dysfunctional family dynamics, and traumatic events in childhood (family maltreatment), and these factors are closely interrelated, highlighting the potential role of family in the development and maintenance of social withdrawal.


Health Disparities in Tobacco Use and Exposure: A Structural Competency Approach.
Fourteen percent of US adults use tobacco products. Because many of those who use tobacco are parents and/or caregivers, children are disproportionately exposed to tobacco smoke. People who use tobacco products often become addicted to nicotine, resulting in tobacco dependence, a chronic, relapsing disease. Tobacco use and exposure are more likely to occur in vulnerable and marginalized groups, including those living in poverty. Although some view tobacco use as a personal choice, evidence suggests that structural forces play an important role in tobacco uptake, subsequent nicotine addiction, and perpetuation of use. Viewing tobacco use and tobacco dependence through a structural competency lens promotes recognition of the larger systemic forces perpetuating tobacco use, including deliberate targeting of groups by the tobacco industry, lack of enforcement of age-for-sale laws, inferior access to health insurance and health care, poor access to cessation resources, and economic stress. Each of these forces perpetuates tobacco initiation and use; in turn, tobacco use perpetuates the user's adverse health and economic conditions. Pediatricians are urged to view family tobacco use as a social determinant of health. In addition to screening adolescents for tobacco use and providing resources and treatment of tobacco dependence, pediatricians are encouraged to systematically screen children for secondhand smoke exposure and support family members who smoke with tobacco cessation. Additionally, pediatricians can address the structural issues perpetuating tobacco use by becoming involved in policy and advocacy initiatives.


Postural perturbations: new insights for treatment of balance disorders.
This article reviews the neural control of posture as understood through studies of automatic responses to mechanical perturbations. Recent studies of responses to postural perturbations have provided a new view of how postural stability is controlled, and this view has profound implications for physical therapy practice. We discuss the implications for rehabilitation of balance disorders and demonstrate how an understanding of the specific systems underlying postural control can help to focus and enrich our therapeutic approaches. By understanding the basic systems underlying control of balance, such as strategy selection, rapid latencies, coordinated temporal spatial patterns, force control, and context-specific adaptations, therapists can focus their treatment on each patient's specific impairments. Research on postural responses to surface translations has shown that balance is not based on a fixed set of equilibrium reflexes but on a flexible, functional motor skill that can adapt with training and experience. More research is needed to determine the extent to which quantification of automatic postural responses has practical implications for predicting falls in patients with constraints in their postural control system.


Physiotherapy treatment experiences of persons with persistent musculoskeletal pain: A qualitative study.
The aim of this study was to explore and describe the physiotherapy treatment experiences of persons with persistent musculoskeletal pain. Eleven participants with persistent musculoskeletal pain in the back, neck, or shoulders were included in the study. Data was collected via semi-structured interviews and were analysed with qualitative content analysis. The analysis resulted in the theme "Towards acceptance and management of pain", comprising four sub-themes: 1) Establishing and maintaining a therapeutic alliance; 2) Being active, taking initiative and facing challenges; 3) Appreciating guidance, incentive and having a sounding board; and 4) Acquired knowledge and new body awareness change behaviours. The theme and sub-themes describe how the participants used increased knowledge, awareness, movements and exercises learned from the physiotherapy treatment to develop strategies for managing pain and the process of acceptance. A trusting relationship and continual dialogue with the physiotherapist was considered to be important. The participants were actively involved in the process as exercises, activities and other treatment modalities were individualized. This was rewarding but also challenging and required effort on their part. The physiotherapist's initiatives and actions were an important incentive and means of support.


Anger arousal and behavioral anger regulation in everyday life among patients with chronic low back pain: Relationships to patient pain and function.
The objective of this study was to determine the degree to which patient anger arousal and behavioral anger regulation (expression, inhibition) occurring in the course of daily life was related to patient pain and function as rated by patients and their spouses. Married couples (N = 105) (one spouse with chronic low back pain) completed electronic daily diaries, with assessments 5 times/day for 14 days. Patients completed items on their own state anger, behavioral anger expression and inhibition, and pain-related factors. Spouses completed items on their observations of patient pain-related factors. Hierarchical linear modeling was used to test concurrent and lagged relationships. Patient-reported increases in state anger were related to their reports of concurrent increases in pain and pain interference and to spouse reports of patient pain and pain behavior. Patient-reported increases in behavioral anger expression were related to lagged increases in pain intensity and interference and decreases in function. Most of these relationships remained significant with state anger controlled. Patient-reported increases in behavioral anger inhibition were related to concurrent increases in pain interference and decreases in function, which also remained significant with state anger controlled. Patient-reported increases in state anger were related to lagged increases in spouse reports of patient pain intensity and pain behaviors. Results indicate that in patients with chronic pain, anger arousal and behavioral anger expression and inhibition in everyday life are related to elevated pain intensity and decreased function as reported by patients. Spouse ratings show some degree of concordance with patient reports.


