Factors affecting visual sensitivity in a hemianopic subject.
A well-studied subject with visual cortex damage (G.Y.) was tested in his hemianopic field with temporally modulated sinusoidal and square-wave gratings. The purpose was to use an extended range of parameters to obtain a detailed spatiotemporal specification of his residual vision and to try to resolve the discrepancy between negative findings of Hess and Pointer (1989) and previous positive claims. Both the spatial and temporal parameters could be Gaussian-weighted. Detection as a function of spatial frequency, contrast, temporal modulation frequency, stimulus size, and slope of the temporal and spatial Gaussian functions was investigated using a two-alternative forced-choice procedure. The most important parameters for this subject were found to be the slope of the temporal Gaussian function and the size and contrast of gratings. With optimum parameters he could reliably achieve a score of 95-100% correct in his 'blind' field. The results are consistent with earlier studies of this subject, especially his ability to respond to moving stimuli, and also may account for why negative results had been reported for him when particular fixed parameters were used.


Social decision-making in the brain: Input-state-output modelling reveals patterns of effective connectivity underlying reciprocal choices.
During social interactions, decision-making involves mutual reciprocity-each individual's choices are simultaneously a consequence of, and antecedent to those of their interaction partner. Neuroeconomic research has begun to unveil the brain networks underpinning social decision-making, but we know little about the patterns of neural connectivity within them that give rise to reciprocal choices. To investigate this, the present study measured the behaviour and brain function of pairs of individuals (N = 66) whilst they played multiple rounds of economic exchange comprising an iterated ultimatum game. During these exchanges, both players could attempt to maximise their overall monetary gain by reciprocating their opponent's prior behaviour-they could promote generosity by rewarding it, and/or discourage unfair play through retaliation. By adapting a model of reciprocity from experimental economics, we show that players' choices on each exchange are captured accurately by estimating their expected utility (EU) as a reciprocal reaction to their opponent's prior behaviour. We then demonstrate neural responses that map onto these reciprocal choices in two brain regions implicated in social decision-making: right anterior insula (AI) and anterior/anterior-mid cingulate cortex (aMCC). Finally, with behavioural Dynamic Causal Modelling, we identified player-specific patterns of effective connectivity between these brain regions with which we estimated each player's choices with over 70% accuracy; namely, bidirectional connections between AI and aMCC that are modulated differentially by estimates of EU from our reciprocity model. This input-state-output modelling procedure therefore reveals systematic brain-behaviour relationships associated with the reciprocal choices characterising interactive social decision-making.


Comparing aftereffects after split-belt treadmill walking and unilateral stepping.
Asymmetric step length is a problem common to many orthopedic and neurologic populations. Herein, we compare step length aftereffects during overground gait after two rehabilitation intervention strategies to combat step length asymmetry: split-belt treadmill (SBT) walking and unilateral stepping. Eighteen healthy young adults (22 ± 3 yr) first performed 10 overground gait trials. Participants then walked for 10 min under three different treadmill conditions in a randomized order: SBT walking, slow unilateral stepping, and fast unilateral stepping. Immediately after each treadmill condition, participants performed ten overground gait trials. Mean step length asymmetry was calculated across the first five strides of the overground gait trials to assess the storage of aftereffects after each treadmill condition. We also explored the lower extremity kinematics during each treadmill condition to investigate movement patterns that lead to greatest aftereffects. Significantly higher step length asymmetry was observed in overground gait trials after SBT walking compared to those after slow and fast unilateral stepping, indicating greater aftereffect/carryover of the SBT walking pattern to overground gait. During fast unilateral stepping, increased flexion in the hip, knee, and ankle of the stationary limb was significantly associated with increased step length aftereffects. The aftereffects observed after acute SBT walking were significantly greater than those after unilateral stepping. Both exercises induce aftereffects of similar kinematic patterns, although likely through different mechanisms. In sum, SBT walking induces the greatest aftereffects, although unilateral stepping also induces a change in gait behavior. During unilateral stepping, the largest aftereffects occur when the walker does not simply fully extend the stationary limb and allow the treadmill to passively move the stepping limb during stance.


Beta 2-integrin and L-selectin are obligatory receptors in neutrophil aggregation.
We have recently found that antibodies to L-selectin, the homing receptor on neutrophils, are as effective as those to beta 2-integrin at blocking formyl peptide-stimulated aggregation. Therefore, we investigated the requirements for expression of L-selectin and beta 2-integrin on adjacent cells during aggregation. Fluorescence flow cytometry allowed characterization of aggregates on the basis of size and color, as well as antibody binding to these two adhesive molecules. Formyl peptide-stimulated aggregate formation was measured for individual populations fluorescently labeled red (LDS-751) or green (CD44-FITC), and interpopulation red-green cell conjugates. Blocking either the beta 2-integrin or L-selectin adhesive epitope with monoclonal antibody on individual cell populations resulted in an approximately 50% reduction in two-color aggregation as compared with that in unblocked samples. Shedding the L-selectin on a cell population by preincubation with complexes of lipopolysaccharide and its plasma membrane binding protein also decreased aggregation to a control population by approximately 50%. We examined the aggregation of neutrophils from patients genetically deficient in beta 2-integrin and clinically leukocyte adhesion deficient (LAD). LAD adhesion to normal neutrophils was dependent on the expression of L-selectin on LAD cells and beta 2-integrin on normal cells. Thus, the minimum requirement for adhesion between two mixed populations of neutrophils was that one population expressed the beta 2-integrin and the other expressed the L-selectin adhesive epitope.


Intervention to improve the quality of antimicrobial prescribing for urinary tract infection: a cluster randomized trial.
Overuse of antimicrobial therapy in the community adds to the global spread of antimicrobial resistance, which is jeopardizing the treatment of common infections. We designed a cluster randomized complex intervention to improve antimicrobial prescribing for urinary tract infection in Irish general practice. During a 3-month baseline period, all practices received a workshop to promote consultation coding for urinary tract infections. Practices in intervention arms A and B received a second workshop with information on antimicrobial prescribing guidelines and a practice audit report (baseline data). Practices in intervention arm B received additional evidence on delayed prescribing of antimicrobials for suspected urinary tract infection. A reminder integrated into the patient management software suggested first-line treatment and, for practices in arm B, delayed prescribing. Over the 6-month intervention, practices in arms A and B received monthly audit reports of antimicrobial prescribing. The proportion of antimicrobial prescribing according to guidelines for urinary tract infection increased in arms A and B relative to control (adjusted overall odds ratio [OR] 2.3, 95% confidence interval [CI] 1.7 to 3.2; arm A adjusted OR 2.7, 95% CI 1.8 to 4.1; arm B adjusted OR 2.0, 95% CI 1.3 to 3.0). An unintended increase in antimicrobial prescribing was observed in the intervention arms relative to control (arm A adjusted OR 2.2, 95% CI 1.2 to 4.0; arm B adjusted OR 1.4, 95% CI 0.9 to 2.1). Improvements in guideline-based prescribing were sustained at 5 months after the intervention. A complex intervention, including audit reports and reminders, improved the quality of prescribing for urinary tract infection in Irish general practice. ClinicalTrials.gov, no. NCT01913860.


3D imaging in forensic odontology.
This paper describes the investigation of a new 3D capture method for acquiring and subsequent forensic analysis of bite mark injuries on human skin. When documenting bite marks with standard 2D cameras errors in photographic technique can occur if best practice is not followed. Subsequent forensic analysis of the mark is problematic when a 3D structure is recorded into a 2D space. Although strict guidelines (BAFO) exist, these are time-consuming to follow and, due to their complexity, may produce errors. A 3D image capture and processing system might avoid the problems resulting from the 2D reduction process, simplifying the guidelines and reducing errors. Proposed Solution: a series of experiments are described in this paper to demonstrate that the potential of a 3D system might produce suitable results. The experiments tested precision and accuracy of the traditional 2D and 3D methods. A 3D image capture device minimises the amount of angular distortion, therefore such a system has the potential to create more robust forensic evidence for use in courts. A first set of experiments tested and demonstrated which method of forensic analysis creates the least amount of intra-operator error. A second set tested and demonstrated which method of image capture creates the least amount of inter-operator error and visual distortion. In a third set the effects of angular distortion on 2D and 3D methods of image capture were evaluated.


Primary care utilization in people who experience imprisonment in Ontario, Canada: a retrospective cohort study.
Access to primary care is an important determinant of health, and data are sparse on primary care utilization for people who experience imprisonment. We aimed to describe primary care utilization for persons released from prison, and to compare utilization with the general population. We linked correctional data for all persons released from provincial prison in Ontario, Canada in 2010 with health administrative data. We matched each person by age and sex with four general population controls. We compared primary care utilization rates using generalized estimating equations. We adjusted rate ratios for aggregated diagnosis groups, to explore this association independent of comorbidity. We examined the proportion of people using primary care using chi squared tests and time to first primary care visit post-release using the Kaplan-Meier method. Compared to the general population controls, the prison release group had significantly increased relative rates of primary care utilization: at 6.1 (95% CI 5.9-6.2) in prison, 3.7 (95% CI 3.6-3.8) in the week post-release and between 2.4 and 2.6 in the two years after prison release. All rate ratios remained significantly increased after adjusting for comorbidity. In the month after release, however, 66.3% of women and 75.5% of men did not access primary care. Primary care utilization is high in prison and post-release for people who experience imprisonment in Ontario, Canada. Increased use is only partly explained by comorbidity. The majority of people do not access primary care in the month after prison release. Future research should identify reasons for increased use and interventions to improve care access for persons who are not accessing care post-release.


Silica-like malleable materials from permanent organic networks.
Permanently cross-linked materials have outstanding mechanical properties and solvent resistance, but they cannot be processed and reshaped once synthesized. Non-cross-linked polymers and those with reversible cross-links are processable, but they are soluble. We designed epoxy networks that can rearrange their topology by exchange reactions without depolymerization and showed that they are insoluble and processable. Unlike organic compounds and polymers whose viscosity varies abruptly near the glass transition, these networks show Arrhenius-like gradual viscosity variations like those of vitreous silica. Like silica, the materials can be wrought and welded to make complex objects by local heating without the use of molds. The concept of a glass made by reversible topology freezing in epoxy networks can be readily scaled up for applications and generalized to other chemistries.