Terfezia boudieri: A Desert Truffle With Anticancer and Immunomodulatory Activities.
Desert truffles have high nutritional value and grow wild in the Mediterranean basin and Western Asia. Although, many studies were performed to evaluate truffles nutritious values and phytochemical composition, studies are limited to evaluate their anticancer and/ or immunomodulatory effects. Our study was conducted to evaluate the anticancer and immunomodulatory effects of Terfezia boudieri (desert truffle). Different solvent extracts were prepared from the truffle and MTT assay was used to measure their anticancer activity against cancer cell lines (T47D, MCF-7, MDA-MB231, HCT-116, and Hela). Total phenolic content in each extract was determined by using Folin-Ciocalteu reagent and qualitative phytochemical screening was performed using standard methods. The degree of apoptosis induction (using caspase 3 assay) and vascular endothelial growth factor expression were detected using standard kits. Also, ELISA was used to measure levels of IFN-γ, IL-2, IL-4, and IL-10 secreted by splenocytes after treatment with the extracts. The effect of the extracts on splenocytes proliferation was measured using MTT assay. Macrophage function was evaluated using nitro blue tetrazolium assay and pinocytosis function was evaluated using neutral red method. Terpenoids, phytosterols, and carbohydrates were present in all the solvent extracts, while tannins, alkaloids and flavonoids were detected only in aqueous/methanol and aqueous extracts. The highest total phenolic content was observed in aqueous and aqueous methanol extracts. The growth of cancer cell lines was inhibited by T. boudieri extracts in a dose dependent manner. N-hexane extract was the most potent against most cell lines. Aqueous/methanol extract showed high apoptosis induction and angiogenesis suppression effects. An increase in TH1 cytokines (IFN-γ, IL-2) level and a decrease in TH2 cytokine (IL-4) level were evident after lymphocytes stimulation by aqueous/methanol, n-hexane and ethyl acetate extracts of T. boudieri. Ethyl acetate extract of T. boudieri were the most potent extracts to stimulate lymphocytes proliferation while all other extracts showed moderate stimulation. Aqueous/methanol extract was the most active extract to stimulate phagocytosis. Ethyl acetate extract was the most active extract to stimulate pinocytosis. The use of T. boudieri provides variable health benefits. N-hexane, ethyl acetate, and aqueous/methanol extracts exhibited anticancer activities and are potent stimulators of innate and acquired immunity. Further testing is needed to identify the biologically active compounds and detect them quantitatively using GC-MS analysis.


Bacterial ferrochelatase turns human: Tyr13 determines the apparent metal specificity of Bacillus subtilis ferrochelatase.
Ferrochelatase catalyzes the insertion of Fe(2+) into protoporphyrin IX. The enzymatic product heme (protoheme IX) is a well-known cofactor in a wide range of proteins. The insertion of metal ions other than Fe(2+) occurs rarely in vivo, but all ferrochelatases that have been studied can insert Zn(2+) at a good rate in vitro. Co(2+), but not Cu(2+), is known to be a good substrate of the mammalian and Saccharomyces cerevisiae ferrochelatases. In contrast, Cu(2+), but not Co(2+), has been found to be a good substrate of bacterial Bacillus subtilis ferrochelatase. It is not known how ferrochelatase discriminates between different metal ion substrates. Structural analysis of B. subtilis ferrochelatase has shown that Tyr13 is an indirect ligand of Fe(2+) and a direct ligand of a copper mesoporphyrin product. A structure-based comparison revealed that Tyr13 aligns with a Met residue in the S. cerevisiae and human ferrochelatases. Tyr13 was changed to Met in the B. subtilis enzyme by site-directed mutagenesis. Enzymatic measurements showed that the modified enzyme inserted Co(2+) at a higher rate than the wild-type B. subtilis ferrochelatase, but it had lost the ability to use Cu(2+) as a substrate. Thus, the B. subtilis Tyr13Met ferrochelatase showed the same metal specificity as that of the ferrochelatases from S. cerevisiae and human.


Distribution and bioaccumulation of POPs and mercury in the Ga-Selati River (South Africa) and the rivers Gudbrandsdalslågen and Rena (Norway).
Biomagnification of Hg and persistent organic pollutants (POPs: polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs)) in aquatic food chains can lead to high pollutant concentrations in top predators, including humans. Despite this threat to human health, research concerning bioaccumulation is still underrepresented in the southern hemisphere and in (sub)arctic and (sub)tropical areas, emphasizing the need for research in these locations. In this study, samples of water, sediment and aquatic biota were analyzed to determine concentrations of POPs and total mercury (THg) in the Ga-Selati river (South Africa) and two rivers Rena and Gudbrandsdalslågen in Norway. Trophic magnification factors (TMFs) were determined to evaluate and compare the biomagnification and the threat to human health due to consumption of the fish was assessed. Concentrations of POPs in sediment and biota samples were generally low except for relatively high concentrations of ∑DDX (dichlorodiphenyltrichloroethane and metabolites) in aquatic biota from the Ga-Selati river (ranging from 1.9 to 133 ng/g ww in invertebrates and 1.9 to 5643 ng/g ww in fish). Dissolved THg concentrations were high in the Ga-Selati river (ranging from 0.009 to 0.036 μg/l) but THg concentrations in sediment and biota were low in studied rivers compared to other studies. Biomagnification occurred for THg, several DDT-metabolites and PCB compounds, TN and CN. Biomagnification of p,p'-DDT and THg differed significantly between the two countries, supporting existing patterns found in literature, although more data is needed to attribute these differences to climatic or other factors. Concentrations in fish from the rivers Ga-Selati and Rena were under the threshold levels reported for THg and POPs, but caution should be taken when consuming Northern pike (Esox Lucius) from the subarctic river Gudbrandsdalslågen, to avoid harmful effects due to both elevated THg and PBDE exposure.