A new crystal form of human acetylcholinesterase for exploratory room-temperature crystallography studies.
Structure-guided design of novel pharmacologically active molecules relies at least in part on functionally relevant accuracy of macromolecular structures for template based drug design. Currently, about 95% of all macromolecular X-ray structures available in the PDB (Protein Data Bank) were obtained from diffraction experiments at low, cryogenic temperatures. However, it is known that functionally relevant conformations of both macromolecules and pharmacological ligands can differ at higher, physiological temperatures. We describe in this article development and properties of new human acetylcholinesterase (AChE) crystals of space group P31 and a new unit cell, amenable for room-temperature X-ray diffraction studies. We co-crystallized hAChE in P31 unit cell with the reversible inhibitor 9-aminoacridine that binds at the base of the active center gorge in addition to inhibitors that span the full length of the gorge, donepezil (Aricept, E2020) and AChE specific inhibitor BW284c51. Their new low temperature P31 space group structures appear similar to those previously obtained in the different P3121 unit cell. Successful solution of the new room temperature 3.2 Å resolution structure of BW284c51*hAChE complex from large P31 crystals enables us to proceed with studying room temperature structures of lower affinity complexes, such as oxime reactivators bound to hAChE, where temperature-related conformational diversity could be expected in both oxime and hAChE, which could lead to better informed structure-based design under conditions approaching physiological temperature.


Consensus statement on normothermic regional perfusion in donation after circulatory death: Report from the European Society for Organ Transplantation's Transplant Learning Journey.
Normothermic regional perfusion (NRP) in donation after circulatory death (DCD) is a safe alternative to in situ cooling and rapid procurement. An increasing number of countries and centres are performing NRP, a technically and logistically challenging procedure. This consensus document provides evidence-based recommendations on the use of NRP in uncontrolled and controlled DCDs. It also offers minimal ethical, logistical and technical requirements that form the foundation of a safe and effective NRP programme. The present article is based on evidence and opinions formulated by a panel of European experts of Workstream 04 of the Transplantation Learning Journey project, which is part of the European Society for Organ Transplantation.


Chemotaxis away from uncouplers of oxidative phosphorylation in Bacillus subtilis.
In a capillary assay, uncouplers of oxidative phosphorylation and inhibitors of electron transport are repellents for Bacillus subtilis. They also cause transient tumbling in naturally smooth swimming strains. Tumbling strains can be made to swim smoothly by addition of attractant and then immediately returned to tumbling by subsequent addition of repellent. Arsenate does not cause transient tumbling, suggesting that decrease in concentration of adenosine triphosphate does not cause tumbling and that adenosine triphosphate concentration does not govern tumbling frequency. Instead, the evidence suggests that diminution of the energized state of the membrane, or membrane potential, causes tumbling although the level of the energized state itself does not govern tumbling frequency.


Differential messenger RNA distribution of lactate dehydrogenase LDH-1 and LDH-5 isoforms in the rat brain.
The role of lactate in brain energy metabolism has recently received renewed attention. Although blood-borne monocarboxylates such as lactate poorly cross the blood-brain barrier in the adult brain, lactate produced within the brain parenchyma may be a suitable substrate for brain cells. Lactate dehydrogenase is crucial for both the production and utilization of lactate. In this article, we report the regional distribution of the messenger RNAs for lactate dehydrogenase isoforms 1 and 5 in the adult rat brain using in situ hybridization histochemistry with specific [alpha-(35)S]dATP 3' end-labeled oligoprobes. The autoradiographs revealed that the lactate dehydrogenase-1 messenger RNA is highly expressed in a variety of brain structures, including the main olfactory bulb, the piriform cortex, several thalamic and hypothalamic nuclei, the pontine nuclei, the ventral cochlear nucleus, the trigeminal nerve and the solitary tractus nucleus. In addition, the granular and Purkinje cell layers of the cerebellum showed a strong labeling. The neocortex (e.g., cingular, retrosplenial and frontoparietal cortices) often exhibits a marked laminar pattern of distribution of lactate dehydrogenase-1 messenger RNA (layers II/III, IV and VI being most strongly labeled). In contrast, expression of the lactate dehydrogenase-5 messenger RNA generally seemed more diffusely distributed across the different brain regions. Expression was particularly strong in the hippocampal formation (especially in Ammon's horn and dentate gyrus) and in the cerebral cortex, where no laminar pattern of distribution was observed. Overall, these data are consistent with the emerging idea that lactate is an important energy substrate produced and consumed by brain cells.


Using 10-essential-services training to revive, refocus, and strengthen your environmental health programs.
The 10 essential services of environmental health, which are based on the 10 essential public health services, can guide environmental health practitioners in systematically organizing and managing environmental public health programs and activities. The National Center for Environmental Health of the Centers for Disease Control and Prevention has used the 10 essential services of environmental health as a basis for its six goals for the revitalization of environmental health in the 21st century. Nevertheless, studies indicate that very few environmental health practitioners are aware of the 10 essential services. This article discusses how essential-services training has increased the awareness and knowledge of environmental health practitioners about the development, value, and use of the essential services. Examples of training outcomes are offered to illustrate how the use of the essential-services framework has improved environmental health performance and practice.


Happiness Scale Interval Study. Methodological Considerations.
The Happiness Scale Interval Study deals with survey questions on happiness, using verbal response options, such as 'very happy' and 'pretty happy'. The aim is to estimate what degrees of happiness are denoted by such terms in different questions and languages. These degrees are expressed in numerical values on a continuous [0,10] scale, which are then used to compute 'transformed' means and standard deviations. Transforming scores on different questions to the same scale allows to broadening the World Database of Happiness considerably. The central purpose of the Happiness Scale Interval Study is to identify the happiness values at which respondents change their judgment from e.g. 'very happy' to 'pretty happy' or the reverse. This paper deals with the methodological/statistical aspects of this approach. The central question is always how to convert the frequencies at which the different possible responses to the same question given by a sample into information on the happiness distribution in the relevant population. The primary (cl)aim of this approach is to achieve this in a (more) valid way. To this end, a model is introduced that allows for dealing with happiness as a latent continuous random variable, in spite of the fact that it is measured as a discrete one. The [0,10] scale is partitioned in as many contiguous parts as the number of possible ratings in the primary scale sums up to. Any subject with a (self-perceived) happiness in the same subinterval is assumed to select the same response. For the probability density function of this happiness random variable, two options are discussed. The first one postulates a uniform distribution within each of the different subintervals of the [0,10] scale. On the basis of these results, the mean value and variance of the complete distribution can be estimated. The method is described, including the precision of the estimates obtained in this way. The second option assumes the happiness distribution to be described as a beta distribution on the interval [0,10] with two shape parameters (α and β). From their estimates on the basis of the primary information, the mean value and the variance of the happiness distribution in the population can be estimated. An illustration is given in which the method is applied to existing measurement results of 20 surveys in The Netherlands in the period 1990-2008. The results clarify our recommendation to apply the model with a uniform distribution within each of the category intervals, in spite of a better validity of the alternative on the basis of a beta distribution. The reason is that the recommended model allows to construct a confidence interval for the true but unknown population happiness distribution. The paper ends with a listing of actual and potential merits of this approach, which has been described here for verbal happiness questions, but which is also applicable to phenomena which are measured along similar lines.


Complete mitochondrial DNA diversity in Iranians.
Due to its pivotal geographical location and proximity to transcontinental migratory routes, Iran has played a key role in subsequent migrations, both prehistoric and historic, between Africa, Asia and Europe. To shed light on the genetic structure of the Iranian population as well as on the expansion patterns and population movements which affected this region, the complete mitochondrial genomes of 352 Iranians were obtained. All Iranian populations studied here exhibit similarly high diversity values comparable to the other groups from the Caucasus, Anatolia and Europe. The results of AMOVA and MDS analyses did not associate any regional and/or linguistic group of populations in the Anatolia/Caucasus and Iran region pointing to close genetic positions of Persians and Qashqais to each other and to Armenians, and Azeris from Iran to Georgians. By reconstructing the complete mtDNA phylogeny of haplogroups R2, N3, U1, U3, U5a1g, U7, H13, HV2, HV12, M5a and C5c we have found a previously unexplored genetic connection between the studied Iranian populations and the Arabian Peninsula, India, Near East and Europe, likely the result of both ancient and recent gene flow. Our results for Persians and Qashqais point to a continuous increase of the population sizes from ∼24 kya to the present, although the phase between 14-24 kya is thought to be hyperarid according to the Gulf Oasis model. Since this would have affected hunter-gatherer ranges and mobility patterns and forced them to increasingly rely on coastal resources, this transition can explain the human expansion across the Persian Gulf region.


Bioavailability of Quercetin in Humans with a Focus on Interindividual Variation.
After consumption of plant-derived foods or beverages, dietary polyphenols such as quercetin are absorbed in the small intestine and metabolized by the body, or they are subject to catabolism by the gut microbiota followed by absorption of the resulting products by the colon. The resulting compounds are bioavailable, circulate in the blood as conjugates with glucuronide, methyl, or sulfate groups attached, and they are eventually excreted in the urine. In this review, the various conjugates from different intervention studies are summarized and discussed. In addition, the substantial variation between different individuals in the measured quercetin bioavailability parameters is assessed in detail by examining published human intervention studies where sources of quercetin have been consumed in the form of food, beverages, or supplements. It is apparent that most reported studies have examined quercetin and/or metabolites in urine and plasma from a relatively small number of volunteers. Despite this limitation, it is evident that there is less interindividual variation in metabolites which are derived from absorption in the small intestine compared to catabolites derived from the action of microbiota in the colon. There is also some evidence that a high absorber of intact quercetin conjugates could be a low absorber of microbiota-catalyzed phenolics, and vice versa. From the studies reported so far, the reasons or causes of the interindividual differences are not clear, but, based on the known metabolic pathways, it is predicted that dietary history, genetic polymorphisms, and variations in gut microbiota metabolism would play significant roles. In conclusion, quercetin bioavailability is subject to substantial variation between individuals, and further work is required to establish if this contributes to interindividual differences in biological responses.