Prevalence and Variation of Developmental Screening and Surveillance in Early Childhood.
Since 2001, the American Academy of Pediatrics has recommended universal developmental screening and surveillance to promote early diagnosis and intervention and to improve the outcomes of children with developmental delays and disabilities. To examine the current prevalence and variation of developmental screening and surveillance of children by various sociodemographic, enabling, and health characteristics. This cross-sectional analysis of the Health Resources and Services Administration's 2016 National Survey of Children's Health-a nationally representative survey of US children completed between June 2016 and February 2017-examined 5668 randomly selected children 9 through 35 months of age whose parent or caregiver responded to the address-based survey by mail or via a website. All analyses were weighted to account for the probability of selection and nonresponse and to reflect population counts of all noninstitutionalized US children residing in housing units. Developmental screening was measured through a validated set of 3 items indicating receipt in the past year of parent-completed screening from a health care professional with age-appropriate content regarding language development and social behavior. Surveillance was determined by an item capturing verbal elicitation of developmental concerns by a health care professional. Of the estimated 9.0 million children aged 9 through 35 months, an estimated 30.4% (95% CI, 28.0%-33.0%) were reported by their parent or guardian to have received a parent-completed developmental screening and 37.1% (95% CI, 34.4%-39.8%) were reported to have received developmental surveillance from a health care professional in the past year. Characteristics associated with screening and/or surveillance that remained significant after adjustment included primary household language, family structure, household education, income, medical home, past-year preventive visit, child health status, and special health care needs. Having health care that meets medical home criteria was significantly associated with both developmental screening (adjusted rate ratio, 1.34; 95% CI, 1.13-1.57) and surveillance (adjusted rate ratio, 1.24; 95% CI, 1.08-1.42), representing an 8 to 9 absolute percentage point increase. State-level differences spanned 40 percentage points for screening (17.2% in Mississippi and 58.8% in Oregon) and surveillance (19.1% in Mississippi and 60.8% in Oregon), with approximately 90% of variation not explained by child and family characteristics. Despite more than a decade of initiatives, rates of developmental screening and surveillance remain low. However, state-level variation indicates continued potential for improvement. Systems-level quality improvement efforts, building on the medical home, will be necessary to achieve recommended screening and surveillance goals.


Macrophage inflammatory protein-1 alpha activates basophils and mast cells.
Macrophage inflammatory protein-1 (MIP) is a recently cloned cytokine that causes neutrophilic infiltration and induces an inflammatory response. We studied the effect of MIP-1 alpha on histamine secretion from basophils and mast cells. Leukocytes from allergic and normal subjects were studied. MIP-1 alpha caused dose-dependent release of histamine from basophils of 14 of 20 allergic donors at concentrations of 10(-9)-10(-7) M, and the mean release was 13.50 +/- 2.9% at the highest concentration. In the same experiments, the mean histamine release by anti-immunoglobulin E and monocyte chemotactic and activating factor (MCAF) (10(-7) M) was 32 +/- 7% and 31 +/- 3%, respectively. The cells from only 2 of 10 normal subjects released histamine in response to MIP-1 alpha. Histamine release by MIP-1 alpha was rapid, and almost complete within the first 3 min. MIP-1 alpha-induced degranulation was a calcium-dependent noncytotoxic process. MIP-1 alpha showed chemotactic activity for purified basophils that was comparable to MCAF. Both MIP-1 alpha and MCAF at 10(-7) M concentration elicited a chemotactic response that was 40% of the maximal response to C5a (1 microgram/ml). Murine MIP-1 alpha induced histamine release from mouse peritoneal mast cells in a dose-dependent manner. Thus, we have established that MIP-1 alpha is a novel activator of basophils and mast cells.


Diabetes in Population Isolates: Lessons from Greenland.
Type 2 diabetes (T2D) is an increasing health problem worldwide with particularly high occurrence in specific subpopulations and ancestry groups. The high prevalence of T2D is caused both by changes in lifestyle and genetic predisposition. A large number of studies have sought to identify the genetic determinants of T2D in large, open populations such as Europeans and Asians. However, studies of T2D in population isolates are gaining attention as they provide several advantages over open populations in genetic disease studies, including increased linkage disequilibrium, homogeneous environmental exposure, and increased allele frequency. We recently performed a study in the small, historically isolated Greenlandic population, in which the prevalence of T2D has increased to more than 10%. In this study, we identified a common nonsense variant in TBC1D4, which has a population-wide impact on glucose-stimulated plasma glucose, serum insulin levels, and T2D. The variant defines a specific subtype of non-autoimmune diabetes characterized by decreased post-prandial glucose uptake and muscular insulin resistance. These and other recent findings in population isolates illustrate the value of performing medical genetic studies in genetically isolated populations. In this review, we describe some of the advantages of performing genetic studies of T2D and related cardio-metabolic traits in a population isolate like the Greenlandic, and we discuss potentials and perspectives for future research into T2D in this population.


Phase and interface determination in computer simulations of liquid mixtures with high partial miscibility.
Partially miscible solutions can represent a challenge from the computer simulation standpoint, especially if the mutual solubility of the components is so large that their concentrations do not change much from one phase to another. In this case, identifying which molecules belong to which phase becomes a complicated task. Here, we propose a density-based clustering approach with self-tuning capabilities and apply it to the case of the mixture of an ionic liquid with benzene. The almost linear scaling of the algorithm makes it suitable for the analysis of long Molecular Dynamics or Monte Carlo trajectories.


Tolerance to Hololaimus columbus in Glyphosate-Resistant, Transgenic Soybean Cultivars.
Transgenic soybean cultivars, resistant to glyphosate herbicide in maturity groups V and VI, were evaluated for tolerance to the Columbia lance nematode, Hoplolaimus columbus, in field experiments conducted in 1998 and 1999. Treatment with 43 liter/ha of 1,3-dichloropropene was effective in suppressing H. columbus population densities in a split-plot design. Fumigation increased soybean yield, but a significant cultivar x fumigation interaction indicated variation in cultivar response to H. columbus. A tolerance index (yield of nontreated / yield of treated x 100) was used to compare cultivar differences. Two cultivars in maturity group VI and one cultivar in maturity group V had a tolerance index greater than 90, indicating a high level of tolerance.