Enhanced conversion of carbohydrates to the platform chemical 5-hydroxymethylfurfural using designer ionic liquids.
5-Hydroxymethylfurfural (HMF) is a key platform chemical that may be obtained from various cellulosic (biomass) derivatives. Previously, it has been shown that ionic liquids (ILs) facilitate the catalytic conversion of glucose into HMF. Herein, we demonstrate that the careful design of the IL cation leads to new ionic solvents that enhance the transformation of glucose and more complex carbohydrates into HMF significantly. In Situ NMR spectroscopy and computational modeling pinpoint the key interactions between the IL, catalyst, and substrate that account for the enhanced reactivities observed.


1-Pentyl-3-phenylacetylindoles and JWH-018 share in vivo cannabinoid profiles in mice.
Smoking of synthetic cannabinoid-enhanced "herbal incense" is an emerging substance abuse problem. The indole-derived cannabinoids identified in these products were originally developed as research tools and are structurally distinct from cannabinoids in the cannabis plant. Although abused by humans, most published research on this class of compounds has been performed in vitro. The purpose of this study was to evaluate a novel series of 1-pentyl-3-phenylacetylindoles in mice. The potencies of these analogs to produce the cannabinoid agonist effects of antinociception, hypothermia and suppression of locomotion were evaluated in ICR mice. The major structural manipulations in the present series included the type of substituent (i.e., unsubstituted, methyl, methoxy, chloro, bromo, and fluoro) and the position of the substituent on the phenyl ring (i.e., 2-, 3- or 4-position). Potencies of this series of phenylacetylindoles for each cannabinoid effect were highly correlated with CB(1) receptor affinities reported previously. Active compounds produced a profile of effects that resembled that exhibited by Δ(9)-tetrahydrocannabinol (THC). The most critical factor affecting in vivo potency was the position of the substituent. Whereas compounds with 2- and 3-phenylacetyl substituents were efficacious with good potencies, 4-substituents resulted in compounds that had poor potency or were inactive. These results suggest that phenylacetylindoles with good CB(1) binding affinity share pharmacological properties with THC in mice; however, they also emphasize the complexity of molecular interactions of synthetic cannabinoids with CB(1) receptors and suggest that scheduling efforts based solely upon structural features should proceed with caution.


Demonstration of cell types among cone bipolar neurons of cat retina.
We identified all the cone bipolar cells (80) in a small patch of one retina and then studied in detail the complete subset (42) that sends axons to sublamina b of the inner plexiform layer. The point was to learn whether the 'types' suggested previously, based on a few examples from a large population, could be substantiated or whether there would be intermediate forms. Tissue from the area centralis (1 degree eccentricity), was prepared as a series of 279 ultrathin sections and photographed in the electron microscope. Thirteen cells were reconstructed completely and parcelled into five categories (b1-b5) based on external morphology. For nine of these cells (two from categories b1-b4 and one from b5) most of the synaptic inputs and outputs were identified. When these nine cells were parcelled according to their synaptic patterns, they sorted into the same five categories. The remaining 29 cells in the population, though not reconstructed, were studied in detail by tracing their processes through the series. Ten of these cells, those near the margin of the series, were incomplete. The other 19 cells had essentially the same distribution of morphologies and synaptic patterns as the subset studied by total reconstruction: when plotted in multiparametric space, they formed distinct clusters corresponding to the five morphological categories. There was no hint of intermediate forms. That all the neurons in the population sort into some cluster (no intermediate forms), and that each neuron sorts into the same cluster by different criteria, argues that the clusters represent natural types. Each type forms a regular array in the region studied with an axonal 'coverage factor' that is close to one.


In silico QTL mapping of basal liver iron levels in inbred mouse strains.
Both iron deficiency and iron excess are detrimental in many organisms, and previous studies in both mice and humans suggest that genetic variation may influence iron status in mammals. However, these genetic factors are not well defined. To address this issue, we measured basal liver iron levels in 18 inbred strains of mice of both sexes on a defined iron diet and found ∼4-fold variation in liver iron in males (lowest 153 μg/g, highest 661 μg/g) and ∼3-fold variation in females (lowest 222 μg/g, highest 658 μg/g). We carried out a genome-wide association mapping to identify haplotypes underlying differences in liver iron and three other related traits (copper and zinc liver levels, and plasma diferric transferrin levels) in a subset of 14 inbred strains for which genotype information was available. We identified two putative quantitative trait loci (QTL) that contain genes with a known role in iron metabolism: Eif2ak1 and Igf2r. We also identified four putative QTL that reside in previously identified iron-related QTL and 22 novel putative QTL. The most promising putative QTL include a 0.22 Mb region on Chromosome 7 and a 0.32 Mb region on Chromosome 11 that both contain only one candidate gene, Adam12 and Gria1, respectively. Identified putative QTL are good candidates for further refinement and subsequent functional studies.


First Report of Soybean Rust Caused by Phakopsora pachyrhizi on Phaseolus spp. in the United States.
Phakopsora pachyrhizi Syd. & P. Syd., the cause of soybean rust, was first observed in the continental United States in November 2004 (2). During the growing season of 2005, P. pachyrhizi was confirmed on soybean (Glycine max) and/or kudzu (Pueraria montana) in nine states in the southern United States. It is known that P. pachyrhizi has a much larger host range within the Fabaceae family. On 29 September 2005 in Quincy, FL, 45 entries of mostly large-seeded legumes were planted next to soybeans that were infected with P. pachyrhizi. Several seeds of each entry were planted on one hill. Soybean plants growing adjacent to these potential hosts had 15 to 25% of the leaf area affected, 95% incidence, and 73% defoliation on 16 November. On 7 December 2005, all the plants of Phaseolus coccineus L. (scarlet runner bean, PI311827), Phaseolus lunatus L. (lima bean, PI583558), and two Phaseolus vulgaris L. (kidney bean) cvs. Red Hawk and California Early Light Red Kidney (CELRK) were found to have leaves with suspected rust lesions. These plants were at physiological maturity but had not senesced. None of the hosts had been inoculated other than from spores produced by the adjacent rust-infected soybean plants or from unknown locations. On the basis of microscopic examination, suspected infected leaves from plants of the Phaseolus spp. had rust pustules characteristic of P. pachyrhizi uredinia. Uredinia were counted within a randomly selected 2-cm2 area of one leaf of each sample. The mean and range of uredinia per lesion for Phaseolus coccineus was 29 uredinia with a range of 0 to 3 uredinia per lesion, Phaseolus lunatus had 2 uredinia with 0 to 1 uredinium per lesion, Phaseolus vulgaris cv. Red Hawk had 22 uredinia with 0 to 5 uredinia per lesion, and Phaseolus vulgaris cv. CELRK had 43 uredinia with 0 to 4 uredinia per lesion. Polymerase chain reactions using two sets of primers (Ppa1/Ppa2 and Pme1/Pme2) were performed on DNA extracted from leaves of the three species with sporulating rust pustules (1). The results of these tests and further tests conducted by the USDA/APHIS confirmed that P. pachyrhizi was the causal organism for the observed rust. References: (1) P. F. Harmon et al. On-line publication. doi:10.1094/PHP-2005-0613-01-RS. Plant Health Progress, 2005. (2) R. W. Schneider et al. Plant Dis. 89:774, 2005.


Pneumonia and pneumonia related mortality in patients with COPD treated with fixed combinations of inhaled corticosteroid and long acting β2 agonist: observational matched cohort study (PATHOS).
To investigate the occurrence of pneumonia and pneumonia related events in patients with chronic obstructive pulmonary disease (COPD) treated with two different fixed combinations of inhaled corticosteroid/long acting β2 agonist. Observational retrospective pairwise cohort study matched (1:1) for propensity score. Primary care medical records data linked to Swedish hospital, drug, and cause of death registry data for years 1999-2009. Patients with COPD diagnosed by a physician and prescriptions of either budesonide/formoterol or fluticasone/salmeterol. Yearly pneumonia event rates, admission to hospital related to pneumonia, and mortality. 9893 patients were eligible for matching (2738 in the fluticasone/salmeterol group; 7155 in the budesonide/formoterol group), yielding two matched cohorts of 2734 patients each. In these patients, 2115 (39%) had at least one recorded episode of pneumonia during the study period, with 2746 episodes recorded during 19,170 patient years of follow up. Compared with budesonide/formoterol, rate of pneumonia and admission to hospital were higher in patients treated with fluticasone/salmeterol: rate ratio 1.73 (95% confidence interval 1.57 to 1.90; P<0.001) and 1.74 (1.56 to 1.94; P<0.001), respectively. The pneumonia event rate per 100 patient years for fluticasone/salmeterol versus budesonide/formoterol was 11.0 (10.4 to 11.8) versus 6.4 (6.0 to 6.9) and the rate of admission to hospital was 7.4 (6.9 to 8.0) versus 4.3 (3.9 to 4.6). The mean duration of admissions related to pneumonia was similar for both groups, but mortality related to pneumonia was higher in the fluticasone/salmeterol group (97 deaths) than in the budesonide/formoterol group (52 deaths) (hazard ratio 1.76, 1.22 to 2.53; P=0.003). All cause mortality did not differ between the treatments (1.08, 0.93 to 1.14; P=0.59). There is an intra-class difference between fixed combinations of inhaled corticosteroid/long acting β2 agonist with regard to the risk of pneumonia and pneumonia related events in the treatment of patients with COPD. Clinical Trials.gov NCT01146392.