Assessment of coronary artery disease by post-mortem cardiac MR.
Minimally invasive or virtual autopsies are being advocated as alternative to traditional autopsy, but have limited abilities to detect coronary artery disease. It was the objective of this study to assess if the occurrence of chemical shift artifacts (CSA) along the coronary arteries on non-contrast, post-mortem cardiac MR may be used to investigate coronary artery disease. We retrospectively compared autopsy and CT findings of 30 cases with significant (≥75%), insignificant (<75%), or absent coronary artery stenosis to post-mortem cardiac MR findings. The chi-square test was used to investigate if the occurrence of CSA depends on the presence or absence of stenosis. Sensitivity, specificity and predictive values were calculated for each finding. CSA indicates the absence of (significant) stenosis (p<0.001). The occurrence of paired dark bands in lieu of CSA on post-mortem cardiac MR suggests (significant) coronary arteries stenosis (p<0.001). Both findings have a high specificity but low sensitivity. CSA is a marker of vessel patency. The presence of paired dark bands indicates stenosis. These criteria improve the ability of minimally invasive or virtual autopsy to detect coronary artery disease related deaths.


Conjunctival squamous cell carcinoma in patients with human immunodeficiency virus infection seen at two hospitals in Kenya.
To estimate the prevalence and pattern of conjuctival squamous cell carcinoma (CSCC) in patients with HIV infection. A hospital based cross sectional study. Kenyatta National Hospital (KNH) and Kikuyu Eye Unit (KEU) during the period November 2003 and May 2004. Four hundred and nine HIV positive patients. Four hundred and nine HIV positive patients aged 25 to 53 years were screened. Male to Female ratio was 1:1. One hundred and three had conjunctival growths. Thirty two had histologically proven conjunctiva squamous cell carcinoma (CSCC). Estimated prevalence of CSCC among HIV positive patients was 7.8%. The average duration of growth of the conjunctival masses was 21.8 months. The average size of the lesions at the time of presentation was 6.6 mm. Twenty two (68.8%) patients had primary CSCC, while ten (31.2%) had recurrent lesions. The pattern of the histopathology results was: fifteen (46.9%) patients had poorly differentiated squamous cell carcinoma; nine (28%) had moderately differentiated squamous cell carcinoma; five patients (15.6%) had CIN; two patients (6.3%) had dysplasia and one patient (3.1%) had a well differentiated squamous cell carcinoma. Prevalence of CSCC in HIV/AIDS patients was 7.8%. Patients present late with advanced lesions. Recurrence rates from previous surgery are high. The often uncharacteristic complaints and findings on presentation complicate the clinical diagnosis. Active search for early manifestations of CSCC in HIV / AIDS patients, complete surgical excision and close follow up is necessary. Alternative treatment methods and techniques like the topical use of antimetabolites should be explored further.


Outbreak of leptospirosis among triathlon participants in Langau, Austria, 2010.
We report on the first documented outbreak of leptospirosis in Austria. In July 2010, four cases of serologically confirmed leptospirosis occurred in athletes after a triathlon held in Langau. Heavy rains preceded the triathlon (rainfall: 22 mm). The index case (Patient A) was a 41-year-old previously healthy male, who was admitted to hospital A on July 8 with a four-day history of fever up to 40°C that began 14 days after attending the triathlon event. On July 7, patient B, a 42-year-old male, was admitted to the same hospital, with signs and symptoms of kidney failure. Hemodialysis was performed every other day for 3 weeks. While the serum drawn on the day of admission was negative for antibodies against Leptospira, a specimen from July 28 tested positive with Leptospira interrogans. On July 11, patient C, a 40-year-old male, was admitted to hospital B for nephritis. On July 14, patient D, a 44-year-old male, was admitted to hospital C with a ten days history of intermittent fever, mild dry cough and headache. Our report underlines that in Austria recreational users of bodies of freshwater must be aware of an existing risk of contracting leptospirosis, particularly after heavy rains. The suppressive influence of a triathlon on the immune system is well documented and therefore an outbreak in this population group can be seen as a sensitive indicator concerning possible risk for the general population.


The longitudinal association between social functioning and theory of mind in first-episode psychosis.
There is some cross-sectional evidence that theory of mind ability is associated with social functioning in those with psychosis but the direction of this relationship is unknown. This study investigates the longitudinal association between both theory of mind and psychotic symptoms and social functioning outcome in first-episode psychosis. Fifty-four people with first-episode psychosis were followed up at 6 and 12 months. Random effects regression models were used to estimate the stability of theory of mind over time and the association between baseline theory of mind and psychotic symptoms and social functioning outcome. Neither baseline theory of mind ability (regression coefficients: Hinting test 1.07 95% CI -0.74, 2.88; Visual Cartoon test -2.91 95% CI -7.32, 1.51) nor baseline symptoms (regression coefficients: positive symptoms -0.04 95% CI -1.24, 1.16; selected negative symptoms -0.15 95% CI -2.63, 2.32) were associated with social functioning outcome. There was evidence that theory of mind ability was stable over time, (regression coefficients: Hinting test 5.92 95% CI -6.66, 8.92; Visual Cartoon test score 0.13 95% CI -0.17, 0.44). Neither baseline theory of mind ability nor psychotic symptoms are associated with social functioning outcome. Further longitudinal work is needed to understand the origin of social functioning deficits in psychosis.


Improving homology models for protein-ligand binding sites.
In order to improve the prediction of protein-ligand binding sites through homology modeling, we incorporate knowledge of the binding residues into the modeling framework. Residues are identified as binding or nonbinding based on their true labels as well as labels predicted from structure and sequence. The sequence predictions were made using a support vector machine framework which employs a sophisticated window-based kernel. Binding labels are used with a very sensitive sequence alignment method to align the target and template. Relevant parameters governing the alignment process are searched for optimal values. Based on our results, homology models of the binding site can be improved if a priori knowledge of the binding residues is available. For target-template pairs with low sequence identity and high structural diversity our sequence-based prediction method provided sufficient information to realize this improvement.


Challenges of Detecting Lead in Drinking Water Using at-Home Test Kits.
Lead in drinking water remains a significant human health risk. At-home lead in water test kits could provide consumers with a convenient and affordable option to evaluate this risk, but their accuracy and reliability is uncertain. This study examined the ability of at-home lead test kits to detect varying concentrations of dissolved and particulate lead in drinking water. Sixteen brands representing four test kit types (binary color, binary strip, colorimetric vial, and color strip) were identified. Most kits (12 of 16 brands) were not suitable for drinking water analysis, with lead detection limits of 5-20 mg/L. Binary strips detected dissolved lead at drinking water-relevant levels but failed to detect particulate lead. Household acids (lemon juice and vinegar) improved the strip's ability to detect lead by dissolving some of the lead particulates to the point soluble lead exceeded 15 μg/L. These results illustrate the applications of at-home testing kits for drinking water analysis, highlight limitations and areas for possible improvement, and put forth a testing protocol by which new at-home lead test kits can be judged.


It Works Both Ways: The Relationship between Exposure to Sexual Content in the Media and Adolescent Sexual Behavior.
Using a longitudinal web-based survey of adolescents 14-16 years of age, we estimate regression models where self-reported sexual behavior and content analytic-based exposure to sex in the media are related cross-sectionally and longitudinally. We find evidence for both cross-sectional non-recursive and prospective longitudinal relationships even after adjusting for both established predictors of sexual behavior (e.g., physical development, having a romantic partner, parental monitoring, peer and parental norms, respondent's age) and of exposure to sexual media content (e.g., time the respondent goes to bed, extracurricular activities, television in the bedroom, total time spent with television, music, videogames, and magazines). Sexually active adolescents are more likely to expose themselves to sex in the media and those exposed to sex in the media are more likely to progress in their sexual activity. These findings are consistent with others in the literature that demonstrate a causal effect of exposure to sexual content on sexual behavior but extend established results by also looking at the causal effect of sexual behavior on exposure both cross-sectionally and over time.


Linkage mapping in the oilseed crop Jatropha curcas L. reveals a locus controlling the biosynthesis of phorbol esters which cause seed toxicity.
Current efforts to grow the tropical oilseed crop Jatropha curcas L. economically are hampered by the lack of cultivars and the presence of toxic phorbol esters (PE) within the seeds of most provenances. These PE restrict the conversion of seed cake into animal feed, although naturally occurring 'nontoxic' provenances exist which produce seed lacking PE. As an important step towards the development of genetically improved varieties of J. curcas, we constructed a linkage map from four F₂ mapping populations. The consensus linkage map contains 502 codominant markers, distributed over 11 linkage groups, with a mean marker density of 1.8 cM per unique locus. Analysis of the inheritance of PE biosynthesis indicated that this is a maternally controlled dominant monogenic trait. This maternal control is due to biosynthesis of the PE occurring only within maternal tissues. The trait segregated 3 : 1 within seeds collected from F₂ plants, and QTL analysis revealed that a locus on linkage group 8 was responsible for phorbol ester biosynthesis. By taking advantage of the draft genome assemblies of J. curcas and Ricinus communis (castor), a comparative mapping approach was used to develop additional markers to fine map this mutation within 2.3 cM. The linkage map provides a framework for the dissection of agronomic traits in J. curcas, and the development of improved varieties by marker-assisted breeding. The identification of the locus responsible for PE biosynthesis means that it is now possible to rapidly breed new nontoxic varieties.


Allopregnanolone elevations following pregnenolone administration are associated with enhanced activation of emotion regulation neurocircuits.
The neurosteroid allopregnanolone is a potent allosteric modulator of the gamma-aminobutyric acid type A receptor with anxiolytic properties. Exogenous administration of allopregnanolone reduces anxiety, and allopregnanolone blockade impairs social and affective functioning. However, the neural mechanism whereby allopregnanolone improves mood and reduces anxiety is unknown. In particular, brain imaging has not been used to link neurosteroid effects to emotion regulation neurocircuitry. To investigate the brain basis of allopregnanolone's impact on emotion regulation, participants were administered 400 mg of pregnenolone (n=16) or placebo (n=15) and underwent 3T functional magnetic resonance imaging while performing the shifted-attention emotion appraisal task, which probes emotional processing and regulation. Compared with placebo, allopregnanolone was associated with reduced activity in the amygdala and insula across all conditions. During the appraisal condition, allopregnanolone increased activity in the dorsal medial prefrontal cortex and enhanced connectivity between the amygdala and dorsal medial prefrontal cortex, an effect that was associated with reduced self-reported anxiety. These results demonstrate that in response to emotional stimuli, allopregnanolone reduces activity in regions associated with generation of negative emotion. Furthermore, allopregnanolone may enhance activity in regions linked to regulatory processes. Aberrant activity in these regions has been linked to anxiety psychopathology. These results thus provide initial neuroimaging evidence that allopregnanolone may be a target for pharmacologic intervention in the treatment of anxiety disorders and suggest potential future directions for research into neurosteroid effects on emotion regulation neurocircuitry.


Pax7-expressing satellite cells are indispensable for adult skeletal muscle regeneration.
Distinct cell populations with regenerative capacity have been reported to contribute to myofibres after skeletal muscle injury, including non-satellite cells as well as myogenic satellite cells. However, the relative contribution of these distinct cell types to skeletal muscle repair and homeostasis and the identity of adult muscle stem cells remain unknown. We generated a model for the conditional depletion of satellite cells by expressing a human diphtheria toxin receptor under control of the murine Pax7 locus. Intramuscular injection of diphtheria toxin during muscle homeostasis, or combined with muscle injury caused by myotoxins or exercise, led to a marked loss of muscle tissue and failure to regenerate skeletal muscle. Moreover, the muscle tissue became infiltrated by inflammatory cells and adipocytes. This localised loss of satellite cells was not compensated for endogenously by other cell types, but muscle regeneration was rescued after transplantation of adult Pax7(+) satellite cells alone. These findings indicate that other cell types with regenerative potential depend on the presence of the satellite cell population, and these observations have important implications for myopathic conditions and stem cell-based therapeutic approaches.