Smoky indoor cooking fires are associated with elevated hemoglobin concentration in iron-deficient women.
Indoor air pollution from the burning of such biomass fuels as wood and agricultural waste is associated with a higher risk of a number of respiratory problems. The effect on other health outcomes, such as fetal growth, has not yet been adequately documented. The objective of this study was to determine whether, among women who burn biomass fuels for cooking indoors, the use of "smoky" fires is associated with elevated hemoglobin concentration in comparison to women using "smokeless" stoves, that is, stoves that are designed to reduce indoor air pollution. This research was conducted as part of a series of preliminary studies to determine the feasibility and potential health benefits of a randomized stove intervention to reduce indoor air pollution from the burning of biomass fuels for cooking. A cross-sectional observational study was conducted in rural highland communities of Guatemala from March to August 1994. Venous blood samples were collected and analyzed for hemoglobin and ferritin. All the women studied burned biomass fuels and cooked indoors, and none of the women was pregnant. Eighty-nine indigenous women using smokeless stoves (designated as the not-exposed group) and 185 indigenous women from the same communities using smoky fires (the exposed group) were studied. Multiple linear regression analyses were used to investigate the relationship between exposure (smokeless stove or smoky fire) and hemoglobin concentration, with adjusting for potential confounding factors. No effect of exposure (smokeless stove or smoky fire) on hemoglobin concentration was found in univariate or multivariate analyses. In routine post hoc analysis to determine whether hemoglobin elevation is observed in some particular subgroup, we found that the use of a smoky fire was associated with a 5.2 g/L elevation in hemoglobin concentration among women with low ferritin stores (P < 0.10). The elevation of hemoglobin concentration through exposure to indoor air pollution resulting from the burning of biomass fuels in smoky fires for cooking could have important implications for the diagnosis of anemia. However, considering the observational nature of this study, further research using more rigorous measures of exposure to carbon monoxide as well as additional measures of iron status are needed to confirm the relationships among iron status, exposure to smoke from the burning of biomass fuels indoors, and hemoglobin concentration of women living at moderately high altitude. Further study of this matter could help to assure that appropriate adjustments to anemia cutoffs are made, if warranted, and could assist in clarifying potentially negative outcomes of exposure to smoke from biomass fuels burned indoors.


Trends in senescent life expectancy.
The distinction between senescent and non-senescent mortality proves to be very valuable for describing and analysing age patterns of death rates. Unfortunately, standard methods for estimating these mortality components are lacking. The first part of this paper discusses alternative methods for estimating background and senescent mortality among adults and proposes a simple approach based on death rates by causes of death. The second part examines trends in senescent life expectancy (i.e., the life expectancy implied by senescent mortality) and compares them with trends in conventional longevity indicators between 1960 and 2000 in a group of 17 developed countries with low mortality. Senescent life expectancy for females rises at an average rate of 1.54 years per decade between 1960 and 2000 in these countries. The shape of the distribution of senescent deaths by age remains relatively invariant while the entire distribution shifts over time to higher ages as longevity rises.


Ancient and modern colonization of North America by hemlock woolly adelgid, Adelges tsugae (Hemiptera: Adelgidae), an invasive insect from East Asia.
Hemlock woolly adelgid, Adelges tsugae, is an invasive pest of hemlock trees (Tsuga) in eastern North America. We used 14 microsatellites and mitochondrial COI sequences to assess its worldwide genetic structure and reconstruct its colonization history. The resulting information about its life cycle, biogeography and host specialization could help predict invasion by insect herbivores. We identified eight endemic lineages of hemlock adelgids in central China, western China, Ulleung Island (South Korea), western North America, and two each in Taiwan and Japan, with the Japanese lineages specializing on different Tsuga species. Adelgid life cycles varied at local and continental scales with different sexual, obligately asexual and facultatively asexual lineages. Adelgids in western North America exhibited very high microsatellite heterozygosity, which suggests ancient asexuality. The earliest lineages diverged in Asia during Pleistocene glacial periods, as estimated using approximate Bayesian computation. Colonization of western North America was estimated to have occurred prior to the last glacial period by adelgids directly ancestral to those in southern Japan, perhaps carried by birds. The modern invasion from southern Japan to eastern North America caused an extreme genetic bottleneck with just two closely related clones detected throughout the introduced range. Both colonization events to North America involved host shifts to unrelated hemlock species. These results suggest that genetic diversity, host specialization and host phylogeny are not predictive of adelgid invasion. Monitoring non-native sentinel host trees and focusing on invasion pathways might be more effective methods of preventing invasion than making predictions using species traits or evolutionary history.


Comparison of dual-mobility cup and unipolar cup for prevention of dislocation after revision total hip arthroplasty.
Background and purpose - Revision total hip arthroplasty (THA) is associated with higher dislocation rates than primary THA. We compared the risk of dislocation within 6 months and all-cause re-revision during the whole study period using either the dual-mobility cup or the unipolar cup. Methods - We used a prospective hospital registry-based cohort including all total and cup-only revision THAs performed between 2003 and 2013. The cups used were either dual-mobility or unipolar; the choice was made according to the preference of the surgeon. 316 revision THAs were included. The mean age of the cohort was 69 (25-98) years and 160 THAs (51%) were performed in women. The dual-mobility group (group 1) included 150 THAs (48%) and the mean length of follow-up was 31 (0-128) months. The unipolar group (group 2) included 166 THAs (53%) and the mean length of follow-up was 52 (0-136) months. Results - The incidence of dislocation within 6 months was significantly lower with the dual-mobility cup than with the unipolar cup (2.7% vs. 7.8%). The unadjusted risk ratio (RR) was 0.34 (95% CI: 0.11-1.02) and the adjusted RR was 0.28 (95% CI: 0.09-0.87). The number of patients needed to treat with a dual-mobility cup in order to prevent 1 case of dislocation was 19. The unadjusted incidence rate ratio for all-cause re-revision in the dual-mobility group compared to the unipolar group was 0.6 (95% CI: 0.3-1.4). Interpretation - Use of a dual-mobility rather than a unipolar cup in revision THA reduced the risk of dislocation within 6 months.


Geographical variation in rates of common mental disorders in Britain: prospective cohort study.
There is little geographical variation in the prevalence of the common mental disorders. However, there is little longitudinal research. To estimate variance in rates of common mental disorders at individual, household and electoral ward levels prospectively. A 12-month cohort study of 7659 adults aged 16-74 years in 4338 private households, in 626 electoral wards. Data were collected as part of the British Household Panel Survey. Common mental disorders were assessed using the 12-item General Health Questionnaire (GHQ). Ward-level socio-economic deprivation was measured using the Carstairs index. Less than 1% of total variance, in onset and maintenance of common mental disorders and change in GHQ score between waves, occurred at ward level. However, 12% of variance, which is a statistically significant difference, was found at household level (a much smaller geographical unit) and this difference remained after further analyses. Ward level socio-economic deprivation does not influence the onset and maintenance of common mental disorders in Britain but local factors at the household level do. Reasons for this remain unclear.


Anti-anxiety efficacy of Sudarshan Kriya Yoga in general anxiety disorder: A multicomponent, yoga based, breath intervention program for patients suffering from generalized anxiety disorder with or without comorbidities.
Surdashan Kriya Yoga (SKY) is a procedure that in various studies, has shown evidences of efficacy in alleviating Depression and Anxiety disorders, but in Europe and USA it has not been studied yet on a Caucasian population as an adjunct therapy for psychiatric Disorders. The study involved a sample of consenting women and men (n = 69) who received SKY therapy for a six-month time period. They were assessed at recruitment, after two weeks, after three months and after six months using Hamilton Rating Scale for Anxiety (HRSA), Hamilton Rating Scale for Depression (HRSD), Zung Self-Rating Anxiety Scale (ZSAS), Zung Self-Rating Depression Scale (ZSDS) and Symptom Checklist-90 (SCL-90). All the analyses have shown that SKY therapy significantly reduces the scores of Anxiety and Depression. This is plain, especially after the initial SKY treatment, which is followed by a long plateau phase that seems to verge on no Anxiety/Depression scores. It was found that SKY effects lead to a significant convergence between the self-assessment (Zung Self-Rating Scale) and hetero-assessment (Hamilton Rating Scale). The study should be replicated on a larger clinical sample in a controlled trial to learn more about the effectiveness of SKY Protocol. Participation in SKY adjunct therapy ten days intense workshop and follow-ups, coupled with daily individual and independent practice of a simplified protocol of breathing techniques (30 min), can lead to significant reduction in levels of Anxiety and Depression.


Trends in underlying causes of death in people with HIV from 1999 to 2011 (D:A:D): a multicohort collaboration.
With the advent of effective antiretroviral treatment, the life expectancy for people with HIV is now approaching that seen in the general population. Consequently, the relative importance of other traditionally non-AIDS-related morbidities has increased. We investigated trends over time in all-cause mortality and for specific causes of death in people with HIV from 1999 to 2011. Individuals from the Data collection on Adverse events of anti-HIV Drugs (D:A:D) study were followed up from March, 1999, until death, loss to follow-up, or Feb 1, 2011, whichever occurred first. The D:A:D study is a collaboration of 11 cohort studies following HIV-1-positive individuals receiving care at 212 clinics in Europe, USA, and Australia. All fatal events were centrally validated at the D:A:D coordinating centre using coding causes of death in HIV (CoDe) methodology. We calculated relative rates using Poisson regression. 3909 of the 49,731 D:A:D study participants died during the 308,719 person-years of follow-up (crude incidence mortality rate, 12.7 per 1000 person-years [95% CI 12.3-13.1]). Leading underlying causes were: AIDS-related (1123 [29%] deaths), non-AIDS-defining cancers (590 [15%] deaths), liver disease (515 [13%] deaths), and cardiovascular disease (436 [11%] deaths). Rates of all-cause death per 1000 person-years decreased from 17.5 in 1999-2000 to 9.1 in 2009-11; we saw similar decreases in death rates per 1000 person-years over the same period for AIDS-related deaths (5.9 to 2.0), deaths from liver disease (2.7 to 0.9), and cardiovascular disease deaths (1.8 to 0.9). However, non-AIDS cancers increased slightly from 1.6 per 1000 person-years in 1999-2000 to 2.1 in 2009-11 (p=0.58). After adjustment for factors that changed over time, including CD4 cell count, we detected no decreases in AIDS-related death rates (relative rate for 2009-11 vs 1999-2000: 0.92 [0.70-1.22]). However, all-cause (0.72 [0.61-0.83]), liver disease (0.48 [0.32-0.74]), and cardiovascular disease (0.33 [0.20-0.53) death rates still decreased over time. The percentage of all deaths that were AIDS-related (87/256 [34%] in 1999-2000 and 141/627 [22%] in 2009-11) and liver-related (40/256 [16%] in 1999-2000 and 64/627 [10%] in 2009-11) decreased over time, whereas non-AIDS cancers increased (24/256 [9%] in 1999-2000 to 142/627 [23%] in 2009-11). Recent reductions in rates of AIDS-related deaths are linked with continued improvement in CD4 cell count. We hypothesise that the substantially reduced rates of liver disease and cardiovascular disease deaths over time could be explained by improved use of non-HIV-specific preventive interventions. Non-AIDS cancer is now the leading non-AIDS cause and without any evidence of improvement. Oversight Committee for the Evaluation of Metabolic Complications of HAART, with representatives from academia, patient community, US Food and Drug Administration, European Medicines Agency and consortium of AbbVie, Boehringer Ingelheim, Bristol-Myers Squibb, Gilead Sciences, ViiV Healthcare, Merck, Pfizer, F Hoffmann-La Roche, and Janssen Pharmaceuticals.