Characterization of Hematopoiesis in Sickle Cell Disease by Prospective Isolation of Stem and Progenitor Cells.
The consequences of sickle cell disease (SCD) include ongoing hematopoietic stress, hemolysis, vascular damage, and effect of chronic therapies, such as blood transfusions and hydroxyurea, on hematopoietic stem and progenitor cell (HSPC) have been poorly characterized. We have quantified the frequencies of nine HSPC populations by flow cytometry in the peripheral blood of pediatric and adult patients, stratified by treatment and control cohorts. We observed broad differences between SCD patients and healthy controls. SCD is associated with 10 to 20-fold increase in CD34dim cells, a two to five-fold increase in CD34bright cells, a depletion in Megakaryocyte-Erythroid Progenitors, and an increase in hematopoietic stem cells, when compared to controls. SCD is also associated with abnormal expression of CD235a as well as high levels CD49f antigen expression. These findings were present to varying degrees in all patients with SCD, including those on chronic therapy and those who were therapy naive. HU treatment appeared to normalize many of these parameters. Chronic stress erythropoiesis and inflammation incited by SCD and HU therapy have long been suspected of causing premature aging of the hematopoietic system, and potentially increasing the risk of hematological malignancies. An important finding of this study was that the observed concentration of CD34bright cells and of all the HSPCs decreased logarithmically with time of treatment with HU. This correlation was independent of age and specific to HU treatment. Although the number of circulating HSPCs is influenced by many parameters, our findings suggest that HU treatment may decrease premature aging and hematologic malignancy risk compared to the other therapeutic modalities in SCD.


Nodular Fasciitis of the Breast Previously Misdiagnosed as Breast Carcinoma.
BACKGROUND: Nodular fasciitis of the breast is a rare benign pathology that can mimic breast cancer clinically, radiologically, and histopathologically. CASE REPORT: An 18-year-old female patient had first visited a physician in a different center with the complaint of a lump in her left breast. Breast examination had revealed a palpable mass located in the left upper outer quadrant. Ultrasonography had demonstrated a hypoechoic lesion. Excisional biopsy of the lump had been performed and histopathologic examination misdiagnosed this lump as a mesenchymal tumor. The patient was then referred to our clinic for further investigations. Pathologic revision was performed and the diagnosis of nodular fasciitis of the breast was established. CONCLUSION: Awareness of this rare clinical entity, nodular fasciitis, in the breast eliminates the misdiagnosis of breast cancer.


Converting Predation Cues into Conservation Tools: The Effect of Light on Mouse Foraging Behaviour.
Prey face a conflict between acquiring energy and avoiding predators and use both direct and indirect cues to assess predation risk. Illumination, an indirect cue, influences nocturnal rodent foraging behaviour. New Zealand holds no native rodent species but has introduced mice (Mus musculus) that severely impair native biodiversity. We used Giving-Up Densities (GUDs) and observations of foraging frequency and duration to assess if artificial light induces risk avoidance behaviour in mice and could limit their activity. We found both captive (wild strain) mice in outdoor pens and wild mice within a pest fenced sanctuary (Maungatautari, New Zealand) displayed avoidance behaviour in response to illumination. In captivity, total foraging effort was similar across lit and unlit pens but mice displayed a strong preference for removing seeds from dark control areas (mean: 15.33 SD: +/-11.64 per 3.5 hours) over illuminated areas (2.00 +/-3.44). Wild mice also removed fewer seeds from illuminated areas (0.42 +/-1.00 per 12 hours) compared to controls (6.67 +/-9.20). Captive mice spent less than 1.0% of available time at illuminated areas, versus 11.3% at controls; visited the lit areas less than control areas (12.00 +/- 9.77 versus 29.00 +/-21.58 visits respectively); and spent less time per visit at illuminated versus control areas (8.17 +/-7.83 versus 44.83 +/-87.52 seconds per visit respectively). Illumination could provide protection at ecologically sensitive sites, damaged exclusion fences awaiting repair, fence terminus zones of peninsula sanctuaries and shipping docks that service offshore islands. We promote the hypothesis that the tendency of mice to avoid illumination could be a useful conservation tool, and advance knowledge of risk assessment and foraging under perceived danger.


Digital colposcopy for the diagnosis of cervical adenocarcinoma using a narrow band imaging system.
Although the colposcopic features of cervical glandular disease and cervical adenocarcinoma are not widely well known, unique microvascular patterns are reportedly useful for identifying such diseases. The narrow band imaging (NBI) system used in endoscopy can be used to obtain high-contrast vascular images. Therefore, we examined the utility of NBI colposcopy and compared the results with those of conventional colposcopy. Twenty-one patients with adenocarcinoma in situ or early invasive adenocarcinomas were examined using digital NBI colposcopy, and the photo records were compared with those of conventional colposcopy. The histological examination and immunohistochemistry with anti-CD31 antibody confirmed the microvascular pattern. Digital NBI colposcopy depicted the fine vascular texture on the surface of the cervix more clearly than conventional colposcopy. The vascular pattern was depicted in 86% (18/21) of glandular disease cases. The characteristic fine vascular patterns were critical for identifying cervical glandular diseases. Digital NBI colposcopy was useful for identifying early cervical adenocarcinoma as well as adenocarcinoma in situ. This system yields cervical glandular disease-related colposcopic findings that may be useful for both clinical and educational purposes.


Validation of the Spanish Addiction Severity Index Multimedia Version (S-ASI-MV).
This study aimed to develop and test the reliability and validity of a Spanish adaptation of the ASI-MV, a computer administered version of the Addiction Severity Index, called the S-ASI-MV. Participants were 185 native Spanish-speaking adult clients from substance abuse treatment facilities serving Spanish-speaking clients in Florida, New Mexico, California, and Puerto Rico. Participants were administered the S-ASI-MV as well as Spanish versions of the general health subscale of the SF-36, the work and family unit subscales of the Social Adjustment Scale Self-Report, the Michigan Alcohol Screening Test, the alcohol and drug subscales of the Personality Assessment Inventory, and the Hopkins Symptom Checklist-90. Three-to-five-day test-retest reliability was examined along with criterion validity, convergent/discriminant validity, and factorial validity. Measurement invariance between the English and Spanish versions of the ASI-MV was also examined. The S-ASI-MV demonstrated good test-retest reliability (ICCs for composite scores between .59 and .93), criterion validity (rs for composite scores between .66 and .87), and convergent/discriminant validity. Factorial validity and measurement invariance were demonstrated. These results compared favorably with those reported for the original interviewer version of the ASI and the English version of the ASI-MV.


Administration of galacto-oligosaccharide prebiotics in the Flinders Sensitive Line animal model of depression.
Major depressive disorder is the leading source of disability globally and current pharmacological treatments are less than adequate. Animal models such as the Flinders Sensitive Line (FSL) rats are used to mimic aspects of the phenotype in the human disorder and to characterise candidate antidepressant agents. Communication between the gut microbiome and the brain may play an important role in psychiatric disorders such as depression. Interventions targeting the gut microbiota may serve as potential treatments for depression, and this drives increasing research into the effect of probiotics and prebiotics in neuropsychiatric disorders. Prebiotics, galacto-oligosaccharides and fructooligosaccharides that stimulate the activity of gut bacteria have been reported to have a positive impact, reducing anxiety and depressive-like phenotypes and stress-related physiology in mice and rats, as well as in humans. Bimuno, the commercially available beta-galacto-oligosaccharide, has been shown to increase gut microbiota diversity. Here, we aim to investigate the effect of Bimuno on rat anxiety-like and depressive-like behaviour and gut microbiota composition in the FSL model, a genetic model of depression, in comparison to their control, the Flinders Resistant Line (FRL) rats. Sixty-four male rats aged 5-7 weeks, 32 FSL and 32 FRL rats, will be randomised to receive Bimuno or control (4 g/kg) daily for 4 weeks. Animals will be tested by an experimenter unaware of group allocation on the forced swim test to assessed depressive-like behaviour, the elevated plus maze to assess anxiety-like behaviour and the open field test to assess locomotion. Animals will be weighed and food and water intake, per kilogram of bodyweight, will be recorded. Faeces will be collected from each animal prior to the start of the experiment and on the final day to assess the bacterial diversity and relative abundance of bacterial genera in the gut. All outcomes and statistical analysis will be carried out blinded to group allocation, group assignments will be revealed after raw data have been uploaded to Open Science Framework. Two-way analysis of variance will be carried out to investigate the effect of treatment (control or prebiotic) and strain (FSL or FRL) on depressive-like and anxiety-like behaviours.


Putting the community back in community ecology and education: the role of field schools and private reserves in the ethical training of primatologists.
In 1993 and 1999, with the assistance of a Nicaraguan family, we founded La Suerte Biological Research Station in northeastern Costa Rica and Ometepe Biological Research Station in southern Nicaragua as a privately owned conservation-oriented business. Our goal was to develop a program of sustainable community ecology focused on education, research, and the conservation of primates and tropical forests. In order to accomplish this we developed field courses in which undergraduate and graduate students conduct scientific research, experience local cultures, and learn about conservation. Over 120 of these students have received doctoral degrees or are currently in graduate programs. Four doctoral dissertations, several MA theses, and some 20 scientific articles have been published based on research conducted at our field stations. In order to achieve our long-term goals of preserving the environment, we also needed to engage directly with local communities to address their needs and concerns. To this end, we developed a series of community-based initiatives related to health care, bilingual education, and conservation education using traditional and on-line teaching tools. In this article, we describe our efforts in Costa Rica and Nicaragua teaching conservation-oriented field courses and working with the local human communities. Building upon these experiences, we outline a set of ethical considerations and responsibilities for private reserves, conservation-oriented businesses, NGOs, and conservancies that help integrate members of the local community as stakeholders in conservation.


Traumatic dental injuries as reported during school hours in Bergen.
To identify existing guidelines for managing traumatic dental injuries (TDIs) in the schools of Bergen, to ascertain the frequency of occurrence of such injuries, and to estimate the need for further information among teachers and school administrators. The study, undertaken among teachers and school administrators of elementary and lower secondary schools in Bergen municipality, was questionnaire-based and included a cross section of staff. The structured short questionnaire included items registering TDIs during 2009, existence of routines or guidelines for managing TDIs, previous relevant training, and request for TDI education or information. The statistical methods included frequency tables and logistic regression analysis. The response rate was 73%. The incidence proportion of TDIs was measured to 0.74% of children at risk, varying according to children's classes (peak at third class: 1.68% of children in the population). No schools had adequate written guidelines for handling TDIs. Previous education on the subject was scarce. In 20 schools, there was no perceived need for TDI-related education or information. The schools' routines for TDI reporting, who was in charge of the reporting, acquired TDI education and expressed need for TDI information or education, did not influence the number of reported TDI cases. This study has produced reliable information that schools in the municipality of Bergen could improve ways of reporting and managing TDIs. As teachers with skills in handling TDIs could help to improve the prognosis for injured teeth, some types of educational intervention in schools should be launched.


Thermococcus kodakarensis DNA replication.
DNA replication plays an essential role in all life forms. Research on archaeal DNA replication began approximately 20 years ago. Progress was hindered, however, by the lack of genetic tools to supplement the biochemical and structural studies. This has changed, however, and genetic approaches are now available for several archaeal species. One of these organisms is the thermophilic euryarchaeon Thermococcus kodakarensis. In the present paper, the recent developments in the biochemical, structural and genetic studies on the replication machinery of T. kodakarensis are summarized.