The molecular epidemiology of ocular Chlamydia trachomatis infections in Western Australia: implications for trachoma control.
We studied the serovar distribution of Chlamydia trachomatis in patients with clinical eye disease in Western Australia. Most disease occurred in indigenous communities and was caused by trachoma serovars Ba and C. Serovar Ba was genetically homogeneous throughout Western Australia and identical to strains previously described in indigenous communities in Northern Territory. This finding probably results from movement of these populations, and suggests that a widely coordinated, rather than local or regional, approach is needed to control trachoma in mobile populations. Serovar C strains within Western Australia were homogeneous but distinct from those in Northern Territory, possibly because of inherent differences in transmissibility or differences in population movements among communities carrying the different serovars. Genital serovars were occasional causes of eye diseases in infants, adolescents, and adults in trachoma-endemic areas. These serovars should be considered in the differential diagnosis of acute follicular conjunctivitis in these groups.


Assessing equity in access to health care provision in the UK: does where you live affect your chances of getting a coronary artery bypass graft?
Equity should be monitored routinely for all health care services, but ideal studies for each service would be prohibitively expensive and time consuming. A simple, quick, and cheap method for the preliminary exploration of equity in health care provision using routine data was devised. This method was illustrated by examining whether coronary artery bypass graft (CABG) operations reflect socioeconomic differences in ischaemic heart disease (IHD) mortality. Ecological comparison of operation rates was undertaken for CABG for 1991 and IHD mortality for 1981-85 by quartiles of Townsend deprivation score. North East Thames Regional Health Authority, London, UK. All residents of this region aged 35-74 were the denominator population. Numerators were 26,834 IHD deaths and 1041 CABG operations for the defined time periods. IHD mortality showed a steady, significant increase with increasing area deprivation scores for both men and women. CABG rate ratios increased linearly for women, while for men there was a U shaped pattern, being lowest for the second and third quartiles. This pattern was attenuated, but not abolished, when adjusted for geographical proximity to cardiothoracic surgical units. The ratio of CABG operations to IHD mortality by deprivation was relatively constant in women suggesting equitable provision. In men, this ratio was significantly lower for the third quartile. Inequities may exist in the provision of CABG operations for men in this region and this finding should be the stimulus for further detailed studies. Other health care systems should also examine equity in provision.


Making sense of it all: bacterial chemotaxis.
Bacteria must be able to respond to a changing environment, and one way to respond is to move. The transduction of sensory signals alters the concentration of small phosphorylated response regulators that bind to the rotary flagellar motor and cause switching. This simple pathway has provided a paradigm for sensory systems in general. However, the increasing number of sequenced bacterial genomes shows that although the central sensory mechanism seems to be common to all bacteria, there is added complexity in a wide range of species.


Improved treatment of the Stevens-Johnson syndrome.
The Stevens-Johnson syndrome (SJS), a disease resulting in greater than 30% body surface area epidermal and mucosal desquamation, is reported to have a mortality of greater than 50%. Recommended treatment has included steroids, prophylactic antibiotics, nutritional support, and application of topical antibacterials. Since the lesions represent only epidermal loss without dermal necrosis, rapid reepithelialization should occur if the dermis is protected from desiccation with a biological dressing. Five nonburned patients with severe SJS were treated in the University of Washington burn center, Seattle, with urgent operative debridement of their wounds and application of porcine xenograft immediately after admission. Steroid administration was stopped. Antibiotics were used only for culture-proved infections. All patients completely re-epithelialized within a mean of 18 days. There was one death, a 9-year-old boy who was completely re-epithelialized when sudden fatal sepsis that originated from a ventriculovenous shunt placed when he was an infant developed. In this group of patients, rapid wound closure with xenograft and supportive care permitted rapid reepithelialization and decreased mortality and morbidity.


Effects of parsley (Petroselinum crispum) on the liver of diabetic rats: a morphological and biochemical study.
Parsley is used by diabetics in Turkey to reduce blood glucose. The present study aims to investigate both the morphological and biochemical effects of parsley on liver tissue. Rat hepatocytes were examined by light and electron microscopy. Degenerative changes were observed in the hepatocytes of diabetic rats. These degenerative changes were significantly reduced or absent in the hepatocytes of diabetic rats treated with parsley. Blood glucose levels, alanine transaminase and alkaline phosphatase were observed to be raised in diabetic rats. Diabetic rats treated with parsley demonstrated significantly lower levels of blood glucose, alanine transaminase and alkaline phosphatase. The present study suggests that parsley demonstrates a significant hepatoprotective effect in diabetic rats.


Expression of Pax-3 is initiated in the early neural plate by posteriorizing signals produced by the organizer and by posterior non-axial mesoderm.
Pax-3 is a paired-type homeobox gene that is specifically expressed in the dorsal and posterior neural tube. We have investigated inductive interactions that initiate Pax-3 transcript expression in the early neural plate. We present several lines of evidence that support a model where Pax-3 expression is initiated by signals that posteriorize the neuraxis, and then secondarily restricted dorsally in response to dorsal-ventral patterning signals. First, in chick and Xenopus gastrulae the onset of Pax-3 expression occurs in regions fated to become posterior CNS. Second, Hensen's node and posterior non-axial mesoderm which underlies the neural plate induce Pax-3 expression when combined with presumptive anterior neural plate explants. In contrast, presumptive anterior neural plate explants are not competent to express Pax-3 in response to dorsalizing signals from epidermal-ectoderm. Third, in a heterospecies explant recombinant assay with Xenopus animal caps (ectoderm) as a responding tissue, late, but not early, Hensen's node induces Pax-3 expression. Chick posterior non-axial mesoderm also induces Pax-3, provided that the animal caps are neuralized by treatment with noggin. Finally we show that the putative posteriorizing factors, retinoic acid and bFGF, induce Pax-3 in neuralized animal caps. However, blocking experiments with a dominant-inhibitory FGF receptor and a dominant-inhibitory retinoic acid receptor suggest that Pax-3 inductive activities arising from Hensen's node and posterior non-axial mesoderm do not strictly depend on FGF or retinoic acid.


Photodynamic properties of vital dyes for vitreoretinal surgery.
The purpose of this study was to evaluate photodynamic properties of indocyanine green (ICG), brilliant blue G (BBG) and trypan blue (TB) as currently used vital dyes for chromovitrectomy. Under consideration of intraoperative illumination intensities and dye concentrations, a simulative in vitro investigation was set up. Therefore, standardized dilutions of original ICG, BBG and TB vials were irradiated at a wavelength of 366 nm with an intensity of 14 µW/cm2 between 0 and 48 h. After this, all samples were measured spectroscopically in a 220- to 750-nm bandwidth. Analyzing the vital dyes over the time course, an exponential photolysis was observed for ICG, whereas BBG and TB presented photostable properties. Regarding ICG, 5% of the concentration was degraded to toxic metabolites every 20 min. For this reason, our study provides evidence that intraocular dye concentrations and modern endoillumination systems alone cannot fully prevent ICG photodegradation.


Educational attainment & quitting smoking: A structural equation model approach.
In the United States, disparities in smoking prevalence and cessation by socioeconomic status are well documented, but there is limited research on reasons why and none conducted in a national sample assessing multiple potential mechanisms. We identified smoking and cessation-related behavioral and environmental variables associated with both educational attainment and quitting success. We used a structural equation model of cross-sectional data from respondents ≥25 years from the United States 2010-2011 Tobacco Use Supplement-Current Population Survey. Quitting success was defined as former (n = 2607) versus continuing smokers (n = 7636); categories of educational attainment were ≤high school degree, some college/college degree, and advanced degree. Results indicated that using nicotine replacement therapy (NRT) >1 month and having a home smoking restriction were associated with both educational attainment and quitting success. Those with lower educational attainment versus those with an advanced degree were less likely to report using NRT >1 month (≤high school: β = -0.50, p < 0.001; college: β = -0.24, p = 0.019). Use of NRT >1 month, in turn, was positively associated with quitting success (β = 0.25, p < 0.001). Those with lower educational attainment were also less likely to report a home smoking restriction (≤high school: β = -0.42, p < 0.001; college: β = -0.21, p = 0.009). Having a home smoking restriction was positively associated with quitting success (β = 0.50, p < 0.001). Results were similar with income substituted for education. Using NRT >1 month and having a home smoking restriction are two strategies that may explain the association between low education and lower cessation success; these strategies should be further tested for their potential ability to mitigate this association.


Improved heuristics for minimum-flip supertree construction.
The utility of the matrix representation with flipping (MRF) supertree method has been limited by the speed of its heuristic algorithms. We describe a new heuristic algorithm for MRF supertree construction that improves upon the speed of the previous heuristic by a factor of n (the number of taxa in the supertree). This new heuristic makes MRF tractable for large-scale supertree analyses and allows the first comparisons of MRF with other supertree methods using large empirical data sets. Analyses of three published supertree data sets with between 267 to 571 taxa indicate that MRF supertrees are equally or more similar to the input trees on average than matrix representation with parsimony (MRP) and modified min-cut supertrees. The results also show that large differences may exist between MRF and MRP supertrees and demonstrate that the MRF supertree method is a practical and potentially more accurate alternative to the nearly ubiquitous MRP supertree method.


Do depressive symptoms "blunt" effort? An analysis of cardiac engagement and withdrawal for an increasingly difficult task.
Research on depression and effort has suggested "depressive blunting"-lower cardiovascular reactivity in response to challenges and stressors. Many studies, however, find null effects or higher reactivity. The present research draws upon motivational intensity theory, a broad model of effort that predicts cases in which depressive symptoms should increase or decrease effort. Because depressive symptoms can influence task-difficulty appraisals-people see tasks as subjectively harder-people high in depressive symptoms should engage higher effort at objectively easier levels of difficulty but also quit sooner. A sample of adults completed a mental effort challenge with four levels of difficulty, from very easy to difficult-but-feasible. Depressive symptoms were assessed with the CESD and DASS; effort-related cardiac activity was assessed via markers of contractility (e.g., the cardiac pre-ejection period [PEP]) obtained with impedance cardiography. The findings supported the theory's predictions. When the task was relatively easier, people high in depressive symptoms showed higher contractility (shorter PEP), consistent with greater effort. When the task was relatively harder, people high in depressive symptoms showed diminished contractility, consistent with quitting. The results suggest that past research has been observing a small part of a larger trajectory of trying and quitting, and they illustrate the value of a theoretically grounded analysis of depressive symptoms and effort-related cardiac activity.


Notochord morphogenesis in Xenopus laevis: simulation of cell behavior underlying tissue convergence and extension.
Cell intercalation and cell shape changes drive notochord morphogenesis in the African frog, Xenopus laevis. Experimental observations show that cells elongate mediolaterally and intercalate between one another, causing the notochord to lengthen and narrow. Descriptive observations provide few clues as to the mechanisms that coordinate and drive these cell movements. It is possible that a few rules governing cell behavior could orchestrate the shaping of the entire tissue. We test this hypothesis by constructing a computer model of the tissue to investigate how rules governing cell motility and cell-cell interactions can account for the major features of notochord morphogenesis. These rules are drawn from the literature on in vitro cell studies and experimental observations of notochord cell behavior. The following types of motility rules are investigated: (1) refractory tissue boundaries that inhibit cell motility, (2) statistical persistence of motion, (3) contact inhibition of protrusion between cells, and (4) polarized and nonpolarized protrusive activity. We show that only the combination of refractory boundaries, contact inhibition and polarized protrusive activity reproduces normal notochord development. Guided by these rules, cells spontaneously align into a parallel array of elongating cells. Self alignment optimizes the geometric conditions for polarized protrusive activity by progressively minimizing contact inhibition between cells. Cell polarization, initiated at refractory tissue boundaries, spreads along successive cell rows into the tissue interior as cells restrict and constrain their neighbors' directional bias. The model demonstrates that several experimentally observed intrinsic cell behaviors, operating simultaneously, may underlie the generation of coordinated cell movements within the developing notochord.


A new model for Hendra virus encephalitis in the mouse.
Hendra virus (HeV) infection in humans is characterized by an influenza like illness, which may progress to pneumonia or encephalitis and lead to death. The pathogenesis of HeV infection is poorly understood, and the lack of a mouse model has limited the opportunities for pathogenetic research. In this project we reassessed the role of mice as an animal model for HeV infection and found that mice are susceptible to HeV infection after intranasal exposure, with aged mice reliably developing encephalitic disease. We propose an anterograde route of neuroinvasion to the brain, possibly along olfactory nerves. This is supported by evidence for the development of encephalitis in the absence of viremia and the sequential distribution of viral antigen along pathways of olfaction in the brain of intranasally challenged animals. In our studies mice developed transient lower respiratory tract infection without progressing to viremia and systemic vasculitis that is common to other animal models. These studies report a new animal model of HeV encephalitis that will allow more detailed studies of the neuropathogenesis of HeV infection, particularly the mode of viral spread and possible sequestration within the central nervous system; investigation of mechanisms that moderate the development of viremia and systemic disease; and inform the development of improved treatment options for human patients.


Subthreshold PTSD in primary care: prevalence, psychiatric disorders, healthcare use, and functional status.
The purpose of this study is to determine the prevalence of subthreshold posttraumatic stress disorder (PTSD) and its association with specific traumas, other psychiatric diagnoses, healthcare use, and functional status among 669 veterans in four VA Medical Centers. A cross-sectional, epidemiological design incorporating self-report measures, structured interviews, and chart reviews was used to obtain relevant information for analyses. Comparisons across three trauma-exposed groups (PTSD, subthreshold PTSD, no PTSD) revealed that veterans in the subthreshold PTSD group did not use mental health services more often than those in the no PTSD group despite the presence of additional mental health diagnoses and worse functional status. These data indicate that clinicians may be overlooking a subset of individuals suffering from subsyndromal PTSD, suggesting the need to detect and serve these individuals better within healthcare settings.


Control of expression of insulin resistance and hyperglycemia by different genetic factors in diabetic C57BL/6J mice.
The inheritance of the tendency to develop diet-induced non-insulin-dependent (type II) diabetes was analyzed in crosses between diabetes-prone C57BL/6J (BL/6) mice and diabetes-resistant A/J mice. The effects of a diabetogenic diet on blood glucose and insulin levels, insulin sensitivity, and weight were evaluated in F1 and both (BL/6 X A/J) F1 X BL/6 and (BL/6 X A/J) F1 X A/J backcross mice. These results suggest that diet-induced hyperglycemia is largely determined by a recessive gene and diet-induced insulin resistance by a dominant gene. Analyses of both backcrosses indicated that insulin sensitivity and blood glucose levels were unrelated, suggesting that they are controlled by different genetic factors. This conclusion was supported by data from nine recombinant inbred BXA strains in which no correlation was observed between these variables. Furthermore, insulin sensitivity and body weight correlated differently in the two backcross groups, suggesting that insulin resistance is not simply a function of obesity. The number of genes that predominantly influence diabetic traits was estimated by comparing the variance observed in (BL/6 X A/J) F1 X BL/6 backcross mice with that observed in parental mice. The data suggest that relatively few genes predominantly affect the diabetic phenotype in this murine model.


Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment.
Atopic dermatitis is chronically relapsing pruritic eczema and prevails around the world especially in developed countries. Complex interactions between genetic and environmental factors are known to play an important role in the pathophysiology of atopic dermatitis. However, we still lack a detailed picture of the pathogenesis of this disease. Thus, it is of importance to develop appropriate animal models for elucidating the progression of atopic dermatitis. Moreover, investigating the effect of environmental factors such as air pollutants on atopic dermatitis expands understanding of the disease. Here, we describe a method for inducing atopic dermatitis in rats with neonatal capsaicin treatment and a protocol for exposure of a constant concentration of formaldehyde to rats to reveal effects on the development of atopic dermatitis in infantile and adolescent periods. These protocols have been successfully applied to several experiments and can be used for other substances.


The vitamin D receptor gene as a determinant of survival in pancreatic cancer patients: Genomic analysis and experimental validation.
Advanced pancreatic cancer is a highly refractory disease almost always associated with survival of little more than a year. New interventions based on novel targets are needed. We aim to identify new genetic determinants of overall survival (OS) in patients after treatment with gemcitabine using genome-wide screens of germline DNA. We aim also to support these findings with in vitro functional analysis. Genome-wide screens of germline DNA in two independent cohorts of pancreatic cancer patients (from the Cancer and Leukemia Group B (CALGB) 80303 and the Mayo Clinic) were used to select new genes associated with OS. The vitamin D receptor gene (VDR) was selected, and the interactions of genetic variation in VDR with circulating vitamin D levels and gemcitabine treatment were evaluated. Functional effects of common VDR variants were also evaluated in experimental assays in human cell lines. The rs2853564 variant in VDR was associated with OS in patients from both the Mayo Clinic (HR 0.81, 95% CI 0.70-0.94, p = 0.0059) and CALGB 80303 (HR 0.74, 0.63-0.87, p = 0.0002). rs2853564 interacted with high pre-treatment levels of 25-hydroxyvitamin D (25(OH)D, a measure of endogenous vitamin D) (p = 0.0079 for interaction) and with gemcitabine treatment (p = 0.024 for interaction) to confer increased OS. rs2853564 increased transcriptional activity in luciferase assays and reduced the binding of the IRF4 transcription factor. Our findings propose VDR as a novel determinant of survival in advanced pancreatic cancer patients. Common functional variation in this gene might interact with endogenous vitamin D and gemcitabine treatment to determine improved patient survival. These results support evidence for a modulatory role of the vitamin D pathway for the survival of advanced pancreatic cancer patients.


Diagnostic tests for acute pancreatitis.
Serum amylase continues to be the most widely used test to diagnose acute pancreatitis; however, its popularity does not appear to be justified. The serum amylase test has a poor sensitivity and specificity. Furthermore, it has an extremely low sensitivity in detecting acute alcoholic pancreatitis, which is the most common cause of acute pancreatitis in city hospitals. Older assay techniques for serum lipase were cumbersome and time-consuming. The newer methods seem to have overcome the disadvantages of the previous techniques. They are quick, reliable, and inexpensive. Recent studies indicate that serum lipase may be a better test to diagnose acute pancreatitis. Therefore, serum lipase should be used more frequently in the diagnosis of acute pancreatitis. Serum trypsin, although sensitive, is difficult to estimate and is not routinely available. Serum elastase offers no additional benefit over the serum amylase or lipase tests. Markers such as alpha 2-macroglobulin, RNase, phospholipase, and polymorphonuclear elastase predict severity of disease, but assay techniques for these agents are still experimental and confined to specialized centers. C-reactive protein is a reasonably reliable indicator of severity and, because it is universally available, should be used more frequently. Of the imaging techniques, computerized tomography scanning is the best method to delineate the pancreas; however, ultrasound is more cost-effective in clinical practice.