Selection and co-selection of antibiotic resistances among Escherichia coli by antibiotic use in primary care: An ecological analysis.
The majority of studies that link antibiotic usage and resistance focus on simple associations between the resistance against a specific antibiotic and the use of that specific antibiotic. However, the relationship between antibiotic use and resistance is more complex. Here we evaluate selection and co-selection by assessing which antibiotics, including those mainly prescribed for respiratory tract infections, are associated with increased resistance to various antibiotics among Escherichia coli isolated from urinary samples. Monthly primary care prescribing data were obtained from National Health Service (NHS) Digital. Positive E. coli records from urine samples in English primary care (n = 888,207) between April 2014 and January 2016 were obtained from the Second Generation Surveillance System. Elastic net regularization was used to evaluate associations between prescribing of different antibiotic groups and resistance against amoxicillin, cephalexin, ciprofloxacin, co-amoxiclav and nitrofurantoin at the clinical commissioning group (CCG) level. England is divided into 209 CCGs, with each NHS practice prolonging to one CCG. Amoxicillin prescribing (measured in DDD/ 1000 inhabitants / day) was positively associated with amoxicillin (RR 1.03, 95% CI 1.01-1.04) and ciprofloxacin (RR 1.09, 95% CI 1.04-1.17) resistance. In contrast, nitrofurantoin prescribing was associated with lower levels of resistance to amoxicillin (RR 0.92, 95% CI 0.84-0.97). CCGs with higher levels of trimethoprim prescribing also had higher levels of ciprofloxacin resistance (RR 1.34, 95% CI 1.10-1.59). Amoxicillin, which is mainly (and often unnecessarily) prescribed for respiratory tract infections is associated with increased resistance against various antibiotics among E. coli causing urinary tract infections. Our findings suggest that when predicting the potential impact of interventions on antibiotic resistances it is important to account for use of other antibiotics, including those typically used for other indications.


Comparative global immune-related gene profiling of somatic cells, human pluripotent stem cells and their derivatives: implication for human lymphocyte proliferation.
Human pluripotent stem cells (hPSCs), including embryonic stem cells (ESCs) and induced PSCs (iPSCs), represent potentially unlimited cell sources for clinical applications. Previous studies have suggested that hPSCs may benefit from immune privilege and limited immunogenicity, as reflected by the reduced expression of major histocompatibility complex class-related molecules. Here we investigated the global immune-related gene expression profiles of human ESCs, hiPSCs and somatic cells and identified candidate immune-related genes that may alter their immunogenicity. The expression levels of global immune-related genes were determined by comparing undifferentiated and differentiated stem cells and three types of human somatic cells: dermal papilla cells, ovarian granulosa cells and foreskin fibroblast cells. We identified the differentially expressed genes CD24, GATA3, PROM1, THBS2, LY96, IFIT3, CXCR4, IL1R1, FGFR3, IDO1 and KDR, which overlapped with selected immune-related gene lists. In further analyses, mammalian target of rapamycin complex (mTOR) signaling was investigated in the differentiated stem cells following treatment with rapamycin and lentiviral transduction with specific short-hairpin RNAs. We found that the inhibition of mTOR signal pathways significantly downregulated the immunogenicity of differentiated stem cells. We also tested the immune responses induced in differentiated stem cells by mixed lymphocyte reactions. We found that CD24- and GATA3-deficient differentiated stem cells including neural lineage cells had limited abilities to activate human lymphocytes. By analyzing the transcriptome signature of immune-related genes, we observed a tendency of the hPSCs to differentiate toward an immune cell phenotype. Taken together, these data identify candidate immune-related genes that might constitute valuable targets for clinical applications.


Alcohol identification and intervention in English emergency departments.
In the ED, alcohol identification and brief advice is an effective method of reducing consumption and related harms. Our objective was to conduct a national survey of English EDs to determine current practice regarding alcohol identification and provision of brief advice and to compare changes in activity to a previous National Survey conducted in 2011. This was a cross-sectional survey of all consultant-led EDs in England. Of 180 departments, 147 (81.6%) responded. All departments may question adult patients about their alcohol consumption, with many (63.6%) asking all patients aged over 18 years as part routine care and using a formal screening tool (61.4%). The majority of departments asked young people (aged 11-17 years) about their consumption (83.8%), but only 11.6% did so as a part of routine practice. Compared with the 2011 survey, there have been significant increases in routine screening among adults (15.9%, CI 4.16% to 27.18%; p=0.006), general practitioners being informed about patients'alcohol-related presentations (10.2%, CI 0.64% to 19.58%; p=0.028) and access to an alcohol health worker or a clinical nurse specialist (13.4%, CI 3.64% to 22.91%, p=0.005). Modest (non-significant) changes were also found in access to training on brief advice (9.7%) and the use of formal screening questions on adult patients (9.7%). Alcohol screening together with referral or intervention is becoming part of routine practice in England. Compared with our previous national survey, increases in alcohol screening and intervention activity are demonstrated, with improvements in routine questioning (among adults), the number of general practitioners being informed about alcohol-related attendances, provision of training, access to specialist services and the use of formal screening tools.


A Culinary Misadventure: A Case Report of Shiitake Dermatitis.
Shiitake dermatitis is a rare cutaneous reaction that can occur after consumption of shiitake mushrooms. It is characterized by highly pruritic lesions with a distinct whip lash appearance. Management of this condition includes reassurance and providing symptomatic relief. Although most reported cases have occurred in Asia, this rash has made appearances in the Western world. Here, we report a case of a 23-year-old female in the United States who experienced this itchy ailment. A diagnosis was made based on the presence of the recognizable lesions and dietary history. She was discharged with advice to continue symptomatic care and given reassurance about her condition. Her symptoms resolved within a few weeks. As the use of shiitake mushrooms become more widespread, it is important for clinicians to be familiar with this rash and remember to take a detailed dietary history. Rash, dermatology, toxicology, shiitake mushrooms.