Return to Play After Multiligament Knee Injuries in National Football League Athletes.
Return to play (RTP) of National Football League (NFL) athletes after isolated anterior cruciate ligament (ACL) tears has been reported. However, no studies have reported on RTP of NFL athletes after multiligament knee injuries. NFL athletes with multiligament knee injuries have lower RTP rates and longer recoveries than athletes with isolated ACL tears. Second, athletes with ACL and medial collateral ligament (MCL) injuries will have higher RTP rates and shorter time to RTP than athletes with an ACL tear and posterolateral corner involvement. Retrospective cohort study. Level 4. Publicly available NFL injury data were reviewed for all multiligament knee injuries incurred between 2000 and 2016 with RTP information. Athletes were excluded if RTP was limited for reasons unrelated to the injury. Extracted data included type of injury, RTP, time to RTP (days), number and percentage of games played, and performance. A total of 50 NFL athletes with multiligament knee injuries met inclusion and exclusion criteria. The overall RTP rate was 64.0%. Athletes with ACL/MCL tears had an RTP rate of 70.8%, whereas athletes with ACL and posterior collateral ligament/lateral collateral ligament (PCL/LCL) tears had an RTP rate of 55.6% ( P = 0.26). Mean time to RTP for all 50 athletes was 388.71 ± 198.52 days. The mean time to RTP for athletes with ACL/MCL injuries was 305.1 ± 58.9 days, compared with 459.2 ± 245.1 days ( P = 0.004) and 609.3 ± 183.1 days ( P < 0.0001) for those with combined ACL and PCL/LCL injuries and frank knee dislocations, respectively. Athletes with ACL/MCL injuries were more likely to return to prior performance level (43.5%) than those with ACL and PCL/LCL injuries (18.5%) ( P < 0.001). The RTP rate for athletes with multiligament knee injuries is significantly less than the RTP rate for athletes with isolated ACL tears. In addition, athletes with ACL and MCL tears have a higher RTP rate, a significantly shorter time to RTP, and a greater likelihood of returning to prior performance than athletes with ACL and PCL/LCL tears. Multiligament knee injuries significantly affect the ability of a football player to return to sport. ACL and MCL tears are associated with better RTP prognosis compared with ACL and PCL/LCL tears.


Genome and low-iron response of an oceanic diatom adapted to chronic iron limitation.
Biogeochemical elemental cycling is driven by primary production of biomass via phototrophic phytoplankton growth, with 40% of marine productivity being assigned to diatoms. Phytoplankton growth is widely limited by the availability of iron, an essential component of the photosynthetic apparatus. The oceanic diatom Thalassiosira oceanica shows a remarkable tolerance to low-iron conditions and was chosen as a model for deciphering the cellular response upon shortage of this essential micronutrient. The combined efforts in genomics, transcriptomics and proteomics reveal an unexpected metabolic flexibility in response to iron availability for T. oceanica CCMP1005. The complex response comprises cellular retrenchment as well as remodeling of bioenergetic pathways, where the abundance of iron-rich photosynthetic proteins is lowered, whereas iron-rich mitochondrial proteins are preserved. As a consequence of iron deprivation, the photosynthetic machinery undergoes a remodeling to adjust the light energy utilization with the overall decrease in photosynthetic electron transfer complexes. Beneficial adaptations to low-iron environments include strategies to lower the cellular iron requirements and to enhance iron uptake. A novel contribution enhancing iron economy of phototrophic growth is observed with the iron-regulated substitution of three metal-containing fructose-bisphosphate aldolases involved in metabolic conversion of carbohydrates for enzymes that do not contain metals. Further, our data identify candidate components of a high-affinity iron-uptake system, with several of the involved genes and domains originating from duplication events. A high genomic plasticity, as seen from the fraction of genes acquired through horizontal gene transfer, provides the platform for these complex adaptations to a low-iron world.


Validity of Borg Ratings of Perceived Exertion During Active Video Game Play.
During physically interactive video game play (e.g., Nintendo Wii), users are exposed to potential distracters (e.g., video, music), which may decrease their ratings of perceived exertion (RPE) throughout game play. The purpose of this investigation was to determine the association between RPE scores and heart rate while playing the Nintendo Wii. Healthy adults (N = 13, 53.5 ± 5.4 years old) participated in two exercise sessions using the Nintendo Wii Fit Plus. During each session participants played a five-minute warm-up game (Basic Run), two separate Wii Fit Plus games (Yoga, Strength Training, Aerobics or Balance Training) for fifteen minutes each, and then a five-minute cool down game (Basic Run). Borg RPE and heart rate were assessed during the final 30 seconds of the warm up and cool down, as well during the final 30 seconds of play for each Wii Fit Plus game. Correlation analysis combining data from both exercise sessions indicated a moderate positive relationship between heart rate and RPE (r = 0.32). Mixed-effects model regression analyses demonstrated that RPE scores were significantly associated with heart rate (p < 0.001). The average percentage of age-predicted heart rate maximum achieved (58 ± 6%) was significantly greater (p = 0.001) than the percentage of maximum RPE indicated (43 ± 11%). Borg RPE scores were positively associated with heart rate in adults during exercise sessions using the Wii Fit Plus. However, this relationship was lower than observed in past research assessing RPE validity during different modes of exercise (e.g. walking, running) without distracters.


Neurones in the supraoptic nucleus of the rat are regulated by a projection from the suprachiasmatic nucleus.
1. In the rat, projections from the suprachiasmatic nucleus (SCN) to the supraoptic nucleus (SON) of the hypothalamus were characterized in vivo using extracellular recordings and in slice preparations using both extracellular and whole-cell patch clamp recording. 2. Of 117 magnocellular neurones recorded in the SON in vivo, fifteen (13%) displayed a short latency excitation, sixty-eight (58%) a short latency inhibition, six (5%) were unresponsive and twenty-eight (24%) gave long latency responses following SCN stimulation. 3. The responses of putative vasopressin cells in the SON to SCN stimulation in vivo (4 out of 61 cells, 7% excited; 49 out of 61 cells, 80% inhibited) were significantly different from those of putative oxytocin cells (10 out of 50 cells, 20% excited and 16 out of 50 cells, 32% inhibited; P < 0.02, test for differences between proportions). 4. Recordings in vitro using patch technology in whole-cell mode showed both inward and outward currents in SON cells at holding potentials near resting membrane potential following stimulation of the SCN region. The outward currents could be blocked by bicuculline (10 microM; n = 7) and the inward currents were blocked by the non-NMDA antagonist 6-nitro-7-sulphamoylbenzo(f)quinoxaline-2,3-dione (5 microM; n = 4). 5. We conclude that there is a strong projection from the SCN to the SON with both inhibitory (GABAergic) and excitatory (glutamatergic) components which may regulate the daily changes in neurohypophysial hormone secretion.


Investigation of the stallion sperm proteome by mass spectrometry.
Stallion spermatozoa continue to present scientific and clinical challenges with regard to the biological mechanisms responsible for their survival and function. In particular, deeper understanding of sperm energy metabolism, defence against oxidative damage and cell-cell interactions should improve fertility assessment and the application of advanced reproductive technologies in the equine species. In this study, we used highly sensitive LC-MS/MS technology and sequence database analysis to identify and characterise the proteome of Percoll-isolated ejaculated equine spermatozoa, with the aim of furthering our understanding of this cell's complex biological machinery. We were able to identify 9883 peptides comprising 1030 proteins, which were subsequently attributed to 975 gene products. Gene ontology analysis for molecular and cellular processes revealed new information about the metabolism, antioxidant defences and receptors of stallion spermatozoa. Mitochondrial proteins and those involved in catabolic processes constituted dominant categories. Several enzymes specific to β-oxidation of fatty acids were identified, and further experiments were carried out to ascertain their functional significance. Inhibition of carnitine palmitoyl transferase 1, a rate-limiting enzyme of β-oxidation, reduced motility parameters, indicating that β-oxidation contributes to maintenance of motility in stallion spermatozoa.


MHC class II transactivator CIITA is a recurrent gene fusion partner in lymphoid cancers.
Chromosomal translocations are critically involved in the molecular pathogenesis of B-cell lymphomas, and highly recurrent and specific rearrangements have defined distinct molecular subtypes linked to unique clinicopathological features. In contrast, several well-characterized lymphoma entities still lack disease-defining translocation events. To identify novel fusion transcripts resulting from translocations, we investigated two Hodgkin lymphoma cell lines by whole-transcriptome paired-end sequencing (RNA-seq). Here we show a highly expressed gene fusion involving the major histocompatibility complex (MHC) class II transactivator CIITA (MHC2TA) in KM-H2 cells. In a subsequent evaluation of 263 B-cell lymphomas, we also demonstrate that genomic CIITA breaks are highly recurrent in primary mediastinal B-cell lymphoma (38%) and classical Hodgkin lymphoma (cHL) (15%). Furthermore, we find that CIITA is a promiscuous partner of various in-frame gene fusions, and we report that CIITA gene alterations impact survival in primary mediastinal B-cell lymphoma (PMBCL). As functional consequences of CIITA gene fusions, we identify downregulation of surface HLA class II expression and overexpression of ligands of the receptor molecule programmed cell death 1 (CD274/PDL1 and CD273/PDL2). These receptor-ligand interactions have been shown to impact anti-tumour immune responses in several cancers, whereas decreased MHC class II expression has been linked to reduced tumour cell immunogenicity. Thus, our findings suggest that recurrent rearrangements of CIITA may represent a novel genetic mechanism underlying tumour-microenvironment interactions across a spectrum of lymphoid cancers.


Progress in design and architecture of metal nanoparticles for catalytic applications.
Over the past few years, nanometer-sized transition metal particles have been intensively pursued as potentially advanced catalysts because their special properties lie between those of single metal atoms and bulk metal. Achieving the accurate control of particle size and overall particle size distribution is one of the most crucial challenges to provide unique chemical and physical properties. We highlight herein our recent progress in the exploitation of promising nanoparticle (NP)-based catalysts designed by precise architecture that enable efficient and selective chemical transformations and can be completely separated and are recyclable. This perspective article consists of the following two specific topics: (i) multifunctional catalysts based on magnetic NPs and (ii) new routes for the preparation of supported metal NPs catalysts. The synthetic strategies described here are simple and general for practical catalyst design, thus allowing a strong protocol for creating various nanostructured catalysts.


Outbreak of paralytic poliomyelitis in Oman: evidence for widespread transmission among fully vaccinated children.
From January, 1988, to March, 1989, a widespread outbreak (118 cases) of poliomyelitis type 1 occurred in Oman. Incidence of paralytic disease was highest in children younger than 2 years (87/100,000) despite an immunisation programme that recently had raised coverage with 3 doses of oral poliovirus vaccine (OPV) among 12-month-old children from 67% to 87%. We did a case-control study (70 case-patients, 692 age-matched controls) to estimate the clinical efficacy of OPV, assessed the immunogenicity of OPV and extent of poliovirus spread by serology, retrospectively evaluated the cold chain and vaccine potency, and sought the origin of the outbreak strain by genomic sequencing. 3 doses of OPV reduced the risk of paralysis by 91%; vaccine failures could not be explained by failures in the cold chain nor on suboptimum vaccine potency. Cases and controls had virtually identical type 1 neutralising antibody profiles, suggesting that poliovirus type 1 circulation was widespread. Genomic sequencing indicated that the outbreak strain had been recently imported from South Asia and was distinguishable from isolates indigenous to the Middle East. Accumulation of enough children to sustain the outbreak seems to have been due to previous success of the immunisation programme in reducing spread of endemic strains, suboptimum efficacy of OPV, and delay in completing the primary immunisation series until 7 months of age. Additionally, the estimated attack rate of infection among children aged 9-23 months exceeded 25% in some regions, suggesting that a substantial proportion of fully vaccinated children had been involved in the chain of transmission.


Adaptation of the agyrophil technique for nucleolar organizer regions to canine peripheral blood smears.
Nucleolar organizer regions (NORs) are chromosomal segments which are selectively stained by silver methods and evaluated as agyrophilic nucleolar organizer regions (AgNORs). The evaluation of AgNORs provides an insight into the level of cellular proliferation. This technique has extensively and sparsely been used in human and veterinary histopathology respectively. However, two major drawbacks have been irreproducible results and excessive staining precipitates. This study seeks to adapt the technique to canine peripheral blood smears in order to establish a routine staining protocol. Standardized volumes and concentrations of silver nitrate, gelatin, and formic acid were applied to smears at different temperatures for varying lengths of time. The technique was applied to unfixed and fixed smears. In some cases, a reducing agent (1% potassium iodide) was applied. It was shown that the optimum staining protocol was achieved by applying standardized solutions to a fixed smear at 46°C for 50 minutes. It is concluded that the staining protocol outlined in this study is practicable, and produces excellent and reproducible results that would enhance evaluation of AgNORs in canine peripheral blood cells.


Thrombosis in patients with paroxysmal nocturnal hemoglobinuria.
Paroxysmal nocturnal hemoglobinuria is a disorder associated with hemolysis, pancytopenia, and thrombosis due to the loss of the glycosylphosphatidylinositol (GPI) anchored complement regulatory proteins. The mechanism of thrombosis is multifactorial. Although intravascular hemolysis has been implicated as the etiology, the effect of complement on GPI anchor-deficient platelets, granulocytes, monocytes, and endothelial cells contributes significantly to the risk of thrombosis. Moreover, there appears to be an underlying inflammatory state that is linked to hemostatic activation that may induce thrombosis through a pathway independent of hemolysis.


Candida glabrata: review of epidemiology, pathogenesis, and clinical disease with comparison to C. albicans.
Until recently, Candida glabrata was considered a relatively nonpathogenic commensal fungal organism of human mucosal tissues. However, with the increased use of immunosuppressive agents, mucosal and systemic infections caused by C. glabrata have increased significantly, especially in the human immunodeficiency virus-infected population. A major obstacle in C. glabrata infections is their innate resistance to azole antimycotic therapy, which is very effective in treating infections caused by other Candida species. Candida glabrata, formerly known as Torulopsis glabrata, contrasts with other Candida species in its nondimorphic blastoconidial morphology and haploid genome. C. glabrata currently ranks second or third as the causative agent of superficial (oral, esophageal, vaginal, or urinary) or systemic candidal infections, which are often nosocomial. Currently, however, there are few recognized virulence factors of C. glabrata and little is known about the host defense mechanisms that protect against infection. Two established animal models (systemic and vaginal) have been established to study treatment, pathogenesis, and immunity. Treatment of C. glabrata infections can include azoles but often requires amphotericin B or flucytosine. This review summarizes all known clinical and experimental information about C. glabrata infections with comparisons to C. albicans as a means of contrasting the two species commonly observed and emphasizing the many recognized differences.


Genome sequence of Thermotoga sp. strain RQ2, a hyperthermophilic bacterium isolated from a geothermally heated region of the seafloor near Ribeira Quente, the Azores.
Thermotoga sp. strain RQ2 is probably a strain of Thermotoga maritima. Its complete genome sequence allows for an examination of the extent and consequences of gene flow within Thermotoga species and strains. Thermotoga sp. RQ2 differs from T. maritima in its genes involved in myo-inositol metabolism. Its genome also encodes an apparent fructose phosphotransferase system (PTS) sugar transporter. This operon is also found in Thermotoga naphthophila strain RKU-10 but no other Thermotogales. These are the first reported PTS transporters in the Thermotogales.


Ligand-induced activation of human TRPM2 requires the terminal ribose of ADPR and involves Arg1433 and Tyr1349.
TRPM2 (transient receptor potential channel, subfamily melastatin, member 2) is a Ca2+-permeable non-selective cation channel activated by the binding of adenosine 5'-diphosphoribose (ADPR) to its cytoplasmic NUDT9H domain (NUDT9 homology domain). Activation of TRPM2 by ADPR downstream of oxidative stress has been implicated in the pathogenesis of many human diseases, rendering TRPM2 an attractive novel target for pharmacological intervention. However, the structural basis underlying this activation is largely unknown. Since ADP (adenosine 5'-diphosphate) alone did not activate or antagonize the channel, we used a chemical biology approach employing synthetic analogues to focus on the role of the ADPR terminal ribose. All novel ADPR derivatives modified in the terminal ribose, including that with the seemingly minor change of methylating the anomeric-OH, abolished agonist activity at TRPM2. Antagonist activity improved as the terminal substituent increasingly resembled the natural ribose, indicating that gating by ADPR might require specific interactions between hydroxyl groups of the terminal ribose and the NUDT9H domain. By mutating amino acid residues of the NUDT9H domain, predicted by modelling and docking to interact with the terminal ribose, we demonstrate that abrogating hydrogen bonding of the amino acids Arg1433 and Tyr1349 interferes with activation of the channel by ADPR. Taken together, using the complementary experimental approaches of chemical modification of the ligand and site-directed mutagenesis of TRPM2, we demonstrate that channel activation critically depends on hydrogen bonding of Arg1433 and Tyr1349 with the terminal ribose. Our findings allow for a more rational design of novel TRPM2 antagonists that may ultimately lead to compounds of therapeutic potential.


Science for a wilder Anthropocene: Synthesis and future directions for trophic rewilding research.
Trophic rewilding is an ecological restoration strategy that uses species introductions to restore top-down trophic interactions and associated trophic cascades to promote self-regulating biodiverse ecosystems. Given the importance of large animals in trophic cascades and their widespread losses and resulting trophic downgrading, it often focuses on restoring functional megafaunas. Trophic rewilding is increasingly being implemented for conservation, but remains controversial. Here, we provide a synthesis of its current scientific basis, highlighting trophic cascades as the key conceptual framework, discussing the main lessons learned from ongoing rewilding projects, systematically reviewing the current literature, and highlighting unintentional rewilding and spontaneous wildlife comebacks as underused sources of information. Together, these lines of evidence show that trophic cascades may be restored via species reintroductions and ecological replacements. It is clear, however, that megafauna effects may be affected by poorly understood trophic complexity effects and interactions with landscape settings, human activities, and other factors. Unfortunately, empirical research on trophic rewilding is still rare, fragmented, and geographically biased, with the literature dominated by essays and opinion pieces. We highlight the need for applied programs to include hypothesis testing and science-based monitoring, and outline priorities for future research, notably assessing the role of trophic complexity, interplay with landscape settings, land use, and climate change, as well as developing the global scope for rewilding and tools to optimize benefits and reduce human-wildlife conflicts. Finally, we recommend developing a decision framework for species selection, building on functional and phylogenetic information and with attention to the potential contribution from synthetic biology.


Fabrication of 3D hepatic tissues by additive photopatterning of cellular hydrogels.
We have fabricated a hepatic tissue construct using a multilayer photopatterning platform for embedding cells in hydrogels of complex architecture. We first explored the potential of established hepatocyte culture models to stabilize isolated hepatocytes for photoencapsulation (e.g., double gel, Matrigel, cocultivation with nonparenchymal cells). Using photopolymerizable PEG hydrogels, we then tailored both the chemistry and architecture of the hydrogels to further support hepatocyte survival and liver-specific function. Specifically, we incorporated adhesive peptides to ligate key integrins on these adhesion-dependent cells. To identify the appropriate peptides for incorporation, the integrin expression of cultured hepatocytes was monitored by flow cytometry and their functional role in cell adhesion was assessed on full-length extracellular matrix (ECM) molecules and their adhesive peptide domains. In addition, we modified the hydrogel architecture to minimize barriers to nutrient transport for these highly metabolic cells. Viability of encapsulated cells was improved in photopatterned hydrogels with structural features of 500 microm in width over unpatterned, bulk hydrogels. Based on these findings, we fabricated a multilayer photopatterned PEG hydrogel structure containing the adhesive RGD peptide sequence to ligate the alpha5beta1 integrin of cocultured hepatocytes. Three-dimensional photopatterned constructs were visualized by digital volumetric imaging and cultured in a continuous flow bioreactor for 12 d where they performed favorably in comparison to unpatterned, unperfused constructs. These studies will have impact in the field of liver biology as well as provide enabling tools for tissue engineering of other organs.


Needs assessment and current practice of alcohol risk assessment of pregnant women and women of childbearing age by primary health care professionals.
Assessing the current practices and learning and resource needs of primary health care professionals in regards to their alcohol risk assessment practices is an important step in providing optimal training and educational methods. Needs and current practices in alcohol risk assessment of pregnant women and women of child bearing years may vary according to practitioner demographics. To appraise alcohol risk assessment current practices and learning and resource needs among Saskatchewan primary health care professionals, a mail and online survey was distributed in the spring of 2006 to family physicians/general practitioners and nurse practitioners. In total, 876 surveys were distributed and 386 were returned for an overall response rate of 44.1%. The majority of survey respondents reported either rarely or never using a standardized screening tool in assessing alcohol risk in women or reported using a standardized screening tool that is less sensitive. Current practices varied according to gender, length of time in practice and practice location, while learning and resource needs were more likely to be identified by nurse practitioners, female physicians, and physicians from rural areas. Physicians who had practiced for less than 5 years were more likely to want an online course. Knowing the needs and practices of health care professionals may assist learning and resource training and could assist in teaching best practices in alcohol risk assessment. Assessing alcohol risk in pregnant women and women of childbearing age is critical for prevention of FASD.